An

Assignment
On
“NEAR-INFRARED SPECTROSCOPY
AND FLAME PHOTOMETRY”

SUBMITTED TO: SUBMITTED BY:

DR. H. H. Savsani R. k. bloch
Reg. No. 2040417002
Associate Professor & M.V.Sc Scholar
Head Animal Nutrition
Dept. of Animal
Nutrition

College of Veterinary Sci. & Animal Husbandry

Junagadh Agricultural University
Junagadh-362001

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NEAR-INFRARED SPECTROSCOPY:
Introduction:
Measurements in the near-IR (NIR) spectral region (0.7–2.5μm, equal to 700–2500 nm) are
more widely used for quantitative analysis. Several commercial instruments are available for
compositional analysis of foods using NIR spectroscopy. A major advantage of NIR
spectroscopy is its ability to measure directly the composition of solid food products by use of
diffuse reflection techniques.

 NIR absorption spectra are less useful for identification and more useful for quantitative
analysis of compounds containing functional groups made up of hydrogen bonded to
carbon, nitrogen, and oxygen.
 NIR spectroscopy is used most extensively for quantitative applications, using either
transmission or diffuser flection measurements that can be taken directly from solid
foods. By using multivariate statistical techniques.
 NIR instruments can be calibrated to measure the amounts of various constituents in a
food sample based on the amount of IR radiation absorbed at specific wavelengths.

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Principle:
 When radiation strikes a solid or granular material, part of the radiation is reflected
from the sample surface. This mirror-like reflection is called specular reflection, and
gives little useful information about the sample.
 Most of the specularly reflected radiation is directed back toward the energy source.
Another portion of the radiation will penetrate through the surface of the sample and be
reflected off several sample particles before it exits the sample.
 This is referred to as diffuse reflection, and this diffusely reflected radiation emerges
from the surface at random angles through 180◦. Each time the radiation interacts with
a sample particle, the chemical constituents in the sample can absorb a portion of the
radiation. Therefore, the diffusely reflected radiation contains information about the
chemical composition of the sample, as indicated by the amount of energy absorbed at
specific wavelengths.
 The amount of radiation penetrating and leaving the sample surface is affected by the
size and shape of the sample particles. Compensation for this effect may be achieved
by grinding solid or granular materials with a sample preparation mill to a fine, uniform
particle size, or by applying mathematical corrections when the instrument is calibrated.

How Does Near Infrared Spectroscopy Work?

 NIR works because specific organic molecules absorb specific wavelengths of NIR
(near infrared) light energy. The absorptions are directly correlated with the
concentration of the organic molecules.
 This relationship must be scaled by a primary technique, otherwise known as the wet
chemistry laboratory. Therefore, the NIR is dependent upon the wet chemistry methods
to attain the linear relationship between the molecular absorptions and the actual
constituent concentrations.

NEAR-INFRARED SPECTROSCOPY (NIRS) APPLICATION

 Near infrared spectroscopy is routinely deployed in food, agricultural, dairy,

pharmaceutical, chemical, petrochemical and feed industries. The application of the
technology varies in each industry, and many applications are custom designed to suit
different companies and their specific products and needs.

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 NIR spectroscopy has been successfully used in the production of cheese, meat and
meat products, fish, egg products, milk, butter, beer, wine, fruit juices, baked goods,
cereal, produce, jams, jellies, nutritional supplements and more.
 Some of the common applications include measuring:
1. Fat levels
2. Sugar content
3. Moisture content
4. Blend uniformity analysis
5. Starch content
6. Protein
7. Oil content

1. To evaluate composition and quality of red meats and processed meat products poultry
and fish.
2. Measuring specific chemical constituents in a food that affect its end-use quality, for
monitoring changes that occur during processing or storage, and for directly predicting
processing characteristics of a commodity that are related to its chemical composition.
3. Incoming raw materials can be analysed by NIR to verify that suppliers are sending a
quality material.
4. The product mix (dough, etc.) can be analysed by NIR to ensure that all ingredients
were added correctly.
5. Finished product can be analysed by NIR to ensure product quality and to verify label
claims.
6. Raw fruit can be analysed by NIR to determine optimum oil and moisture content
before harvest
7. Expenditure and dependence on expensive Wet Chemical analysis can be significantly
reduced
8. Real time results can be achieved for industries requiring fast process analysis.
9. Determination of property concentrations
10. Identification of raw materials in intermediate and finished products (qualitative
analysis)
11. Specific properties concentrations (quantitative analysis) can be analysed based on their
importance in the process (for example):

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 12. Moisture control (important because it affects the taste of the product and has
microbiological impacts on shelf life)
13. Fat or oil (affects the taste of the product)
14. Control of fat added to product, in the form of spray oil, etc., (an important cost savings
for food plants).
Absorption Bands in the NIR
Region
 The absorption bands observed in the NIR region are primarily overtones and
combinations.
 The bonds involved are usually C—H, N—H, and O—H.
 The amount of radiation penetrating and leaving the sample surface is affected by the
size and shape of the sample particles. Compensation for this effect may be achieved
by grinding solid or granular materials with a sample preparation mill to a fine, uniform
particle size, or by applying mathematical corrections when the instrument is calibrated.
 NIR region arise primarily from functional groups that have a hydrogen atom attached
to a carbon, nitrogen or oxygen, which are common groups in the major constituents of
food such as water, proteins, lipids, and carbohydrates.
 Note that strong absorption bands associated with the −OH groups of water are centered
at ca. 1450 and 1940 nm.
 These bands are the dominant features in the spectrum of cheese, which contains 30–
40% moisture, and they are still prominent even in the lower moisture wheat and egg
white samples. Bands arising from the −NH groups in protein.

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Advantages:
NIR spectroscopy is a proven and accepted method of food analysis and is used to monitor
product quality parameters within the food, beverage, and dairy industries and offers significant
advantages:
1. -Real-time analysis
2. -Measure multiple constituents simultaneously
3. -Non-destructive testing
4. -No chemical waste stream, environmentally friendly
5. -Less sample waste
6. -Decrease operation costs, increase yield!
Disadvantages:
1. -Must still measure reduced set of samples with wet chemistry
2. -Calibration is less accurate than wet chemistry
3. -Measurement outside of range of calibration samples is invalid
4. -Small calibration sample sizes can lead to over confidence.

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FLAME PHOTOMETRY
Principle:
• 1-flame photometry, a branch of atomic spectroscopy is used for determining the
concentration of certain metal ions such as sodium, potassium, lithium, calcium,
Cesium, etc.
• 2-The basis of flame photometric working is that, the types of metals are dissociated
due to the thermal energy provided by the flame source. Due to this thermal excitation,
some of the atoms are excited to a higher energy level where they are not stable.
• 3-The subsequent loss of energy will result in the movement of excited atoms to the
low energy ground state with emission of some radiations as a wavelengths .The
emitted wavelengths are specific for specific elements.

• 1. The solution containing metal to be measure is first aspirated into the burner.
• 2. The solvent then evaporated leaving fine divided solid particles.
• 3. This solid particles move towards the flame, where the gaseous atoms and ions are
produced.
• 4. The ions absorb the energy from the flame and excited to high energy levels.
• 5. When the atoms return to the ground state radiation of the characteristic element is
emitted.
• 6. The intensity of emitted light is related to the concentration of the element.

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FP Component:
• 1- Flame: A burner that provides flame and can be maintained in a constant form and
at a constant temperature.
• 2- Nebuliser and mixing chamber: Helps to transport the homogeneous solution of
the substance into the flame at a steady rate.
• 3- Monochromator: helps in isolating the wavelength to be measured from that of any
other extraneous emissions.
• 4. Photo detector: Detect the emitted light and measure the intensity of radiation
emitted by the flame. That is, the emitted radiation is converted to an electrical signal
with the help of photo detector. The produced electrical signals are directly proportional
to the intensity of light.

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Flame (burner)
 Burner used in FP should have the following properties:
The flame should have ability to evaporate the solvent to give a residue (deposit)
It should convert this residue to gases state atom and finally into individual atoms
 The effect of flame depend on the temperature of flame and this temperature can be
monitored by following method:
- Fuel to air ratio
- Type of solvent for preparing sample solution
- Amount of solvent which is entering to flame
- Type of burner used in FP

Flame photometry employs a variety of fuels mainly air, oxygen or nitrous oxide (N2O) as
oxidant.
- Burners are of two types:
Total Consumption Burner and Premix or Laminar flow Burner.

The various processes in the flame are discussed below:

• Desolvation: The liquid solvent is evaporated, and the metal particles are dehydrated
by the flame
• Vapourisation: The sample vaporises to a gas.
• Atomization: Reduction of metal ions in the solvent to metal atoms by the flame heat.
• Excitation: The electrostatic force of attraction between the electrons and nucleus of
the atom helps them to absorb a particular amount of energy. The atoms then jump to
the exited energy state.

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 • Emission process: Since the higher energy state is unstable the atoms jump back to the
stable low energy state with the emission of energy in the form of radiation of
characteristic wavelength, which is measured by the photo detector.

Type of FP
• 1- Direct type:
Standard solution of Na & K are atomised or aspirated into flame to provide a series of meter
readings in which our unknown is going to compared with.
Direct type disadvantages
• 1- Any minor fluctuations in air, gas pressure might change response of instrument and
then cause errors.
• 2- Separate analyses and sometimes separate solutions must be made for Na and K.

• 2- Internal standard method:

Another element (Li) is added to all standards, blanks and unknowns in equal concentration.
This element has a criteria of:
• High emission intensity.
• Is absent from biological fluid.

Advantages:

1. Simple quantitative analytical test based on the flame analysis.

2. Inexpensive.
3. The determination of elements such as alkali and alkaline earth metals is performed
easily with most reliable and convenient methods.
4. Quite quick, convenient, and selective and sensitive to even parts per million (ppm) to
parts per billion (ppb) range.

Disadvantages:
1. Alteration of light emission because of altered flame temp.
2. It needs perfect control of flame temperature.
3. Interference by other elements is not easy to be eliminated
4. Heavy and transition metals , the number of absorption and emission lines is enormous
and the spectra are complex
5. Inadequate selectivity of WL.
6. Differences in viscosity between standards and sample.

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