Foodcatalog 2013 Finalpdf PDF

Food & Dietary
Supplements
2013 Directory
of Services
Introduction
Advancements in the way food and dietary supplements are developed, packaged, and prepared
require flexibility, innovation, and a broad scientific understanding of those implications on analytical
data. Covance has over 75 years of experience in food analysis and has built a reputation for
scientific and regulatory expertise.
Understanding the key drivers in analytical testing — placing the customer first, creating processes to
drive efficiency, and providing high quality data — has positioned Covance as a leader in outsourced
food and dietary supplement testing services.
This directory provides a description of our analytical chemistry assays, quality processes,
service descriptions, and ordering information. We invite you to visit our facilities, speak with
our staff, and experience a level of personal commitment to helping you reach your product
development goals while remaining focused on regulatory compliance and food safety.
Table of Contents
Quick Reference . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-16
Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17-68
Quality . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69-72
ISO Accreditation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 69
Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
Certifications/Accreditations/Registrations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
Proficiency Testing . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
Staff & Facilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73-82

Battle Creek, Michigan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
Greenfield, Indiana . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
Madison, Wisconsin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Singapore . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Site Visitor Guides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75-81
Battle Creek, Michigan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Greenfield, Indiana . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
Madison, Wisconsin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Singapore . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .80
Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 83-84
Sampling . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Importing Samples to Covance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 84
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85-97
Ordering Please order using our online food catalog at

Information http://www.covance.com/foodcatalog
Sample Submittal/Shipping
[ assay ]
quick reference
Nutrition Labeling Mandatory Nutrient List
Regulatory initiatives such as the Nutrition Labeling and Education Act have had a profound impact on the food industry.
The adoption of standardized content and format guidelines for nutrition labeling have provided consumers with the tools
to plan a balanced diet as well as given producers data to distance their product from the competition.
NL-Proximate (moisture, ash, protein)
Calories (by calculation)
Calories from fat (by calculation)
Total fat [sum of fatty acids (saturated, monounsaturated, polyunsaturated, trans)]
Cholesterol
Total carbohydrates (by calculation)
Total dietary fiber
Sugars
HPLC
GC
Either HPLC or GC will be used, depending on the sample matrix. HPLC is appropriate for most, but not all, samples.
Vitamin A
beta Carotene
If there is a source of beta carotene, the assay for beta carotene should be used instead of (or in addition to) vitamin A
to obtain total vitamin A activity for nutrition labels.
Vitamin C
Calcium, iron, sodium (ICP)

These inorganics can also be run by atomic absorption (AA).
Mandatory Nutrient Package

This package includes all nutrients, with sugars assayed by HPLC, minerals by ICP, and total fat as the sum of the
fatty acids calculated as triglycerides.
Priority service is available for an additional fee.
608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 1

[ q u ic k re f e re nce ]
Vitamin Analysis
Biotin Tocotrienols (alpha, beta, gamma, and delta)
Carotenoids Vitamin A (retinol)
Option 1: alpha Carotene, beta Carotene, Lycopene Vitamin A for dietary supplements
Option 2: Cryptoxanthin, Lutein, Zeaxanthin Vitamin B profiles (HPLC)
Option 3: Other carotenoids
Vitamin B6 (pyridoxine)
Choline (chemical)
Vitamin B12 (cyanocobalamin)
Choline (enzymatic)
Vitamin C
Inositol
Vitamin D
Folate
Vitamin E (alpha tocopherol)
Niacin
Vitamin E for dietary supplements
Pantothenic acid
Vitamin K1
Riboflavin (vitamin B2)
Thiamin (vitamin B1)
Tocopherols (alpha, beta, gamma, and delta)
Lipids
Cholesterol Glycerol
Conjugated linoleic acid (CLA) Hexanal
Fatty acid profile (C8 - C22) Iodine value
Fatty acid profile (C8 - C24) (including Omega-3 and p-Anisidine
Omega-6 isomers) Peroxide value
Fatty acid profile (C4 - C24) (including Omega-3 and Sterols
Omega-6 isomers) TBA value
Free fatty acids (total by titration)
Carbohydrates
Acid detergent fiber Neutral detergent fiber (enzymatic)

Aspartame, Acesulfame K, DKP and saccharine Polydextrose
beta Glucan Resistant starch
Crude fiber Starch
Dietary fiber containing supplemented resistant Sucralose (finished products)
maltodextrin (fibersol) Sucralose (packets, pure material)
Galactooligosaccharides (GOS) content in Infant Formula Sugar alcohols
Galactooligosaccharide (GOS) content in Raw Material Sugars by acid hydrolysis
Insoluble, soluble, and total dietary fiber (CODEX Sugars by GC
definition) by enzymatic-gravimetric method and liquid Sugar by HPAEC-PAD, low level
chromatography (McCleary)
Sugars by HPLC
Insoluble and soluble fiber
Sugars by IC
Total by calculation
Total dietary fiber (Prosky)
Inulin (fructan), HPLC
Total dietary fiber (CODEX definition) by enzymatic-
Inulin (fructan), spectrographic
gravimetric method and liquid chromatography
Lactulose (McCleary)
Lignin
2 Pricing available from a Covance representative. For updates to assays, refer to our online catalog at www.covance.com/foodcatalog.
Amino Acids
Total amino acid profile (AOAC)
Alanine, arginine, aspartic acid (including asparagine), cystine, glutamic acid (including glutamine), glycine, histidine,
isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, valine
Total amino acid profile (HPLC)
Alanine, arginine, aspartic acid (including asparagine), cystine (including cysteine), glutamic acid (including glutamine),
glycine, histidine, hydroxyproline, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine,
valine
Free amino acid profile (AAA)
5-hydroxytryptophan (5HTP), L-ornithine, L-theanine, L-cysteine, beta-alanine, N-acetyl cysteine
Free amino acid profile (HPLC)
L-aspartic acid, L-glutamic acid, L-serine, L-histidine, glycine, L-threonine, L-arginine, L-alanine, L-tyrosine, L-cystine,
L-valine, L-methionine, L-phenylalanine, L-isoleucine, L-leucine, L-lysine, proline, L-asparagine, L-glutamine, L-citrulline,
L-tryptophan, and hydroxyproline
5-Hydroxytrytophan (5-HTP)
Acetyl Cysteine
beta Alanine
Carnitine
Cysteine
Cystine and methionine (AOAC)
GABA (gamma aminobutyric acid)
Monosodium glutamate (as glutamic acid)
Ornithine
Taurine
Theanine
Tryptophan (AOAC)
Tryptophan (HPLC)
Organic Acids
Organic Acid Profiles
Option 1: Benzoic acid (sodium benzoate), sorbic acid (potassium sorbate)
Option 2: Citric acid, malic acid, lactic acid, acetic acid
Option 3: Tartaric acid, formic acid, succinic acid, fumaric acid
Option 4: Propionic acid
Option 5: Oxalic acid
Option 6: Quinic acid, malic acid, citric acid (cranberry)
Option 7: Pyruvic acid

Proximate
Acid insoluble ash Fat (sum of fatty acids)
Ash Loss on drying (LOD)
Available carbohydrates (direct determination) Moisture (Karl Fischer)
< 0.5% Moisture (vacuum oven)
> 0.5% pH
Bomb calorimetry Protein (Kjeldahl)
Density Protein (Dumas)
Fat extractions Proximate package (moisture, ash, protein, and fat)
Acid hydrolysis Total carbohydrates (by calculation)
Soxhlet Calories (by calculation)
Roese-Gottlieb
Residue on ignition (ROI)
Methanol/chloroform
Other Assays
Alcohol (added) Ethanol and Methanol
Alcohol (residual) Nucleosides
Allergens Nucleotides
Almond Protein by ELISA Osmolality
Egg Protein by ELISA Parabens (methyl, ethyl, propyl, butyl, isopropyl, isobutyl)
Gluten/Gliadan by ELISA Polysorbates
Hazelnut protein by ELISA Soy isoflavones (genistein, daidzein, glycitein)
Peanut Protein by ELISA Species Identification by ELISA (beef, poultry or pork)
Sesame Protein by ELISA Sulfites (Monier-Williams)
Soy Protein by ELISA TBHQ
Total Milk Protein by ELISA Titratable acidity
BHA/BHT (antioxidants) Viscosity (Brookfield or Bostwick)
Caffeine, theobromine, and theophylline Water activity
Capsacin/capsacinoid (Scoville heat)
Dissolution
Inorganic Analysis
The method used for the analysis of various elements in food and dietary supplement samples is dependent on both the
concentration and the sample matrix. For most elements the precision using any of these techniques is less than 5% RSD.
Inductively Coupled Plasma Spectroscopy (ICP) is used for macro-levels inorganic constituents in all matrices. Usually
10g of sample is either dry or wet ashed, or samples are analyzed directly. The limit of quantitation is based on the
individual analyte and matrix, but generally yields detection in the PPM range. A common scan is for Calcium, Iron and
Sodium used for nutritional labeling.
Inductively Coupled Plasma-Mass Spectrometry (ICP-MS) is the method of choice when analyzing trace level
analytes. It is applicable to all matrices. Usually 0.5g of sample is wet ashed in a closed-vessel microwave digestion,
although other ashing techniques are used. Limits of quantitation are lower than ICP and are dependent on the analyte
and matrix, but generally yield detection in the PPB range. A common scan is for Arsenic, Cadmium, Lead and Mercury
used for heavy metal analysis.
Atomic Absorption Spectrospcopy (AA) is used for single analyte measurements using flame, Hydride-generation, or
graphite furnace techniques. This technology, in general, has been surpassed by the advances made in ICP and ICP-MS
techniques. Please see the details in the assay section of the catalog for specifics.
Individual Elements
ICP Flame AA
Aluminum Magnesium Silicon
Barium Manganese Titanium
Beryllium Molybdenum Other analytes available on request include calcium, chromium (in
Boron Nickel fortified supplements), cobalt, copper, iron, lead, magnesium, manganese,
Cadmium Phosphorus molybdenum, potassium, selenium, sodium, and zinc.
Calcium Potassium
Chromium Sodium Graphite Furnace AA
Cobalt Strontium Aluminum
Copper Vanadium Other analytes, such as chromium, available on request
Iron Zinc
Miscellaneous AA
Selenium (hydride)
ICP-MS
Antimony Manganese
Arsenic Mercury
Barium Molybdenum
Beryllium Nickel
Bismuth Palladium
Cadmium Platinum
Caesium Rubidium
Calcium Ruthenium
Chromium Selenium
Cobalt Silver
Copper Strontium
Gallium Sulfate*
Germanium Sulfur
Iodine Thallium
Iron Tin
Lanthanum Titanium
Lead Tungsten
Lithium Vanadium
Magnesium Zinc
* Calculated from total sulfur by ICP-MS.

Inorganic Analysis
Inorganics by ICP Emission Spectrometry
Packages
Option 1: ICP3–calcium, iron, sodium (for nutrition labeling)
Option 2: ICPL–calcium, copper, iron, magnesium, manganese, phosphorus, potassium, sodium, zinc
Option 3: ICPN–Option 2 plus aluminum, barium, boron, chromium*, molybdenum*, strontium
Option 4: ICPS–Option 3 plus beryllium, cadmium, cobalt, nickel, vanadium
* May need to perform an alternate test for certain matrices.
Inorganics by Wet Chemistry

Chloride
Fluoride (chemical)
Fluoride (ISE)
Iodine (chemical)
Iodine (ISE)
Nitrate/nitrite by HPLC
Nitrate/nitrite by IC and VIS
Heavy metals as lead by USP <231>
Microbiology
Services
Covance understands the challenges your company faces in Covance has the further capability to offer assistance in
today’s marketplace. Consumer awareness of food safety is developing your Hazard Analysis and Critical Control Points
heightening. Regulatory efforts such as the US Food Safety (HACCP) plan and HACCP validation services. Certified
Modernization Act continue to focus on prevention of, rather HACCP instructors have been responsible for assisting in the
than response to, food contamination. Covance has responded to development of over 200 HACCP plans pertaining to meat,
your needs by expanding its microbiological capabilities. We now dairy, agricultural products, paper, and non-food products.
offer you the breadth of services you need to stay competitive.
Having confidence that your resources are being allocated
Covance brings you the highest level of food safety testing wisely is important. You will gain this confidence knowing
through our dedicated microbiology laboratories in Battle Creek, that all client plans assisted by our scientific staff have passed
Michigan, and Madison, Wisconsin. In addition to these high- governmental and client inspections. Our training programs
throughput testing capabilities, expert microbiology consulting have been developed to be flexible to your unique needs. This is
and training services are available. why Covance offers two options—HACCP training classes at
one of our facilities or customized training specific to your plant
If one of your products, product contact surfaces or environment and product delivered at your location. All HACCP certification
would be found to be positive for presence of a pathogen, Covance courses offered by Covance are accredited by the International
can conduct emergent microbial harborage point investigations. HACCP Alliance.
We can also assist you with responses to regulatory actions such
as a FDA 483 or a USDA notice of suspension or offer an expert Contact us today at 1-855-83MICRO and discover why
witness in legal cases relating to food safety claims. Covance is your optimum partner in food safety services.
Microbiology Consulting and Research Services

Food safety program development or review Process validations
Prerequisite program development or review Microbial harborage point investigations
GFSI audit preparation Responses to regulatory actions
Challenge studies Expert witness services
Training Services
From individualized and customized training to classroom offerings, Covance offers training in:
HACCP
Laboratory
- Pathogen and spoilage organisms
- Laboratory techniques
- Equipment validation
Plant employee good manufacturing practice (GMP)/prerequisites
Other topics upon request
All training may be customized to plant, product and delivered on location.

Microbiology Assays List

Food & Environmental Swabs
Aerobic plate count Listeria species
Anaerobic plate count Listeria species cultural confirmation
Bacillus cereus count Mesophilic aerobic spore count
Chronobacter sakazakii Mesophilic anaerobic spore count
Clostridium perfringens Microbiological identification - gram negative
Coagulase positive S. aureus count Microbiological identification - gram positive
Coliform count Gram stain
Coliform count - MPN Osmophilic yeast and mold
E. coli count Probiotics
E. coli count - MPN Salmonella
E. coli O157:H7 Salmonella cultural confirmation
E. coli O157:H7 cultural confirmation Staphylococcus aureus
Enterobacteriaceae count Thermophilic aerobic bacterial sporeformers
Lactic acid bacteria count (spoilage) Thermophilic anaerobic spore count
Lactobacillus - heterofermentative (spoilage) Total aciduric plate count
Listeria monocytogenes Yeast and mold count
Listeria monocytogenes cultural confirmation
Additional Charges for Qualitative Assays (E. coli O157:H7, Listeria monocytogenes, Listeria species, Salmonella, etc.)
Samples size >100g Compositing of individual samples
Additional Charges for Quantitative Assays

Additional dilutions beyond 1:10, 1:100, 1:1000
Dietary Supplements & Nutritional Products

Suitability for enumeration methods Salmonella
Suitability for specified microorganism methods Staph aureus
Total aerobic microbial count (TAMC) Pseudomonas aeruginosa
Total combined yeasts and molds count (TYMC) Clostridia spp.
Bile-tolerant gram-negative bacteria Candida albicans
E. coli
Additional Charges for USP Assays

Additional dilutions beyond 1:10 Sample neutralization
Contaminants in Foods, Feeds

and Dietary Supplements
Global distribution of food, feed and dietary supplement Our Center of Excellence for Contaminants in Greenfield,
products into new regions requires a broad understanding of the Indiana, offers a broad portfolio of contaminant testing services.
interplay between growers, manufacturers, and regulators. The 10,000 sq ft state-of-the-art facility in Greenfield provides
First-hand experience developing new testing methods, global clients the most up-to-date analysis of chemical residues and
regulatory knowledge, and the scientific expertise to develop an contaminants as well as the development of new analytical methods.
appropriate testing program is critical to meeting your specific
We approach the analysis of contaminants based on analytical
product needs. Our scientists have designed and managed
quantitation using highly selective technology including ICP-MS,
hundreds of testing programs for heavy metals, acrylamide,
LC- and GC-MS/MS. As a true partner in food and dietary
melamine and pesticide residues in support of Proposition 65,
supplement analysis, we have the resources to meet your
response to economic adulteration, routine supplier verification
requirements for quality, flexibility, value and customer service.
and Infant Formula EU Directive 2006/141/EC, respectively.
Multi-Residue Analysis (MRA) by GC-MS/MS and LC-MS/MS (300+ compounds)

Botanicals, Spices and Other Specialty Commodities and Products
All analyses reported to 10 ppb or LOQ , whichever is higher.
Finished Food and Raw Ingredients

All analyses reported to 10 ppb or LOQ , whichever is higher.
This offering includes:
Fruits and vegetables
Grains and cereals
Fats and oils
High-fat food products
Finished processed foods, including meals
Infant Formula
All analytes are reported to 10 ppb (or lower if required). EU Directive 2006/141/EC (10 ng/g or less) offering is available.
Matrix-Based Quantitation
Additional analyses (after initial analysis) to perform Method of Standard Addition, which provides the most accurate
quantitation.
Multi-Residue Analysis for Quintozene and other specific compounds for ginseng
Individual Pesticides or Subgroups

Individual pesticides, subgroups of or additions to the MRA
Carbamates by LC-MS/MS
Pyrethrins and Piperonyl Butoxide by LC-MS/MS

USP/EP Analysis (99 Analytes)

USP 561 and EP Pesticides Full Reporting Package
Includes: GC-MS/MS and LC/MS/MS Multi-Residue Analysis, EBDCs (dithiocarbamates) by GC, inorganic bromide by GC,
matrix-based quantitation*, and pass/fail report
*Matrix-Based Quantitation: additional analyses (after initial analysis) to perform Method of Standard Addition, which provides the
most accurate quantitation.
Ala Cart Analysis

GC-MS/MS and LC-MS/MS Multi-Residue Analysis, EBDCs (dithiocarbamates) by GC, and matrix-based quantitation
GC-MS/MS and LC-MS/MS Multi-Residue Analysis (90+ analytes)
Standard Addition (matrix-based quantitation)
EBDCs (dithiocarbamates) by GC
Bromide, inorganic by GC
Classical Pesticide Methods by GC

Chlorinated/organophosphorus pesticides
FDA PAM 302, 303, or 304
Chlorophenoxy acid herbicides
Nitrogen-containing pesticides
Other individual pesticides or groups (e.g., phenyl urea herbicides)
Other Contaminants
4-Methylimidazole (4-MEI)
Heavy metals by ICP-MS (arsenic, cadmium, lead, mercury)
Aflatoxins by HPLC (B1, B2, G1, G2)
Aflatoxins by HPLC (B1, B2, G1, G2, Ochratoxin A, and Zearalenone)
Aflatoxins (M1)
Acrylamide by LC-MS/MS
Residual solvents
Residual Solvents (USP<467> Class 1, 2a, 2b, and 3)
Melamine and cyanuric acid by LC-MS/MS
Bisphenol A (BPA)
Polycyclic aromatic hydrocarbons (PAH)
Compendial Testing Services

Raw Material/Ingredient Analysis
At Covance, we have been performing monograph analyses of raw materials and ingredients for over 30 years. In
accordance with the USP, EP, BP, JP, CP, JECFA and many others, our database of over 400 different raw materials and
capacity to set up more provides you with the analytical services you need for your raw materials testing.
To expedite your analysis, please provide the following information in advance:

• Quarterly or yearly list of raw materials
• Notice of shipment
• Analysis request form (available on-line or from Client Services)
This testing is compliant with GMP regulations (21 CFR Part 111) for ingredients in dietary supplements. For assays
compliant with 21 CFR Part 211 for human drug ingredients, please contact us.
Stability Studies: Stability and Shelf Life Evaluation

Covance provides comprehensive analytical services related to stability and shelf-life of products for the food and dietary
supplement industries. Stability is defined as the extent to which a product retains, within specified limits throughout its
period of storage and use, the same properties and characteristics possessed at the time of packaging. Understanding
product stability is an essential measure of the quality of a food or dietary supplement. Covance can custom design your
stability study from start to finish.
Study Design - Essential Information for Stability Goals of the Stability Study
Studies • Ensure the product is stable over its shelf-life
• Packaging materials • Establish or verify the shelf-life of the product
• Region the product is being sold • Aid in product research and development
• Product specifications • Provide data to support the quality of the product over
• Type of study (accelerated or real time) time
• Shelf-life anticipated • Fulfill any regulatory requirements
• Regulatory requirements (if applicable)
• Method applicability Due to the custom nature of designing a stability study,
please inquire on pricing and storage availability.
Testing Services for Stability Studies
• Chemical analysis (nutrients, active compounds)
Typical Stability Studies
• Physical properties (moisture, water activity, Testing Intervals
disintegration, hardness, etc.) Storage Study Condition (ICH guidelines)
• Organoleptic evaluation (appearance, taste, smell, color) Real Time 25°C/60% RH 3, 6, 9, 12, 18, 24 months
• Microbiological evaluation (safety assessment, Accelerated 40°C/75% RH 3, 6 months
antimicrobial preservative effectiveness) Alternate 30°C/65% RH 6, 9, 12 months
• Monograph testing (USP or International)
Phytochemicals
The terms phytochemicals and phytonutrients have been adopted as descriptors for certain organic components of plants
that are thought to promote human health. Unlike the traditional nutrients, these compounds are not “essential” for life from
a traditional standpoint but are being increasingly recognized as having beneficial health characteristics. As the popularity of
these products has increased with mainstream consumers, the use of a variety of dietary supplements, herbals/botanicals,
and functional foods has expanded and resulted in an intense and competitive marketplace. The ability to scientifically
substantiate claims for quality, purity, consistency, stability, safety and efficacy is an essential marketing tool and frequently
a regulatory requirement.

Method Development, Validation and Transfer

Responding to changes in market or regulatory requirements demands new or improved high quality scientific methods
engineered for greater efficiency, lower limits of detection, and improved reproducibility. As a partner in developing
solutions to method challenges, Covance offers the services of our Technical Development team on a contract basis. You
can leverage the team’s expertise across a wide range of matrices and technologies, including LC-MS/MS and UHPLC,
to address your analytical challenges. Projects are priced on a time/materials or project basis.
Typical projects we have completed include:

• New method development • Client method validation • New technology application
• Compendial/journal method set up • Existing method modernization • Client method variability resolution
• Matrix extensions (incl. validation)
Special Services
Colors
Artificial Colors
FD&C Red No. 2 (Amaranth) FD&C Green No. 3 (Fast Green FCF) FD&C Yellow No. 5 (Tartrazine)
FD&C Red No. 3 (Erythrosin B) FD&C Blue No. 1 (Erigluacine) FD&C Yellow No. 6 (Sunset Yellow FCF)
FD&C Red No. 4 (Ponceau SX) FD&C Blue No. 2 (Indigo Carmine) FD&C Yellow No. 10 (Quinoline Yellow)
FD&C Red No. 40 (Allura Red) Acid Orange 12 (Crocein Orange G)
Japanese Export Testing
Sennosides (UV)
Fenfluramine, N-nitroso-fenfluramine (HPLC)
p-Hydroxybenzoates (methyl, ethyl, propyl, butyl, isoproyl, isobutyl)
Thryoxine, triiodothyronine (HPLC)
Benzoic and sorbic acids
Prices for these tests are all product-dependent—specific pricing for your supplement product can be obtained by calling us at
608-242-2712, Ext. 8311-4170 or by faxing your ingredient list and request to 608-242-7903.
Botanicals
For centuries, people have used herbs and other botanicals for their healthful effects, possibly due to their phytochemical
composition, and in recent years, the use of these products has experienced a rise in popularity among consumers. This has
resulted in corresponding regulatory initiatives and the accelerated need for analytical methods for botanical products.
Several regulatory initiatives, including changes to current Good Manufacturing Practices (cGMP), address identification
testing for botanical ingredients.
Food Contact Materials (Migration Compliance)

The potential for migration of indirect additives and contaminants from food contact material is a concern for both food
products and dietary supplements. As the design and use of innovative packaging methods change, there is a corresponding
effect on the regulatory and experimental guidelines for determining the impact of packaging components on the safety of
food and dietary products. In the case of dietary supplements and botanicals, migration could potentially have a negative
effect on the efficacy of the active ingredient. When evaluating the safety of packaging materials there are three key aspects
to be considered: the source of the materials, the nature of the manufacturing process, and the conditions of use.
• Total and chloroform-soluble extractables from food • End use testing verifying compliance with 21 CFR
packaging Parts 172-181
• Recycled and degradable packaging • Conditions of use A-J
• Retort pouch barrier integrity • Experience with FCN and EU recommendations
• Pigment and dye migration from consumer products
• Benzene in polymeric packaging materials For guidance, please refer to: www.packaginglaw.com
• Microwave packaging
Dietary Supplements
Covance has over 75 years of experience in food analysis and has built a reputation for scientific and regulatory
expertise, responsiveness to customer needs and the capacity to turn around samples quickly and efficiently. As a full-service
laboratory, we analyze food, dietary supplement and biotechnology products.
Product/Analyte Testing Method

5-Hydroxytryptophan (5-HTP)................................................................................................................................ HPLC
Acetyl Cysteine......................................................................................................................................................... HPLC
Allicin (garlic)........................................................................................................................................................... HPLC
alpha Lipoic Acid (USP)........................................................................................................................................... HPLC
Anthocyanins (bilberry, elderberry, cranberry, fruits)....................................................................................................VIS
Anthocyanins............................................................................................................................................................ HPLC
beta Glucan..................................................................................................................................................................VIS
Bilberry (Anthocyanins)...............................................................................................................................................VIS
Black Currant Oil (linoleic acid and GLA).................................................................................................................GC
Caffeine, Theobromine, Theophylline....................................................................................................................... HPLC
Capsaicin/Capsaicinoid (Scoville Heat).................................................................................................................... HPLC
Catechins (EGCG, GCG, catechin, EGC, EC, ECG, GC, CG) + gallic acid........................................................ HPLC
Chlorogenic Acid...................................................................................................................................................... HPLC
Chondroitin Sulfate by CPC (USP)........................................................................................................................Titration
Coenzyme Q10......................................................................................................................................................... HPLC
Conjugated Linoleic Acid (CLA)................................................................................................................................GC
Cranberry (quinic, malic, citric acids)........................................................................................................................ HPLC
Creatine..................................................................................................................................................................... HPLC
Cryptoxanthin........................................................................................................................................................... HPLC
DHEA or 7-KETO DHEA
(Dehydroepiandrosterone) or (3-acetyl-7-oxo-dehydropiandrosterone)................................................................ HPLC
Dong Quai (ferulic acid)........................................................................................................................................... HPLC
Echinacea spp. (caftaric, chlorogenic, echinacoside, chicoric)..................................................................................... HPLC
Echinacea spp. [total phenolics as gallic acid equivalents (GAE)].................................................................................VIS
Elagic Acid................................................................................................................................................................ HPLC
Ephedrine Alkaloids
Ephedra or Ma Huang......................................................................................................................................LC-MS/MS
Evening Primrose Oil (linoleic acid and GLA)...........................................................................................................GC
Fo-ti Powder (trans-resveratrol)................................................................................................................................ HPLC
Fructan (AOAC 999.03)
Oligofructans, Fructo-Oligosaccharides (FOS), Inulin from Chicory, Dahlia, Raftilose..........................................VIS
Fructan (AOAC 997.08).....................................................................................................................................HPAEC-PAD
Furfural...................................................................................................................................................................... HPLC
gamma Aminobutyric Acid (GABA)........................................................................................................................ HPLC
gamma Linoleic Acid (GLA).......................................................................................................................................GC
Garcinia cambogia (hydroxycitric acid)....................................................................................................................... HPLC

Dietary Supplements (continued)

Garlic (allicin)........................................................................................................................................................... HPLC
Ginkgo Biloba Flavonoids........................................................................................................................................ HPLC
Ginkgo Biloba Terpenoids........................................................................................................................................ HPLC
Ginseng, Panax or Korean (ginsenosides)................................................................................................................. HPLC
Glucosamine..................................................................................................................................................HPLC/HPAEC-PAD
Glycerol (glycerine)......................................................................................................................................................GC
Goldenseal (hydrastine and berberine)...................................................................................................................... HPLC
Grape Seed Extract
Catechins............................................................................................................................................................... HPLC
Total Polyphenols (Folin-Ciocalteu Method) ..........................................................................................................VIS
Proanthocyandins (polyphenols - catechins)...................................................................................................... VIS/HPLC
trans-Resveratrol.................................................................................................................................................... HPLC
Green Tea Catechins................................................................................................................................................. HPLC
Griffonia Seed Extract (5-Hydroxytryptophan) 5-HTP........................................................................................... HPLC
Guarana (theobromine, theophylline, caffeine)......................................................................................................... HPLC
Hydroxycitric Acid (garcinia extract)........................................................................................................................ HPLC
Isoflavones (daidzein, glycitein, genistein, daidzin, glycitin, genistin)....................................................................... HPLC
Kudzu (isoflavones)................................................................................................................................................... HPLC
Lutein........................................................................................................................................................................ HPLC
Lycopene................................................................................................................................................................... HPLC
Marine Lipids (EPA/DHA)........................................................................................................................................GC
Methylsulfonylmethane (MSM)..................................................................................................................................GC
Milk Thistle (silychristin, silydianin, silibibin A & B).............................................................................................. HPLC
Omega-3 Fatty Acids...................................................................................................................................................GC
ORAC, Total (includes hydrophilic and lipophilic)..................................................................................................... FL
Oxalic acid................................................................................................................................................................. HPLC
PABA (para-aminobenzoic acid)............................................................................................................................... HPLC
Phenolic Acid (Phenylproponoids) Caffeic, p-Coumaric, Sinapic, and Ferulic Acids.............................................. HPLC
Phospholipids (Lecithin Phospholipids)
Phosphatidic acid, phosphatidylethanolamine, phosphatidylcholine, phosphatidylserine, phosphatidylinositol.... HPLC
Phytic Acid................................................................................................................................................................ HPLC
Phytosterols (stigmasterol, beta sistosterol, campesterol, brassicasterol).......................................................................GC
Policosanol (octacosanol, triacontanol, tetracosanol, hexacosanol)...............................................................................GC
Polygonum cuspidatum (trans-resveratrol)................................................................................................................... HPLC
Polydextrose...................................................................................................................................................HPLC/HPAEC-PAD
Polyphenols (Total Polyphenols) Folin-Ciocalteu Method..........................................................................................VIS
Polyphenols (Total) with % Solids
Tea and tea products for Tea Association members only..........................................................................................VIS
Pueraria (isoflavones)................................................................................................................................................ HPLC
Punicalagins.............................................................................................................................................................. HPLC
Dietary Supplements (continued)

Pygeum (beta sitosterol)...............................................................................................................................................GC
Pyruvate (pyruvic acid).............................................................................................................................................. HPLC
Quercetin.................................................................................................................................................................. HPLC
Rose Hips (ascorbic acid)............................................................................................................................................. FL
Royal Jelly 10-HDA (10-hydroxy-2-decanoic acid)................................................................................................. HPLC
Rutin......................................................................................................................................................................... HPLC
trans-Resveratrol....................................................................................................................................................... HPLC
SAMe (S-adenosylmethionine)................................................................................................................................. HPLC
Saw Palmetto (phytosterols + fatty acids).....................................................................................................................GC
Scoville Heat (capsaicin and capsaicinoid)................................................................................................................ HPLC
Soy Isoflavones.......................................................................................................................................................... HPLC
Stinging Nettles (5-hydroxytryptophan)................................................................................................................... HPLC
Theobroma cacoa (caffeine, theobromine, theophylline).............................................................................................. HPLC
Tocotrienols (alpha, beta, gamma, delta)................................................................................................................... HPLC
Tomato (lycopene)..................................................................................................................................................... HPLC
Ubiquinone (Coenzyme Q10)................................................................................................................................... HPLC
Valerian Extract (valerenic acid)................................................................................................................................ HPLC
Yerba Mate (caffeine, theobromine, theophylline)..................................................................................................... HPLC
Zeaxanthin................................................................................................................................................................ HPLC

[ assay ]
assays
5-Hydroxytryptophan (5-HTP) Aflatoxins
Analyzed with free amino acid procedure Purpose: Applicable for the determination of aflatoxins B1,
B2, G1, and G2 in a wide range of matrices including
premixes, raw materials, food ingredients and finished feeds.
Acesulfame K
Analyzed with aspartame: see Aspartame Method facts:
• Sample size: 50 g
Acetic acid • Limit of quantitation: 0.5 ppb for each aflatoxin compound
see Organic acid profiles — option 2 • Precision: NA
• Method reference: AOAC 991.31
Acid detergent fiber Description: The sample is extracted with a mixture of
see Detergent fiber, acid methanol:water. The extract is diluted with water and a portion
is applied to an antibody affinity column. The column is
washed first with water to remove major interferences. The
Acrylamide aflatoxins are eluted with acetonitrile and the eluent is then
Purpose: Applicable for the determination of acrylamide in dried under a stream of nitrogen. The aflatoxins are derivitized
various foods. with acid to form the highly fluorescent hemi-acetal
compounds. The sample extract is then quantified utilizing
Method facts:
HPLC by comparison to standards of known concentration.
• Sample size: 5g
• Limit of quantitation: 10ppb Aflatoxin (M1)
• Precision: Varies with matrix Purpose: This method is applicable to the analysis of
• Method reference: United States Food and Drug aflatoxin M1 in fluid milk, infant formulas and other
Administration, Center for Food Safety and Applied matrixes
Nutrition Office of Plant & Dairy Foods and Beverages,
Method facts:
“Detection and Quantitation of Acrylamide in Foods”
(2002) • Sample size: 20 g of powder or 100 mL of liquid
Description: Acrylamide is extracted from food samples • Limit of Quantitation: 0.1 ppb for M1 compound
with 0.1% formic acid. If product contains fat, petroleum • Precision: NA
ether is used prior to extraction with 0.1% formic acid. The
extract is purified through a solid phase extraction (SPE • Method Reference: Official Methods of Analysis for
cartridge. Acrylamide in the sample extract is determined AOAC International 2000.08
using LC-MS/MS. The acrylamide is determined using Description: The sample is extracted with a buffer solution.
least square linear regression with 13C3-labeled acrylamide The extract is diluted and a portion is applied to an
as an internal standard. Ions monitored for acrylamide are antibody affinity column. The column is washed first with
m/z 55, 44, and 27 and for the internal standard m/z 58. water to remove major interferences. The aflatoxins are
The ratio of peak areas for m/z 55 (acrylamide) and m/z 58 eluted with acetone and the eluent is then dried under a
(internal standard) are compared to those for standards over stream of nitrogen. The aflatoxins are derivatized acid to
the standard curve range. form the highly fluorescent hemi-acetal compounds. The
sample extract is then quantified utilizing high performance
liquid chromatography by comparison to standards of
known concentration.

[ assay ]
Aflatoxin, Ochratoxin A and Zearalenone by HPLC test is read with a Stat Fax reader. Optical densities of the
Purpose: Applicable for the analysis of aflatoxins B1, B2, controls form a standard curve which samples are plotted
G1, G2, ochratoxin A and zearalenone in a wide range of against to obtain concentrations of allergen protein in ppm.
foods, feeds and nutriceutical products.
Method facts: alpha Carotene

• Sample size: 50g see beta Carotene
• Limit of quantitation: 0.5 ppb for each aflatoxin
compound, 1.0 ppb for ochratoxin A and 10 ppb for alpha Lipoic acid (USP)
zearalenone Purpose: Applicable for the determination of alpha lipoic
• Precision: NA acid in tablets, capsules, premixes, and raw material.
• Method reference: AOAC 2005.08, AOAC 993.17, Method facts:
AOAC 999.07 • Sample size: 5 g
Description: The mycotoxins are extracted from the sample • Limit of quantitation: Most matrices - 0.025%
with a methanol/sodium bicarbonate mix. Following
extraction, the samples are cleaned with liquid-liquid • Precision: On a drink preparation, the RSD is 6.37%
partition and antibody affinity solid phase extraction • Method reference: The United States Pharmacopoeia
columns. The mycotoxins are determined by HPLC-
fluorescence detection, utilizing post column photochemical Description: Sample extraction is performed by mechanical
derivitization to determine aflatoxin B1 and G1. shaking. A portion of the extract is analyzed by HPLC and
UV detection, then quantitated by comparing the response
Alachlor to a standard curve of known concentration.
see USP 561 and EP pesticides
alpha Tocopherol
Aldrin and Dieldrin Purpose: Applicable to the determination of tocopherols
see USP 561 and EP pesticides and tocotrienols in foods, feeds, and nutritional products.
This method may also be used at the supervisor’s discretion
for matrices outside of the intended scope.
Allicin (garlic)
see Garlic Method facts:
• Sample size: 30g in food products, 10g in supplements and
premixes, 50g in feeds
Almond Protein by ELISA
Purpose: Allergens are proteins in food that can create an • Limit of quantitation: 0.500mg/100g, but may vary with
immune response in sensitive individuals. Clear ingredient sample matrix
labeling of food products, by manufacturers, aids in • Precision: On an infant formula matrix, the RSD is 6.10%
protection from accidental ingestion by those individuals. • Method reference: Speek, A.J., Schijver, J., and Schreurs,
Testing for the presence of these proteins ensures the food is W.H.P., Journal of Food Science, 50: 121-124 (1985)
free of the allergen at a potentially harmful level. (Modified)
Method facts: Cort, W.M., Vincente, T.S., Waysek, E.H., and Williams,
• Sample Size: 50g B.D., Journal of Agricultural Food Chemistry, 31: 1330-
• Limit of Quantitation: 2.5 ppm 1333 (1983) (Modified)
• Method Reference: Veratox ® Quantitative Almond McMurray, C.H., Blanchflower, W.J., and Rice, D.A.,
Allergen Kits Neogen Corporation Journal of the Association of Official Analytical Chemists,
63: 1258-1261 (1980) (Modified)
Description: Allergen Protein is extracted with a buffered
Description: The product is typically saponified to break
salt solution, allowed to settle then placed in an antibody
down the fat and release the vitamins. The digest is then
(capture) coated microwell to bind if present. All unbound
extracted with organic solvent. Additionally, alternate
protein is washed away before a detector antibody (enzyme
extraction procedures which do not require saponification
labeled) is added. The detector antibody then binds to
may be utilized for specified matrices. The tocopherols and
bound protein if present. After another wash, and the
tocotrienols are quantitated by ultra or high performance
addition of a substrate, color develops as a result of bound
liquid chromatography (UHPLC or HPLC) using
detector antibodies. After the addition of a stop reagent, the
fluorescence detection.
[ assay ] A
Aluminum (ICP) Amino acid profile, free by AAA

see Inorganic analysis by ICP Purpose: Applicable to all samples for the determination
of amino acid content of 5-hydroxytryptophan (5HTP),
L-ornithine, L-theanine, L-cysteine, beta-alanine, and
Aluminum (graphite furnace)
N-acetyl cysteine.
Purpose: Applicable for the determination of aluminum in
foods, feeds, dietary supplements, and biological materials. Method facts:
Method facts:
• Sample size: 10g • Limit of quantitation: Most matrices - 0.1mg/g
• Limit of quantitation: Most matrices - 0.5 ppm • Precision: NA
• Precision: On a rice flour sample matrix with a certified • Method reference: AOAC Method 982.30
value of 4.4ppm, the RSD is 5.85% Description: The sample is extracted in a mild acid.
• Method reference: Standard Methods for the Examination Determination is by an amino acid analyzer.
of Water and Wastewater, 20th Ed., Method 3113: 3-20-
3-27, American Public Health Association, Washington, Amino acid profile, free by HPLC
DC (1998) (modified) Purpose: Applicable to all samples for the following
amino acids: L-aspartic acid, L-glutamic acid, L-serine,
Description: An appropriately sized sample is wet-ashed with L-histidine, glycine, L-threonine, L-arginine, L-alanine,
nitric acid, hydrofluoric acid, and 30% hydrogen peroxide using L-tyrosine, L-cystine, L-valine, L-methionine,
open vessel microwave digestion. The amount of aluminum L-phenylalanine, L-isoleucine, L-leucine, L-lysine, proline,
is determined by comparing the signal of the unknown L-asparagine, L-glutamine, L-citrulline, L-tryptophan, and
sample, measured by the graphite furnace atomic absorption hydroxyproline.
spectrophotometer, with the signal of the standard solutions.
A rhodium/magnesium matrix modifier is used in the analysis. Method facts:
• Limit of quantitation: Most matrices - 0.1mg/g
Amino acid profile (AOAC), total
• Precision: On a corn sample, examples of typical RSDs are
Purpose: Applicable to all samples for the following amino
as follows: proline 4.8%, glutamic acid 5.0%, histidine
acids: Alanine, arginine, aspartic acid (including asparagine),
4.6%, aspartic acid 3.1%.
cystine, glutamic acid (including glutamine), glycine,
histidine, isoleucine, leucine, lysine, methionine, • Method reference: Henderson, J.W., Ricker, R.D.,
phenylalanine, proline, serine, threonine, tyrosine, Bidlingmeyer, B.A., Woodward, C., “Rapid, Accurate,
tryptophan, valine. (An accurate protein value is Sensitive, and Reproducible HPLC Analysis of Amino
recommended to perform these analyses.) Acids, Amino Acid Analysis Using Zorbax Eclipse-AAA
columns and the Agilent 1100 HPLC,” Agilent
Method facts: Publication, 2000
• Sample size: 5g (total) or 10g (free)
R. Schuster, “Determination of Amino Acids in Biological,
• Limit of quantitation: Most matrices - 0.1mg/g Pharmaceutical, Plant and Food Samples by Automated
• Precision: On an infant formula matrix, examples of Precolumn Derivitization and HPLC”, J. Chromatogr.,
typical RSDs are as follows: leucine - 1.9%, aspartic - 1.6%, 1988, 431, 271-284
glutamic - 1.5%, alanine - 1.7%
Description: The sample is extracted in acid. Determination
is by high-performance liquid chromatography (HPLC) with
Description: The sample is hydrolyzed in hydrochloric acid fluorescence or diode array detection. The primary amino
(HCl) and adjusted to pH 2.2 for all amino acids except acids are derivitized with fluorenylmethyl chloroformate
tryptophan. Tryptophan samples are hydrolyzed in sodium before injection.
hydroxide and adjusted to pH 5.2. Individual amino acids
are determined using an automated amino acid analyzer. Amino acid profile (HPLC), total
Cystine and methionine (AOAC) only Purpose: Applicable to all samples for the following amino
Tryptophan (AOAC) only acids: alanine, arginine, aspartic acid (including asparagine),
cystine (including cysteine), glutamic acid (including

[ assay ]
glutamine), glycine, histidine, hydroxyproline, isoleucine, Description: The sample is diluted in isooctane and read on
leucine, lysine, methionine, phenylalanine, proline, serine, a spectrophotometer. The sample is then spiked and
threonine, tyrosine, tryptophan, and valine. re-read on a spectrophotometer to determine the amount of
aldehydes in the sample.
Method facts:
• Sample size: 10g
• Limit of quantitation: Most matrices - 0.1 mg/g Anthocyanins, total
Purpose: Applicable for the determination of total anthocyanin
• Precision: On an infant formula, examples of typcial RSDs
content in blueberry, cranberry, bilberry, and other fruit
are as follows: leucine 1.4%, aspartic acid 1.8%, tyrosine
extracts in tablets, capsules, premixes, and raw material.
2.5%, methionine 2.9%.
• Method reference: Henderson, J.W., Ricker, R.D., Method facts:
Bidlingmeyer, B.A., Woodward, C., “Rapid, Accurate, • Sample size: 5 g
Sensitive, and Reproducible HPLC Analysis of Amino • Limit of quantitation: <0.01%
Acids, Amino Acid Analysis Using Zorbax Eclipse-AAA • Precision: Varies with matrix
columns and the Agilent 1100 HPLC,” Agilent
• Method reference: Givisti, M.M., Molar Absorptivity and
Publication, 2000
Color Characteristics of Acylated and Non-Acylated
Barkholt and Jenson, “Amino Acid Analysis: Pelrgonidin-based Anthocyanins, J. Agri and Food
Determination of Cystine plus Half-Cystine in Proteins Chemistry, 47(11):4631-4637 (1999)
after Hydrochloric Acid Hydrolysis with a Disulfide
Compound as Additive,” Analytical Biochemistry, 1989, Description: Sample extraction is performed with water and
177, 318-322 sonication. Portions of that extract are subjected to buffers
with differing pH. Aliquots are analyzed for absorbance on
R. Schuster, “Determination of Amino Acids in Biological,
a spectrophotometer. The difference in absorbance and the
Pharmaceutical, Plant and Food Samples by Automated
molar absorbtivity coefficient of cyanidin-3-glucoside
Precolumn Derivitization and HPLC”, J. Chromatogr.,
determine total anthocyanin content.
1988, 431, 271-284
Description: The samples are hydrolyzed in 6 N hydrochloric Anthocyanins by HPLC

acid for 24 hours at approximately 110°C. Phenol is added to Purpose: Applicable for the determination of anthocyanin
the 6N hydrochloric acid to prevent halogenation of tyrosine. content in blueberry, cranberry, bilberry, and other fruit
Cystine and cysteine are converted to S-2- extracts in tablets, capsules, premixes, and raw material.
carboxyethylthiocysteine by the addition of dithiodipropionic
acid. Tryptophan is hydrolyzed from proteins by heating at Method facts:
approximately 110°C in 4.2 N sodium hydroxide. • Sample size: 5 g
The samples are analyzed by HPLC after pre-injection • Limit of quantitation, most matrices: <10ppm
derivitization. The primary amino acides are derivitized with • Precision: Varies with matrix
o-phthalaldehyde (OPA) and the secondary amino acids are • Method reference: Indena Method 29/04/LRA1-00
derivitized with fluorenylmethyl chloroformate (FMOC)
before injection. Description: Samples are extracted in an acidic methanol
solution with sonication. Separation is achieved with
reversed-phase HPLC and a C18 column with detection set
p-Anisinide value at 535 nm. Twenty-one anthocyanins and anthocyanidins
Purpose: Applicable for the determination of p-anisinide are identified and quantified by comparing to cyanidin
value in all normal fats and oils. This method determines chloride and cyanidin-3-O-glucoside, respectively. The final
the amount of aldehydes in animal and vegetable oils. result for each compound is then converted by the molecular
weight ratio.
Method facts:
Arsenic
• Limit of quantitation: NA
see Inorganic analysis by ICP-MS
• Precision: NA
• Method reference: AOCS Cd 18-90 Ascorbic acid
see Vitamin C
[ assay ] A-B
L-ascorbyl-2-phosphate Description: Samples are weighed, then dissolved in a

(2-phospho-L-ascorbic acid) phosphate buffer/methanol solution. The solutions are
Purpose: Applicable to the quantitation of L-ascorbyl-2- injected on a HPLC equipped with a UV detector. Samples
phosphate in foods, feeds, and premixes. Also applicable are calculated against standards of known concentration.
to the various salts of the acid. This method does not
determine the di- or tri-phosphate forms of ascorbic acid. BHA/BHT (antioxidants)
Method facts: Purpose: Purpose: Applicable for the determination of
• Sample size: 25g Butylated Hydroxyanisole (BHA)/Butylated Hydroxytoluene
(BHT)/TBHQ.
• Limit of quantitation: 50mg/kg
• Precision: 2.25% on a dry dog food control Method facts:
• Method reference: Simultaneous HPLC Analysis of
L-Ascorbic Acid, L-Ascorbyl-2-Sulfate, and L-Ascobyl-2- • Limit of quantitation: Varies with matrix
Polyphosphate, Journal of Liquid Chromatography and • Precision: 5% for most matrices
Related Technology, 19(19):3105-3118 (1996), modified • Official Methods of Analysis of AOAC
Description: The sample is extracted with an aqueous INTERNATIONAL (2000) 17th Ed., AOAC
phosphate buffer solution. The extract is then filtered, pH INTERNATIONAL, Gaithersburg, MD, USA, Official
adjusted, and injected onto an HPLC system equipped with Method 968.17 (Modified)
a Phenomenex, Inertsit ODS-2, 250 mm x 4.6 mm, 5 µL Description: The lipid is extracted from food and feed
column, held in an oven set at 30.0°C. The analyte is eluted samples and diluted. BHA, BHT and TBHQ are quantified
with a phosphate buffer, acetonitrile, and ethanol mobile by gas liquid chromatography (GLC).
phase. The amount of L-ascorbyl-2-phosphate is detected
using an ultraviolet absorbance detector.
Benzene
Purpose: Applicable for the determination of benzene in
Ash most matrices.
Purpose: Applicable for the determination of ash in most
foods and feeds. Method facts:
Method facts:
• Limit of quantitation: Most matrices - 0.02 mg/100g
• Sample size: Sample size: 5g
• Precision: Varies with matrix
• Limit of quantitation: Most matrices - 0.1%
• Method Reference: Bertsch, Brian, “Developing One
• Precision: On a dog food matrix, the RSD is 5.0% Universal Method for Residual Solvents Using the New
• Method reference: AOAC 923.03 Teledyne Tekmar HT3 Headspace Sample Introduction
System,” Application Note (Teledyne Tekmar), Document
Description: Organic matter is burned off by igniting the #HT3-001, September 2005
sample at 550°C in an electric furnace. The remaining material
is determined gravimetrically and referred to as ash. Description: The samples are introduced to a gas
chromatograph mass spectrometer (GCMS) via a Tekmar
Aspartame and Acesulfame K, DKP HT3 purge-and-trap headspace autosampler.
and Saccharin
Purpose: Applicable for the determination of aspartame, Benzoic acid (sodium benzoate), sorbic acid
diketopiperazine (DKP), acesulfame K and saccharine in (potassium sorbate)
food and beverages. It may not be applicable for samples see Organic acid profiles - option 1
with a high protein content.
Berberine
Method facts: see Goldenseal
• Limit of quantitation: Most matrices - 50ppm beta Carotene
• Precision: Varies with matrix Purpose: Applicable for the determination of alpha and beta
carotene in food, feeds, and dietary supplements.
• Method reference: Prodolliet, J., and Bruelhart, M.,
Determination of Aspartame and its Major Decomposition Method facts:
Products in Foods. J. of AOAC, 76(2) (1993) • Sample size: 10 g

[ assay ]
• Limit of quantitation: Most matrices - 0.02 mg/100 g Description: The sample is extracted with either water,
• Precision: On an infant formula matrix, the RSD is 7.4% dilute sodium hydroxide (NaOH), or sulfuric acid (H2SO4).
• Method reference: AOAC 941.15 The amount of biotin is determined by comparing the growth
response of the sample, using the bacteria Lactobacillus
Q uackenbush, F. W., Journal of Liquid Chromatography,
plantarum, with the growth response of a biotin standard.
10:643-653 (1987)
This response is measured turbidimetrically.
Description: Food products are de-esterified with potassium
hydroxide and extracted with hexane. Juice products are
Bisphenol A
blended with alcohol and extracted. Nutraceutical products
Purpose: Applicable to analysis of Bisphenol A (BPA) in
are enzymatically treated and then extracted. The extracts infant formula, infant formula concentrates, baby food and
are injected on a reverse-phase HPLC system equipped with various food matrices. BPA as part of a packaging material
UV detection and compared to a standard curve. analysis is available upon request.
beta Glucan Method facts:

Purpose: Applicable for the determination of beta glucan • Sample size: 10 g
in food products, with the exception of yeast and mushroom • Limit of quantitation: 0.5ppb in infant formula; LOQ
products. varies by sample matrix
Method facts: • Precision: NA
• Sample size: 2g • Method reference: Determination of Bisphenol A in
• Limit of quantitation: Most matrices - 0.5% Liquid Infant Formula by Solid Phase Extraction with
• Precision: On a cereal sample matrix, the RSD is 4.81% Acetic Anhydride Derivatization and Gas Chromatography-
• Method reference: AOAC 995.16 Mass Spectrometry, Minister of Health Canada (2008).
Description: beta Glucan is hydrolyzed to oligosaccharides Description: The sample is diluted with water and
by lichenase. These oligosaccharides are reduced to glucose by acetonitrile, then mixed and centrifuged. The supernatant
beta glucosidase. The glucose is reacted with a glucose is diluted with a phosphate buffer and passed through a
oxidase/peroxidase reagent mixture, and the reaction product solid phase extraction column (Varian 1210-2052, 500mg
is determined spectrophotomerically at 510 nm. C18, 6mL). The column is eluted with an aqueous solution
of acetonitrile and the extract concentrated. The sample is
derivatized and extracted into iso-octane and MTBE, then
Bilberry concentrated to near dryness and diluted with toluene. The
see Anthocyanins, total BPA is quantified and confirmed by GC/MS on an HP-5ms
see Anthocyanins by HPLC column.
Biotin Black currant oil (linolenic acid and GLA)

Purpose: Applicable for the determination of biotin in most see Fatty acid profiles
foods, feeds and dietary supplements.
Method facts: Bomb calorimetry

• Sample size: 2g Purpose: Applicable for the determination of total calories
• Limit of quantitation: Most matrices - 0.005 mcg/g in foods and feeds.
• Precision: On an infant formula matrix, the RSD is 7.1% Method facts:
• Method reference: Wright & Skeggs, Procedures of the • Sample size: 2g for solids, 7g for liquids
Society of Experimental Biology and Medicine, 56:95 (1944) • Limit of quantitation: Most matrices - 5 calories/g
Schiner & Deritter, “Biotin Content of Feed Stuffs,” Journal • Precision: On a benzoic acid matrix, the RSD is 0.67%
of Agricultural Food Chemistry, 23:1157-1162 (1975)
• Method reference: Metals Energy Mining Agriculture
Methods of Analysis for Infant Formulas, Infant Formula Geology, AC-350 Instruction Manual, LECO Corporation,
Council (1985) St. Joseph, MI, pp 5-26 through 5-29 (2002)
Journal of the AOAC, 49:882, (1996)
Description: Calories are determined by burning a weighed
[ assay ] B-C
sample in an atmosphere of oxygen inside a bomb submerged Description: The sample is extracted with boiling water.
in a measured quantity of water. The temperature rise of the A portion of the extract is injected onto an HPLC system
water, resulting from the combustion of the sample, is used with UV detection set at 272 nm. The areas of the peaks
to calculate the number of calories liberated. are determined and compared with those obtained from
injected standards.
Bromide, Inorganic Calcium (ICP)

Bromine Containing Fumigants Determined as Total see Inorganic analysis by ICP
Inorganic Bromide
Purpose: Applicable to the determination of inorganic Calories, by calculation

bromide in dietary supplements (raw materials, ingredients Purpose: Used to calculate calories in foods and feeds.
and soft gels), high fat matrices and plant materials. Method facts:
• Sample size: NA
Method facts:
• Sample size: 25g • Limit of quantitation: <1.7 calorie
• Limit of Quantitation: Dry samples, LOQ is 5 μg/g. Moist • Precision: NA
or liquid samples, LOQ is 1 μg/g • Method reference: 21 CFR 101.9
• Method Reference: Community Reference Laboratory for Description: Calories = (Protein(g) x 4) +
Single Residue Methods, CVUA, Stuttgart, Schaflandstr. (Carbohydrates(g) x 4) + (Fat(g) x 9)
3/2, 70736 Fellbach, Germany
T. Stijve, Gas Chromatographic Determination of Calories from fat, by calculation

Purpose: Used to calculate calories from fat.
Inorganic Bromide Residues - a Simplified Procedure,
Dtsch. Lebenm. Rundsch. 77 99-101 (1981) Method facts:
• Sample size: NA
Deutsche Forschungsgemeinschaft (DFG), Manual of
• Limit of quantitation: <0.9 calorie from fat
Pesticide Residue Analysis, Volume I, by Verlag Chemie,
1987. The bromide method has the code S 18. ISBN • Precision: NA
3-527-27010-8 • Method reference: 21 CFR 101.9
Description: The samples are homogenized and suspended Description: Calories from fat = Fat(g) x 9
in an acidified aqueous solution of propylene oxide, with
bromide being simultaneously extracted and derivatized into
Capsaicin/Capsaicinoid (Scoville heat)
1-bromo-2-propanol and 2-bromo-1-propanol. The
Purpose: Applicable for the determination of pungency levels
derivatives are partitioned into ethyl acetate and determined
in capsicums and their oleoresins.
by GC-ECD without further cleanup.
Method facts:
Cadmium (ICP or ICP-MS)
see Inorganic analysis by ICP or ICP-MS • Limit of quantitation: Most matrices ~10ppm
• Precision: On a powder matrix, the RSD is 5.40%
Caffeic acid • Method reference: AOAC 995.03
see Phenolic acid
Description: The sample is extracted in warm ethanol.
The extract is filtered and then injected using HPLC
Caffeine, theobromine, theophylline
equipped with an ultraviolet detector. Results are typically
Purpose: Applicable for the determination of caffeine,
theobromine, and theophylline in various food and supplements. expressed in Scoville units.
Method facts: Carbamate pesticides (LC-MS/MS)

Purpose: Applicable but not limited, to the analysis of
• Limit of quantitation: Most matrices - 50 ppm selected carbamate pesticides and related metabolites in food
• Precision: On a cocoa powder matrix, the RSD is 5.42% and dietary supplement products and raw materials.
for caffeine and 3.62% for theobromine
Method facts:
• Method reference: Journal of Food Science,
48:745-747 (1983) • Sample size: 25g

[ assay ]
• Limit of quantitation: Listed in the table below Dioxacarb 0.01

• Precision: Varies with matrix Ethiofencarb 0.01
• Method reference: Fenobucarb 0.01
1AOAC Official Method 2007.01, Pesticide Residues in Fenoxycarb 0.01

Foods by Acetonitrile Extraction and Partitioning with Indoxacarb 0.02
Magnesium Sulfate Iprovalicarb 0.01
2
CEN Standard Method EN 15662: Food of plant origin Isoprocarb 0.01
– Determination of pesticide residues using GC-MS and/ Methiocarb 0.01

or LC-MS/MS following acetonitrile extraction/ Methiocarb sulfone 0.01
partitioning and clean-up by dispersive SPE – Methiocarb sulfoxide 0.01
QuEChERS method Methomyl 0.02
3
Anastassiades, M.; Lehotay, S.J.; Stajnbaher, D.; Schenck, Metolcarb 0.01
F.J. Fast and easy multiresidue method employing Oxamyl 0.01
acetonitrile extraction/partitioning and “dispersive solid- Promecarb 0.01
phase extraction” for the determination of pesticide Propham 0.01
residues in produce. Journal of AOAC Propoxur 0.01
INTERNATIONAL 2003, 86, 412-431 Thiofanox 0.05
4
Lehotay, S.J.;Mastovska, K; Lightfield, A.R. Use of
buffering and other means to improve results of Carbohydrates, total, by calculation
problematic pesticides in a fast and easy method for Purpose: Applicable for the determination of carbohydrates
residue analysis of fruits and vegetables. Journal of AOAC in foods and feeds.
INTERNATIONAL 2005, 88, 615-629
5 Method facts:
Lehotay, S.J.;Mastovska, K.; Yun, S. J. Evaluation of two
• Sample size: Sample size: NA.
fast and easy methods for pesticide residue analysis in
fatty food matrixes. Journal of AOAC • Limit of quantitation: Most matrices - <0.4%
INTERNATIONAL 2005, 88, 630-638 • Precision: NA
6
Mastovska, K.; Dorweiler, K.; Lehotay, S.J.; Wegscheid, J.; • Method reference: 21 CFR 101.9
Szpylka, K. Pesticide multiresidue analysis in cereal grains
using modified QuEChERS method combined with Description:
automated direct sample introduction GC-TOFMS and Carbohydrate = 100 - [protein(g) + fat(g) + ash(g) + moisture(g)]
UPLC-MS/MS techniques. J. Agric. Food Chem., 2010,
58, 5959-5972 L-carnitine
Description: The sample extraction and clean-up are based Purpose: Applicable for the quantitation of L-carnitine in
on the QuEChERS method.1-6 The final extract is analyzed infant formulas, powders, and premixes.
using liquid chromatography-tandem mass spectrometry
Method facts:
(LC-MS/MS). Typical reporting limits are listed in the
table below. Other reporting limits (at or above LOQ for a
given carbamate-matrix combination) can be used based on • Limit of quantitation: 0.50mg/100g
client requests: • Precision: 4.15% on an infant formula control
• Method reference: Starey et al.; Journal of AOAC
Carbamate Pesticide LOQ (mg/kg)
INTERNATIONAL vol. 91, No. 1, 2008 (modified)
Aldicarb 0.01
Aldicarb sulfone 0.01 Description: The sample is diluted in water and a
Aldicarb sulfoxide 0.01 deuterated analog of carnitine is added as an internal
Bendiocarb 0.01 standard. It is then filtered and injected onto a Waters
Butocarboxim 0.02 Symmetry C8, 50 mm x 2.1 mm, 3.5 µm column, held in an
Butoxycarboxim 0.01 oven set at 30.0°C. Three mobile phases are used in a
Carbaryl 0.01 gradient consisting of 0.1% heptafluorobutyric acid (HFBA)
Carbofuran 0.01 in water, 0.1% HFBA in methanol, and 80% water / 20%
Carbofuran, 3-hydroxy- 0.01
methanol. The amount of L-carnitine is determined using
HPLC-MS/MS.
Chlorpropham 0.01
[ assay ] C
Carotenoids • Method reference: Sakakibara, H., Honda, Y., Nakagawa, S.,

Option 1: alpha Carotene, beta Carotene, Lycopene Ashida, H., and Kanazawa, K., “Simultaneous Determination
Option 2: Cryptoxanthin, Lutein, Zeaxanthin of All Polyphenols in Vegetables, Fruits, and Teas,” Journal
Option 3: Other carotenoids of Ag Food Chem., 51(3):572-580 (2003)
Purpose: Applicable to the determination for the following
Description: Sample extraction is performed using a
carotenoids in foods, feeds, premixes, pharmaceutical and
buffered solution. A portion of that extract is analyzed on a
nutrition supplements: lycopene, α and β-carotene, free
HPLC using UV detection. Analytes are then quantitated
lutein, lutein esters, β-cryptoxanthin, or zeaxanthin. Vitamin
by comparing the response of a standard of known
A from carotenes may be calculated.
concentration.
Method facts:
• Sample size: 25g for food or feed products; 2g for
Chloride
supplements and premixes
Purpose: Applicable for the determination of acid-soluble
• Limit of quantitation: 0.0200 mcg/100g, but can vary by chloride in feeds, plants, food products, and tissues.
sample size
• Precision: Varies with matrix Method facts:
• Method reference: Official Methods of Analysis of AOAC
INTERNATIONAL, Current Ed., Method 2005.07, • Limit of quantitation: Most matrices - 200 ppm
AOAC INTERNATIONAL, Gaithersburg, MD, USA • Precision: On a cereal matrix, the RSD is 1.01%
(Modified). • Method reference: AOAC 963.05, 969.10, 971.27 (modified)
Official Methods of Analysis of AOAC
INTERNATIONAL, Current Ed., Methods 941.15, Description: Samples are weighed, double-deionized water
2005.07, AOAC INTERNATIONAL, Gaithersburg, MD, is added, and the solution is mixed thoroughly and made
USA (Modified) acidic with nitric acid. Chloride is determined
potentiometrically by titrating with a silver nitrate solution
Quackenbush, F. W., Reverse Phase HPLC Separation of to a predetermined endpoint.
cis- and trans-Carotenoids and it’s Application to Beta
Carotenes in Food Materials,” Journal of Liquid
Chromatography, 10: 643-653 (1987) (Modified) Chlorinated/Organophosphate pesticides
see Pesticides, chlorinated/organophosphate
Description: Low fat samples are extracted with alcohol and/
or tetrahydrofuran. Samples with a higher level of fat are
saponified and extracted with hexane. Each sample is then Chlorogenic acid
injected on a reverse phase high performance liquid Purpose: Applicable for the determination of chlorogenic acid
chromatography system (HPLC) with ultraviolet (UV) in raw material, extracts, and dietary supplement preparations.
detection. Quantitation is achieved with a linear regression
analysis. Method facts:
• Sample size: 5g
Catechins • Limit of quantitation: 50 ppm, 1ppm for liquids

Epigallocatechin, catechin, epigallocatechin gallate, • Precision: 5.02%
gallocatechin gallate, gallic acid, epicatechin, epicatechin • Method reference: Journal of Ag Food Chem, 51(3):572-580
gallate, gallocatechin, catechin gallate.
Description: The sample is extracted in a variety of ways
Purpose: Applicable for the determination of catechins depending upon the matrix. A portion of the extract is
in various matrices including extracts (e.g. grape seed), tea, injected into an HPLC using UV detection. Chlorogenic acid
fortified products, and premixes. is quantified by comparing the response to a known standard.
Method facts:
• Sample size: 5g Chlorophenoxy acid herbicides
• Limit of quantitation: Most matrices-100 ppm, liquids-1ppm see Herbicides, chlorophenoxy acid
• Precision: On a green tea matrix, the RSD is 4.09%

[ assay ]
Chlorophyll • Method reference: Glick, Journal of Biological Chemistry,

Purpose: Applicable to the determination of total chlorophyll 156:643 (1944)
content derived from plants.
Description: The sample is hydrolyzed with mild acid.
Method facts: The resulting solution is purified on an ion exchange
• Sample size: 25g column. The concentration of choline is determined
spectrophotometrically from a reineckate coupling reaction.
• Limit of quantitation: most matrices - 0.01mg/g
• Precision: NA
• Method reference: AOAC Official Methods of Analysis Choline (enzymatic)
of AOAC International (2006) 18th Edition, AOAC Purpose: Applicable for the determination of choline in
INTERNATIONAL, Gaithersburg, MD, USA, Official milk, infant formula, foods and beverages.
Method 942.04. (Modified)
Method facts:
Description: Chlorophyll is extracted from the sample with • Sample size: 5g
acetone. The sample extract is then washed with ethyl ether • Limit of quantitation: Most matrices - 25mg/100g
in a separatory funnel. The chlorophyll migrates to the ethyl • Precision: On an infant formula matrix, the RSD is 3.78%
ether layer and the remaining acetone is removed through a • Method reference: AOAC 999.14
series of water rinses. The ethyl ether extract is then read at
642.5 and 660 nm on a UV-VIS spectrophotometer to Description: The product is hydrolyzed at 70°C to release
determine the total chlorophyll content. bound choline. Following pH adjustment, residual choline
phospholipids are cleaved with phospholipase D and free
choline is subjected to choline oxidase with liberation
Cholesterol of peroxide. In the presence of peroxidase, phenol is
Purpose: Applicable for the determination of cholesterol in oxidized and a quinoneimine chromiphore is formed with
most matrices including foods, feeds, and fecal samples. 4-aminoantipyrine. Absorbance is measured and choline
content calculated by interpolation from a multilevel calibration.
Method facts:
• Sample size: 5g
• Limit of quantitation: 1mg/100g Chondroitin sulfate by CPC
• Precision: 2-5% depending upon matrix Purpose: Applicable for the determination of chondroitin
sulfate in tablets, premixes, capsules and other dietary
• Method reference: AOAC 994.10 supplements. Sources may include porcine, bovine, or shark.
Method facts:
Description: The sample is saponified using ethanolic • Sample size: 5g
potassium hydroxide. The unsaponifiable fraction that • Limit of quantitation: 25mg/g
contains cholesterol and other sterols is extracted with • Precision: on a tablet, the RSD is 1.87%
toluene. The toluene is evaporated to dryness and the • Method reference: USP
residue is dissolved in dimethylformamide (DMF). The
samples are derivitized to form trimethylsilyl ethers. The Description: Sample extraction is performed with water and
derivitized cholesterol is quantitatively determined by gas agitation. Extracts are analyzed on an auto-titrator equipped
chromatography using 5 a-cholestane as an internal standard. with a light emitting phototriode. Samples are calculated
against a standard curve of known concentrations.
Choline (chemical)
Purpose: Applicable for the determination of choline in Chromium (atomic absorption)
premixes, infant formula, dietary supplements, and feeds. Purpose: Applicable for the determination of chromium
in feeds, animal tissues, food products, plants, dietary
Method facts: supplements, water, soils, and minerals.
• Limit of quantitation: most matrices - 15.1mg/100g; Method facts:
liquid samples - 3.3mg/100g • Sample size: 15g
• Precision: On a powdered infant formula matrix, the • Limit of quantitation: Most matrices - 2 ppm
RSD is 6.31% • Precision: Based on a powdered drink control, the RSD is 1.90%
[ assay ] C
• Method reference: Analytical Methods for Atomic • FD&C Green No. 3 (Fast Green FCF)
Absorption Spectrophotometry, Perkin-Elmer: Norwalk, CT • FD&C Blue No. 1 (Erigluacine)
(2000)
• FD&C Blue No. 2 (Indigo Carmine)
AOAC 974.27
• Acid Orange 12 (Crocein Orange G)
Description: The sample can either be dry-ashed, wet-ashed, • FD&C Yellow No. 5 (Tartrazine)
or read directly. If dry-ashed, the sample is ashed at 500°C
• FD&C Yellow No. 6 (Sunset Yellow FCF)
±50° until ashing is complete. The resulting ash is treated with
concentrated hydrochloric acid, dried and redissolved in • FD&C Yellow No. 10 (Quinoline Yellow)
hydrochloric acid solution. If wet-ashed, the sample is Purpose: This method is designed to demonstrate the
digested on a hot plate with nitric acid, hydrochloric acid, absence of food coal tar colorants.
and/or hydrogen peroxide.
The amount of chromium is determined by comparing the Method facts:
signal of the unknown sample, measured by the atomic • Sample size: 10g
absorption (AA) spectrophotometer, with the signal of the • Limit of quantitation: 5 ppm
standard solutions. • Precision: NA
• Method reference: Canadian Food Inspection Agency;
Chromium (ICP or ICP-MS) HPLC Identification and Quantification of Synthetic
see Inorganic analysis by ICP or Inorganic analysis by Food Colours in Aqueous Media; LCAQ-016-05; 2009
ICP-MS (Modified)
Description: Samples are extracted in a basic solution and then

Citric acid injected on an HPLC equipped with a UV detector capable of
see Organic acid profiles - option 2 reading at multiple wavelengths. Samples are then calculated
against standards of a known concentration. Analytes detected:
FD&C Red No. 2, FD&C Red No. 3, FD&C Red No. 4,
Cobalt (ICP or ICP-MS) FD&C Red No. 40, FD&C Green No. 3, FD&C Blue No. 1,
see Inorganic analysis by ICP or Inorganic analysis by FD&C Blue No. 2, Acid Orange 12, FD&C Yellow No. 5,
ICP-MS FD&C Yellow No. 6, FD&C Yellow No. 10.
Coenzyme Q10 Conjugated linoleic acid (CLA)

Purpose: Applicable for the determination of coenzyme Purpose: This procedure is used to quantitate conjugated
Q10 in dietary supplements and raw material. linoleic acids in foods and feeds.
Method facts: Applicable for foods and feed containing at least 0.2% lipids.
• Sample size: 10g The method detects fatty acids having 8 to 22 carbon atoms
• Limit of quantitation: Most matrices - 5 mg/100 g as methyl esters. The conditions specified in this method are
• Precision: Based on a powdered drink control, the RSD is not suitable for determining epoxy or oxidized fatty acids that
1.90% have been polymerized.
• Method reference: AOAC 2008.07 Method facts:
Description: Coenzyme Q10 is extracted using alcohol and • Sample size: 4g, 1g if oil
hexane. The extract is then injected on a reverse-phase HPLC • Limit of quantitation: Most matrices - 0.1%
system using UV detection for quantitation. • Precision: The RSD value for 18:1 conjugated in an oil
control sample is 2.76%.
Colors, artificial • Method reference: AOAC 996.06 /AOCS Ce 1k-07/
• FD&C Red No. 2 (Amaranth) Ce 2-66
• FD&C Red No. 3 (Erythrosin B) Description: The lipid is extracted, saponified with 0.5N
• FD&C Red No. 4 (Ponceau SX) methanolic sodium hydroxide, and methylated with 14%
BF3-methanol. The resulting methyl esters of the fatty acids
• FD&C Red No. 40 (Allura Red) are extracted with heptane containing an internal standard.

[ assay ]
The methyl esters of the fatty acids are analyzed by gas Description: Cyanuric acid is extracted from tissue and
chromatography using external standards for quantitation. infant formula with a 50:50 acetonitrile:water extraction
solution, followed by centrifugation and SPE cleanup. The
compound is analyzed using a zwitterionic HILIC LC
Copper (ICP or ICP-MS) column. Electrospray ionization is used in the negative ion
see Inorganic analysis by ICP or ICP-MS
(cyanuric acid) mode. This procedure has been validated by
the FDA for the determination of cyanuric acid in tissues.
Coumaric acid The method has been validated in infant formula powder by
see Phenolic acids Covance. Two multiple reaction monitoring (MRM)
transitions are monitored for the compound. Isotope internal
standards are used to correct for any matrix effects. Fortified
Cranberry (quinic, malic, citric acids) test portions were within 85-105% recovery for the infant
see Organic acid profiles formula powder validation.
Cranberry Cysteine, free

see Anthocyanins, total see Amino acid profile, free by AAA
see Anthocyanins by HPLC
Cystine (AOAC)
Creatine see Amino acid profile (AOAC)
Purpose: Applicable for the determination of creatine and
creatinine in dietary supplements, sports nutrition products, etc.
Cystine, free
Method facts: see Amino acid profile, free
• Limit of quantitation: 0.5% DHEA or 7-KETO DHEA
• Precision: On a creatine capsule, the RSD is 1.87% Dehydroepiandrosterone or
• Method reference: Analytical Biochemistry, 214, pp278-283 3-acetyl-7-oxo-dehydropiandrosterone
Description: The sample is extracted with water. The extract Purpose: Applicable for the determination of DHEA or
is injected into an HPLC using UV detection. Results are 7 KETO DHEA in premixes and dietary supplements.
calculated against a standard of known concentration.
Method facts:
• Sample size: 5g
Cryptoxanthin • Limit of quantitation: 50 ppm
see Carotenoids
• Precision: 4.2%
• Method reference: Journal of AOAC INTERNATIONAL,
Cyanocobalamin 83(4):847-857
see Vitamin B12
Description: Extraction is conducted by sonicating with
Cyanuric acid (LC-MS/MS) methanol. Samples are analyzed by HPLC and quantified
Purpose: Applicable for the determination of cyanuric acid by comparison with a known standard.
using LC-MS/MS in food products.
Density
Method facts: Purpose: Applicable for the determination of specific gravity
• Sample size: 25g in liquid samples.
• Limit of quantitation: 0.5ppm Cyanuric Acid
Method facts:
• Precision: 5% RSD at 0.5ppm
• Sample size: 100mL
• Method reference: United States Food and Drug • Limit of quantitation: NA
Administration, Interim Method for Determination of
• Precision: On a water matrix, the RSD is 0.3%
Melamine and Cyanuric Acid Residues in Food using
LC-MS/MS: Version 1.0 Laboratory Information Bulletin • Method reference: USP <841>
No. 4422 (October 2008)
[ assay ] D-E
Description: A known volume of sample is weighed. The Description: Samples are extracted with chloroform-
weight of the sample per unit volume is calculated. methanol solution, yielding a lipid extract that contains the
total fat and olestra. The extracted lipid is hydrolyzed by
lipase, yielding unreacted olestra and fatty acid from the fat.
Detergent fiber, acid (ADF)
The fatty acids are precipitated as calcium soaps. Olestra is
Purpose: Applicable for the determination of acid detergent extracted from the insoluble soaps with hexane and
fiber in forages and feeds with low carbohydrate contents. discarded. The isolated soaps are converted back to fatty
acids with hydrochloric acid and extracted into hexane.
Method facts:
These fatty acids are converted to methyl esters with boron
• Sample size: 5g trifluoride-methanol solution and quantified by gas
• Limit of quantitation: Most matrices - 0.1% chromatography using an internal standard.
• Precision: On a dog food matrix, the RSD is 14.3%
• Method reference: United States Department of Agriculture Dissolution preparation (USP)
Forage and Fiber Analysis, Handbook #379.8, United States Purpose: This test is provided to determine compliance with
Department of Agriculture, Washington, D.C. (1970) the dissolution requirements where stated in the individual
monograph for a tablet or capsule dosage form, except where
Description: The protein, carbohydrate, and ash content is the label states that the tablets are to be chewed.
removed by treating samples with a boiling detergent solution
Method facts:
and filtering. The fats and pigments are removed via an
acetone wash leaving lignocellulose fraction in a frit, which • Sample size: 25 tablets or capsules
is determined gravimetrically. • Method reference: United States Pharmacopeia, <2040>,
United States Pharmacopeial Convention, Inc.: Rockville,
Detergent fiber, neutral (NDF) Maryland (current edition)
Purpose: Applicable for the determination of neutral Description: Unless otherwise stated in the individual
detergent fiber in forages and feeds. monograph, 6 dosage units are tested as directed under
Method facts: Dissolution <2040>. A stated volume of Dissolution
• Sample size: 5g Medium is placed into each vessel of the dissolution
apparatus specified in the individual monograph. Once the
• Limit of quantitation: 0.1%
Dissolution Medium is equilibrated to 37 ± 0.5°C, 1 tablet
• Precision: On commercial dog food matrix, the RSD is 11.1% or 1 capsule is placed into each of 6 vessels. The apparatus is
• Method reference: AACC 32.20 immediately operated at the rate specified in the individual
United States Department of Agriculture, Forage and monograph. Within the time interval specified, a specimen
Fiber Analysis, Agriculture Handbook #379.8, United is withdrawn from a specific zone of each vessel. Equal
States Department of Agriculture, Washington, D.C. (1970) volumes from each vessel are filtered and pooled into a
single container.
Description: The protein, carbohydrate, enzyme, and ash
content is dissolved by a boiling detergent solution and
filtered off. The fats and pigments are removed via an Dong quai (ferulic acid)
acetone wash leaving hemicellulose, cellulose, and lignin see Phenolic acids
fractions in a frit and determined gravimetrically.
EBDCs
Digestible fat Purpose: Applicable for the determination of EBDC
Purpose: Applicable to fat and saturated fat in Olestra residues in milk, eggs, nuts, feeds, crops, and formulations.
products, pure Olestra, and blended Olestra. This method screens for EBDC compounds using standards
from mancozeb, maneb, metiram, thiram, nabam, ziram, and
Method facts: zineb.
• Sample size: 5g
Method facts:
• Precision: On a potato chip the RSD is 9.28%
• Limit of quantitation: Most matrices - 2000 ppb
• Method reference: “Capillary Gas Chromatographic
• Precision: Based on spike recovery; most matrices are
Determination of Fat in Olestra Savory Snack Products,”
70-120%
Guererra III, Frank P., Pharmline Inc., personal
correspondence, April 1996 • Method reference: The Analyst, 6:782-787 ( July 1981)

[ assay ]
Description: EBDCs are quantitatively decomposed to Description: The ephedrine-type alkaloids are extracted
carbon disulfide using a headspace solvent layer procedure. from dietary supplements with methanol:water (80:20). The
The sample is weighed into a 125mL serum bottle and a amount of ephedrine-type alkaloids present in dietary
hydrochloric acid-stannous chloride solution is added. supplements is determined by liquid chromatography using
Iso-octane is volumetrically pipetted onto the sample and tandem mass selective detection (LC-MS/MS).
the serum bottle is sealed with a crimp top cap fitted with a
Teflon-coated silicone septum. The sample is heated at
approximately 80°C in a water bath for 1 hour and allowed Egg Protein by ELISA
to cool to room temperature. The sample is then centrifuged Purpose: Allergens are proteins in food that can create an
to separate it from the iso-octane layer. The iso-octane layer immune response in sensitive individuals. Clear ingredient
is then pipetted off the top of the sample and analyzed on a labeling of food products, by manufacturers, aids in
gas chromatograph using a flame photometric detector in protection from accidental ingestion by those individuals.
sulfur mode. Testing for the presence of these proteins ensures the food is
free of the allergen at a potentially harmful level.
Echinacea spp. Method facts:

Caftaric, chlorogenic, echinacoside, chicoric • Sample Size: 50g
• Limit of Quantitation: 2.5 ppm
Purpose: Applicable for the determination of echinacosides
in raw material or dietary supplements. • Method Reference: Veratox ® Quantitative Egg Allergen
Kits Neogen Corporation
Method facts:
• Sample size: 10g Description: Allergen protein is extracted with a buffered
• Limit of quantitation: 50 ppm (1ppm for liquids)
• Precision: Varies with matrix protein is washed away before a detector antibody (enzyme
• Method reference: Journal of Ag Food Chem., 51(3): 572- labeled) is added. The detector antibody then binds to
580 bound protein if present. After another wash, and the
Description: Extraction technique may vary with matrix. The detector antibodies. After the addition of a stop reagent, the
extract is injected into an HPLC with UV detection. The test is read with a Stat Fax reader. Optical densities of the
echinacosides are quantified by comparison to known standards. controls form a standard curve which samples are plotted
against to obtain concentrations of allergen protein in ppm.
Elderberry
see Anthocyanins, total Erythritol
see Anthocyanins by HPLC
see Sugar alcohols
Ephedrine alkaloids Evening primrose oil

Ephedra or Ma Huang
Linolenic acid and GLA
Purpose: Applicable to the analysis of the ephedrine-type see Fatty acid profiles
alkaloids norephedrine, norpseudoephedrine, ephedrine,
pseudoephedrine, methylephedrine, and
methylpseudoephedrine. This method is applicable to
Ethanol and Methanol
dietary supplements, raw ephedra herb, ephedra extracts, Purpose: This method is applicable to the determination of
ephedra capsules, and high-protein drink mix. residual alcohols in most matrices. Other analytes
commonly requested are acetone, 2-propanol and ethyl
Method facts: acetate.
• Sample size: 20 tablets, 20 capsules or 10g
• Limit of quantitation: 1ppm (most matrices) Method facts:
• Precision: NA • Sample size: 10g
• Method reference: Sullivan D. et al., “Determination • Limit of Quantitation: 10 ppm in most matrices
of Ephedra Alkaloids by Liquid Chromatography/ • Method Reference: Anthony, Robert M.; Sutheimer, Craig
Tandem Mass Spectrometry” Journal of AOAC A.; Sunshine, Irving, Acetaldehyde, Methanol, and
INTERNATIONAL, 3:86 Ethanol Analysis by Headspace Gas Chromatography,
[ assay ] E-F
Journal of Analytical Toxicology, Volume 4, Number 1, Description: A known amount of sample is enzymatically
January 1980, pp. 43-45(3) (Modified) hydrolyzed and extracted using chloroform and methanol.
An aliquot of the extract is transferred to a tared flask and
Description: Liquid samples can be analyzed directly or evaporated to dryness under a stream of nitrogen gas. The lipid
diluted with water prior to analysis. Solid samples should be residue is dried and weighed and the percent lipid is calculated.
weighed and diluted to a volume similar to that of the
standards. A mass of 0.1g to 1.0 gram may be necessary.
With increased sample mass, a matrix effect is more likely to Fat (Soxhlet)
interfere with spike recoveries. Purpose: Applicable for the determination of fat in meats,
seeds, and nuts. It may also be used for other foods and
Samples are incubated at 80°C for approximately 18 feeds by client request.
minutes. An injection is made with a syringe temperature of
Method facts:
85°C using a LEAP auto sampler with a 150C injection
temperature, 2000µL injection volume and 18.5 minute run • Sample size: 6g
time. The column is a DB-WAXetr 30m x 0.53 mm with a • Limit of quantitation: Most matrices - 0.1%
film thickness of 2 µm or another column of similar • Precision: On a peanut matrix, the RSD is 0.47%; on a
dimensions. Carrier gas is Helium at a pressure of 5psi and meat matrix, the RSD is 3.69%
flow of 8mL/min. Detector is a FID at 250°C and an air
• Method reference: AOAC 960.39, 948.22
flow of 400mL/min. H2 flow of 40mL/min. Makeup gas is
Helium with a gas flow of 40mL/min. Description: The sample is weighed into a cellulose thimble
containing sand or sodium sulfate. The thimble is dried to
The amount of each residual alcohol is determined by remove excess moisture. Pentane is dripped through the
comparing the signal of the unknown sample, measured by sample to remove the fat. The extract is then evaporated,
the gas chromatograph and FID, with the signal of reference dried, and weighed.
standard solutions.
Fat, total (Roese-Gottlieb)
Fat (acid hydrolysis) Purpose: Applicable for the determination of fat in milk
Purpose: Applicable for the determination of fat in (liquid or powder), buttermilk, half and half, cream, and
most products. ice cream.
Method facts: Method facts:

• Sample size: 2g • Sample size: 5g
• Limit of quantitation: Most matrices - 0.1% • Limit of quantitation: 0.1%
• Precision: On a dog food matrix, the RSD is 1.55% • Precision: 4.7% on a dry milk matrix
• Method reference: AOAC 922.06, 925.32, 933.05, 954.02 • Method reference: AOAC 989.05, 920.11, 932.06
Description: The sample is hydrolyzed with hydrochloric Description: The sample is hydrolyzed in a water bath using
acid. The fat is extracted using ether and hexane. The extract concentrated ammonium hydroxide. The fat is extracted using
is filtered through a sodium sulfate column. The solvent is ether and hexane. The extract is evaporated, dried, and weighed.
evaporated from the remaining extract and the fat is dried
and weighed.
Fat, total (NLEA)
see Fatty acid profiles
Fat (methanol/chloroform)
Purpose: Applicable for the determination of lipids in foods Fatty acid profiles
and feed. This method is preferred when the sample contains
These tests are based on AOCS method Ce1b-89 with the
fish, shellfish, or their products.
following options:
Method facts: • Fatty acids C8-C22: Applicable to most
• Sample size: 5g food products
• Limit of quantitation: Most matrices - 0.1% • Fatty acids C8-C24: Applicable to products
• Precision: On a dog food matrix, the RSD is 4.52% containing marine lipids EPA/DHA
• Method reference: AOAC 983.23 • Fatty acids C4-C24: Applicable to products containing
dairy and those with EPA/DHA

[ assay ]
Purpose: This procedure is used for traditional fatty acid analyzed by gas chromatography using external standards
profile analysis. The method detects fatty acids having 4 to for quantitation.
24 carbon atoms as methyl esters. The conditions specified
in this method are not suitable for determining epoxy or
Fatty acids, free (total by titration)
oxidized fatty acids that have been polymerized.
Purpose: Applicable for the determination of free fatty acids
Method facts: in vegetable oils and marine oils, animal fats, dark sulfonated
• Sample size: 2g oils, and other extracted lipids.
• Limit of quantitation: Most matrices - 0.01% Method facts:
• Precision: On a butter matrix, the RSDs for fatty acids are • Sample size: 10g of food/feed, 5g of oil
1-5%
• Method references: AOCS methods Ce 1b-89, Ce 1-62,
Ce 1e-91, Ce 1e-07 and Ce 2-66 • Precision: NA
• Method reference: AOCS Ca 5a-40
Description: The extraction can vary depending upon the
type of product being analyzed. For some products, the lipid USP <4017> fats and fixed oils
is extracted, saponified, and subsequently derivitized. For Description: Lipid is extracted from food and feed samples;
other products a direct saponification is performed followed
oils are sampled directly. The free (uncombined) fatty acids
by derivatization. The methyl esters of the fatty acids are
are determined by titration with sodium hydroxide solution.
analyzed by gas chromatography using external standards
for quantitation.
Fatty acids, free (by GC)
Purpose: Applicable for the determination of any free
Fatty acid profiles (NLEA) or extracted anhydrous fat. It is not applicable to acid
These tests are based on AOAC method 996.06 with the hydrolysis fat.
following options:
Method facts:
• Fatty acids C8-C22: Applicable to most
• Sample size: Sample size: 10g
food products
• Fatty acids C8-C24: Applicable to products
• Precision: Varies for each fatty acid
containing marine lipids EPA/DHA
• Method reference: AOCS Ce 1-62
• Fatty acids C4-C24: Applicable to products containing
dairy and those with EPA/DHA Description: Free fatty acids in a fat are absorbed on anhydrous
alkaline ion exchange resin. Lipid is washed from the resin
Purpose: This procedure is used for quantitation of NLEA with petroleum ether. The free fatty acids are converted to
total fat using fatty acid profile analysis to include trans fat, methyl esters directly on the resin by treatment with HCl
polyunsaturated fatty acids, and monosaturated fatty acids. methanol. The methyl esters of the free fatty acids are then
The method detects fatty acids having 4 to 24 carbon atoms determined by gas-liquid chromatography.
as methyl esters. The conditions specified in this method are
not suitable for determining epoxy or iodized fatty acids that
FDA PAM 302
have been polymerized.
Purpose: Applicable for screening of up to 375 pesticide and
Method facts: herbicide residues in fruits, vegetables and low fat
• Sample size: 2g sample matrices.
• Limit of quantitation: Most matrices - 0.01% Method facts:

• Precision: On a hydrogenated vegetable oil matrix, the • Sample size: 100g
RSDs for fatty acids are 1-5%
• Method reference: AOAC 996.06, AOCS Ce 1j-07 and
Ce 1h-05
Description: The extraction can vary depending upon the

type of product being analyzed. For some products, the lipid
is extracted, saponified, and subsequently derivitized. For • Precision: Determined on a spiked recovery basis
other products, a direct saponification is performed followed • Method reference: FDA PAM
by derivatization. The methyl esters of the fatty acids are
[ assay ] F
Description: Residues are extracted from samples by blending Ferulic acid

with acetone or water/acetone, then transferred from the filtered see Phenolic acids
aqueous extract into organic solvent and examined by various
determinative steps, SPE clean-up with GC and HPLC analysis.
Fiber, crude
Also see pages 50 and 62 for other pesticide screens. Purpose: Applicable for the determination of crude fiber in
foods, feeds, grains, meals, flours, plant material, and other
fiber-bearing material from which the fat can be extracted,
FDA PAM 303 leaving a workable residue.
Complex matrices requiring extensive cleanup
Method facts:
Purpose: Applicable for screening of up to 118 pesticide • Sample size: 5g
residues in crops, feeds, herbal extracts, and other appropriate
• Limit of quantitation: Varies with matrix
commodities.
• Precision: On a dog food matrix, the RSD is 18.0%
Method facts: • Method reference: AOAC 962.09
Description: Crude fiber is the loss on ignition of dried
residue remaining after digestion of sample with 1.25%
sulfuric acid and 1.25% sodium hydroxide solutions under
specific conditions. Note: for products containing fructan
sources, a fructan analysis may need to be performed.
• Precision: Determined on a spiked recovery basis
Fiber, insoluble and soluble dietary (LEE)
• Method reference: FDA PAM
Purpose: Applicable to most foods and feeds.
Description: The sample is blended with a mixture of
acetonitrile and water, extracted via liquid:liquid partitioning, Method facts:
and cleaned up by gel permeation chromatography (GPC) • Sample size: Solid matrices - 5g, liquid matrices-10g
followed by florisil column chromatography. The extract is • Limit of quantitation: Most matrices - 1.0%
injected for organophosphate, chlorinated, and nitrogen
• Precision: On a commercial cereal sample, the RSD is
pesticide residues on a GC system.
6.83% (IDF) and 13.9% (SDF)
Also see pages 50 and 62 for other pesticide screens. • Method reference: AOAC 991.43
Description: Duplicate food samples are gelatinized with
FDA PAM 304 alpha-amylase and digested with enzymes in a Mes-Tris buffer
M304 screen, 285 compounds to break down starch and some protein. The digest is then
filtered and the filtrate retained for soluble dietary fiber analysis.
Purpose: Applicable for the screening of up to 285 residues
The residue (insoluble dietary fiber) is washed with ethanol
in foods.
and acetone to remove starch and protein degradation products
Method facts: and moisture. Residue is dried and weighed. Protein content is
• Sample size: 100g determined for one of the duplicates; ash content is determined
for the other. The soluble dietary fiber (SDF) and insoluble
dietary fiber (IDF) contents of the sample are then calculated
after adjustment for the protein and ash values. Note: for
products containing fructan sources, a fructan analysis may
need to be performed.
• Method reference: FDA PAM Fiber, total dietary (LEE)

Purpose: Applicable to most foods.
Description: The sample is extracted with ethyl acetate,
concentrated, cleaned with gel permeation chromatography Method facts:
(GPC), and injected on an HPLC and various GCs to • Sample size: Solid matrices - 5g / liquid matrices - 10g
determine the absence of measurable levels of pesticides.
Also see pages 50 and 62 for other pesticide screens. • Precision: On a cereal matrix, the RSD is 3.93%

[ assay ]
Description: Duplicate food samples are gelatinized with solubilized and hydrolyzed to glucose and maltose by the
alpha-amylase and digested with enzymes in a Mes-Tris combined action of the two enzymes. Protein in the sample
buffer to break down starch and some protein. Soluble is digested with protease. For the measurement of high
dietary fiber in the aqueous enzyme digest is precipitated by molecular weight dietary fiber, ethanol is added and the
treatment with ethanol. The digest is then filtered and the insoluble and high molecular weight soluble dietary fiber is
residue is washed with ethanol and acetone to remove captured, washed with ethanol and acetone, dried and
starch, protein degradation products and moisture. The weighed. One of the duplicate residues is analyzed for
residue is dried and weighed. Protein content is determined protein and the other for ash. The low molecular weight
for one of the duplicates; ash content is determined for the soluble dietary fiber in the filtrate is concentrated on a
other. The total dietary fiber content of the sample is rotary evaporator, desalted through ion exchange resins,
calculated after adjustment for the protein and ash values. further concentrated and finally analyzed by size exclusion
Note: for products containing fructan sources, a fructan HPLC equipped with a refractive index detector. Results are
analysis may need to be performed. reported for total dietary fiber (CODEX definition), total
resistant oligosaccharides, and sum of measured fibers.
Fiber, total dietary (Prosky)

Purpose: Applicable to most foods. Fiber, insoluble, soluble, and total dietary (CODEX
definition) by enzymatic-gravimetric method and
Method facts: liquid chromatography (McCleary)
• Sample size: solid matrices - 2 g / liquid matrices - 10 g Purpose: This method is applicable for the determination of
• Limit of quantitation: Most matrices - 0.75% Insoluble, Soluble, and Total Dietary Fiber (IDF, SDF, and
• Precision: On a cereal matrix, the RSD is 7.44% TDF, respectively) inclusive of resistant starch (RS) and
dietary fiber that is not precipitated in 4 parts alcohol, 1 part
• Method reference: AOAC 985.29 water (SDFS/LMWSDF) of degree of polymerization (DP)
Description: Duplicate samples are gelatinized with > 3 consistent with the CODEX definition adopted in 2009
alpha-amylase and digested with enzymes in a phosphate (ALINORM 09/32/REP). This method is applicable to
buffer to break down starch and some protein. Ethanol is plant material, foods, and food ingredients.
added to each sample to precipitate the soluble fiber. The
samples are filtered, and the residue is rinsed with ethanol Method facts:
and acetone to remove starch and protein degradation • Sample size: 10g
products and moisture. Protein content is determined for • Limit of Quantitation: Most matrices – 0.1%
one of the duplicates; ash content is determined for the
• Precision: On a fortified breakfast cereal, the RSD for
other. The total dietary fiber in the sample is calculated
Total Dietary Fiber is 5.09%
after adjustment for the protein and ash values. Note: for
products containing fructan sources, a fructan analysis may • Method Reference:
need to be performed. Official Methods of Analysis of AOAC
INTERNATIONAL (2012) 19th Ed., AOAC
INTERNATIONAL, Gaithersburg, MD, USA, Official
Fiber, total dietary (CODEX definition) by enzymatic- Method 2011.25. (Modified)
gravimetric method and liquid chromatography
(McCleary) Description: Duplicate test portions are incubated with
Purpose: Applicable for the determination of total dietary pancreatic α-amylase and amyloglucosidase (AMG) for 16
fiber and low molecular weight soluble fiber (LMWSF) in hr at 37oC in sealed 250 mL bottles in a shaking water bath
food products containing dietary fiber, resistant starch, and while mixing with sufficient vigor to maintain continuous
resistant oligosaccharides. suspension. During this step, non-resistant starch is
solubilized and hydrolyzed to glucose and maltose by the
Method facts: combined action of the two enzymes. The reaction is
• Sample size: Solid matrices - 2g / liquid matrices - 10 g terminated by pH adjustment and temporary heating.
• Limit of quantitation: Most matrices - 1.0% Protein in the sample is digested with protease. For the
• Precision: Available on request measurement of IDF, the digestate is filtered and the IDF is
determined gravimetrically after correction for any protein
or ash in the residue. For the measurement of the water
Description: Duplicate test portions are incubated with soluble, but water: alcohol insoluble dietary fiber (SDFP/
pancreatic α-amylase and amyloglucosidase (AMG) for 16 HMWSF), ethanol is added to the filtrate of the IDF; the
hr at 37°C. During this step, non-resistant starch is precipitated SDFP is captured by filtration and determined
[ assay ] F
gravimetrically after correction for any protein or ash in the Description: Fluoride is determined potentiometrically by
precipitate. Nonprecipitable, water:alcohol soluble dietary the use of an ion selective electrode. An ionic strength buffer
fiber (SDFS/LMWSF) in the filtrate is recovered by is used to eliminate the affects of other ionic species present
concentrating the filtrate, deionizing through ion exchange in the sample. The concentration of fluoride is calculated
resins, concentrating, and quantitating by LC. using a standard curve.
Fibersol Folate
see Resistant maltodextrin see Folic acid
Flavonoids in tea
Folic acid/Folate
With % solids tea and tea products for Tea Association
members only Purpose: Applicable for the determination of folic acid in
most foods, dietary supplements, and feeds.
see Polyphenols, total
Method facts:
• Sample size: 2g
Fluoride (chemical) • Limit of quantitation: Most matrices - 0.06 mcg/g
Purpose: Applicable for the determination of total fluoride
in a variety of samples, including oils, raw materials, plant • Precision: On an infant formula matrix, the RSD is 8.89%
tissues, bones, water, and many types of foods. • Method reference: AOAC 960.46 & 992.05
Method facts: Description: The sample is hydrolyzed in potassium phosphate

• Sample size: 10g buffer. Following hydrolysis, the sample is treated with a
• Limit of quantitation: Most matrices - <2.0ppm chicken-pancreas enzyme and incubated for approximately
18 hours. Utilizing the bacteria Lactobacillus rhamnosus
• Precision: On a feed sample matrix, the RSD is 6.19%
(casei), the amount of folic acid is determined
• Method reference: AOAC 944.08 (modified) turbidimetrically by comparing the growth response of a
Description: The sample is weighed into a platinum dish sample against the growth response of a folic acid standard.
along with a small amount of calcium hydroxide and
phenolphthalein to ensure the sample is alkaline. The sample
Formic acid
is mixed with double-deionized water to form a slurry. It is
see Organic acid profiles — option 3
then dried on a hot plate, and ashed for at least 4 hours at
550°C in a muffle furnace. The ashed portion is then
transferred into a distillation apparatus where the fluoride is Fo-ti powder
steam distilled with perchloric acid. Fluoride levels are then see trans-Resveratrol
determined titrametrically.
Free amino acids

Fluoride (Ion Selective Electrode — ISE)
see Amino acid profile, free by HPLC
Purpose: Applicable for the determination of fluoride in
see Amino acid profile, free by AAA
drinking water, surface water, saline water, and domestic and
industrial wastes. This method is also applicable to some
foods, infant formula, and pharmaceutical grade reagents.
Method facts: Free fatty acids

• Sample size: 20mL see Fatty acids, free (by titration)
• Limit of quantitation: Most matrices - 0.2 ppm see Fatty acids, free (by GC)
• Precision: On a commercially manufactured mouth formula
rinse, the RSD is 5.50% Fructan (HPLC)
• Method reference: Standard Methods for the Examination Purpose: Applicable for the determination of fructans,
of Water and Wastewater, 17th Edition, Method 4500-F-C., which includes inulin and fructo-oligosaccharides (FOS).
APHA, AWWA, WPCF, Washington, DC (1989) (modified) Well suited for high-level fructan samples.
Food Chemical CODEX, Sixth Ed. pp 1077-1078,
National Academy Press, Washington, DC (2009) Method facts:
(modified) • Sample size: 20g

[ assay ]
• Limit of quantitation: Most matrices - 0.5% Fury (zeta cypermethrin)

• Precision: Varies with matrix Information available on request
Description: Fructans are extracted from the matrix with Galactooligosaccharides (GOS) content in Infant
water. The extract is centrifuged, filtered and an appropriate Formula
dilution is injected for the analysis of free fructose and Purpose: This method is applicable for the determination of
sucrose. An aliquot of the filtrate is also subjected to treatment galactooligosaccharides (GOS) in infant formula by Intact
by enzymes to liberate fructose from the fructans. The net Fingerprint Analysis using High Performance Anion
fructose content is determined on a HPAEC using PAD Exchange Chromatography with Pulsed Amperometric
and compared against known standards. Detection (HPAEC-PAD)
Method Facts:
Fructan (spectrophotometric) • Level of Quantitation: 0.1%
Purpose: Applicable for the determination of fructans, • %RSD: 1.7
which includes: fructo-oligosaccharides (FOS), and inulin.
• Sample Required: 25 g
Method facts: • Method Reference: Covance Internal Method
• Sample size: 5g
• Limit of quantitation: Most matrices – 0.5% Description: Powdered infant formula is extracted in water
with low heat. The extract from the powdered infant
• Precision: On a spiked flour matrix, the RSD is 2.04% formula or liquid (ready to feed) infant formula is then
• Method reference: AOAC 999.03 diluted appropriately with water taking into account the
final dilution with acetonitrile. An aliquot is finally treated
Description: The sample is extracted with water. The extract
with acetonitrile to precipitate protein, centrifuged, and
is then treated with enzymes to hydrolyze and remove sucrose,
filtered into an injection vial to be injected onto a high
starch, and maltosaccharides. An aliquot of the filtrate is also
subjected to treatment by enzymes to liberate fructose and performance anion exchange chromatography (HPAEC)
glucose from fructan. The amount of these reducing sugars system to separate and quantify five specific oligomer peaks
from fructan is determined by spectrophotometric methods. that represent the total GOS concentration within the
sample. The actual lot used to fortify the product is used as
the reference material to generate a calibration curve for
Fructose
quantification. A reference material is made from a mixture
see Sugar profile by GC
of GOS reference material lots from a specific supplier to
see Sugar profile by HPLC
see Sugar profile by IC generate a calibration curve for quantification if the supplier
see Sugar profile, low levels is known, but the actual lot of reference material used to
fortify the product is unknown. The associated error
between peak profile differences between individual lots is
Fruits decreased with this procedure. The same concentration of
see Anthocyanins, total the reference material is used to represent each of the five
oligomer peaks separately. The final GOS content of the
product is calculated from the average result of the five
Fumaric acid peaks.
Galactooligosaccharide (GOS) content in Raw

Furfural Material
Purpose: Applicable to the determination of free furfurals. Purpose: This method is applicable for the determination of
Method facts: galactooligosaccharide (GOS) purity in the raw material
• Sample size: 5g used to fortify a variety of products in the food industry by
• Limit of quantitation: 0.5ppm High Performance Anion Exchange Chromatography with
Pulsed Amperometric Detection (HPAEC-PAD)
• Precision: 1.7%
Description: The sample is extracted with 4% TCA and Method Facts:
injected into an HPLC. Furfural is quantified against • Range of Quantitation: The range of quantitation for this
standards of known concentration. assay is 50-75% on a dry basis.
[ assay ] G
• %RSD: 1.2 gamma Linolenic acid (GLA)

• Sample Required: 10 g Can be analyzed as part of the fatty acid profile. See Fatty
acid profiles.
• Method Reference: [1] Official Methods of Analysis of
AOAC INTERNATIONAL (2006) 18th Ed., AOAC
INTERNATIONAL, Gaithersburg, MD, USA, Official Garcinia cambogia
Method 2001.02 (Modified) see Hydroxycitric acid
[2] Dionex Application Note 155: Determination of
Trans-Galactooligosaccharides in Foods by AOAC Garlic
Method 2001.02 (Modified) Purpose: Applicable for the determination of allicin potential
Description: A buffered extract of the GOS is analyzed in powdered garlic and powdered garlic tablets or capsules.
directly for the inherent amounts of galactose, glucose, and
Method facts:
lactose. A separate aliquot of the buffered extract is treated • Sample size: 5 g
with the enzyme β-galactosidase in order to hydrolyze the
• Limit of quantitation: Most matrices - 1.0 mg/g
GOS oligomers and lactose to galactose and glucose.
Appropriate dilutions of the untreated and treated extracts • Precision: On a garlic tablet matrix, the RSD is 8.9%
are injected onto a high performance anion exchange • Method reference: Simultaneous Determination of Alliin
chromatography (HPAEC) system to separate and quantify and Allicin in Allium Plants and Their Products by Liquid
the galactose, glucose and lactose which are detected and Chromatography, Journal of AOAC International,
quantified by pulsed amperometric detection (PAD). The 80(5) (1997)
GOS content is calculated by subtracting the amount of free Description: Garlic powder is dissolved in a buffer solution.
galactose and the galactose released from lactose from the Alliin present in the sample reacts with alliinase enzyme to form
total galactose (from free, lactose, and GOS). A factor allicin. The extract is then filtered and analyzed using HPLC.
determined from the ratio of glucose and galactose released
from GOS is applied that corrects for hydrolysis and the
contribution from glucose. Ginkgo biloba flavonoids
Purpose: Applicable for the determination of the ginkgo
flavones, quercetin, isorhamnetin, and kaempferol, in ginkgo
gamma Aminobutyric acid (GABA) biloba or ginkgo biloba extracts.
Purpose: Applicable to all samples prepared for the analysis
Method facts:
of gamma aminobutyric acid (GABA).
Method facts:
• Limit of quantitation: varies with matrix
• Precision: 4-7%
• Limit of quantitation: Varies within matrix
• Method reference: Journal of Chromatographic Science,
• Precision: 1.7%
605: 41-48
• Method reference: Hischenhuber, C., Deutsche
Lebensmittel-Rundschau, “High Performance Liquid Description: Samples are extracted with methanol:water.
Chromatographic and Thin-Layer Chromatographic The extract is injected into an HPLC with ELSD
Determination of Taurine in Infant Formulas,” /84, detection. The flavones are quantified by comparison with
JAHRG /HEFT 4/1988 (modified) known standards.
Stuart, J.D., Wilson, T.D., Hill, D.W., Walters, F.H., Feng,
S.Y., “High Performance Liquid Chromatographic
Ginkgo biloba terpenoids
Separation and Fluorescent Measurement of Taurine, a
Key Amino Acid,” Journal of Liquid Chromatography, Purpose: Applicable for the determination of ginkgo terpenes
2(6), 809-821 (1979) in ginkgo biloba or ginkgo biloba extracts.
Description: The sample is dissolved in warm water with Method facts:

trichloroacetic acid to precipitate proteins. GABA is then • Sample size: 30g
derivatized with orthophthalaldehyde and quantitated on a • Limit of quantitation: Varies with matrix
reverse-phase high performance liquid chromatography
• Precision: 6% for the major compounds
(HPLC) system.
• Method reference: Journal of Chromatographic Science,
vol 36, (April 1998)

[ assay ]
Description: Samples are extracted with methanol/water. individuals with this intolerance must avoid gluten, and rely
The extract is injected into an HPLC system equipped upon the correct labeling of food to make appropriate food
with an ELSD. Results are quantified by comparison to choices. Testing for the presence of gluten ensures food
known standards. manufacturers that an unlabeled, and potentially dangerous,
ingredient did not make its way into a food product.
Ginseng, Panax or Korean (ginsenosides) Method facts:

Purpose: Applicable for the determination of ginsenosides • Sample Size: 50g
in powdered ginseng root and ginseng extracts. • Limit of Quantitation: 5 ppm gluten (2.5 ppm gliadin)
Method facts: • Method Reference: RIDASCREEN® Gliadin Kit
• Sample size: 30g r-biopharm AOAC RI License Number 120601, Official
Method (SM) number 2012.01
• Limit of quantitation: Varies with ginsenoside compound
and matrix Description: The basis of this test is the antigen-antibody
• Precision: 1-6% depending upon the ginsenoside compound reaction. Samples are extracted using a RIDA® Extraction
and matrix Solution specific to this ELISA kit. After extraction and
• Method reference: Journal of Chromatographic Science, sample dilution, standards and samples are placed in
736: 77-81 microwells coated with specific antibodies against gliadins.
Present gliadin will bind to the capture antibodies resulting
Description: Samples are extracted with methanol/water. in an antibody-antigen-complex. Components not bound
The extract is injected into an HPLC system with ELSD. are removed with a washing step. Antibody conjugated to
Results are quantified by comparison to known standards. peroxidase is added. This antibody-conjugate is bound to the
Ginsenosides tested include Rf, Rg1, Re, Rb1, Rc, Rb2, and Rd. Ab-Ag-complex (antibody-antigen) forming the “sandwich”.
Any unbound enzyme conjugate is removed with another
washing. Enzyme substrate and chromogen are added to the
Glucosamine wells and incubated. A color reaction occurs where protein is
Purpose: Applicable for the determination of glucosamine present and results are measured with a Stat Fax reader.
compounds in premixes, tablet preparations, raw materials, Optical densities of the standard control forms a curve
dietary supplements, and pet foods. which samples are plotted against to obtain concentrations
Method facts: of gluten/gliadin protein in ppm/ppb.
• Sample size: 5g – 20 tablets
• Limit of quantitation: Most matrices - 0.01% Glycerol (glycerine)
• Precision: 5.7% Purpose: Applicable for the determination of glycerol in
detergents, cosmetics, foods, feeds products, and raw material.
Method facts:
Description: Glucosamine is extracted with water. • Sample size: 25g
Triethylamine is used to release glucosamine free base. The
• Limit of quantitation: Most matrices - <1.0%
solution is derivitized and injected into an HPLC with UV
detection. Results are quantified against known standards. • Precision: On a cereal bar matrix, the RSD is 5.58%
• Method reference: Sweely, Bentley, Makita and Wells,
J.A.C.S. 85: 2495-2507 (1963)
Glucose Cierce, A. E., Silylation of Organic Compounds,
see Sugar profile by GC Pierce Chemical
see Sugar profile by HPLC
see Sugar profile by IC Description: The sample is extracted with methanol then
see Sugar profile, low levels filtered. Aliquots of the sample and of the standard solutions
are derivitized with Tri-Sil followed by extraction into hexane.
Samples and standards are then injected on a GC System
Gluten/Gliadin equipped with a flame ionization detector (FID).
Purpose: Gluten is an allergen that is a mixture of prolamin Quantitation of glycerol is performed by comparing sample
and glutelin proteins from wheat (gliadin), rye (secalin), and responses against standards of known concentrations.
barley (hordein). Celiac disease is a permanent intolerance
to gluten that results in damage to the small intestine. Those
[ assay ] G-H
Goldenseal detector antibodies. After the addition of a stop reagent, the

Purpose: Applicable for the determination of hydrastine and test is read with a Stat Fax reader. Optical densities of the
berberine in goldenseal extracts, tablets, capsules and other controls form a standard curve which samples are plotted
dietary supplement preparations. against to obtain concentrations of allergen protein in ppm.
Method facts:
• Sample size: 5g Heavy metals as lead (USP <231>)
• Limit of quantitation: Most matrices - 0.1% Purpose: Applicable for the determination of metallic
• Precision: Varies with matrix impurities (Ag, As, Bi, Cd, Cu, Hg, Mo, Pb, Sb, Sn) that are
colored by the sulfide ion. Primarily applicable to foods,
• Method reference: Sakakibara, H., Honda, Y., Nakagawa, S.,
food additives, feeds, and raw materials.
Ashida, H., and Kanazawa, K., “Simultaneous Determination
of All Polyphenols in Vegetables, Fruits, and Teas,” Method facts:
Journal of Ag Food Chem, 51(3): 572-580 (2003) • Sample size: 10g
Description: Samples are extracted using the appropriate • Limit of quantitation: Most matrices - 5 ppm
technique for the sample matrix. Aliquots are analyzed using • Method reference: USP
HPLC with UV detection and quantified by comparison to
known standards. Description: The sample is digested with nitric acid and
sulfuric acid on a hot plate, then placed in a muffle furnace at
500°C. Dilute hydrochloric acid is added to the cooled sample
Grape seed extract
and taken to dryness on a hot plate. The pH is adjusted to
see Catechins
remove iron impurities. The sample is filtered into a culture
see Polyphenols, total
tube, thioacetomide solution is added and the color that
see trans-Resveratrol
develops is compared with that of lead standards.
Green tea
see Catechins Heavy metals by ICP-MS
see Polyphenols, total see Inorganic analysis by ICP-MS
Griffonia seed extract Heptachlor

see 5-Hydroxytryptophan (free amino acid profile) see USP 561 and EP pesticides
Hazelnut Protein by ELISA Herbicides

Purpose: Allergens are proteins in food that can create an Chlorophenoxy acid herbicides
immune response in sensitive individuals. Clear ingredient
Purpose: Applicable for the determination of processed
labeling of food products, by manufacturers, aids in
foods, feeds, and oil products. Includes:
protection from accidental ingestion by those individuals.
Testing for the presence of these proteins ensures the food is • 2,4-D (Agrotect) • Dicamba (Metambane)
free of the allergen at a potentially harmful level. • 2,4-DB • Dimethoate (Stinger)
• 2,4-DP • MCPA (Mapica)
Method facts: • 2,4,5-T (Weedone) • MCPP (Malerbane)
• Sample Size: 50g • 2,4,5-TP (Silvex) • Triclopyr
• Clopyralid
• Method Reference: Veratox ® Quantitative Hazelnut Method facts:
Allergen Kits Neogen Corporation • Sample size: 50g
• Limit of quantitation: Varies by compound
Description: Allergen Protein is extracted with a buffered • Precision: Varies by compound
• Method reference: FDA PAM
protein is washed away before a detector antibody (enzyme Description: Phenoxy acids are extracted from an acidified
labeled) is added. The detector antibody then binds to water/methanol mixture with nonpolar organic solvents. The
bound protein if present. After another wash, and the acids are then converted to their methyl ester form with
addition of a substrate, color develops as a result of bound diazomethane. The derivitized sample extract is purified by

[ assay ]
gel permeation chromatography (GPC) and the resulting Covance currently has the following reference materials
methyl esters are quantitated on a GC system utilizing a available:
halogen-specific detector.
Common Name Species Name Plant Part
Herbicides American Ginseng Panax quinquefolium Root

Triazine and chloracetamide herbicides Bilberry Vaccinium myrtillus Fruit
Bitter Melon Momordica charantia Leaf
Purpose: Applicable for the determination of processed foods, Bitter Melon Momordica charantia Fruit
feeds, and oil products. Includes alachlor (Lasso), atrazine,
Black Cohosh Actaea cimicifuga/racemosa Root
cyanazine (Bladex), metolachlor (Dual), simazine.
Carrot Daucus carota Root
Method facts: Chaste Tree Vitex agnus-castus Berry
• Sample size: 100g Chinese Ginseng Panax ginseng Root
• Limit of quantitation: Varies with matrix Chinese Goldthread Coptis chinesis Root
• Precision: Varies with matrix Chinese Licorice Glycyrrhiza uralensis Root
• Method reference: FDA PAM Cinnamon Cinnamon cassia Bark

Fenugreek Trigonella foenum-graecum Seed
Description: The herbicides are extracted with ethyl Gentian Root Gentiana lutea Root
acetate. The sample extract is purified by gel permeation
Ginkgo Ginkgo biloba Leaf
chromatography and the resulting fraction is quantitated on
Grape Seed Vitis vinifera Seed
a GC system equipped with a electrolytic conductivity
detector (ELCD) operated in halogen mode. Green Tea Camellia sinensis Leaf
Hibiscus Hibiscus rosa Flower
Hibiscus Hibiscus sabdariffa Flower
Hexachlorbenzene
see USP 561 and EP pesticides Hops Humulus lupulus Strobile
Indian Frankincense Boswellia serrata Resin
Hexachlorocyclohexane isomers Isatis Root Isatis inidgotica Root

see USP 561 and EP pesticides Licorice Glycyrrhiza glabra Root
Motherwort Leonura cardiaca Herb
Hexanal Nettles Urtica dioica Root
Purpose: Applicable for the determination of hexanal in Pumpkin Cucurbita pepo Seed
foods and ingredients. Pygeum Bark Prunus africana Bark
Method facts: Red Sage Salvia miltiorrhiza Root
• Sample size: 5g Reishi Mushroom (Black) Ganoderma sinense Fruiting body

Rosemary Rosmarinus officinalis Leaf
• Limit of quantitation: 1ppm
Saw Palmetto Serenoa repens Fruit
• Precision: Varies by matrix
Stevia Stevia rebaudiana Leaf
• Method reference: Journal of Food Science, 67: 1212-1218
Turmeric Curcuma longa Rhizome
Description: Samples are analyzed by headspace using
gas chromatography. Description: Methods vary by matrix. Currently there are
over 100 methods available at Covance for HPTLC identity
HPTLC verification testing. Proper documentation of the sample
Identity verification testing by high-performance material by genus, species, part, extract and any additional
thin-layer chromatography information that can be provided to the lab is critical in
providing HPTLC verification. In general, the method
Purpose: HPTLC is a versatile, high-throughput, and cost- involves an extraction, application to a thin-layer stationary
effective tool used to verify the quality and identity of phase plate, application of a mobile phase, derivitization and
botanical materials. exposure to the appropriate wavelength of light. Each
sample separation is monitored for banding (Rf ) which
Method facts: indicates the migration of active ingredients in the raw
• Sample size: 25g material. To ensure the fastest possible turn around time of
• Limit of quantitation: HPTLC for identity verification is samples, please provide a certificate of analysis with each
not quantitative sample to verify the method that will be needed for the
sample.
• Precision: NA
• Method reference: Varies by matrix
[ assay ] H-I
Hydrastine Inorganic analysis by ICP-MS

see Goldenseal Typical heavy metal scan: arsenic, cadmium, lead, mercury
(antimony included upon request). Additional available
Hydroxycitric acid (garcinia extract) elements: Ag, As, Ba, Be, Bi, Ca, Cd, Co, Cr, Cs Cu, Fe, Ga,
Purpose: Applicable for the determination of hydroxycitric Ge, Hg, I, La, Li, Mg, Mn, Mo, Ni, Pb, Pd, Pt, Rb, Ru, S,
acid in dietary supplement products. Sb, Sn, Sr, Ti, Tl, U, V, W, Zn (other non-routine elements
available upon request. Sulfate (calculated from total sulfur
Method facts: by ICP-MS).
• Sample size: 5g Purpose: Applicable for the determination of low level
• Limit of quantitation: 0.05% metals (ppb levels) in foods, food ingredients, feeds,
• Precision: 1.54% biological materials, and various other matrices.
Description: Samples are extracted with acid and clarified
• Limit of quantitation: Most matrices – 10 ppb
with filtration/centrifugation. The extract is injected into an
HPLC system with UV detection. Results are quantified by • Precision: For most elements, the RSD is <5%
comparison with known standards. • Method reference: AOAC 993.14
Description: The sample is wet-ashed with nitric acid using
Hydroxyproline closed-vessel microwave digestion. Depending on matrix,
see Amino acid profile, free or (HPLC), total alternate digestion techniques may include open-vessel
microwave digestion or dry ashing. Samples that are readily
Inorganic analysis by ICP soluble in water may be dissolved, acidified, and then
Purpose: Applicable for the determination of up to 20 analyzed directly. Using ICP-MS, the amount of each
elements (ppm levels) in virtually all matrices. See below for element is determined by comparing the counts generated
options. Options 3 and 4 are matrix dependent. Please call by the unknowns to those generated by standard solutions of
to verify if the package price is applicable to your sample. known concentration.
Option 1:
Calcium, iron, sodium (for nutrition labeling) Inositol
Option 2: Purpose: Applicable to the determination of inositol in foods,
Calcium, copper, iron, magnesium, manganese, phosphorus, feeds, infant formulas, premixes, and nutritional supplement
potassium, sodium, zinc products.
Option 3: Method facts:
Option 2 plus aluminum, barium, boron, chromium, • Sample size: 10g
molybdenum, strontium • Limit of quantitation: Most matrices – 12.5mcg/g
Option 4: • Precision: On an infant formula matrix, the RSD is 2.29%
Option 3 plus beryllium, cadmium, cobalt, nickel,
vanadium • Method reference: Methods of Analysis for Infant
Formulas, Infant Formula Council, ‘Inositol’ (modified)
Individual elements can be performed on request. Atlanta, Georgia, Section C-7, (1985)
Method facts: Tagliaferri, E.G.; Bonetti, G.; Casella, G.; Blake, C.J. Ion
• Sample size: 10g Chromatographic Determination of Inositol in Infant
• Limit of quantitation: Element and matrix dependent Formulae and Clinical Products for Enteral Feeding. J.
Chromatogr. A. 2000, 879, 129-135. (Modified)
• Precision: For most elements, the RSD is <5%
Dionex Technical Note 20: Analysis of Carbohydrates by
High Performance Anion Exchange Chromatography with
Description: The sample can be either dry-ashed, wet-ashed, or Pulsed Amperometric Detection (HPAEC-PAD
read directly. If dry-ashed, the sample is ashed at 500˚C ±50˚ (Modified)
until ashing is complete. The resulting ash is treated with
concentrated hydrochloric acid, dried and redissolved in Description: The inositol sample is extracted with water or
hydrochloric acid solution. If wet-ashed, the sample is acid hydrolyzed at a high temperature and filtered. Samples
digested on a hot plate with nitric acid, hydrochloric acid, containing known levels of glycerol undergo enzyme
and/or hydrogen peroxide. The amount of each element is phosphorylation with glycerol kinase. An appropriate
determined with an ICP spectrometer by comparing the dilution is injected onto a high-performance anion exchange
emission of the unknown sample against the emission of each chromatography (HPAEC) system equipped with a pulsed
element from standard solutions. amperometric detector (PAD) for determination.

[ assay ]
Inulin • Precision: NA
see Fructan (spectrophotometric) • Method reference: AOCS Cd 1b-87
see Fructan (HPLC)
Description: The sample is dissolved in cyclohexane and a
Wijs solution, then titrated to an end point.
Iodide (Ion Selective Electrode — ISE)
Purpose: Applicable for the determination of iodide in dairy
products, infant formulas, and dietary supplements. Iodine (ICP-MS)
Purpose: Applicable for the determination of total iodine in
Method facts: a variety of matrices including foods (i.e., powders, liquids,
• Sample size: 10g (solids) /50 mL (liquids) dairy products, infant formulas), nutritional supplements, and
• Limit of quantitation: Most matrices - 0.10 ppm mineral premixes at levels ranging from low ppm to percent.
• Precision: On a powdered infant formula, the RSD is 8.15%
Method facts:
• Method reference: AOAC 992.24 • Sample size: 10g
Description: Samples are weighed and proteins are precipitated • Limit of quantitation: Most matrices - 0.1 ppm
with addition of dilute acid. Samples are filtered and iodide is
• Precision: On a milk powder matrix, the RSD is <5%
determined using an ion selective electrode and method of
standard addition. Nickel nitrate is added to reduce interferences. • Method reference: “Determination of Iodine in Nutritional
Supplements by ICP-MS,” presented at the 123rd AOAC
Annual Meeting, Philadelphia, PA (2009)
Iodine, high level
“Determination of Iodine by ICP-MS,” presented at the
Purpose: Applicable for the determination of total iodine in
premixes, dietary supplements, salts, and various raw materials. 122nd AOAC Annual Meeting, Dallas, TX (2008)
Method facts: Description: The sample is weighed into a single-use

• Sample size: 10g polypropylene (pp) centrifuge tube. A purified water
solution with a basic pH is added. The sample is digested by
• Limit of quantitation: Most matrices - 40 ppm
warming in a microwave to ensure complete extraction of
• Precision: On a multivitamin matrix, the RSD is 11.1% any iodine present in the sample. A stabilizer is added to
• Method reference: AOAC 935.14, 932.21 maintain iodine in the appropriate form. The digested
USP sample is taken to volume with purified water, filtered, then
diluted appropriately prior to analysis by ICP or ICP-MS.
Description: The sample is digested and fused. The sample
is then filtered using a hot water wash into a 500 mL,
wide-mouth Erlenmeyer flask until the volume is 300 mL. Iron (ICP or ICP-MS)
The sample is neutralized to methyl orange with 85% see Inorganic analysis by ICP or ICP-MS
phosphoric acid (H3PO4), with 1 mL in excess. A slight
excess of bromine (as saturated bromine water) is then added,
Isoflavones
and the solution is boiled for 5 minutes after it has become
colorless. Any excess bromine that remains is removed by the Daidzein, glycitein, genistein, daidzin, glycitin, genistin
addition of salicylic acid. When the solution is cool, 1mL of
Purpose: Applicable for the determination of the isoflavones
85% H3PO4 and about 0.5g of potassium iodide (KI) are
daidzein, glycitein, and genistein and their glucoside
added. The resulting iodine is titrated with thiosulfate
(Na2S2O3) (0.005N or 0.10N) to a clear end point. derivatives found in a variety of food products.
Method facts:
Iodine value (titration) • Sample size: 20g
Purpose: The iodine value is used to measure the amount of • Limit of quantitation: Most matrices - 10 ppm
unsaturation of fats and oils and is expressed in terms of the
number of centigrams of iodine absorbed per gram of • Precision: On an infant formula matrix, the RSDs for
sample. It is a applicable to all normal fats and oils with daidzin, glycitin, genistin, and the total glucosides are 4.7,
iodine values in the range of 15 to 70. It is not applicable to 10.3, 8.3, and 6.2, respectively
products containing conjugated double bonds. • Method reference: Seo, A., and Morr, C. V., “Improved
Method facts: High-Performance Liquid Chromatographic Analysis of
• Sample size: 20g Phenolic acids and Isoflavonoids from Soybean Protein
Products”, Journal of Agricultural and Food Chemistry,
32(3):530-533 (1984)
[ assay ] I-L
Petterson, H., and Kiessling, K.H., “Liquid Chromatographic • Sample size: 25g
Determination of the Plant Estrogens Coumestrol and • Limit of quantitation: Levels as low as 0.005% can be
Isoflavones in Animal Feed,” Association of Official detected, large quantities of some sugars can interfere with
Analytical Chemists Journal, 67(3):503-506, (1984) the proper determination of neighboring sugars.
Description: The samples are extracted using a solution of • Precision: Varies with matrix
hydrochloric acid and reagent alcohol heated on steam baths • Method reference: Journal of AOAC INTERNATIONAL,
or hot plates. The extract is brought to volume, diluted, and Vol. 88, No. 3, 2005
centrifuged. An aliquot of the supernatant is placed onto a
Description: The sample is extracted with warm water.
C18 solid-phase extraction column. The column is washed
Lipids are removed with chloroform extraction and proteins
with 20% methanol and isoflavones are eluted with 80%
are removed by centrifugation. The extract is filtered and
methanol. The samples are analyzed by HPLC with UV
appropriate dilutions are injected onto a high-performance
detection against an external standard curve.
anion exchange chromatograph (HPAEC) equipped with a
pulsed amperometric detector (PAD).
Isomalt
see Sugar alcohols Lead (by ICP-MS)
see Inorganic Analysis by ICP-MS
Japanese export testing
• Sennosides (UV) Lignin
• Fenfluramine, HPLC Purpose: Applicable for the determination of lignin in
• p-Hydroxybenzoates (methyl, ethyl, propyl, butyl, forages and feeds with low carbohydrate contents.
isopropyl, isobutyl)
Method facts:
• Thyroxine, Triiodothyronine HPLC
• Sample size: 5g
• Benzoic acid, sorbic acid
• Artificial colors
• Precision: NA
• Polysorbate
• Method reference: United States Department of
Information available on request Agriculture, Forage and Fiber Analysis, Agriculture
Handbook #379.8, United States Department of
Agriculture, Washington, DC (1970)
Kudzu
see Isoflavones Description: The protein, carbohydrate, and ash content is
removed by treating samples with a boiling detergent solution
and filtered. The fats and pigments are removed via an
Lactic acid acetone wash leaving the lignocellulose fraction in a frit.
see Organic acid profiles — option 2 The cellulose is then dissolved with sulfuric acid leaving
the lignin fraction, which is determined gravimetrically.
Lactitol
see Sugar alcohols Lindane
Lactose
Loss on drying (LOD)
see Sugar profile by GC
see Sugar profile by HPLC Purpose: Applicable for the determination of loss on drying
see Sugar profile by IC in refined, reagent-grade materials.
see Sugar profile, low levels Method facts:
• Sample size: 1g
Lactulose • Limit of quantitation: 0.1%
Purpose: Applicable for the determination of lactulose in • Precision: NA
foods and infant formulas at levels below 0.1%. • Method reference: USP or FCC
Method facts:

[ assay ]
Description: The sample is dried to a constant weight at a • Limit of quantitation: 0.05 ppm Melamine
temperature specified by the individual monograph. • Precision: 6% RSD at 0.05 ppm
• Method reference: United States Food and Drug
Lutein Administration, Interim Method for Determination of
see Carotenoids Melamine and Cyanuric Acid Residues in Foods Using
LC-MS/MS: Version 1.0 Laboratory Information
Bulletin No. 4422 (October 2008)
Lycopene
see Carotenoids Description: Melamine is extracted from tissue and infant
formula with a 50:50 acetonitrile:water extraction solution,
followed by centrifugation and SPE cleanup. The compound
Ma Huang is analyzed using a zwitterionic HILIC LC column.
see Ephedrine alkaloids
Electrospray ionization is used in the positive ion
(melamine) mode. This procedure has been validated by the
Magnesium (ICP) FDA for the determination of melamine in tissues. The
see Inorganic analysis by ICP method has been validated in infant formula powder by
Covance. Two multiple reaction monitoring (MRM)
transitions are monitored for the compound. Isotope internal
Malic acid standards are used to correct for any matrix effects. Fortified
see Organic acid profiles — option 2 test portions were within 85-105% recovery for the infant
formula powder validation.
Maltitol
see Sugar alcohols Mercury (ICP-MS)
see Inorganic Analysis by ICP-MS
Maltodextrin, resistant
see Resistant maltodextrin Methionine (AOAC)
see Amino acid profile (AOAC), total
Maltose
see Sugar profile by GC Methylsulfonylmethane (MSM)
see Sugar profile by HPLC Purpose: Applicable for the determination of MSM in
see Sugar profile by IC tablets, capsules, premixes, or raw materials.
see Sugar profile, low levels
Method facts:
• Sample size: 5g raw material, 20 tablets or capsules
Mandatory nutrient package
This package includes all mandatory labeling nutrients.
Information available on request. • Precision: Based on tablet preparation matrix, the RSD
is 3.77%
Manganese (ICP or ICP-MS) • Method reference: Guedes De Pinho, et al., Journal of
see Inorganic analysis by ICP or ICP-MS Agricultural and Food Chemistry, 51:727-732, (2003)
Menta, et al., Journal of Chromatography, 383:400-404 (1986)
Mannitol Description: The sample is extracted with methanol containing
see Sugar alcohols an internal standard. Samples and standards are then injected
on a GC System equipped with a flame ionization detector
Marine lipids (EPA/DHA) (FID). Quantitation of MSM is done by comparing sample
responses against standards of known concentrations.
Melamine (LC-MS/MS) Milk Protein, Total by ELISA

Purpose: Applicable for the determination of melamine Purpose: Allergens are proteins in food that can create an
using LC-MS/MS in food products. immune response in sensitive individuals. Clear ingredient
labeling of food products, by manufacturers, aids in protection
Method facts:
from accidental ingestion by those individuals. Testing for the
[ assay ] M
presence of these proteins ensures the food is free of the • Limit of quantitation: Most matrices - 0.1%
allergen at a potentially harmful level. • Precision: On a syrup matrix, the RSD is 2.70%
• Sample Size: 50g
Description: The sample is dried in a vacuum oven at 60˚C
• Limit of Quantitation: 2.5 ppm to a constant weight.
• Method Reference: Veratox ® Quantitative Milk Allergen
Kits Neogen Corporation Moisture, 70˚C (vacuum oven)
Purpose: Applicable for the determination of moisture in
Description: Allergen Protein is extracted with a buffered salt
fruits and vegetables.
solution, allowed to settle then placed in an antibody (capture)
coated microwell to bind if present. All unbound protein is Method facts:
washed away before a detector antibody (enzyme labeled) is • Sample size: 20g
added. The detector antibody then binds to bound protein if • Limit of quantitation: Most matrices - 0.1%
present. After another wash, and the addition of a substrate, • Precision: On a ground carrot matrix, the RSD is 1.00%
color develops as a result of bound detector antibodies. After
the addition of a stop reagent, the test is read with a Stat Fax
reader. Optical densities of the controls form a standard curve Description: The sample is dried in a vacuum oven at 70˚C
which samples are plotted against to obtain concentrations of to a constant weight.
allergen protein in ppm.
Moisture, 100˚C (vacuum oven)
Milk thistle Purpose: Applicable for the determination of moisture in
Silychristin, silydianin, silibibin A & B foods and feeds, except for fruit, fruit products, vegetables,
vegetable products, sugar, and sugar products.
Purpose: Applicable for the determination of silymarins in Method facts:
dietary supplement products. • Sample size: 10g
Method facts: • Limit of quantitation: Most matrices - 0.1%
• Sample size: 5g • Precision: On a dog food matrix, the RSD is 2.9%
• Limit of quantitation: 0.05% • Method reference: AOAC 925.09, 926.08
• Precision: Varies with matrix Description: The sample is dried in a vacuum oven at 100˚C
• Method reference: USP (modified) to a constant weight.
Description: Samples are extracted with methanol, filtered,

centrifuged, and are separated using reverse-phase HPLC Moisture (Karl Fischer)
and UV detection. Silychristin, silydianin, silybin A&B, Purpose: Applicable for the determination of water content
in most matrices involving a USP or FCC monograph.
isosilybin A&B, and dehydrosilybin are identified, the sum
of these peak areas is considered total silymarin content of Method facts:
the sample (as silybin equivalents). • Sample size: 10g
Mineral profiles (ICP or ICP-MS) • Precision: Varies with matrix
see Inorganic analysis by ICP or Inorganic analysis by • Method reference: USP
ICP-MS
Description: The sample is dissolved or suspended in 50mL
of solvent and titrated with Karl Fischer moisture reagent.
Moisture, 60˚C (vacuum oven) Milli-Q® water is used to calibrate the instrument.
Purpose: Applicable for the determination of moisture in
sugars, syrups, honey, and high sugar products. Molybdenum (atomic absorption)
Method facts: Purpose: Applicable for the determination of molybdenum
in feeds, animal tissues, food products, plants, vitamin tablets
and supplements, water, soils and minerals.

[ assay ]
Method facts: n-Methyl Carbamates

• Sample size: 15g see Carbamate pesticides (LC-MS/MS)
• Limit of quantitation: Most matrices - 2ppm
• Precision: Determined on a spike recovery basis Neutral detergent fiber
• Method reference: Analytical Methods for Atomic see Detergent fiber, neutral
Absorption Spectrophotometry, Perkin-Elmer: Norwalk,
Connecticut, (2000).
Niacin
Official Methods of Analysis of AOAC Purpose: Applicable for the determination of niacin in foods,
INTERNATIONAL, 18th Ed., Method 974.27, AOAC feeds, infant formulas, premixes, and pharmaceutical products.
INTERNATIONAL, Gaithersburg, MD, USA,
(Modified) Method facts:
Description: The sample can be either dry ashed, wet ashed,
or dissolved and read directly. If dry ashed, the sample is • Limit of quantitation: Most matrices - 0.3 mcg/g
dried, pre-charred, and ashed at approximately 500° C for 5 • Precision: On a infant formula matrix, the RSD is 4.08%
to 16 hours. The sample is removed from the muffle furnace,
• Method reference: AOAC 944.13, 960.46, 985.34
cooled, treated with nitric acid, re-ashed, and dissolved in
dilute hydrochloric acid solution. If wet ashed, the sample USP, and Methods of Analysis for Infant Formulas, Infant
is digested on a hot plate with nitric acid, hydrochloric acid, Formula Council (1985)
and/or hydrogen peroxide.
Description: The sample is hydrolyzed with sulfuric acid
Water samples are evaporated to dryness on a hot plate with (H2SO4) and the pH is adjusted to remove interferences.
nitric acid and repeated as necessary to complete ashing The amount of niacin is determined by comparing the
prior to dissolution with dilute hydrochloric acid. growth response of the sample, using the bacteria
The amount of molybdenum is determined by comparing Lactobacillus plantarum, with the growth response of a
the signal of the unknown sample, measured by the atomic niacin standard. This response is measured turbidimetrically.
absorption (AA) spectrophotometer, with the signal of the
standard solutions.
Nickel (ICP-MS)
see Inorganic analysis by ICP-MS
Molybdenum (ICP or ICP-MS)
Nitrate/nitrite by HPLC
Purpose: Applicable for the determination of nitrates and
Monosodium glutamate (as glutamic acid)
nitrites in premixes, foods, and dietary supplements.
see Amino acid profile, free
Method facts:
n-Acetylcysteine • Sample size: 15g
Purpose: Applicable for the determination of • Limit of quantitation: Most matrices – 1ppm NO2,
n-Acetylcysteine in raw material. 1ppm NO3
Method facts: • Precision: On a recovery basis
• Sample size: 10g • Method reference: Journal of Chromatography, 804:157-160
• Limit of quantitation: Varies with matrix (1998)
• Precision: Varies with matrix Description: Samples are extracted with water. Lipids are
• Method reference: USP (Modified) removed with chloroform extraction and proteins
precipitated by centrifugation. The extract is filtered and
Description: Prepare sample using a sodium bisulfate
solution, dilute to volume and mix. Inject sample into an
anion exchange (HPAE) system equipped with an
HPLC system equipped with UV detection.
electrochemical detector using pulsed amperometric
detection (PAD). Nitrates and nitrites are quantitated
NL-Proximate (moisture, ash, protein) against standards of known concentration.
see individual tests for descriptions
[ assay ] N
Nitrate/nitrite by IC and VIS Method facts:

Purpose: This method is applicable for the determination of • Sample size: 100 g
nitrite and nitrate in food and beverages. • Limit of quantitation: Using a 10 -g sample, the lowest
confidence level for this assay is 1.0 ppb
Method facts:
• Precision: On a recovery basis
• Sample size: 5g
• Method reference: Nitrosamines/One Trap GC-TEA
• Limit of Quantitation: 0.50 ppm for nitrite and 1.00 ppm Mineral Oil Vacuum Distillation Analysis, USDA FSIS
for nitrate. The LOQ for protein samples is 1.00 ppm for Method NTR-1 ( July 1991)
nitrite and 2.50 ppm for nitrate.
Description: The sample is vacuum distilled and collected in a
• Method Reference: Casanova, J., Gross, L., McMullen, S.,
liquid nitrogen trap. After distillation, the trap is allowed to
and Schenck, F. “Use of Griess Reagent Containing
warm to room temperature, then extracted with methylene
Vanadium (III) for Post-Column Derivatization and
chloride. The methylene chloride extract is concentrated to
Simultaneous Determination of Nitrite and Nitrate in
1mL and analyzed on a GC system equipped with a thermal
Baby Food,” J. AOAC Int., 89(2): 447-451 (2006)
energy analyzer.
(Modified)
Gapper, L., Fong, B., Otter, D., Indyk, H., and Woollard,
D. “Determination of Nitrite and Nitrate in Dairy Nitrosamines — Volatile in rubber products
Products by Ion Exchange LC with Spectrophotometric 5-batch minimum for analysis.
Detection,” Int Dairy J., 14: 881-887 (2004) (Modified)
Purpose: Applicable for the determination of
George, S., Ofitserova, M., and Pickering, M.,
N-nitrosodimethylamine (NDMA), N-nitrosodiethylamine
“Simultaneous Determination of Nitrite and Nitrate in
(NDEA), N-nitrosodibutylamine (NDBA),
Processed Foods,” Method Abstract for Post-column
N-nitrosodipropylamine (NDPA), N-nitrosopiperidine
Liquid Chromatography 123, Pickering Laboratories, Inc.
(NPIP), N-nitrosopyrrolidine (NPYR),
(2011) http://www.pickeringlabs.com (accessed 06 Mar
N-nitrosomorpholine (NMOR) in rubber products.
2013).
Griess Reaction Diagram, www.biotek.com (Modified) Method facts:
Description: Samples are extracted or dissolved with water • Sample size: 5g
or buffer. Protein samples are treated with an acetonitrile • Limit of quantitation: Using a 10-g sample, the lowest
precipitation and clarified using centrifugation to remove confidence level for this assay is 1.0 ppb
most protein. The clarified protein extract is diluted with • Precision: On a recovery basis
60% (v/v) acetonitrile if necessary and vialed for injection. • Method reference: AOAC 987.05
All other samples are extracted with chloroform and D.C., Havery, T. Fazio, Fd. Chem. Toxic., 20:939-944 (1982)
centrifuged to remove lipids. A reverse phase solid phase
cartridge is then used to remove any remaining hydrophobic Description: Volatile N-nitrosamines are extracted from
components that may shorten the lifetime of the column. rubber products with methylene chloride. The solution is
The extract is diluted with water and vialed for injection. made alkaline. The methylene chloride and water are distilled
Nitrite and nitrate are separated by anion exchange with and N-nitrosamines in aqueous phase are partitioned back
post column pump and reaction coil. The post column into methylene chloride. The extract is concentrated to 1mL
reagent contain Griess reagent and vanadium (III). Nitrite and analyzed on a GC system equipped with a thermal
elutes first and is derivatized by the Griess reagent to an azo energy analyzer.
compound that is detected at 540 nm. When nitrate elutes,
it is first reduced to nitrite by vanadium at 95°C. Once
converted to nitrite it reacts with the Griess reagent to form Nucleosides
the chromophore detected at 540 nm. Purpose: Applicable for the determination of the nucleosides
cytidine, uridine, inosine, guanosine and adenosine in infant
formula and some food products.
Nitrosamines — Volatile in foods
5-batch minimum for analysis. Method facts:
• Sample size: 5g
N-nitrosodiemethylamine (NDMA), N-nitrosodiethylamine • Limit of quantitation: Most matrices - 0.1mg/g
(NDEA), N-nitrosodibutylamine (NDBA), • Precision: On a control sample the RSDs are as follows:
N-nitrosopiperidine (NPIP), N-nitrosopyrrolidine (NPYR), cytidine - 5.2%, uridine - 8.1%, inosine - 6.9%,
N-nitrosomorpholine (NMOR) in food products. guanosine - 14.5%, adenosine - 4.5%

[ assay ]
• Method reference: Olivartes, J. and Verdys, M., “Isocratic Description: ORAC analysis is conducted by subjecting a
High-Performance Liquid Chromatographic Method for portion of extracted sample into a system that consists of a
Studying the Metabolism of Blood Plasma Pyrimidine fluorescent probe and a free radical generator. This system
Nucleosides and Bases: Concentration and Radioactivity stimulates the in vivo conditions and quantitatively evaluates
Measurements,” Journal of Chromatography, pp 434 (1988) the potential of antioxidant protection against cellular damage
from the free radical. This is accomplished by monitoring
Description: The proteins are removed from the sample the fluorescence of the probe over time. The extraction of
matrix by acid precipitation and filtration. The filtrate is antioxidants can be performed on a hydrophilic basis by
further cleaned on a phenylboronic acid (PBA) solid-phase means of a single extraction. Results are expressed in terms
extraction column and then eluted with a formic acid of Trolox equivalents.
solution. The nucleosides are quantified on an HPLC
system equipped with a UV absorbance detector against a
Omega 3 fatty acids
known standard.
Nucleotides Organic acid profiles

Purpose: Applicable for the determination of the nucleotides Purpose: Applicable for the analysis of organic acids in foods
cytidine-5’-monophosphate (5’CMP), uridine-5’-monophosphate and food products. Please refer to the following options:
(5’UMP), guanosine-5’-monophosphate (5’GMP), inosine-
5’-monophosphate (5’IMP) and adenosine-5’-monophosphate Option 1: Sorbic and benzoic acid
(5’AMP) in infant formula and some food products. Option 2: Citric acid, malic acid, lactic acid and acetic acid
Option 3: Succinic acid, formic acid, fumaric acid and
Method facts: tartaric acid
• Sample size: 5g Option 4: Propionic acid
• Limit of quantitation: Most matrices - 3.0 mg/100g Option 5: Oxalic acid
• Precision: On an infant formula sample, the RSDs are as Option 6: Q uinic acid, malic acid and citric acid in
follows: 5’AMP - 1.85%, 5’IMP - 4.66%, 5’GMP - 2.24%, cranberry products
5’CMP - 1.70%, 5’UMP - 4.58%
Option 7: Pyruvic acid
• Method reference: Paubert-Braquet M., Dupont CH.
Paoletti R, Foods, Nutrition and Immunity, “Quantification Method facts: Option 1
of Nucleotides in Human Milk and their effects on • Sample size: 2g
Cytokine Production by Murine Fibroblasts, J774A1 • Limit of quantitation: Option 1: 5ppm
Macrophages and Human Monocytes,” DYN Nutr Res
• Precision: 5.11%
Basel, Karger, 1:22-34 (1992)
• Method reference: Journal of the AOAC
Description: The nucleotides are extracted into an aqueous, INTERNATIONAL, 70(5) (1987)
protein-free filtrate; collected on a solid-phase extraction (SPE) Description: Samples are extracted with water and
strong anion exchange (SAX) column; and subsequently eluted methanol. The organic acids are separated using reverse-
with phosphate buffer. The eluted nucleotides are quantified phase HPLC and measured with UV detection.
using HPLC equipped with UV detection against a standard
of known concentration. Method facts: Options 2-7
• Sample size: 5g
ORAC
• Precision for Option 2: 3.65%
Total (includes hydrophilic and lipophilic)
• Precision for Options 3 & 4: determined on a recovery basis
Purpose: Applicable for the determination of oxygen radical • Method reference: AOAC 986.13
absorbance capacity in a variety of food products.
Description: Samples are extracted with a weak H2SO4
Method facts: solution. Organic acids are separated using reverse-phase
• Sample size: 5g HPLC and measured with UV detection. Results are
• Limit of quantitation: 5 micromoles TE/g quantified by comparing with a known reference standard.
• Method reference: Journal of Agricultural and Food Organo nitrogen pesticide screen (NIPE)
Chemistry (2003) 3273-3279 Information available on request
[ assay ] O-P
Organophosphate pesticides PAM screens

Purpose: Applicable to samples containing between 5% and see FDA PAM screens
70% fat.
Method facts: Pantothenic acid
• Sample size: 100g Purpose: Applicable for the determination of pantothenic
• Limit of quantitation: Matrix dependent acid in most foods, feeds, and dietary supplements.
Method facts:
• Method reference: FDA Pesticide Analytical Manual,
Volume 1, Section 121.3 through 121.33 and 252 • Sample size: 2g
Journal of the Association of Official Agricultural • Limit of quantitation: Most matrices - 0.4 mcg/g
Chemists, Volume 57, pp. 168-172 (1974) • Precision: On an infant formula matrix, the RSD is 3.51%
Journal of the Association of Official Agricultural • Method reference: AOAC 945.74, 960.46
Chemists, Volume 70, pp. 724-726 (1987)
Description: The sample is diluted with water or treated with
Journal of the Association of Official Agricultural an enzyme mixture to liberate the pantothenic acid from
Chemists, Volume 48, pp. 1158-1160 (1965) co-enzyme A and the pH is adjusted to remove interferences.
Bulletin of Environmental Contamination and Toxicology, The amount of pantothenic acid is determined by comparing
Volume 12, No. 6, pp. 717-720 the growth response of the sample, using the bacteria
Lactobacillus plantarum with the growth response of a calcium
Description: If the sample has a high moisture content,
pantothenate standard. This growth response is measured
sodium sulfate is added; if dry, no sodium sulfate is required.
turbidimetrically.
The sample is extracted with ethyl acetate, concentrated,
cleaned-up with gel permeation chromatography (GPC), and
injected for organophosphate insecticides on a gas Parabens
chromatograph (GC) utilizing a flame photometric detector Methyl, ethyl, propyl, butyl, isopropyl, isobutyl,
(FPD). para-hydroxybenzoates
Purpose: Applicable for the determination of parabenzoates

Organophosphate pesticides (chlorinated) in most samples.
see Pesticides, chlorinated/organophosphate
Method facts:
Ornithine • Limit of quantitation: 5 ppm
see Amino acid profile, free by AAA • Precision: Varies with matrix
• Method reference: Journal of the AOAC
Oxalic acid INTERNATIONAL (1985)
see Organic acid profiles — option 5 Description: Samples are extracted in a 50% methanol and
50% Millipore water solution. The samples are filtered and
then injected into an HPLC equipped with a UV detector.
PABA (para-aminobenzoic acid)
Purpose: Applicable for the determination of p-aminobenzoic PCNB and DDT
acid in fortified products, premixes, and various preparations. Purpose: Applicable for the determination of PCNB and
DDT in ginseng products.
Method facts:
• Sample size: 5g Method facts:
• Limit of quantitation: Most matrices-150 ppm, liquids-5 ppm • Sample size: 50g
• Precision: On a tablet matrix, the RSD is 6.87% • Limit of quantitation: 10 ppb
• Method reference: USP • Precision: Varies with matrix
• Method reference: FDA PAM 303
Description: Sample extraction is performed using an
aqueous/acetonitrile/acetic acid solution. A portion of that extract Description: The pesticides are extracted with a mixture
is analyzed on a HPLC equipped with UV detection set at of acetonitrile and water. The sample extract is purified by
270nm. P-aminobenzoic acid is then quantitated by comparing gel permeation chromatography followed by florisil
chromatography and the resulting fraction is quantitated on
the sample response to standards of known concentration.
a GC using halogen-specific detectors.

[ assay ]
Peanut Protein by ELISA Method facts:

Purpose: Allergens are proteins in food that can create an • Sample size: 100g
immune response in sensitive individuals. Clear ingredient • Limit of quantitation: Varies with matrix
labeling of food products, by manufacturers, aids in protection
from accidental ingestion by those individuals. Testing for the
presence of these proteins ensures the food is free of the • Method reference: FDA PAM
allergen at a potentially harmful level.
Description: The sample is extracted with ethyl acetate,
Method facts: concentrated, cleaned with gel permeation chromatography,
• Sample Size: 50g and injected for organophosphate insecticides on a GC
system utilizing a flame photometric detector. Cleanup for
chlorinated insecticides is done using florisil column
• Method Reference: Veratox ® Quantitative Peanut Allergen chromatography. The sample is concentrated and injected on
Kits Neogen Corporation AOAC RI License Number a GC using a Hall detector or an electron capture detector.
030403
Also see pages 33 and 62 for other pesticide screens.
Description: Allergen Protein is extracted with a buffered salt
solution, allowed to settle then placed in an antibody (capture) Chlorinated compounds Organophosphate compounds
coated microwell to bind if present. All unbound protein is
Aldrin Acephate
washed away before a detector antibody (enzyme labeled) is
added. The detector antibody then binds to bound protein if BHC (Benzene hexachloride) Azinphos – methyl
present. After another wash, and the addition of a substrate, alpha BHC Chlorfenvinphos
color develops as a result of bound detector antibodies. After beta BHC Chlorpyrifos – ethyl
the addition of a stop reagent, the test is read with a Stat Fax gamma BHC (lindane) Chlorpyrifos – methyl
reader. Optical densities of the controls form a standard curve delta BHC Coumaphos
which samples are plotted against to obtain concentrations of
Captafol Demeton – S
allergen protein in ppm.
Captan (Orthocide) Diazinon
Chlorothalonil (Bravo) Dichlofenthion
Peroxide value Cypermethrin Dichlorvos
Purpose: Applicable for the determination of peroxide value DCPA (Dacthal) Dimethoate
in all normal fats and oils and extracted fats and oils. This p,p’-DDD Disulfoton
method is highly empirical and any variation in procedure p,p’-DDE EPN
may result in variation in results. p,p’-DDT Ethion
Dichloran (DCNA, Botran) Fenitrothion
Method facts:
• Sample size: 20g of food/feeds, 5g of oil Dicofol (Kelthane) Fonophos
Dieldrin Malathion
Endosulfan I (Thiodan) Methamidophos
• Precision: NA Endosulfan II Methidathion
• Method reference: AOCS method Cd 8-53 Endosulfan sulfate Mevinphos
AOAC extraction 983.23 Endrin Omethoate
USP <401> fats and fixed oils Folpet (Folpan, Phaltan) Parathion – ethyl
Heptachlor Parathion – methyl
Description: This method determines all substances, in terms Heptachlor epoxide Phosalone
of milli-equivalents of peroxide per kilogram of sample, that Hexachlorobenzene (HCB) Phosmet
oxidize potassium iodide under the conditions of the test. Methoxychlor (Marlate) Pirimiphos – methyl
These are assumed to be peroxides or other similar products Mirex Propetamphos
of fat oxidation.
Oxadiazon Prothiophos
PCNB (Quintozene, Avic) Ronnel
Pesticides Permethrin Thimet
Chlorinated/organophosphate Propyzamide Trithion
Tecnazene
Purpose: Applicable for the determination of residues in Tetradifon
samples containing between 5 and 70% fat.
Vinclozolin
[ assay ] P
Pesticide multiresidue analysis (300+ compounds) on retention time and specific Multiple Reaction
Purpose: Applicable for the determination of pesticides in Monitoring (MRM) transitions, one for quantification, and
various food, dietary supplement, infant formula and other one or more for confirmation of identity. Compounds
related matrices. analyzed include the following:
Method facts: • Butylbenzyl phthalate (BBP)

• Sample size: 25 g (50 g for high-moisture samples, such as • Dibutyl phthalate (DBP)
fruits and vegetables) • Bis(2-ethylhexyl)phthalate (DEHP)
• Limit of quantitation: 0.01 mg/kg for most analytes and • Di-n-octyl phthalate (DNOP)
matrices
• Diisononyl phthalate (DINP)
• Diisodecyl phthalate (DIDP)
• Method reference:
1AOAC Official Method 2007.01
2CEN Standard Method EN 15662 Phenolic acids
3 Purpose: Applicable for the determination of ferulic,
Anastassiades, M.; Lehotay, S.J.; Stajnbaher, D.; Schenck,
p-coumaric, sinapic acid, and caffeic acids in grains and
F.J. Journal of the AOAC INTERNATIONAL, 2003, 86,
412-431 forages.
4 Lehotay, S.J.; Mastovska, K; Lightfield, A.R. Journal of Method facts:
the AOAC INTERNATIONAL, 2005, 88, 615-629 • Sample size: 10g
5 Lehotay, S.J.; Mastovska, K.; Yun, S. J. Journal of the • Limit of quantitation: Most matrices - 17ppm
AOAC INTERNATIONAL, 2005, 88, 630-638
6 Mastovska, K.; Dorweiler, K.; Lehotay, S.J.; Wegscheid, J.;
• Method reference: Hagerman, A. E., Nicholson, R. L.,
Szpylka, K. J. Agric. Food Chem., 2010, 58, 5959-5972
High-Performance Liquid Chromatographic Determination
Description: The sample extraction and clean-up are based of Hydroxycinnamic Acid in Maize Mesocotyl, Journal of
on the QuEChERS method.1-6 The final extract is split Agriculture Food Chemistry, 3(6): 1098-1102 (1982)
for the analysis of GC- and LC-amenable pesticides using
Description: The ground sample is extracted using
gas chromatography-tandem mass spectrometry (GC-MS/
methanol followed by alkaline hydrolysis and buffering prior
MS) and liquid chromatography-tandem mass spectrometry
to injection on an analytical high performance liquid
(LC-MS/MS), respectively. If requested, the results can
chromatography (HPLC) system for quantification of
be corrected to potential matrix effects using the method
sinapic acid, p-coumaric acid, caffeic acid, and ferulic acid by
of standard addition, which provides the most accurate
ultraviolet (UV) detection.
quantification.
Phosalone
Phthalates
Purpose: Applicable for the determination of phthalates in
food or food supplement matrices.
Phospholipids
Method facts: Lecithin phospholipids, phosphatidic acid,
• Sample size: 5g phosphatidylethanolamine, phosphatidylcholine,
• Limit of quantitation: 1 ppm, can vary based on matrix. If phosphatidylserine, phosphatidylinositol
matrix suppression is observed, results will be reported as
“estimated”, and can be confirmed using method of Purpose: Applicable for the determination of lecithin
Standard Addition. phospholipids in oil-containing lecithins, deoiled lecithins,
• Precision: Varies with matrix and lecithin fractions. This method is not applicable for the
determination of lyso-PC and lyso-PE.
• Method reference: Test Method for Assay of Di(2-
ethylhexyl) phthalate (DEHP) in Food, Taiwan Food and Method facts:
Drug Administration (modified) • Sample size: 25g
Description: Phthalates are extracted by sonication with • Limit of Quantification: Most matrices – 1%
methanol and dilution to volume with water. The methanol • Precision: Varies with matrix
solution is analyzed and each phthalate is identified based • Method reference: AOCS Ja 7b-91

[ assay ]
Description: The sample is extracted with 65:25:1 Method facts:

chloroform:2-propanol:water. The extract is then analyzed on • Sample size: 100g
an HPLC system equipped with an evaporative light- • Limit of quantitation: 2 to 16 ppb, can vary based on
scattering detector (ELSD). A calibration curve is used for matrix
quantification.
• Method reference: NOAA Technical Memo
Phosphorus (ICP)
see Inorganic analysis by ICP Description: Samples are extracted with petroleum
ether:ethyl ether and injected into a GC. Residues are
quantitated using a GC-MS/MS. Compounds analyzed are
p-Hydroxybenzoates
as follows:
Methyl, ethyl, propyl, butyl, isopropyl, isobutyl
see Parabens 3-Methylcholanthrene Coronene
5-Methylchrysene Cyclopenta(c,d)pyrene
Phytic acid 7,12-Dimethylbenz(a)anthracene Dibenz(a,h)anthracene
Purpose: Applicable for the determination of phytic acid in Acenaphthene Dibenzo(a,h)pyrene
food products. Acenaphthylene Dibenzo(a,i)pyrene

Anthanthrene Dibenzo(a,l)pyrene
Method facts: Anthracene Dibenzo(a,e)pyrene
• Sample size: 5g Benz(a)anthracene Parathion-methyl
• Limit of quantitation: Most matrices - 0.125% Benzo(a)pyrene Fluoranthene
• Precision: Varies with matrix Benzo(b)fluoranthene Fluorene
• Method reference: Lehrfeld, J., HPLC Separation and Benzo(b)fluorene Dichlorvos
Quantitation of Phytic Acid and Some Inositol Benzo(b)naphthol(2,1-d)thiophene Phosalone
Phosphates in Foods, Journal of Agriculture and Food Benzo(c)phenanthrene Indeno(1,2,3-c,d)pyrene
Chemistry, 42:2726-2731 (1994) Benzo(e)pyrene Perylene
Lehrfeld, J., HPLC Analysis of Phytic Acid on a pH Benzo(g,h,i)perylene Phenanthrene
Stable, Macroporous Polymer Column, Journal of Cereal Benzo(j)fluoranthene Pyrene
Chemists, 66:510-515 (1989) Benzo(k)fluoranthene Naphthalen
Chysene
Description: The sample is extracted using ultrasonication.
Purification and concentration is done on a silica based
anion exchange (SAX) column. Sample analysis is done Polydextrose (high level)
on a macroporous polymer HPLC column and a refractive Purpose: Applicable for the determination of polydextrose
index detector. in premixes, foods, and raw materials.
Method facts:
Phytosterols • Sample size: 5g
see Sterols • Limit of quantitation: Most matrices – 1.0%
• Precision: Based on a spike recovery
Piperonyl butoxide • Method reference: Journal of Chromatography,
see USP 561 and EP pesticides 299: 288-292 (1984)
Description: The sample is extracted with a warm dilute

Pirimiphos-methyl sulfuric acid solution. The sample is then filtered, and an
see USP 561 and EP pesticides appropriate dilution is injected in an HPLC equipped with a
resin column and a refractive index detector. Samples are
then quantitated by comparing responses against a standard
Polyaromatic hydrocarbons of known concentration.
Purpose: Determination of PAH and similar compounds in
vegetable oil, fish and seafood as food, foods, and feeds. Polydextrose, with enzymatic pre-treatment
Purpose: Applicable for the determination of polydextrose
in foods, food products, pet foods and feeds.
[ assay ] P
Method facts: Description: Polysorbates are extracted from the sample

• Sample size: 5g with dichloromethane and ethanol. The extract is
centrifuged and filtered, and a portion is subjected to a
• Limit of quantitation: Most matrices – 0.5%
cleanup procedure using a silica column. After exchanging
• Precision: On a recovery basis the sample into water, the polysorbates are shaken with
• Method reference: Official Methods of Analysis of dichloromethane and a complexing agent to produce a blue
AOAC INTERNATIONAL (2005) 18th Ed., AOAC color, which is read on a spectrophotometer.
INTERNATIONAL, Gaithersburg, MD, USA, Official
Method 2000.11 (Modified) Potassium (ICP)
Description: Interfering fructans such as inulin, starches, see Inorganic analysis by ICP
and maltodextrins are removed by treating the sample with
the enzymes iso-amylase, amyloglucosidase, and fructanase.
Potassium sorbate
An aliquot of the final extract is filtered through a 0.45 μm
filter into an injection vial and then applied to a high-
performance anion exchange chromatography (HPAEC)
system with pulsed-amperometric detection (PAD). The Proanthocyanidins
amount of polydextrose present is determined by comparing Total polyphenols and catechins
the response of a known standard to that of the sample.
see Individual tests
Polyphenols, total
Propionic acid
Folin-Ciocalteu method
Purpose: Applicable for the determination of total polyphenol
content of tea leaves and other vegetable products. Vitamin
C will interfere with this assay. Protein (Dumas)
Purpose: Applicable for the determination of protein in
Method facts: most foods and feeds.
Method facts:
• Limit of quantitation: Most matrices – 1.0 mg/g
• Sample size: 1g
• Precision: On a green tea sample, the RSD is 2.43%
• Method reference: Methods in Enzymology, Oxidants and
• Precision: On a cereal matrix, the RSD is 0.9%
Antioxidants Part A, 299: 152-178 (1999)
Description: Polyphenols are extracted from the sample with
boiling methanol. The sample is reacted with Folin- Description: Nitrogen is determined by a combustion-detection
Ciocalteu reagent (FCR) to produce a color that can be technique. Nitrogen is released at high temperatures into a pure
measured spectrophotometrically at 760 nm. Results are oxygen atmosphere and measured by thermal conductivity. The
reported in units of mg/g gallic acid equivalents (GAE). percent nitrogen is converted to the protein using a factor
of 6.25.
Polysorbates
Purpose: Applicable for the analysis of polysorbates in foods
Protein (Kjeldahl)
and supplements.
Purpose: Applicable for the determination of protein in
Method facts: most foods and feeds.
Method facts:
• Limit of quantitation: Most matrices – 0.20% • Sample size: 5g
• Precision: Based on a spike recovery • Limit of quantitation: Most matrices - 0.1%
• Method references: KATO, H., Nagai, Y., Yamamoso, K., • Precision: On a dog food matrix, the RSD is 1.25%
and Sakabe, Y., “Determination of Polysorbates in Food by • Method reference: AOAC 955.04, 979.09 (Modified)
Colorimetry with Confirmation by Infrared
Spectrophotometry, Thin-Layer Chromatography and Gas Description: The protein and other organic nitrogen in the
Chromatography,” Journal of the Association of Analytical sample is converted to ammonia by digesting the sample
Chemistry, 72(1): 27-29 (1989) with sulfuric acid containing a catalyst mixture. The acid

[ assay ]
digest is made alkaline, and the ammonia is distilled and Please specify analytes of interest and required limit of
titrated with standardized acid. The percent nitrogen is detection when submitting samples.
determined and converted to protein using the factor 6.25.
Method facts:
Pueraria (isoflavones) • Limit of quantitation: Most matrices - 1 to 5ppm
see Isoflavones • Precision: Varies with matrix
• Method reference: Bertsch, Brian, “Developing One
Pygeum (beta-sitosterol) Universal Method for Residual Solvents Using the New
see Sterols Teledyne Tekmar HT3 Headspace Sample Introduction
System,” Application Note (Teledyne Tekmar), Document
#HT3-001 (September 2005)
Pyridoxine
see Vitamin B6 Description: The samples are introduced to a gas
chromatography mass spectrometry system via a Tekmar
HT3 purge-and-trap headspace autosampler. A single point
Pyruvate (pyruvic acid) standard at the tolerance level is normally used.
Residual Solvents (USP<467> Class 1, 2a, 2b,
Quinic acid, malic acid, citric acid (cranberry) and 3)
see Organic acid profiles — option 6 Purpose: This method is applicable to the determination of
residual solvents in most matrices. They include USP <467>
class 1, 2a, 2b and most class 3 compounds. Class 3 solvents
Reducing sugars that are not analyzed by this method are; acetic acid, formic
Purpose: Applicable for the determination of reducing sugars acid and dimethyl sulfoxide and all class 2c analytes. The
in vegetables, sugar products, beverages, dairy products solvents that are a part of this method are tetralin, anisole,
(except cheese), and food mixtures. It is not applicable to cumene, xylenes, ethylbenzene, chlorobenzene, butyl acetate,
samples with large amounts of ascorbic acid or free amino acids. 2-hexanone, 1-pentanol, isobutyl acetate, toluene, isoamyl
alcohol, pyridine, methylisobutylketone, propyl acetate,
Method facts: 1,4-dioxane, methylcyclohexane, trichloroethylene,
• Sample size: 25g 1-butanol, n-heptane, isopropyl acetate, 1,2-dichloroethane,
• Limit of quantitation: Most matrices - 0.5% 1,2-dimethoxyethane, benzene, carbon tetrachloride,
cyclohexane, 1,1,1-trichloroethane, chloroform,
• Precision: On a recovery basis tetrahydrofuran, isobutanol, 2-butanol, ethyl acetate,
• Method reference: AOAC 906.03, 929.09 2-butanone, 1,2-dichloroethene, nitromethane, 1-propanol,
n-hexane, tert-butylmethyl ether, methylene chloride, methyl
Description: The sugars are extracted with water or alcohol, acetate, acetonitrile, isopropanol, ethyl formate, methanol,
and deproteinized. Reducing sugars are gravimetrically acetone, 1,1-dichloroethene, diethyl ether, ethanol, and
determined by the reaction with copper sulfate-alkaline n-pentane.
tartrate, and the remaining copper oxide. An extract may also
be treated with hydrochloric acid to invert the non-reducing Method facts:
agents for a total sugar determination. Sugars are calculated • Sample size: 1g
using the Hammond table from AOAC Official Methods • Limit of quantitation: Equal or less than USP <467>
of Analysis. tolerance for most matrices.
• Precision: Pass / Fail
• Method reference: Residual solvents by USP <467> class 1,
Residual solvents (USP <467> Class 1, 2a, 2b) 2a, 2b and 3
Purpose: Applicable for the screening of residual solvents in
most matrices. They include, but are not limited to Description: Solid and liquid samples are weighed into
methanol, ethanol, diethyl ether, acetone, dichloromethane, tared 20 mL headspace vials at a nominal mass of 0.05 g.
methyl tert butyl ether (MTBE), hexane, 2-butanone, ethyl The samples are dissolved/mobilized in 1 mL DMSO, then
acetate, 1,1,1-trichloroethane, cyclohexane, benzene, diluted with 5 mL of water.
1,2-dichloroethane, heptane, toluene, 1,1,2-trichloroethane,
n-butyl ether and xylenes(o,m,p).
[ assay ] R-S
Resistant maltodextrin (fibersol) casei, against the growth response of riboflavin standard.
with total dietary fiber This growth response is measured turbidimetrically.
Purpose: Applicable for the determination of food products

containing resistant maltodextrins. Rose hips (ascorbic acid)
Purpose: Applicable for the determination of ascorbic acid
Method facts: as an indicator of rose hips. It measures both ascorbic acid
• Sample size: Powder samples 5g, liquid samples 20g and iso-ascorbic acid if present.
• Limit of quantitation: Most matrices – 1.0%
Method facts:
• Precision: On a cereal matrix, the RSD is 3.36%
• Sample size: 8g
• Method reference: AOAC 2001.03 (modified)
• Limit of quantitation: Most matrices - 1 mg/100g
Description: Duplicate samples are gelatinized with alpha- • Precision: On a cereal sample matrix, the RSD is 7.33%
amylase and digested with enzymes to break down starch and • Method reference: AOAC 967.22
protein. Ethanol is added to each sample to precipitate the
soluble fiber. The samples are filtered, and the filtrate is Description: The vitamin C in the sample is extracted, oxidized,
purified and then concentrated. The sample is then analyzed and reacted with o-phenylenediamine to produce a fluorophor.
using an HPLC system with refractive index detection. The vitamin C content is determined by comparing the sample
extract fluorescence to the fluorescence of known standards.
Resistant starch
Royal jelly 10-HDA
Purpose: Applicable for the determination of resistant starch
10-Hydroxy-2-decenoic acid
in food products.
Method facts: 10-Hydroxy-2-decenoic acid in royal jelly and some
• Sample size: 0.1g (most matrices) products containing royal jelly.
Method facts:
• Precision: Available on request
• Limit of quantitation: Varies with matrix
Description: Nonresistant starch is eliminated by enzyme • Precision: Varies with matrix
treatment. Resistant starch (RS) is recovered and hydrolyzed • Method reference: Guererra III, Frank P., Pharmline Inc.,
to glucose, which is then quantified spectrophotometrically “HPLC Determination of 10-Hydroxy-2-decenoic Acid
at 510 nm. From this, the RS content is calculated. (10-HAD) in Royal Jelly,” personal correspondence (April 1996)
Description: Samples are dissolved in mobile phase using
Retinol sonication and filtered into autosampler vials. The samples are
see Vitamin A then analyzed by HPLC equipped with UV detection and
compared against an external standard.
Riboflavin (vitamin B2)

Purpose: Applicable for the determination of riboflavin in Rutin
most foods and feeds. Information available on request
Method facts:
• Sample size: 20g SAMe (S-adenosylmethionine)
Purpose: Applicable for the determination of S-adenosyl
• Limit of quantitation: Most matrices - 0.2mcg/g
methionine in dietary supplements, concentrates, and some
• Precision: On an infant formula matrix, the RSD is 9.06% food products.
Method facts:
The United States Pharmacopeia, 29th Ed., P. 1913 • Sample size: 10g
Description: The sample is hydrolyzed with dilute hydrochloric • Limit of quantitation: Varies with matrix
acid (HCl) and the pH is adjusted to remove interferences. • Precision: 8.9%
The amount of riboflavin is determined by comparing the • Method reference: d’Avignon, A., Reepmeyer, J., Revelle, L.,
growth response of the sample, using the bacteria Lactobacillus Zerfine, C., “Stability-Indicating Proton Nuclear Magnetic

[ assay ]
Resonance Spectroscopic Method for Determination of INTERNATIONAL, Gaithersburg , MD, USA, Official
S-Adenosyl-L-Methionine in Tablets,” 78(2):353 (1995) First Action Method 2011.19 (2011) (Modified)
Edwards, R., “Determination of S-Adenosyl-L-Methionine Description: The sample is digested with concentrated
and S-Adenosyl-L-Homocysteine in Plants,” 6:25-30 (1995) nitric acid and water using a closed-vessel microwave
digestion system. After digestion, the sample is transferred
Description: The product is extracted with a solution of to a single-use disposable plastic vessel, then brought to a
ammonium phosphate buffer mixed with methanol and final volume with water. To normalize the organic
analyzed for SAMe content utilizing reverse-phase HPLC contribution between sample and standard, a dilution is
system with UV detection. prepared for analysis containing methanol. The amount of
selenium is determined with inductively coupled plasma-
Saw palmetto (sterols + fatty acids) mass spectrometry (ICP-MS) by comparing the counts
generated by the unkowns to those generated by standard
Purpose: This involves the use of two methods, one for
solutions.
sterols and one for fatty acids, which serve as indicators for
the presence of saw palmetto.
Selenium (hydride generation)
Method facts:
Purpose: Applicable for the determination of selenium in
• Sample size: 7g
infant formulas, food products and supplements.
• Limit of quantitation: Sterols - 0.001%; fatty acids - 0.1%
• Precision: Sterols - 3.5%; fatty acids - 2-5% Method facts:
• Sample size: 5g
• Method reference: AOAC 994.10 (sterols),
AOCS Ce 1-62, Ce 1e-91, Ce 1a-07 • Limit of quantitation: Most matrices - 30 ppb
• Precision: On a rice flour matrix with a certified value of
Description: 0.38ppm, the RSD is 3.78%
Sterols: The sample (campesterol, betasitosterol,
stigmasterol) is saponified using ethanolic potassium • Method reference: AOAC 986.15, 996.16, 996.17
hydroxide. The unsaponifiable fraction that contains (Modified)
cholesterol and other sterols is extracted with toluene. The Perkin Elmer, Flow Injection mercury/Hydrite Analyses,
toluene is evaporated to dryness and the residue is dissolved Reccommended Analytical Conditions and General
in dimethylformamide (DMF). The samples are derivitized Information, Norwalk, CT (1994) (modified)
to form trimethylsilyl ethers. The derivitized sterols are
quantitatively determined by GC using 5α-cholestane as an Description: The sample is digested with a mixture of nitric
internal standard. and perchloric acids prior to a reduction with hydrochloric
acid. Following reduction, the samples are reacted with sodium
Fatty acids: The lipid is extracted, saponified with 0.5N borohydride to produce the volatile selenium hydride, which
methanolic sodium hydroxide, and methylated with 14% is measured by atomic absorption spectroscopy. The amount
BF3-methanol. The resulting methyl esters of the fatty acids of selenium is determined by comparing the signal of the
are extracted with heptane containing an internal standard. unknown to the signal of standard solutions.
The methyl esters of the fatty acids are analyzed by GC
using external standards for quantitation.
Sennosides
Purpose: Applicable to the determination of sennosides in
Selenium by ICP-MS food products, laxatives, weight loss pills, and other
Purpose: Applicable for the determination of selenium in a nutritional and medicinal products.
variety of matrices including, but not limited to, infant and
adult nutritional formulas, rice, wheat, soy and seeds (e.g., Method facts:
canola). • Sample size: 1g
Method facts: • Limit of quantitation: Most matrices - 100 ppm
• Sample size: 10g • Precision: On a vegetable laxative sample the RSD is
1.70%
• Limit of quantitation: 0.030 ppm
• Method reference: USP
• Precision: The RSD is 3.32% on a non-fat milk powder,
2.08% on a wheat flour, 1.83% on a rice flour, 2.39% on a Description: A standard of a known concentration of
powdered infant formula, 1.80% on an infant/adult sennosides and samples are extracted in a pH 7.0 buffer
nutritional formula, 7.76% on soy meal and 1.24% on solution with sonication. An aliquot is diluted with a sodium
canola meal borate solution. An aliquot of this solution is then reacted
• Method reference: Official Methods of Analysis of AOAC with a sodium dithionite solution in a steam bath in a
INTERNATIONAL 18th Ed., AOAC nitrogen gas atmosphere to form a fluorophor with the
[ assay ] S
sennosides. The intensities of the solutions are measured on measured by the atomic absorption spectrophotometer, to the
a fluorometer at an λex = 390, λem = 500. A blank solution absorbance of the standard solutions.
is treated similarly, but not reacted with sodium dithionite.
Its fluorescence intensity is subtracted from that of the
sample solution to correct for background fluorescence in Sinapic acid
the sample. Results are calculated in reference to the see Phenolic acids
calibration standard
Silver (ICP-MS)
Sesame Protein by BioKits see Inorganic Analysis by ICP-MS
Purpose: Allergens are proteins in food that can create an
immune response in sensitive individuals. Clear ingredient
Sodium (ICP)
labeling of food products, by manufacturers, aids in see Inorganic Analysis by ICP
protection from accidental ingestion by those individuals.
Testing for the presence of these proteins ensures the food is
free of the allergen at a potentially harmful level. Sodium benzoate
Method facts:
• Sample Size: 50g
Sorbic acid
• Limit of Quantitation: 6.25 ppm Sesame
• Method Reference: BioKits Sesame Assay; Neogen
Corporation
Sorbitol
Description: Allergen Protein is extracted with high salt see Sugar alcohols
Tris buffer solution, allowed to settle then placed in
antibody (capture) coated micowell to bind if present. All
unbound protein is washed away then a biotin specific to the Soy isoflavones
allergen is added and incubated. After another wash step, see Isoflavones
Avidin Peroxidase Conjugate is added and incubated. It
binds to already bound protein. After another wash, and the
Soy Protein by ELISA
Purpose: Allergens are proteins in food that can create an
detector antibodies. After the addition of a stop reagent, the immune response in sensitive individuals. Clear ingredient
test is read with a Stat Fax reader. Optical densities of the labeling of food products, by manufacturers, aids in
controls form a standard curve which samples are plotted protection from accidental ingestion by those individuals.
against to obtain concentrations of allergen protein in ppm. Testing for the presence of these proteins ensures the food is
free of the allergen at a potentially harmful level.
Silicon (atomic absorption) Method facts:
Purpose: Applicable for the determination of silicon in most • Sample Size: 50g
matrices including feeds, food products, minerals, animal • Limit of Quantitation: 2.5 ppm
tissues, plants, and water samples.
• Method Reference: Veratox ® Quantitative Almond
Method facts: Allergen Kits Neogen Corporation
Description: Allergen Protein is extracted with a buffered
• Limit of quantitation: Most matrices - 300 ppm salt solution, allowed to settle then placed in antibody
• Precision: Varies with matrix (capture) coated micowell to bind if present. All unbound
• Method reference: Perkin-Elmer, Analytical Methods for protein is washed away then a detector antibody (enzyme
Atomic Absorption Spectrophotometry, Norwalk, CT labeled) is added. It binds to already bound protein. After
(2000) another wash, and the addition of a substrate, color develops
as a result of bound detector antibodies. After the addition
Description: Most samples are dried, precharred, ashed of a stop reagent, the test is read with a Stat Fax reader.
overnight at 500˚C ±50˚, then fused with a mixture of sodium Optical densities of the controls form a standard curve
borate and sodium carbonate. Finally, the sample is dissolved in which samples are plotted against to obtain concentrations
dilute hydrochloric acid. The amount of silicon is determined of allergen protein in ppm.
by comparing the absorbance of the unknown sample,

[ assay ]
Species identification by ELISA Sucralose (finished products)

Beef, Poultry, Pork Purpose: Applicable for the determination of sucralose in
dietary supplements and food products.
Purpose: Applicable for the determination of the presence
of a particular meat species. Method facts:
• Sample size: 5g
Method facts:
• Sample size: 5g • Limit of quantitation: Most matrices - 0.02%
• Limit of quantitation: Economic limit is 1%; scientific • Precision: Based on a spike recovery
limit is 150-250ppm • Method reference: Journal of the AOAC
• Precision: NA INTERNATIONAL 71(5): 934-937 (1988)
• Method reference: ELISA-TEK Cooked Meat Species ELISA
Description: Sucralose is extracted from the sample with water.
Description: The sample is extracted with a saline solution. A portion of the extract is subjected to solid-phase extraction
The extract is carried into the ELISA test. (SPE) clean up. The extract is filtered and appropriate
dilutions are injected onto a high-performance anion exchange
(HPAE) system equipped with an electrochemical detector
Starch using pulsed amperometric detection (PAD). Sucralose is
Purpose: Applicable for the determination of starch in food quantitated against standards of known concentration.
products, pet foods, and feeds.
Method facts: Sucralose (packets, pure material)
• Sample size: 1g Purpose: Applicable for the determination of raw material
• Limit of quantitation: Most matrices - 0.05% or table top packets of sucralose.
• Precision: On a starch recovery basis, the RSD is 3.73% Method facts:
• Method reference: AOAC 996.11 (modified) • Sample size: 100mg
Description: The sample is extracted with alcohol to remove • Limit of quantitation: NA
carbohydrates other than starches. The starches are hydrolyzed • Precision: NA
to glucose with α-amylase and amyloglucosidase. Glucose is
oxidized with glucose oxidase to form a peroxide, which reacts • Method reference: USP, FCC
with a dye in the presence of peroxidase to give a stable Description: Sample is diluted in water/acetonitrile and
color proportional to glucose concentration. The glucose analyzed by HPLC equipped with RI detection.
concentration is quantitated spectrophotomerically at 510nm
and used to calculate starch content.
Sucrose
Sterols see Sugar profile by GC
beta Sitosterol, campesterol, stigmasterol, brassicasterol see Sugar profile by HPLC
see Sugar profile by IC
Purpose: Applicable for the determination of sterols in most see Sugar profile, low levels
matrices including foods, feeds and dietary supplements.
Method facts:
Sugar alcohols
• Sample size: 5g
Purpose: Applicable to quantify sorbitol, xylitol, mannitol,
• Limit of quantitation: 1 mg/100 g maltitol, lactitol, erythritol, and iso-maltitol.
• Precision: On a ground waffle matrix, the RSD is 3.5%
• Sample size: 5g
Description: The sample is saponified using ethanolic
• Limit of quantitation: 200ppm
potassium hydroxide. The unsaponifiable fraction that contains
cholesterol and sterols is extracted with toluene. The toluene • Precision: On a recovery basis
is evaporated to dryness and the residue is dissolved in • Method reference: Dionex Technical Note 20. Analysis of
dimethylformamide (DMF). The sample is derivitized to form Carbohydrates by High-Performance Anion Exchange
trimethysily ethers. The derivitized sterols are quantitatively Chromatography with Pulsed Amperomeric Detection
determined by GC using 5α-cholestane as an internal standard. (HPAE-PAD)
Cataldi, T.R.I.; Campa, C.: Casella, I.G.: Bufo, S.A.
Succinic acid Determination of Maltitol, Isomaltitol, and Lactitol by
[ assay ] S
High-pH Anion-Exchange Chromatography with Pulsed Chemistry, 19(3): 551-554 (1971)

Amperomeric Detection. J. Agric. Food Chem. 1999, 47, Brosbt, K., “Gas Liquid Chromatography of Trimethylsilyl
157-163. Derivations,” Methods in Carbohydrate Chemistry,
Academic Press, New York, NY, 6:3-8 (1972)
Description: A water extraction with low heat is used to extract
the sugar alcohols. An appropriate dilution is injected onto a Description: Sugars are extracted from the sample with
high-performance anion exchange chromatograph (HPAEC) water and methanol. Aliquots are dried under inert gas and
system equipped with a pulsed amperomeric detector (PAD) reconstituted with a hydroxylamine hydrochloride solution
for determination. in pyridine containing phenyl-β-D-glucoside as the internal
standard. The resulting oximes are converted to silyl
derivatives with
Sugar profile by acid hydrolysis hexamethyldisilazane (HMDS) and trifluoracetic acid
Purpose: Applicable to the determination of fructose, (TFA) treatment and analyzed by gas chromatography using
galactose, glucose, sucrose, lactose, maltose, raffinose and a flame ionization detector.
stachyose in food products, beverages, syrups, feeds and grains.
Method facts: Sugar profile by HPLC
Purpose: Applicable for the determination of fructose, glucose,
• Limit of quantitation: Most matrices - 0.1% sucrose, maltose, and lactose in a variety of matrices. It may
• Precision: On a cereal matrix, the RSDs for fructose, glucose, not be applicable for samples that are high in salts (especially
sucrose and maltose are 5.8%, 9.0%, 3.6%, and 4.3%, sodium chloride) or samples with high protein content.
respectively. On a soybean matrix, the RSDs for raffinose and
stachyose are 2.7 % and 4.5%, respectively. Method facts:
• Method reference: Mason, B.S. and Slover, H.T., “A Gas
Chromatographic Method for the Determination of • Limit of quantitation: Most matrices - 0.1%
Sugars in Foods,” Journal of Agricultural and Food • Precision: On a cereal matrix, the RSDs for fructose, glucose,
Chemistry, 19(3): 551-554, (1971) (Modified) and sucrose are 2.74%, 2.77% and 1.68%, respectively.
Brosbt, K., “Gas Liquid Chromatography of Trimethylsilyl • Method reference: AOAC 982.14
Derivations,” Methods in Carbohydrate Chemistry, 6:3-8,
Academic Press, New York, NY (1972) (Modified) Description: Sugars are extracted from the sample with water
and methanol. Aliquots are dried under inert gas and
Description: Sugars are extracted from the sample with reconstituted with an acetonitrile-water solvent. The
sulfuric acid and heat. Aliquots are dried under inert gas and solutions are injected on a HPLC system equipped with a
reconstituted with a hydroxylamine hydrochloride solution in refractive index (RI) detector.
pyridine containing phenyl-β-D-glucoside as the internal
standard. The resulting oximes are converted to silyl
derivatives with hexamethyldisilazane (HMDS) and Sugar Profile by IC
trifluoracetic acid (TFA) treatment and analyzed by gas Purpose: The method is applicable to the determination of
fructose, glucose, sucrose, maltose, and lactose in food
chromatography using a flame ionization detector.
products, beverages, syrups, feeds, and grains. It is not
applicable for samples containing galactose and lactulose.
Sugar profile by GC Method facts:
Purpose: Applicable for the determination of fructose, • Sample size: 10g
galactose, glucose, sucrose, lactose, and maltose in a variety • Limit of Quantitation: Most matrices - 0.1%
of matrices. • Precision: On a cereal matrix the RSD’s for fructose,
glucose, sucrose, maltose are 1.88%, 3.41%, 2.50%, and
Method facts: 5.61% respectively. On a milk powder matrix the RSD for l
• Sample size: 10g actose is 0.76%.
• Limit of quantitation: Most matrices - 0.1% • Method Reference: Official Method No. 982.14, Official
• Precision: On a cereal matrix, the RSDs for fructose, Methods of Analysis of AOAC INTERNATIONAL
glucose, sucrose, and maltose are 2.22%, 1.99%, 1.19%, and (modified), [Online] 2012, AOAC INTERNATIONAL
6.01%, respectively http://www.eoma.aoac.org (accessed 02 Jan 2013)
Dionex Technical Note 20: Analysis of Carbohydrates by
• Method reference: Mason, B.S. and Slover, H.T., “A Gas High-Performance Anion-Exchange Chromatography with
Chromatographic Method for the Determination of Pulsed Amperometric Detection (HPAEC-PAD)
Sugars in Foods,” Journal of Agricultural and Food

[ assay ]
Description: Sugars in the sample are extracted with a Taurine

mixture of equal parts of water and methanol, with the Purpose: Applicable for the determination of taurine in
addition of an internal standard. Samples are diluted within most food, dietary supplements and feeds.
the calibration range with water and filtered prior to analysis.
Method facts:
Sugar profile, low levels
Purpose: Applicable for the determination of galactose, • Limit of quantitation: Most matrices - 0.015mg/g
glucose, isomaltulose, fructose, sucrose, lactose and maltose • Precision: On a cat food matrix, the RSD is 3.5%
in foods and infant formulas at levels below 0.1%. • Method reference: Henderson, J.W., Ricker, R.D.,
Method facts: Bidlingmeyer, B.A., Woodward, C., “Rapid, Accurate,
• Sample size: 25g Sensitive, and Reproducible HPLC Analaysis of Amino
• Limit of quantitation: Levels as low as 0.005% can be Acids, Amino Acid Analysis Using Zorbax Eclipse-AAA
detected, large quantities of some sugars can interfere with columns and the Agilent 1100 HPLC,” Agilent
the proper determination of neighboring sugars. The presence Publication, 2000
of lactulose may interfere with the determination of extremely R. Schuster, “Determination of Amino Acids in Biological,
low levels of lactose. Pharmaceutical, Plant and Food Samples by Automated
• Precision: Varies with matrix Precolumn Derivitization and HPLC”, Journal of
• Method reference: Dionex Chromatography Database Chromatography, 1988, 431, 271-284
4.2.0, Record 755 AOAC 999.12
Description: The sample is extracted with warm water. Description: The sample is hydrolyzed in hydrochloric acid
Lipids are removed with chloroform extraction and proteins and is quantitated using an automated amino acid analyzer.
are removed by centrifugation. The extract is filtered and
Theobroma cacoa
anion exchange chromatograph (HPAEC) equipped with a
see Caffeine, theobromine, theophylline
pulsed amperometric detector (PAD).
Theobromine
Sulfites (Monier-Williams) see Caffeine, theobromine, theophylline
Purpose: Applicable for the determination of total sulfite in
food products, plant tissue, and most other materials in the Theophylline
presence of other volatile sulfur compounds. see Caffeine, theobromine, theophylline
Method facts:
• Sample size: 50g Thiamin (vitamin B1)
• Limit of quantitation: 8 ppm Purpose: Applicable for the determination of thiamin in
• Precision: 4.59% most foods, dietary supplements, and feeds.
• Method reference: AOAC 990.28 (modified)
Method facts:
Description: The sulfite is extracted from the sample in a • Sample size: 10g
distillation flask containing hydrochloric acid, to which a
• Limit of quantitation: Most matrices - 0.01mg/100g
condenser is connected. As the distillation progresses, nitrogen
gas is used to carry the sulfite out the top of the condenser • Precision: On a commercial egg noodle sample, the RSD
into receptacles containing hydrogen peroxide. The resultant is 2.44%
sulfuric acid solution is titrated with sodium hydroxide to • Method reference: AOAC 942.23, 953.17, 957.17
determine the sulfite content.
Description: The sample is autoclaved in dilute acid to
extract the thiamin. The resulting solution is incubated with
Sulfur (ICP-MS) a buffered enzyme solution to release bound thiamin and
see Inorganic analysis by ICP-MS hydrolyze starch. The solution is purified on an ion-exchange
column. An aliquot is taken and reacted with potassium
ferricyanide to convert thiamin to thiochrome. The thiochrome
Tartaric acid
is extracted into isobutyl alcohol and read on a fluorometer
against a known standard.
[ assay ] T
Thiobarbituric acid (TBA) value Tocopherols

Purpose: Applicable for the determination of oxidation products alpha, beta, gamma, delta
of unsaturated fatty acids as a measure of rancidity in foods.
Purpose: Applicable for the determination of alpha, beta,
Method facts: gamma, and delta tocopherol in oils, foods, feeds and
• Sample size: Sample size: 10g nutritional products.
• Limit of quantitation: Most matrices - 1mg/kg
Method facts:
• Precision: Varies with matrix • Sample size: 10g
• Method reference: American Oil Chemists’ Society, ‘Cd • Limit of quantitation: Most matrices - 0.5mg/100g
19-90 2-Thiobarbituric Acid Value Direct Method’, Official
Methods and Recommended Practices of the AOCS, Fifth • Precision: On a infant formula matrix, the RSD is 6.10%
Ed., American Oil Chemists’ Society, Champaign, IL (1997) • Method reference: Cort, W.M., Vicente, T.S., and
Williams, B.D., Journal of Agricultural Food Chemistry,
Description: The sample is homogenized and distilled at 31:1330-1333 (1983)
pH 1.5. The three carbon fraction (malonaldehyde) derived
from oxidation of monoeonic or polyeonic fatty acids is released Description: The product is extracted with an organic solvent
from rancid foods through acidification and distillation. The followed by separation on an HPLC silica column, and
malonaldehyde is collected and reacted with TBA to produce quantitation using fluorescence detection.
a red pigment. The quantity of pigment found is determined
by measuring absorbance at 538 nm on a spectrophotometer.
Tocotrienols
alpha, beta, gamma, delta
Thiabendazole
see PAM 304 pesticide screen Purpose: Applicable for the determination of alpha, beta,
gamma and delta tocotrienols in oils, food, premixes and
other dietary supplement preparations.
Tin (ICP-MS)
see Inorganic Analysis by ICP-MS Method facts:
Tin (graphite furnace) • Limit of quantitation: Most matrices - 0.500mg/100g
Information available on request • Precision: Varies with matrix
• Method reference: Cort, W.M., Vincente, T.S., Waysek,
Titanium (atomic absorption) E.H., and Williams, B.D., Journal of Agricultural Food
Purpose: Applicable for the determination of titanium in Chemistry, 31:1330-1333 (1983)
most materials including feeds, animal tissue, food products, Speek, A.J., Schijver, J., and Schreurs, W.H.P., Journal of
plants, vitamin tablets and supplements, water, soils, minerals, Food Science, 50:121-124 (1985)
and coals. McMurray, C.H., Blanchflower, W.J., and Rice, D.A.,
Journal of the Association of Official Analytical Chemists,
Method facts: 63:1258-1261 (1980)
Description: The product is extracted with organic solvent,
separated on an HPLC silica column and measured using
• Precision: Varies with matrix fluorescence detection.
• Method reference: Perkin-Elmer, Analytical Methods for
Atomic Absorption Spectrophotometer, Norwalk, CT,
(2000) Trans fat and fatty acids, by GC
see Fatty acids
Description: Most samples are dried, pre-charred, ashed
overnight at 500˚C ±50˚, then fused with a mixture of
sodium borate and sodium carbonate. Finally, the sample trans-Resveratrol
is dissolved in dilute hydrochloric acid. The amount of Purpose: Applicable for the determination of trans-
titanium is determined by comparing the absorbance of Resveratrol in raw material, extracts, capsules, tablets or
the unknown sample, measured by the atomic absorption other dietary supplement preparations.
spectrophotometer, to the absorbance of the standard
solutions.

[ assay ]
Method facts: 5Anastassiades, M.; Lehotay, S.J.; Stajnbaher, D.;

• Sample size: 5g Schenck, F.J. Fast and easy multiresidue method
• Limit of quantitation: Most matrices – 50 ppm employing acetonitrile extraction/partitioning and
• Precision: Varies with matrix “dispersive solid-phase extraction” for the determination
of pesticide residues in produce. Journal of AOAC
• Method reference: Sakakibara, H., Honda, Y., Nakagawa, S., INTERNATIONAL 2003, 86, 412-431
Ashida, H., and Kanazawa, K.,“Simultaneous Determination 6
Lehotay, S.J.; Mastovska, K; Lightfield, A.R. Use
of All Polyphenols in Vegetables, Fruits, and Teas,” J. Agric.
of buffering and other means to improve results of
Food Chem., 51(3):572-580 (2003)
problematic pesticides in a fast and easy method for
Description: Sample extraction is performed utilizing the residue analysis of fruits and vegetables. Journal of AOAC
appropriate technique. A portion of that extract is analyzed by
7
Lehotay, S.J.; Mastovska, K.; Yun, S. J. Evaluation
HPLC with fluorescence detection; results are then quantified by
comparing the response to a standard of known concentration. of two fast and easy methods for pesticide residue
analysis in fatty food matrixes. Journal of AOAC
Triazine 8
Mastovska, K.; Dorweiler, K.; Lehotay, S.J.; Wegscheid, J.;
see Herbicides Szpylka, K. Pesticide multiresidue analysis in cereal grains
using modified QuEChERS method combined with
automated direct sample introduction GC-TOFMS and
Trifluralin UPLC-MS/MS techniques. J. Agric. Food Chem., 2010,
see PAM 304 pesticide screen 58 5959-5972
9Lehotay, S.J.; Ae Son, K.; Kwon, H.; Koesukwiwat,
Tryptophan (AOAC) U.; Fu, W.; Mastovska, K.; Hoh, E.; Leepipatpiboon,
see Amino acid profile (AOAC), total N. Comparison of QuEChERS sample preparation
methods for the analysis of pesticide residues in fruits and
vegetables. J. Chromatogr. A 2010, 1217, 2548-2560
Tryptophan (HPLC)
Description: The sample extraction and clean-up are based
see Amino acid profile (HPLC), total
on the QuEChERS method.3-9 The final extract is split for
the analysis of GC- and LC-amenable pesticides using gas
USP 561 and EP Pesticides chromatography-tandem mass spectrometry (GC-MS/MS)
Purpose: Applicable, but not limited, to dietary supplement and liquid chromatography-tandem mass spectrometry (LC-
products and raw materials for the analysis of pesticides MS/MS), respectively. The reporting limits are based on the
listed in the US and European Pharmacopeias (USP and limits specified in the USP and EP pesticide residue
EP).1,2 monographs:
USP/EP Limit
Method facts: Pesticide (mg/kg)
• Sample size: 25g Acephate 0.1
• Limit of quantitation: Varies by compound; USP and EP Alachlor 0.05
have set limits Aldrin and dieldrin (sum of ) 0.05
• Precision: Varies with matrix Azinphos-ethyl 0.1
• Method reference: Azinphos-methyl 1
1USP34-NF29:561 Articles of Botanical Origin. In: The
Bromide, inorganic (calculated as bromide ion)*** 50
United States Pharmacopeia, 2011 Bromophos-ethyl 0.05
2
EP 07/2008:20813 Pesticide Residues. In: The European Bromophos-methyl 0.05
Pharmacopoeia, 7th Edition, 2011
Bromopropylate 3
3
AOAC Official Method 2007.01, Pesticide Residues in Chlordane (sum of cis- and trans- isomers and 0.05
Foods by Acetonitrile Extraction and Partitioning with oxychlordane)
Magnesium Sulfate Chlorfenvinphos 0.5
4
CEN Standard Method EN 15662: Food of plant Chlorpyrifos-ethyl 0.2
origin – Determination of pesticide residues using Chlorpyrifos-methyl 0.1
GC-MS and/or LC-MS/MS following acetonitrile Chlorthal-dimethyl 0.01
extraction/partitioning and clean-up by dispersive SPE –
Cyfluthrin (sum of isomers) 0.1
QuEChERS method
[ assay ] U
Cyhalothrin, λ- 1 Profenophos 0.1

Cypermethrin (sum of isomers) 1 Prothiophos 0.05
DDT (sum of o,p’-DDT, p,p’-DDT, o,p’-DDE, p,p’- 1 Pyrethrum (sum of cinerin I, cinerin II, jasmolin I, 3
DDE, o,p’-DDD, and p,p’-DDD) jasmolin II, pyrethrin I, and pyrethrin II)
Deltamethrin 0.5 Quinalphos 0.05
Diazinon 0.5 Quintozene [sum of quintozene, pentachloroaniline and 1
methyl pentachlorophenyl sulfide (MPCPS)]
Dichlofluanid 0.1
S-421 0.02
Dichlorvos 1
Tecnazene 0.05
Dicofol 0.5
Tetradifon 0.3
Dimethoate and omethoate (sum of ) 0.1
Vinclozolin 0.4
Dithiocarbamates (expressed as CS2)*** 2
Endosulfan (sum of isomers and endosulfan sulfate) 3 ***Bromide and dithiocarbamates (EBDCs) are determined using different methods.
Endrin 0.05
Ethion 2 Ubiquinone (Coenzyme Q10)
Etrimphos 0.05 see Coenzyme Q10
Fenchlorphos (sum of fenchlorphos and fenchlorphos- 0.1
oxon)
Unsaponifiable matter
Fenitrothion 0.5
Purpose: Applicable for the determination of normal animal
Fenpropathrin 0.03
and vegetable fats and oils. It is not suitable for fats and oils
Fensulfothion (sum of fensulfothion, fensulfothion-oxon, 0.05 containing excessive amounts of unsaponifiable matter such
fensulfothion-oxon sulfone and fensulfothion sulfone)
as marine oils.
Fenthion (sum of fenthion, fenthion-oxon, fenthion-oxon 0.05
sulfone, fenthion-oxon sulfoxide, fenthion sulfone and Method facts:
fenthion sulfoxide)
Fenvalerate 1.5
Flucytrinate 0.05
Fluvalinate, τ- 0.05 • Precision: NA
Fonofos 0.05 • Method reference: American Oil Chemists’ Society, Ca
Heptachlor (sum of heptachlor and cis- and trans- 0.05
6a-40 Unsaponafiable Matter, Official Methods and
heptachlor epoxides) Recommended Practices of the AOCS, Fifth Ed.,
Hexachlorobenzene 0.1 American Oil Chemists’ Society, Champaign, IL (1997)
Hexachlorocyclohexane isomers (other than γ) 0.3 Description: The sample is saponified and then extracted
Lindane (γ-hexachlorocyclohexane) 0.6 multiple times using petroleum ether and ethyl alcohol. The
Malathion and malaoxon (sum of ) 1 solvent is evaporated from the remaining extract into a tared
Mecarbam 0.05 beaker. The residue is then weighed and then titrated to
Methacriphos 0.05 obtain the % unsaponifiable matter. Unsaponifiable matter
Methamidophos 0.05 includes substances that are frequently found dissolved in
Methidathion 0.2 fats and oils that cannot be saponified by the caustic alkalies
Methoxychlor 0.05 but are soluble in the ordinary fat solvents. Included are the
Mirex 0.01
higher aliphatic alcohols, sterols, pigments, and hydrocarbons.
Monocrotophos 0.1
Parathion-ethyl and paraoxon-ethyl (sum of ) 0.5 Valerian extract (valerenic acid)
Parathion-methyl and paraoxon-methyl (sum of ) 0.2
Purpose: Applicable for the determination of acetoxyvalerenic,
hydroxyvalerenic, and valerenic acid in extracts.
Pendimethalin 0.1
Pentachloranisol 0.01 Method facts:
Permethrin (sum of isomers) 1 • Sample size: 2g
Phosalone 0.1 • Limit of quantitation: 0.1%
Phosmet 0.05
Piperonyl butoxide 3
• Method reference: Sakakibara, H., Honda, Y., Nakagawa, S.,
Pirimiphos-ethyl 0.05
Ashida, H., and Kanazawa, K. “Simultaneous Determination
Pirimiphos-methyl (sum of pirimiphos-methyl and 4 of All Polyphenols in Vegetables, Fruits, and Teas,” J.
N-desethyl-pirimiphos-methyl)
Agric. Food Chem., 51(3) :572-580 (2003)
Procymidone 0.1

Description: Samples are extracted using the appropriate • Limit of quanititation: Most matrices - 100 IU/g
technique for the sample matrix. Aliquots are analyzed using • Precision: the RSD is 5.65%
HPLC with UV detection. Standards are compared to
• Method reference: Covance developed method based upon
samples to quantify concentrations.
the chemical principles of:
Vanadium (ICP or ICP-MS) U.S. Pharmacopeial Convention, USP-NF [online], current

see Inorganic analysis by ICP or ICP-MS version, Dietary Supplements, Vitamins, Oil- and Water-
Soluble Vitamins with Minerals Tablets, Assay for Vitamin
A, Method 2. http://www.uspnf.com (accessed 23 Jan 13).
Vitamin A, beta carotene
see beta Carotene U.S. Pharmacopeial Convention, USP-NF [online], current
version, USP Monographs, Vitamin E Preparation
Vitamin A, retinol Description: The samples are prepared by one of the

Purpose: Applicable to the determination of retinol in following procedures: Direct extraction dispersal with
foods, food products, infant formulas, premixes, bicarbonate and ascorbate-pyrogallol solution followed by
pharmaceuticals, pet foods, feeds, and other matrices. The organic extraction or treatment with pancreatin followed by
method measures all-trans and 13-cis retinol isomers. It a subsequent phase extraction. Vitamin A esters and free
does not include contributions of vitamin A activity from and esterified. The esters are then analyzed using normal
pro-vitamin A carotenoids. phase chromatography.
Method facts: Vitamin B profile by HPLC

• Sample size: 30g in food products, 10g in supplements and Purpose: Applicable for the determination of vitamins B1,
premixes, 50g in feeds B2, B3, B5 and B6 fortified products, premixes, and
• Limit of quantitation: 1 IU/g but can vary with sample size various preparations.
• Precision: Most matrices – 100IU/100g
• Method reference: Official Methods of Analysis of AOAC Method facts:
International, Current Ed., Methods 992.04, 992.06 and • Sample size: 5g
2001.13, AOAC INTERNATI0NAL, Gaithersburg, MD, • Limit of quantitation: Most matrices-150ppm, liquids-5ppm
USA (Modified) • Precision: On a tablet matrix, the RSD ranges from 2-8%
• Method reference: The United States Pharmacopeia,
Description: The sample is saponified to break down fat and Twenty-Eighth Revision, p. 2162, United States
release the vitamins. The digest is extracted with an organic Pharmacopeial Convention, Inc.: Rockville, Maryland (2005)
solvent. Vitamin A is quantitated directly as all-trans retinol
and 13-cis retinol by ultra or high performance liquid Description: Sample extraction is done utilizing an aqueous/
chromatography (UHPLC or HPLC). acetonitrile/acetic acid solution. A portion of that extract is
analyzed on a HPLC equipped with UV detection set at 210
and 270 nm. The analyte is then quantitated by comparing
Vitamin A, total, retinol and beta carotene the sample response to a standard of known concentration. The
see beta Carotene and Vitamin A, retinol result is then converted and reported in the appropriate form.
Vitamin B2
Vitamin A in Dietary Supplements see Riboflavin
Purpose: This method is for the determination of vitamin A
esters in dietary supplements, premixes, liquid concentrates,
Vitamin B6 (pyridoxine)
and raw materials. These include, but are not limited to,
Purpose: Applicable for the determination of vitamin B6
tablets, capsules, powders and liquids, which contain oil- and
(pyridoxine) in most foods, dietary supplements, and feeds.
water-soluble vitamins with and without minerals. The
method is not recommended for matrices with Method facts:
concentrations below 50 μg/g. This method quantifies the • Sample size: 5g
following vitamin forms: retinyl acetate and retinyl • Limit of quantitation: Most matrices - 0.07mcg/g
palmitate. • Precision: On an infant formula matrix, the RSD is 9.54%
• Sample size: 50g Description: The sample is hydrolyzed with dilute sulfuric
[ assay ] V
acid in an autoclave, and then pH is adjusted to remove Description: The sample is extracted with a m-phosphoric
interferences. Utilizing the yeast Saccharomyces carlsbergenesis, acid and acetic acid solution. The extract is then analyzed on
the amount of B6 is determined turbidimetrically by an HPLC system equipped with an UV detector. A
comparing the growth response of a sample against the calibration curve is used for quantification.
growth response of a B6 standard.
Vitamin D by HPLC
Vitamin B12 (cyanocobalamin)
Purpose: Applicable to the determination of vitamins D2 or
Purpose: Applicable for the determination of vitamin B12
in most foods, feeds, and dietary supplements. D3 in a wide variety of matrices. These include, but are not
limited to, most food products, pet foods, feeds, infant
Method facts: formulas, premixes, multi vitamins, and nutraceutical
• Sample size: 2g products.
• Limit of quantitation: Most matrices - 0.0012 mcg/g
Method facts:
• Precision: On a cereal matrix, the RSD is 9.71% • Sample size: 30g in food products, 5g in supplements and
• Method reference: AOAC 952.20, 960.46 premixes, 50g in feeds
USP (modified) • Limit of quantitation: 20 IU/100g in food products and
feeds, 14.4 IU/g in supplements and premixes
Methods of Analysis for Infant Formulas, Infant Formula
Council (1985) • Precision on an infant formula matrix, the RSD is 7.29%
• Method reference: Official Methods of Analysis of
Description: Vitamin B12 is extracted from the sample into AOAC INTERNATIONAL (2005) 18th Ed., AOAC
a buffer by heating in an autoclave. Utilizing the bacteria INTERNATIONAL, Gaithersburg, MD, USA, Official
Lactobacillus delbrueckii, the amount of B12 is determined Method 982.29, 2002.05 (Modified)
turbidimetrically by comparing the growth response of a Official Methods of Analysis of AOAC International,
sample against the growth response of a B12 standard. (2000) 17th Ed., Methods 979.24 and 980.26, AOAC
INTERNATIONAL, Gaithersburg, MD, USA (Modified)
Vitamin C Description: Samples with more than a negligible amount

Purpose: Applicable for the determination of vitamin C in of lipids are saponified and extracted with hexane. The
most foods, dietary supplements, and feeds. It measures both digests are fractionated by alumina open column
ascorbic acid and erythorbic acid if present. chromatography and/or normal phase high-performance
liquid chromatography (HPLC) to remove interferences.
Method facts: Once the purification of the extracts is complete, the
• Sample size: 8g samples are injected on a reverse phase HPLC system using
• Limit of quantitation: Most matrices - 1 mg/100g ultraviolet (UV) detection. Some matrices, with adequate
levels of vitamin D2 or D3, may not require purification
• Precision: On a cereal sample matrix, the RSD is 7.33%
prior to analysis.
Description: The vitamin C in the sample is extracted, Vitamin D by LC-MS/MS

oxidized, and reacted with o-phenylenediamine to produce Purpose: Applicable to pet food, cereals, and other matrices.
a fluorophor. The vitamin C content is determined by
comparing the sample extract fluorescence to the fluorescence Method facts:
• Sample size: 10g per assay
of known standards.
• Limit of quantitation: D3 in Pet food is 0.05 IU/g;
D2 in Pet food about 0.2 IU/g
Vitamin C by HPLC • Precision: The RSD is 5%
Purpose: Applicable for the analysis of ascorbic acid in • Method reference: Covance Internal Method
tablets, capsules, pre-mixes, juices and raw materials.
Description: The sample is extracted into hexane using
Method facts: BHT as a preservative. A portion of the hexane is washed
• Sample size: 1g with ethanol/water and centrifuged. The solvent is taken
• Limit of quantitation: Most matrices - 2mg/g to dryness and reconstituted in ACN/water and filtered
• Precision: On a multivitamin sample, the RSD is 2.44% through a 0.45 µm PTFE filter. Isotope internal standard is
added for LC-MS/MS analysis. The response is compared
to that of known standards.
Asami, D.K. (2003). Journal of Agricultural and Food
Chemistry. Vol. 51. p. 1237-1241

[ assay ]
Vitamin D3 by HPLC • Method reference: Cort, W.M., Vincente, T.S., Waysek,

Purpose: This method is applicable to research and E.H., and Williams, B.D., Journal of Agricultural Food
Chemistry, 31: 1330-1333 (1983)
commercial products of infant formulas and nutritional
beverages for the determination of vitamin D3. Speek, A.J., Schijver, J., and Schreurs, W.H.P., Journal of
Food Science, 50:121-124 (1985)
Method facts: McMurray, C.H., Blanchflower, W.J., and Rice, D.A.,
• Sample size: 10g in powdered infant formulas and 30g in Journal of the Association of Official Analytical Chemists,
liquid supplements and liquid infant formulas 63: 1258-1261 (1980)
• Limit of quantitation: 0.1 IU/g
Description: The sample is saponified with alcoholic KOH.
• Precision: On an infant formula matrix, the RSD is 8.49% The digest is extracted with an organic solvent. The vitamin E
• Method reference: Official Methods of Analysis of is quantitated directly as alpha tocopherol by HPLC with
AOAC INTERNATIONAL (2000) 17th Ed., AOAC fluorescence detection.
INTERNATIONAL, Gaithersburg , MD, USA, Official
Method 992.26
Vitamin E in Dietary Supplements
Description: Samples with more than a negligible amount Purpose: This method is for the determination of free and
of lipids are saponified and extracted with hexane. The esterfied vitamin E in dietary supplements, premixes, liquid
digests are fractionated by alumina open column concentrates, and raw materials. These include, but are not
chromatography and/or normal phase high-performance limited to, tablets, capsules, powders and liquids, which
liquid chromatography (HPLC) to remove interferences. contain oil- and water-soluble vitamins with and without
Once the purification of the extracts is complete, the minerals. The method is not recommended for matrices
samples are injected on a reverse phase HPLC system using with concentrations below 3 mg/g for vitamin E. This
ultraviolet (UV) detection. Some matrices, with adequate method quantifies the following vitamin forms:
levels of vitamin D2 or D3, may not require purification α-tocopherol, α-tocopheryl acetate, and α-tocopheryl acid
prior to analysis. succinate.
Method facts:
Vitamin E (alpha tocopherol)
Purpose: Applicable for the determination of vitamin E in
foods, dietary supplements, and premixes. • Limit of quanititation: Most matrices - 0.25mg/g
• Precision: On Multiple Vitamin Tablet matrix the RSD is
Method facts: 1.60%
• Method reference: Covance developed method based upon
• Limit of quantitation: 0.5 IU/100g the chemical principles of:
• Precision: On a corn oil matrix, the RSD is about 5%
U.S. Pharmacopeial Convention, USP-NF [online], current
• Method reference: Covance internal method version, Dietary Supplements, Vitamins, Oil- and Water-
Soluble Vitamins with Minerals Tablets, Assay for Vitamin
Description: The product is saponified with alcoholic KOH.
The tocopherols are extracted with organic solvent, separated E, Method 2. http://www.uspnf.com (accessed 23 Jan 13).
on an HPLC silica column, and measured using fluorescence U.S. Pharmacopeial Convention, USP-NF [online], current
detection. Milligrams of alpha tocopherol are converted to version, USP Monographs, Vitamin E Preparation
international units (IU) by the following calculations:
1mg d-alpha tocopherol = 1.49 IU of vitamin E Description: The samples are prepared by one of the
(natural vitamin E) following procedures: Direct extraction dispersal with
bicarbonate and ascorbate-pyrogallol solution followed by
1 mg dl-alpha tocopherol = 1.10 IU of vitamin E organic extraction or treatment with pancreatin followed by
(synthetic vitamin E)
a subsequent phase extraction. Free and esterified vitamin E
are determined separately using High Performance Liquid
Vitamin E (in feeds and forages) Chromatography (HPLC) with UV detection. Free and
Purpose: Applicable for the determination vitamin E esterified vitamin E are analyzed using reverse phase
in feeds. chromatography on a C18 column.
Method facts: Vitamin K1, lipase

• Sample size: 50g Purpose: Applicable to the determination of total vitamin
• Limit of quantitation: Most matrices - 0.5 IU/100g K1 in infant formula, certain nutritional supplements, and
• Precision: On an infant formula matrix, the RSD is 6.10% high fat oil products.
[ assay ] V-Z
Method facts: Yerba mate

• Sample size: 10g see Caffeine, theobromine, theophylline
• Limit of quantitation: 5.00mcg /100g
• Precision: 5.5%
Zeaxanthin
• Method reference: Official Methods of Analysis of AOAC see Carotenoids
INTERNATIONAL Current Ed., Method 999.15,
AOAC INTERNATIONAL, Gaithersburg, MD, USA
(Modified) Zinc (ICP or ICP-MS)
Description: Following enzymatic digestion of fat and
precipitation of fatty acids, vitamin K1 is extracted with
hexane. The sample is injected on a reverse phase high-
performance liquid chromatography (HPLC) system with
post-column reduction and fluorescence detection.
Vitamin K1 (trans phytonadione)

Purpose: Applicable to the determination of total vitamin
K1 in certain vitamin tablets, food products, agricultural
products, infant formulas, drink mixes, and high level
concentrates.
Method facts:
• Sample size: 5g/20 tablets
• Limit of quantitation: Most matrices - 5mcg /100g
• Precision: On an infant formula matrix, the RSD is 5.50%
Description: The sample is extracted with organic solvents

and injected on reverse phase high-performance liquid
chromatography (HPLC) system with post column
reduction and fluorescence detection.
Water activity
Purpose: Applicable to the determination of water activity
in most food products, which is a critical factor in
determining product safety and quality.
Method facts:
• Sample size: 5g
• Limit of quantitation: <0.030aW
• Precision: On ground corn, the RSD is 2.0%
• Method reference: AOAC International, ‘978.18
Water Activity of Canned Vegetables’ (modified), Official
Methods of Analysis, (ed.) Patricia Cunniff, 18th Ed.,
AOAC International, Gaithersburg, MD (2005)
Description: The sample is placed in an AquaLab Series 3

instrument. The instrument measures the headspace
humidity and calculates the water activity (aw ) of the
sample. The water activity is an indicator of perishability.
Xylitol
see Sugar alcohols

[ assay ]
[ assay ]
QuaLity
ISO Accreditation
Establishing and implementing the processes governed by strict scientific standards and stringent regulatory
mandates requires a thorough knowledge of analytical and regulatory issues. Over the past 25 years Covance
Laboratories has developed and improved our quality programs culminating in accreditation to ISO 17025
General Requirements for the Competence of Testing and Calibration Laboratories, which includes an assessment
of compliance to AOAC INTERNATIONAL Guidelines for Laboratories Performing Microbiological and
Chemical Analyses of Food and Pharmaceuticals. Our accreditation further establishes our commitment to the
food and dietary supplement industries.
The Covance Quality Statement

Covance Laboratories Inc., Nutritional Chemistry and Food Safety is committed to providing the
highest quality service and science to our customers. Our goal is to lead the industry in customer
satisfaction and innovation. We will achieve this goal by implementing a comprehensive Quality
Management System.
Forming partnerships with our customers and exceeding their expectations is at the core of our beliefs.
While each customer has unique needs, all expect performance, service, reliability, on-time delivery and
error-free results. We measure success through our customers’ eyes.
Applying sound science to create solutions for our customers is a critical component of our success. To
that end we have created a diverse and motivated team of the best and brightest individuals. Each team
member is committed to implementing the policies and procedures contained within the Quality
Manual.
Management understands that compliance with ISO 17025 is key to our success in maintaining an
effective Quality Management System and thereby meeting our customers’ needs.
ISO accreditation is used as a vehicle to drive continuous improvement and adherence to regulatory standards
and Good Laboratory Practices. Our system of root cause analysis, corrective action and preventive action
ensures that we learn from our experiences. Procedures for monitoring the performance of instrumentation help
ensure quality test results. Our systems for documentation of analyses help maintain compliance with
regulations and facilitate the investigation of results that do not meet our standards of quality. This spirit of
continuous improvement, commitment to quality and adherence to regulations is not simply an expression;
with ISO accreditation it is a proven part of our culture.

[ q u al it y ]
Quality Control
Chemists are the backbone of Covance quality and credibility. Our analysts undergo formal training specific to
their area of assignment. Training modules are uniquely Covance because they are based on our long history
and experience in the food and dietary supplement industries. Covance is committed to institutionalizing our
expertise and experience so that less-senior employees can learn. The added benefit is shortened learning curves
for new analysts.
Procedures have been established for analytical quality control samples. Approximately one out of 10 samples
analyzed is a quality control sample, typically validated control material, but additionally, duplicate sample
analysis and spike recoveries may be employed. Each are used to evaluate the sample set. Control charts are
used to define the acceptable limits for validated control materials.
Data are reported through Covance’s laboratory information management system (LIMS) and undergo a
number of checks performed in a systematic manner. All data are evaluated by a minimum of two people, a
scientist or peer for the first review and a lead scientist, technical leader and/or supervisor prior to release.
These reviews include the evaluation of control results, and checks for the accuracy of hand calculations,
conversions, and data transfers. We also review for consistency with normal or expected values by comparing to
client claims, handbook data, and previously collected data for that sample type. No final report is generated
until we are sure of the data’s accuracy.
Certifications/ Accreditations/Registrations
• USDA accredited for analysis of Moisture, Protein, Fat, and Salt; Chlorinated Hydrocarbons; and Sulfonamides
in Meat and Poultry Products
• Japanese Ministry of Health, Labor and Welfare certified for product release testing of Food and Dietary Supplements
• State of Wisconsin Department of Health and Family Services Materials License
• US Department of Transportation Hazardous Materials Registration
• US Department of Agriculture, Class R Research Facility, Animal Welfare Act
• US Department of Justice DEA Analytical Laboratory Registration
• State of Wisconsin Controlled Substance Authorization
• FDA Drug Establishment Registration
• Therapeutic Goods Administration (TGA) registration: allows Covance to test products under the jurisdiction of the
TGA
[ q u al it y ]
Proficiency Testing
Proficiency testing can be a useful tool for assessing a laboratory’s on-going competency. Covance participates in a wide variety
of programs both public and private. Below is a list of the public programs in which we participate.
Sponsor Matrix
AACC Cereal
AOAC Cottonseed meal Cheese

Egg Meat powders
Egg yolk Marine oil
FAPAS Mixed fat spread Chili paste

Canned meat Jam
Canned crab meat Chocolate
Canned fish Chocolate cake mix
Fruit juices Liquid vitamin supplement
Sheep feed Dairy ration
Pig ration Milk powder
Poultry ration Condensed milk
Infant formula Wheat flour
Oil and 50% ethanol Porridge oats
Olive oil Breadcrumbs
Cod liver oil Crisp bread
Hydrogenated vegetable oil Snack food
Fish oil Butter
Fish paste Cheese and pasta
Cereal Honey
Chili sugar paste Biscuit
FIACC Infant formula Whole egg powder

Vegetable juice High fiber cookie
Water
USDA Meat products
We have over 80 assays and hundreds of analytes covered by proficiency testing.
There is a constant stream of proficiency samples moving through our lab right alongside our clients’ samples.

[ q u al it y ]
[ assay ]
staff & Facilities
Battle Creek, Michigan

Gross square feet – 42,000
Total employment – 35
Services Offered
- Nutritional analysis
- Stability studies and storage
- Microbiology
- Microbiology consulting
- Probiotics
- Allergens
- HACCP training
Instrumentation
- HPLC
- GC
This facility is ISO/IEC 17025 accredited. Please refer
- ICP
to www.A2LA.org for the exact scope of accreditation.
- PCR
- Walk-in and reach-in stability chambers
Greenfield, Indiana
Services Offered
- Chemical contaminants
- Contaminants consulting
- Residues
- Antibiotics in medicated feed
- Method development, transfer, validation
Instrumentation
- LC-MS/MS
- GC-MS/MS
- HPLC
This facility is ISO/IEC 17025 accredited. Please refer - GC

[ facil it ie s]
Madison, Wisconsin Campus

Services Offered
- Chemical contaminants
- Residues
- Microbiology
- Microbiology consulting
- Probiotics
This facility is ISO/IEC 17025 accredited. Please refer - Phytochemicals
to www.A2LA.org for the exact scope of accreditation. - Method development, transfer, validation
- Stability studies and stability storage
- Product container/enclosure
- Nutritional equivalency
- HACCP training
- Process validation
Instrumentation
- LC-MS/MS
- GC-MS/MS
- HPLC
- UHPLC
- GC
- ICP
- ICP/MS
- IC
Singapore
Services Offered
- Stability studies and stability storage
Instrumentation
- HPLC
- GC
- ICP
This facility is ISO/IEC 17025 accredited. Please refer

[ facil it ie s]
Covance Battle Creek

55 Hamblin Avenue, East
Battle Creek, MI 49017
Tel: 269.565.7700
Thank you for visiting the Covance Battle Transportation

Creek Nutritional Chemistry and Food
Safety Services Laboratory. Known as
Kalamazoo/Battle Creek International Airport (AZO)
“Cereal City,” Battle Creek is located in
www.azoairport.com
Calhoun County off I-94 between Chicago
Covance is located 30 minutes from the airport.
and Detroit. We have a proud history and
promising future of providing the world’s Taxi/Limousine Service
leading food brands, research and training. Leisure Limousine & Sedan 269.343.0848
Battle Creek is home to Kellogg Company’s Car Rental
world headquarters, the W.K. Kellogg Avis 800.230.4898 Hertz 800.654.3131
Foundation, the Global Food Protection Enterprise 800.736.8222 National/Alamo 734.784.2309 ext. 2634
Institute, and the International Food Budget 800.527.0700
Protection Training Institute. Battle Creek
has many other major corporate members
Preferred Accommodations Distance From Site
of our community including Post Cereals,
Ralcorp, Denso, II Stanley, and Duncan
Aviation. The city is revitalizing downtown Fairfield Inn Battle Creek 5.3 mi
to accommodate food science and other 4665 Beckley Road
innovation industries. Battle Creek’s 269.979.8000
Air National Guard Base, commercial marriott.com
airport and industrial park are diversifying Quality Inn & Suites (Next to the Casino) 5.6 mi
our economy and attracting worldwide 11081 E Michigan Avenue
businesses for military, aviation, aerospace 269.964.3000
and alternative energy. Nearby rivers, forests, qualityinn.com
parks, world-class Binder Park Zoo, Full McCamly Plaza Hotel 0.2 mi
Blast Water Park and Firekeepers Casino 50 Capital Avenue SW
make our city a family friendly area for 269.963.7050
outdoor play. Battle Creek is located in the mccamlyplaza.com
Eastern Time Zone.
Restaurant Recommendations
For more information on Battle Creek, visit:
Battle Creek Unlimited

Battle Creek Restaurants Kalamazoo Restaurants
www.bcunlimited.org
http://www.battlecreekvisitors.org/ http://kalamazoo.areaconnect.com/
Battle
Creek Downtown Partnership visitors_dining.cfm restaurants/
www.downtownbattlecreek.com Clara’s on the River Craftsman Chop Company
44 McCamly Street North 6905 Sears Drive
For more information on Kalamazoo, visit: 269.963.0966 269.327.2000
City
of Kalamazoo Official Website www.claras.com www.craftsmanchopcompany.com
www.kalamazoocity.org
Aracadia Brewing Company Carrabba’s Italian Grill
For information on Nutritional Chemistry 103 W Michigan Avenue 5690 Westnedge Avenue
& Food Safety Services, visit: 269.963.9520 269.381.0607
www.archadiaales.com www.carrabbas.com
www.covance.com

Malia Mediterranean Bistro Epic Bistro
34 W Michigan Avenue 359 S Burdick Street
Enjoy your stay in Battle Creek, 269.441.2900 269.342.1300
Michigan! www.maliafoods.com www.milleniumrestaurants.com/epicbistro
Don Pablos Mexican Kitchen Great Lakes Shipping Company
5805 Beckley Road 4525 West KL Avenue
269.979.1004 269.375.3650
www.donpablos.com www.greatlakesshippingco.com
Griffin Grill & Pub Fieldstone Grill
38 W Michigan Avenue 3970 W Centre Street
269.965.7206 269.321.8480
www.griffinbc.com www.milleniumrestaurants.com/fieldstone

[ facil it ie s]
Map from Kalamazoo Airport to Battle Creek
Map of Downtown Battle Creek
[ facil it ie s]
Covance Greenfield
671 S. Meridian St.
Greenfield, IN 46140
(317) 467-3700
Thank you for visiting Covance Transportation

Laboratories Inc. in Greenfield! Situated
Indianapolis International Airport Taxi/Cab Companies
about 25 miles east of Indianapolis, our
Greenfield laboratories have a unique sense www.indianapolisairport.com
Carey Indiana
of culture and history. It was founded in Covance is located 50-60 minutes from
800-888-INDY(4639)
1914 by J.K. Lilly Sr. (son of Colonel Eli the airport.
www.careyindiana.com
Lilly) and originally started as an equine
facility. Over the decades that passed,
the site cultivated important advances in Aristocrat Limousine Service
agricultural, industrial, and pharmaceutical 3353 1/2 Central Ave (317) 923-5351
research. With its distinctive Spanish www.aristocratlimo.com
mission-style architecture that balances Restaurant Options
both function and form, the site is
enshrined on the National Register of Greenfield Restaurants Downtown Restaurants
Historic Places and is one of Greenfield’s www.restaurantica.com/in/greenfield/ http://www.indy.org
most recognizable landmarks.
Carnegies Palomino Restaurant
For more information on Greenfield, 100 West North St (317) 462-8480 49 West Maryland St, #189
visit: www.greenfieldin.org/about www.carnegiesrestaurant.blogspot.com (317) 974-0400
www.palomino.com
For more information on Indianapolis,
Applebees
visit: www.indy.org
1792 N State St (317) 462-3004 14 West
For information about Covance, www.applebees.com 14 West Maryland St (317) 636-1414
please visit: www.covance.com www.14westindy.com
O’Charley’s
Enjoy your stay in Greenfield, Indiana! 1993 North State St (317) 462-9240 St. Elmo’s Steakhouse
www.ocharleys.com 127 S. Illinois St (317) 635-0636
www.stelmos.com
The Oceanaire
30 South Meridian Street, Suite 100
(317) 955-2277
http://www.theoceanaire.com

[ facil it ie s]
9
52 min AIRPORT COVANCE, GREENFIELD
NF
20 min AIRPORT DOWNTOWN INDIANAPOLIS
ortv
32 min DOWNTOWN INDIANAPOLIS COVANCE, GREENFIELD
N
il
le P
ike
465
70
70 70
Fortville Pike Rd
8 mi. WEST
COVANCE CENTRAL 70
9
LABORATORY SERVICES
N 600 W
70 70
70
Covance, GREENFIELD
Greenfield
40
W Main St E Main St
DOWNTOWN 40
INDIANAPOLIS 40
70
70
52
12 mi. WEST
INDIANAPOLIS
465
INTERNATIONAL
AIRPORT 52
52
52
465
2 mi
DOWNTOWN INDIANAPOLIS GREENFIELD
Monument
Circle 70 70
70
9
D
W Washington St
N State St
S Pennsylvania
S Illinois St
Layt
B Claypool
on L
Court 9
n
A F
S Capitol Ave
W Maryland St Circle E Maryland St

Centre Mall
C
N State St
E Martindale Dr
N Martindale Dr
W Georgia St
Conseco E
Fieldhouse
200 ft 200 ft
Hotel Options:
Downtown Indianapolis Greenfield
A Marriott C Canterbury Hotel E Hampton Inn Greenfield F Holiday Inn

350 W Maryland St 123 S Illinois St 2271 William Way 2070 North State St
(317) 822-3500 (317) 634-3000 Greenfield, IN 46140 Greenfield, IN 46140
www.indymarriott.com www.canterburyhotel.com (317) 467-0700 (317) 467-0999
Covance Rate $164 http://hamptoninn1.hilton.com www.holidayinn.com
D Conrad Hotel
Covance Rate $99 Covance Rate $79
B Westin Hotel 50 West Washington Street
50 S. Capitol Avenue (317) 713-5000
(317) 262-8100 www.conradindianapolis.com
www.starwoodhotels.com/westin
Covance Rate $123 standard room
Covance Rate $168 club floor
[ facil it ie s]
Covance Madison
3301 Kinsman Boulevard
Madison, WI 53704
Tel: 608.241.4471
Thank you for visiting the Covance Transportation

Madison Nutritional Chemistry and
Food Safety Services laboratory! Built Dane County Regional Airport (MSN)
on an isthmus between lakes Monona and www.msnairport.com
Mendota, Madison is renowned for its Covance is located 10 minutes from the airport.
beautiful scenery. You will find the best Taxi/Cab Companies
of all worlds in Wisconsin’s vibrant capital Badger Cab: 608.256.5566 Madison Taxi: 608.255.8294
city and picturesque surrounding towns: Union Cab: 608.242.2000
natural beauty and outdoor recreation,
stimulating cultural offerings, distinctive Preferred Accommodations Distance From Site
restaurants and shops, and an irreverent
spirit of fun. Madison offers urban The Madison Concourse Hotel 5.4 mi
culture, natural beauty and small town 1 W. Dayton Street Free shuttle service from the airport and
charm. Madison is located in the Central 608.257.6000 • 800.356.8293 roundtrip to Covance. Mention your
time zone. www.concoursehotel.com visit to Covance to receive a special rate.
Cambria Suites 4.3 mi
5045 Eastpark Boulevard Free shuttle service from the airport and
608.241.7070 roundtrip to Covance, Monday-Friday.
For more information on Madison, visit: www.cambriasuites.com Mention your visit to Covance to receive
a special rate.
Greater Madison Convention &

Visitors Bureau Courtyard by Marriott – Madison East 3.6 mi

www.visitmadison.com 2502 Crossroads Drive Free shuttle service from the airport and
608.661.8100 • 888.236.2427 roundtrip to Covance. Mention your
City of Madison Recreation & Tourism
www.courtyardmadisoneast.com visit to Covance to receive a special rate.
www.cityofmadison.com/recTourism.
html
Restaurant Recommendations Cuisine Type
Wisconsin Tourism Information

www.travelwisconsin.com Bellini Italian Restaurant, Italian

401 E. Washington Ave., 608.250.0097 www.bellinirestaurant.com
The Blue Marlin, 101 N. Hamilton St. Seafood
For information about Nutritional 608.255.2255 www.thebluemarlin.net
Chemistry and Food Safety Services,
please visit: http://nutri.covance.com/ Eldorado Grill, 744 Williamson St. Southwestern/Mexican
608.280.9738 www.eldoradogrillmadison.com
Enjoy your stay Erin’s Snug Irish Pub, 4601 American Parkway Irish Pub
in Madison, Wisconsin! 608.242.7616 www.erinssnug.com
Great Dane Brew Pub, 123 E. Doty St. American Pub
608.284.0000 www.greatdanepub.com
Johnny Delmonico’s, 130 S. Pinckney St. Steakhouse
608.25.STEAK www.johnnydelmonicos.com
Ocean Grill, 117 Martin Luther King Jr. Blvd. Seafood
608.285.2582 www.oceangrillmadison.com
The Old Fashioned, 23 N. Pinckney St. Local Specialty
608.310.4545 www.theoldfashioned.com
Takumi, 4222 East Towne Blvd. Sushi
608.663.3899 www.takumirestaurant.net

[ facil it ie s]
Covance Singapore
1 International Business Park
#01-01 The Synergy
Singapore 609917
Tel: +65 6567 7333
Thank you for visiting Covance Singapore.

Transportation
Covance Singapore is located in the
International Business Park in the Jurong Singapore Changi Airport (SIN)
www.changiairport.com
East district, west side of the country. Covance Singapore site is located about 30 – 40 min from the Changi Airport.
Being the cultural melting pot and Taxi stands are located conveniently outside the Arrival Halls. Alternatively, you
dazzling example of the region’s economic may take the MRT or Mass Rapid Transit (Subway train service) from the Changi
successes, wealthy Singapore assails Airport Station to Jurong East Station where the site is located (transfer train at
the senses of the visitors. This tiny but Tanah Merah Station).
bustling city-state is famous for being The Airport Shuttle Service serves most hotels in Singapore – call the Ground
clean and the being safest city of the Transport Desk for information: +65 6241 3818 (24 hour Hotline).
world. You will be expected to be spoilt Taxi/Cab Companies
for vast options for things to see and do Comfort/CityCab Taxi: +65 6552 1111 (recommended)
– especially food. Singapore is a veritable SMRT Taxis: +65 6555 8888
feast for the senses, a heady mixture of Prime Taxi: +65 6778 0808
the familiar and the exotic. It suits all Yellow-Top Taxi: +65 6293 5545
budgets, too, presenting a happy collision
of opposites - grand and expensive at the Preferred Accommodations Distance From Site
famed Raffles Hotel, cheap but good food
in the markets of Bugis Junction Park Hotel Clark Quay 14km (8.7 mi)
and Clarke Quay. 1 Unity Street, Singapore 237983
Tel: +65 6593 8888
Singapore Marriott Hotel 13km (8.1 mi)
For more information on Singapore, visit: 320 Orchard Road, Singapore 238865
Tel: +65 6735 5800
Singapore Tourism Board Marina Bay Sands Singapore 16 km (9.9 mi)
www.stb.gov.sg 10 Bayfront Avenue, Singapore 018956
Tel: +65 6688 8868
To explore a variety of diverse Grand Copthorne Waterfront Hotel 14.7km (9.1 mi)
experiences, attractions and more 403 Havelock Road, Singapore 169632
www.yoursingapore.com/content/ Tel: +65 6733 0011
traveller/en/experience.html
Singapore Food Guide Restaurant Recommendations Cuisine Type

www.hungrygowhere.com/singapore
The Mushroom Pot (Orchard Point) Steamboat
For information about Nutritional 160 Orchard Road, #04-00 Orchard Point
+65 6733 9910
Chemistry and Food Safety Services,
please visit: http://nutri.covance.com/ The Blue Ginger Restaurant Local delicacy (Nonya/Peranakan)
97 Tanjong Pagar Road
+65 6222 3928
Enjoy your stay
in Singapore! Jumbo (Riverside Point) Seafood
30 Merchant Road # 01-01/02 Riverside Point
+65 6532 3435
Bali Thai (Suntec City Mall) Thai
3 Temasek Boulevard, #B1-037/039 Suntec
City Mall Tower 4
+65 6338 2066
[ facil it ie s]
Restaurant Recommendations Cuisine Type
Nasi Padang River Valley Malay / Indonesian

55 Zion Road
+65 6734 3383
Gajalee Indian
17/19 Cuppage Road, Cuppage Terrace
+65 6733 3278
Hummerstons Western/Fusion
11 Unity Street, #02-14 Robertson Walk
+65 6737 8863
Yakiniku Yazawa Japanese
11 Unity Street, #01-01 Robertson Walk
Map
Covance
Covance Singapore Singapore Changi

1 International Business Park Airport
#01-01 The Synergy
Singapore 609917

[ facil it ie s]
[ assay ]
Ordering
Information
Sample Submittal/Shipping
Please order using our online food catalog at

http://www.covance.com/foodcatalog

[ orde r ing]
Sampling International Broker
The analysis performed in the laboratory is only as good as the The shipment should be routed through Chicago, IL. All
sample which is received at the laboratory for analysis. Care paperwork related to the shipment should be faxed to the import
should be taken to ensure that the sample is representative of broker prior to shipping any samples. Please make certain that
the lot or product you are trying to characterize. Additionally, a complete ingredient list is provided for clearance.
enough sample should be sent in to ensure that an adequate
amount of material is present to perform all of the testing Covance Import Broker:
that is requested. Lastly, the sample must be shipped under Scarbrough International
conditions so that it will not degrade in transit to the laboratory. 1350 Michael Drive, Suite C
Wood Dale, IL 60191
Importing samples to covance Phone: (630) 595-3400
Import Registration Fax: (630) 595-3430
Attn: Covance Import Coordinator
The FDA has made changes to the process for importing food rwhitley@scarbrough-intl.com
products. The FDA requires all facilities, including international
parties, to be registered and to provide prior notice to the
FDA once their shipment is ready to leave for the US. These Wire Transfer Information
changes are a direct result of the Bioterrorism Act of 2002.
Non-compliance to these changes results in the shipment Bank Name: PNC Bank
being sent back to source or destroyed. Bank Address: Route 38 & East Gate Drive
Moorestown, NJ 08057
Please make certain any facility shipping samples to Covance is Sort Code: ABA# 0312-07607
registered with the FDA and has provided the FDA with prior Account No: ACCT# 8005497809
notice for the materials. The registration must be completed by Account Name: Covance
the facility sending the shipment. There is no cost to register Company Name: Covance Laboratories Inc.
and the registration takes 15 to 20 minutes. The Covance Company Address: 3301 Kinsman Blvd.
Madison WI 53704
Madison FDA registration number is 14345226602. This
Company Phone: 608-241-4471
registration number is needed while establishing prior notice.
VAT Number: none
FDA website: http://www.fda.gov
Click on “Register a Facility” or, once registered,
“Prior Notice of Imports."
[ inde x]
Assay Page
5-Hydroxytryptophan 17
5-HTP: see 5-Hydroxytryptophan 17
Acephate: see USP 561 and EP pesticides 62
Acesulfame K (analyzed with aspartame): see Aspartame and Acesulfame K, DKP and Saccharin 21
Acetic acid: see Organic acid profiles — option 2 48
Acid detergent fiber: see Detergent fiber, acid (ADF) 29
Acrylamide 17
Aflatoxins 17
Aflatoxin (M1) 17
Aflatoxin, Ochratoxin A, and Zearalenone by HPLC 18
Alachlor: see USP 561 and EP pesticides 62
Aldrin and Dieldrin: see USP 561 and EP pesticides 62
Allicin (garlic): see Garlic 37
Almond Protein by ELISA 18
alpha Carotene: see beta Carotene 21
alpha Lipoic acid (USP) 18
alpha Tocopherol 18
Aluminum (ICP): see Inorganic analysis by ICP 41
Aluminum (graphite furnace) 19
Amino acid profile (AOAC), total 19
Amino acid profile, free by AAA 19
Amino acid profile, free by HPLC 19
Amino acid profile (HPLC), total 19
p-Anisinide value 20
Anthocyanins, total 20
Anthocyanins by HPLC 20
Arsenic: see Inorganic analysis by ICP-MS 41
Artificial colors: see Colors, artificial 27
Ascorbic acid: see Vitamin C 65
L-ascorbyl-2-phosphate (2-phospho-L-ascorbic acid) 21
Ash 21
Aspartame and Acesulfame K, DKP and Saccharin 21
Azinphos-ethyl: see USP 561 and EP pesticides 62
Azinphos-methyl: see USP 561 and EP pesticides 62
BHA/BHT (antioxidants) 21
Bendiocarb: see Carbamate pesticides (LC-MS/MS) 23
Benzene 21
Benzoic acid, sorbic acid: see Organic acid profiles — option 1 48

[ inde x]
Assay Page
Berberine: see Goldenseal 39
beta Carotene 21
beta Glucan 22
Bilberry: see Anthocyanins, total or by HPLC 20
Biotin 22
Bisphenol A 22
Black currant oil (linolenic acid and GLA): see Fatty acid profiles 31
Bomb calorimetry 22
Bromide, inorganic 23
Bromophos-ethyl: see USP 561 and EP pesticides 62
Bromophos-methyl: see USP 561 and EP pesticides 62
Bromopropylate: see USP 561 and EP pesticides 62
Cadmium (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Caffeic acid: see Phenolic acids 51
Caffeine, theobromine, theophylline 23
Calcium (ICP): see Inorganic analysis by ICP 41
Calcium, iron, sodium (ICP): see Inorganic analysis by ICP 41
Calories, by calculation 23
Calories from fat, by calculation 23
Capsaicin/Capsaicinoid (Scoville heat) 23
Carbamate pesticides (LC-MS/MS) 23
Carbohydrates, total, by calculation 24
L-carnitine 24
Carotenoids 25
Option 1: alpha Carotene, beta Carotene, Lycopene 25
Option 2: Cryptoxanthin, Lutein, Zeaxanthin 25
Option 3: Other carotenoids 25
Catechins 25
Chloracetamide: see Herbicides, triazine and chloracetamide 40
Chlordane: see USP 561 and EP pesticides 62
Chlorfenvinphos: see USP 561 and EP pesticides 62
Chloride 25
Chlorinated/Organophosphate pesticides: see Pesticides, chlorinated/organophosphate 50
Chlorogenic acid 25
Chlorophenoxy acid herbicides: see Herbicides, chlorophenoxy acid 39
Chlorpyrifos: see USP 561 and EP pesticides 62
Chlorophyll 26
[ inde x]
Assay Page
Chlorpropham: see Carbamate pesticides (LC-MS/MS) 23
Chlorpyrifos-ethyl: see USP 561 and EP pesticides 62
Chlorpyrifos-methyl: see USP 561 and EP pesticides 62
Chlorthal-dimethyl: see USP 561 and EP pesticides 62
Cholesterol 26
Choline (chemical) 26
Choline (enzymatic) 26
Chondroitin sulfate by CPC 26
Chromium (atomic absorption) 27
Chromium (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Citric acid: see Organic acid profiles — option 2 48
Cobalt (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Coenzyme Q10 27
Colors, artificial 27
Conjugated linoleic acid (CLA) 28
Copper (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Coumaric acid: see Phenolic acids 51
Cranberry (quinic, malic, citric acids): see Organic acid profiles 48
Cranberry: see Anthocyanins, total 20
Creatine 28
Cryptoxanthin: see Carotenoids 25
Cyanocobalamin: see Vitamin B12 65
Cyanuric acid (LC-MS/MS) 28
Cyfluthrin: see USP 561 and EP pesticides 62
Cyhalothrin, λ-: see USP 561 and EP pesticides 62
Cypermethrin: see USP 561 and EP pesticides 62
Cysteine, free: see Amino acid profile, free by AAA 19
Cystine, free: see Amino acid profile, free 19
Cystine (AOAC): see Amino acid profile (AOAC), total 19
DDT: see USP 561 and EP pesticides 62
DHEA or 7-KETO DHEA (Dehydroepiandrosterone or 3-acetyl-7-oxo-dehydroepiandrosterone) 28
Deltamethrin: see USP 561 and EP pesticides 62
Density 28
Detergent fiber, acid (ADF) 29
Detergent fiber, neutral (NDF) 29
Diazinon: see USP 561 and EP pesticides 62

[ inde x]
Assay Page
Dichlofluanid: see USP 561 and EP pesticides 62
Dichlorvos: see USP 561 and EP pesticides 62
Dicofol: see USP 561 and EP pesticides 62
Digestible fat 29
Dimethoate and omethoate: see USP 561 and EP pesticides 62
Dithiocarbamates: see USP 561 and EP pesticides 62
Dong quai (ferulic acid): see Phenolic acids 51
EBDCs 29
Echinacea spp. (caftaric, chlorogenic, echinacoside, chicoric) 30
Elderberry: see Anthocyanins, total 20
Endosulfan: see USP 561 and EP pesticides 62
Endrin: see USP 561 and EP pesticides 62
Ephedrine alkaloids (Ephedra or Ma Huang) 30
Egg Protein by ELISA 30
Erythritol: see Sugar alcohols 58
Ethanol and Methanol 30
Ethion: see USP 561 and EP pesticides 62
Etrimphos: see USP 561 and EP pesticides 62
Evening primrose oil (linolenic acid and GLA): see Fatty acid profiles 31
Fat (acid hydrolysis) 31
Fat (methanol/chloroform) 31
Fat (Soxhlet) 31
Fat, total (Roese-Gottlieb) 31
Fat, total (NLEA): see Fatty acid profiles 31
Fatty acid profiles 31
Fatty acid profiles (NLEA) 32
Fatty acids, free (total by titration) 32
Fatty acids, free (by GC) 32
FDA PAM 302 32
FDA PAM 303 33
FDA PAM 304 (M304 screen, 285 compounds) 33
Fenchlorphos: see USP 561 and EP pesticides 62
Fenfluramine (HPLC): see Japanese export testing 43
Fenitrothion: see USP 561 and EP pesticides 62
Fenoxycarb: see Carbamate pesticides (LC-MS/MS) 23
Fenpropathrin: see USP 561 and EP pesticides 62
Fensulfothion: see USP 561 and EP pesticides 62
[ inde x]
Assay Page
Fenthion: see USP 561 and EP pesticides 62
Fenvalerate: see USP 561 and EP pesticides 62
Ferulic acid: see Phenolic acids 51
Fiber, crude 33
Fiber, insoluble and soluble dietary (LEE) 33
Fiber, insoluble, soluble, and total dietary (CODEX definition) by enzymatic-gravimetric
34
method and liquid chromatography (McCleary)
Fiber, total dietary (LEE) 33
Fiber, total dietary (Prosky) 34
Fiber, total dietary (CODEX definition) by enzymatic gravimetric method and liquid
34
chromatography (McCleary)
Fibersol: see Resistant maltodextrin 55
Flucytrinate: see USP 561 and EP pesticides 62
Fluoride (chemical) 35
Fluoride (Ion Selective Electrode — ISE) 35
Fluvalinate, τ-: see USP 561 and EP pesticides 62
Folate: see Folic acid/Folate 35
Folic acid /Folate 35
Fonofos: see USP 561 and EP pesticides 62
Formic acid: see Organic acid profiles — option 3 48
Fo-ti powder: see trans-Resveratrol 61
Free amino acids: see Amino acid profile, free 19
Free fatty acids: see Fatty acids, free 32
Fructan (HPLC) 35
Fructan (spectrophotometric) 36
Fructose
see Sugar profile by GC 59
see Sugar profile by HPLC 59
see Sugar profile by IC 59
see Sugar profile, low levels 60
Fruits: see Anthocyanins, total 20
Fumaric acid: see Organic acid profiles — option 3 48
Furfural 36
Fury (zeta cypermethrin): Information available on request 36
Galactooligosaccharides (GOS) content in Infant Formula 36
Galactooligosaccharide (GOS) content in Raw Material 36
gamma Aminobutyric acid (GABA) 37
gamma Linolenic acid (GLA): see Fatty acid profiles 31

[ inde x]
Assay Page
Garcinia cambogia: see Hydroxycitric acid 41
Garlic 37
Ginkgo biloba flavonoids 37
Ginkgo biloba terpenoids 37
Ginseng, Panax or Korean (ginsenosides) 38
Glucosamine 38
Glucose
Gluten/Gliadan 38
Glycerol (glycerine) 38
Goldenseal 39
Grape seed extract: see Catechins 25
Grape seed extract: see Polyphenols, total 53
Grape seed extract: see trans-Resveratrol 61
Green tea: see Catechins 25
Green tea: see Polyphenols, total 53
Griffonia seed extract: see 5-Hydroxytryptophan 17
Hazelnut Protein by ELISA 39
Heavy metals as lead (USP <231>) 39
Heavy metals by ICP-MS: see Inorganic analysis by ICP-MS 41
Heptachlor: see USP 561 and EP pesticides 62
Herbicides (chlorophenoxy acid herbicides) 39
Herbicides (triazine and chloracetamide herbicides) 40
Hexachlorobenzene: see USP 561 and EP pesticides 62
Hexachlorocyclohexane isomers: see USP 561 and EP pesticides 62
Hexanal 40
HPTLC (identity verification testing by high-performance thin-layer chromatography) 40
Hydrastine: see Goldenseal 39
Hydroxycitric acid (garcinia extract) 41
Hydroxyproline: see Amino acid profile, free or (HPLC), total 19
ICP spectrometry: see Inorganic analysis by ICP 41
Indoxacarb: see Carbamate pesticides (LC-MS/MS) 23
Inorganic analysis by ICP 41
[ inde x]
Assay Page
Inorganic analysis by ICP-MS 41
Inositol 41
Inulin: see Fructan (HPLC) 35
Inulin: see Fructan (spectrophotometric) 36
Iodide (Ion Selective Electrode — ISE) 42
Iodine, high level 42
Iodine value (titration) 42
Iodine (ICP-MS) 42
Iprovalicarb: see Carbamate pesticides (LC-MS/MS) 23
Iron (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Isoflavones (daidzein, glycitein, genistein, daidzin, glycitin, genistin) 42
Isomalt: see Sugar alcohols 58
Japanese export testing: Information available on request 43
Kudzu: see Isoflavones 42
Lactic acid: see Organic acid profiles — option 2 48
Lactitol: see Sugar alcohols 58
Lactose
Lactulose 43
Lead (by ICP-MS): see Inorganic analysis by ICP-MS 41
Lignin 43
Lindane: see USP 561 and EP pesticides 62
Loss on drying (LOD) 43
Lutein: see Carotenoids 25
Lycopene: see Carotenoids 25
Ma Huang: see Ephedrine alkaloids 30
Magnesium (ICP): see Inorganic analysis by ICP 41
Malathion: see USP 561 and EP pesticides 62
Malic acid: see Organic acid profiles — option 2 48
Maltitol: see Sugar alcohols 58
Maltodextrin, resistant: see Resistant maltodextrin 55

[ inde x]
Assay Page
Maltose
Mandatory nutrient package 44
Manganese (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Mannitol: see Sugar alcohols 58
Marine lipids (EPA/DHA): see Fatty acid profiles 31
Mecarbam: see USP 561 and EP pesticides 62
Melamine (LC-MS/MS) 44
Mercury: see Inorganic Analysis by ICP-MS 41
Methacriphos: see USP 561 and EP pesticides 62
Methamidophos: see USP 561 and EP pesticides 62
Methidathion: see USP 561 and EP pesticides 62
Methiocarb: see Carbamate pesticides (LC-MS/MS) 23
Methionine (AOAC): see Amino acid profile (AOAC), total 19
Methiocarb sulfone: see Carbamate pesticides (LC-MS/MS) 23
Methiocarb sulfoxide: see Carbamate pesticides (LC-MS/MS) 23
Methoxychlor: see USP 561 and EP pesticides 62
Methylsulfonylmethane (MSM) 44
Milk Protein, Total by ELISA 44
Milk thistle (silychristin, silydianin, silibibin A & B) 45
Mineral profiles, ICP or ICP-MS: see Inorganic analysis by ICP or ICP-MS 41
Mirex: see USP 561 and EP pesticides 62
Moisture, 60˚C (vacuum oven) 45
Moisture (Karl Fischer) 45
Molybdenum (atomic absorption) 45
Molybdenum (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Monocrotophos: see USP 561 and EP pesticides 62
Monosodium glutamate (as glutamic acid): see Amino acid profile, free 19
n-Acetylcysteine 46
NL-Proximate (moisture, ash, protein): see individual tests for descriptions 46
n-Methyl carbamates: see Carbamate pesticides (LC-MS/MS) 23
[ inde x]
Assay Page
Neutral detergent fiber: see Detergent fiber, neutral (NDF) 29
Niacin 46
Nickel (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Nitrate/nitrite by HPLC 46
Nitrate/nitrite by IC and VIS 47
Nitrosamines — Volatile in foods 47
Nitrosamines — Volatile in rubber products 47
Nucleosides 47
Nucleotides 48
ORAC, Total (includes hydrophilic and lipophilic) 48
Omega 3 fatty acids: see Fatty acid profiles 31
Organic acid profiles 48
Organo nitrogen pesticide screen (NIPE): Information available on request 48
Organophosphate pesticides 49
Organophosphate pesticides (chlorinated): see Pesticides, chlorinated/organophosphate 49
Ornithine: see Amino acid profile, free by AAA 19
Oxalic acid: see Organic acid profiles — option 5 48
PABA (para-aminobenzoic acid) 49
PAM screens: see FDA PAM screens 32
Pantothenic acid 49
Parabens (methyl, ethyl, propyl, butyl, isopropyl, isobutyl, para-hydroxybenzoates) 49
Parathion: see USP 561 and EP pesticides 62
Parathion-methyl: see USP 561 and EP pesticides 62
PCNB and DDT 49
Pendimethalin: see USP 561 and EP pesticides 62
Pentachloranisol: see USP 561 and EP pesticides 62
Permethrin: see USP 561 and EP pesticides 62
Peroxide value 50
Pesticides, chlorinated /organophosphate 51
Pesticide multiresidue analysis 50
Phthalates 51
Phenolic acids 51
Phosalone: see USP 561 and EP pesticides 62
Phosmet: see USP 561 and EP pesticides 62

[ inde x]
Assay Page
hospholipids (lecithin phospholipids, phosphatidic acid, phosphatidylethanolamine,
P
51
phosphatidylcholine, phosphatidylserine, phosphatidylinositol)
Phosphorus (ICP): see Inorganic analysis by ICP 41
p-Hydroxybenzoates (methyl, ethyl, propyl, butyl, isopropyl, isobutyl): see Parabens 49
Phytic acid 52
Phytosterols: see Sterols 58
Piperonyl butoxide: see USP 561 and EP pesticides 62
Pirimiphos-ethyl: see USP 561 and EP pesticides 62
Pirimiphos-methyl: see USP 561 and EP pesticides 62
Polyaromatic hydrocarbons 52
Polydextrose (high level) 52
Polydextrose, with enzymatic pre-treatment 52
Polyphenols, total (Folin-Ciocalteu Method) 53
Polysorbates 53
Potassium (ICP): see Inorganic analysis by ICP 41
Potassium sorbate: see Organic acid profiles — option 1 48
Proanthocyanidins (total polyphenols and catechins): see Individual tests 53
Procymidone: see USP 561 and EP pesticides 62
Profenophos: see USP 561 and EP pesticides 62
Promecarb: see Carbamate pesticides (LC-MS/MS) 23
Propham: see Carbamate pesticides (LC-MS/MS) 23
Propionic acid: see Organic acid profiles — option 4 48
Propoxur (baygon): see Carbamate pesticides (LC-MS/MS) 23
Protein (Dumas) 53
Protein (Kjeldahl) 53
Prothiophos: see USP 561 and EP pesticides 62
Pueraria (isoflavones): see Isoflavones 42
Pygeum (beta-sitosterol): see Sterols 58
Pyrethrum: see USP 561 and EP pesticides 62
Pyridoxine: see Vitamin B6 64
Pyruvate (pyruvic acid): see Organic acid profiles — option 7 48
Quinalphos: see USP 561 and EP pesticides 62
Quinic acid, malic acid, citric acid (cranberry): see Organic acid profiles — option 6 48
Reducing sugars 54
Residual solvents (USP<467> Class 1, 2a, 2b) 54
[ inde x]
Assay Page
Residual Solvents (USP<467> Class 1, 2a, 2b, and 3) 54
Resistant maltodextrin (fibersol) with total dietary fiber 55
Resistant starch 55
Retinol: see Vitamin A 64
Riboflavin (vitamin B2) 55
Rose hips (ascorbic acid) 55
Royal jelly 10-HDA (10-hydroxy-2-decenoic acid) 55
Rutin: Information available on request 55
S-421: see USP 561 and EP pesticides 62
SAMe (S-adenosylmethionine) 55
Saw palmetto (phytosterols + fatty acids) 56
Selenium by ICP-MS 56
Selenium (hydride generation) 56
Sennosides 56
Sennosides (UV): see Japanese export testing 43
Sesame Protein by BioKits 57
Silicon (atomic absorption) 57
Sinapic acid: see Phenolic acids 51
Silver (ICP-MS): see Inorganic analysis by ICP-MS 41
Sodium (ICP): see Inorganic analysis by ICP 41
Sodium benzoate: see Organic acid profiles — option 1 48
Sorbic acid: see Organic acid profiles — option 1 48
Sorbitol: see Sugar alcohols 58
Soy isoflavones: see Isoflavones 42
Soy Protein by ELISA 57
Species identification by ELISA 58
Starch 58
Sterols (beta sitosterol, campesterol, stigmasterol, brassicasterol) 58
Succinic acid: see Organic acid profiles — option 3 48
Sucralose (finished products) 58
Sucralose (packets, pure material) 58
Sucrose

[ inde x]
Assay Page
Sugar alcohols 58
Sugar profile by acid hydrolysis 59
Sugar profile by GC 59
Sugar profile by HPLC 59
Sugar profile by IC 59
Sugar profile, low levels 60
Sulfites (Monier-Williams) 60
Sulfur (ICP-MS): see Inorganic analysis by ICP-MS 41
Tartaric acid: see Organic acid profiles — option 3 48
Taurine 60
Tecnazene: see USP 561 and EP pesticides 62
Tetradifon: see USP 561 and EP pesticides 62
Theobroma cacoa: see Caffeine, theobromine, theophylline 23
Theobromine: see Caffeine, theobromine, theophylline 23
Theophylline: see Caffeine, theobromine, theophylline 23
Thiamin (vitamin B1) 60
Thiobarbituric acid (TBA) value 61
Thiabendazole: see FDA PAM 304 pesticide screen 33
Thyroxine, Triiodothyronine (HPLC): see Japanese export testing 43
Tin (ICP-MS): see Inorganic analysis by ICP-MS 41
Tin (graphite furnace): Information available on request 61
Titanium (atomic absorption) 61
Tocopherols (alpha, beta, gamma, delta) 61
Tocotrienols (alpha, beta, gamma, delta) 61
Trans fat and fatty acids, by GC: see Fatty acids 31
trans-Resveratrol 61
Triazine: see Herbicides 40
Trifluralin: see FDA PAM 304 pesticide screen 33
Tryptophan (AOAC): see Amino acid profile (AOAC), total 19
Tryptophan (HPLC): see Amino acid profile (HPLC), total 19
USP-NF pesticides 62
Ubiquinone (Coenzyme Q10): see Coenzyme Q10 27
Unsaponifiable matter 63
Valerian extract (valerenic acid) 63
Vanadium (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41
Vinclozolin: see USP 561 and EP pesticides 62
Vitamin A, beta carotene: see beta Carotene 21
[ inde x]
Assay Page
Vitamin A (retinol) 64
Vitamin A, total, retinol and beta carotene: see beta Carotene and Vitamin A, retinol 64
Vitamin A for dietary supplements 64
Vitamin B profile by HPLC 64
Vitamin B2: see Riboflavin 55
Vitamin B6 (pyridoxine) 64
Vitamin B12 (cyanocobalamin) 65
Vitamin C 65
Vitamin C by HPLC 65
Vitamin D by HPLC 65
Vitamin D by LC-MS/MS 65
Vitamin D3 by HPLC 66
Vitamin E (alpha tocopherol) 66
Vitamin E (in feeds and forages) 66
Vitamin E in dietary supplements 66
Vitamin K1, lipase 66
Vitamin K1 (trans phytonadione) 67
Water activity 67
Xylitol: see Sugar alcohols 58
Yerba mate: see Caffeine, theobromine, theophylline 23
Zeaxanthin: see Carotenoids 25
Zinc (ICP or ICP-MS): see Inorganic analysis by ICP or ICP-MS 41

Covance is an independent, publicly held company with headquarters in Princeton, New Jersey, USA.
Covance is the marketing name for Covance Inc. and its subsidiaries around the world.
The Americas +1.888.COVANCE ( +1.888.268.2623 )

+1.609.452.4440
Europe/Africa +800.2682.2682 +44.1423.500888
Asia Pacific +800.6568.3000 +65.6.5686588
Web Site: www.covance.com E-mail: onlinecatalog.msn@covance.com
© Copyright 2013, Covance Inc.

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Food & Dietary

Staff & Facilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73-82

Ordering Please order using our online food catalog at

NL-Proximate (moisture, ash, protein)

Calories (by calculation)

Calories from fat (by calculation)

Total fat [sum of fatty acids (saturated, monounsaturated, polyunsaturated, trans)]

Total carbohydrates (by calculation)

Total dietary fiber

Calcium, iron, sodium (ICP)

Mandatory Nutrient Package

Priority service is available for an additional fee.

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 1

Acid detergent fiber Neutral detergent fiber (enzymatic)

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 3

* Calculated from total sulfur by ICP-MS.

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 5

Inorganics by Wet Chemistry

Microbiology Consulting and Research Services

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 7

Microbiology Assays List

Additional Charges for Quantitative Assays

Dietary Supplements & Nutritional Products

Additional Charges for USP Assays

Contaminants in Foods, Feeds

Multi-Residue Analysis (MRA) by GC-MS/MS and LC-MS/MS (300+ compounds)

Finished Food and Raw Ingredients

Individual Pesticides or Subgroups

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 9

USP/EP Analysis (99 Analytes)

Ala Cart Analysis

Classical Pesticide Methods by GC

Compendial Testing Services

To expedite your analysis, please provide the following information in advance:

Stability Studies: Stability and Shelf Life Evaluation

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 11

Method Development, Validation and Transfer

Typical projects we have completed include:

Food Contact Materials (Migration Compliance)

Product/Analyte Testing Method

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 13

Dietary Supplements (continued)

Product/Analyte Testing Method

Dietary Supplements (continued)

Product/Analyte Testing Method

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 15

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 17

Method facts: alpha Carotene

Aluminum (ICP) Amino acid profile, free by AAA

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Description: The samples are hydrolyzed in 6 N hydrochloric Anthocyanins by HPLC

L-ascorbyl-2-phosphate Description: Samples are weighed, then dissolved in a

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 21

beta Glucan Method facts:

Biotin Black currant oil (linolenic acid and GLA)

Method facts: Bomb calorimetry

Bromide, Inorganic Calcium (ICP)

Purpose: Applicable to the determination of inorganic Calories, by calculation

T. Stijve, Gas Chromatographic Determination of Calories from fat, by calculation

Method facts: Carbamate pesticides (LC-MS/MS)

608-242-2712, Ext. 8311-4170 • www.covance.com/foodcatalog 23

• Limit of quantitation: Listed in the table below Dioxacarb 0.01

• Method reference: Fenobucarb 0.01

1AOAC Official Method 2007.01, Pesticide Residues in Fenoxycarb 0.01

Carotenoids • Method reference: Sakakibara, H., Honda, Y., Nakagawa, S.,

Chlorophyll • Method reference: Glick, Journal of Biological Chemistry,