New USP

<232/233>
Elemental
Impurity (Metals)
Tests

Improved Accuracy and

Productivity using
Analytical Instrumentations
for the Measurement of
Heavy Metals in
Pharmaceutical Materials

Page 1

USP Webinar
Dec. 08, 2010

2010 Varian Becomes a Part of Agilent

A Heritage of Innovation: Varian & Agilent

Page 2

USP Webinar
Dec. 08, 2010

Todays Agilent: Atomic Spectroscopy

More choices
The addition of the Varian Atomic Absorption (AA) and ICP-OES
products to Agilents ICP-MS products provides a complete
portfolio for routine and research analysis in environmental,
food, agriculture, clinical, pharmaceutical, product safety,
semiconductor, chemical/petrochemical, geochemical/mining,
metals, academic/research and other applications.

Agilent provides the best choice for every lab through:

A full range of atomic spectroscopy instrumentation
Optimal product offering for any budget / application
Continued focus on reliability and performance

Page 3

USP Webinar
Dec. 08, 2010

Todays Agilent: Atomic Spectroscopy

Create the perfect
solution

ICP-MS
ICP-OES

AA

Worlds best, most complete atomic spectroscopy portfolio!

Page 4

USP Webinar
Dec. 08, 2010

Testing for Metal Impurities in

Pharmaceutical Products

Page 5

USP Webinar
Dec. 08, 2010

Trace Metal Analysis in Pharmaceuticals, Why?

The control of impurities has always been a critical issue to the
pharmaceutical industry
Catalysts
Raw material (plants, animal proteins, rDNA, etc.)
Excipients (stabilizers, fillers, binders, release agents, flavors, colors,
coatings, etc)
Production equipment such as reactors, pipes, filters, etc.

Traces of inorganic impurities can reduce drug stability and shelf

life of some pharmaceutical products
The U.S. Food and Drug Administration (FDA) and the British
Pharmacopeia (BP) strongly advise that contamination
problems be fully investigated in a timely fashion

Page 6

USP Webinar
Dec. 08, 2010

Regulated Pharmaceutical and Clinical Applications

Pharmacopeia Testing (API)
Natural products (TCM)
Metal impurities (Catalysts, Extractables/Leachables)
Clinical trials (Metallo-proteins/drugs)

Metals Leached from Pharmaceutical Packaging

Interaction between formulation and packaging material
Storage conditions impact leaching (heat, UV radiation, storage time)

Metal ions can affect the stability of the formulation, catalyze the
degradation of the active pharmaceutical ingredient (API) and cause
unqualified degradates to form, or pose a toxicity threat on their own.
KYLE A. FLlSZAR, DAVID WALKER and LEONARDO ALLAIN, Profiling of Metal Ions Leached from Pharmaceutical
Packaging Materials, PDA Journal of Pharmaceutical Science and Technology, Vol. 60, No.6, November-December 2006

Page 7

USP Webinar
Dec. 08, 2010

Traditional Metals Test for Pharma - USP<231>

USP<231> is a limit test for heavy metals in samples
It is a qualitative (at best semi-quantitative) test that indicates the
content of metallic impurities by colored sulfide precipitate
Elements for which the method can be used are silver, arsenic, bismuth,
cadmium, copper, mercury, molybdenum, lead, antimony, and tin.
- Over 100 years old (circa 1905)
- Colorimetric method
- Sample ignition and ashing at 600C
(800C, EP)
- pH adjustment
- Addition of H2S, thioacetamide
- Compared to 0.001% (10 ppm) Lead
standard
- Visual (subjective) comparison of color
of metal sulfide precipitates

Page 8

USP Webinar
Dec. 08, 2010

USP<231> Sample Preparation

Three different sample preparation techniques can be utilized
for USP<231>, depending on the sample type
Each specified in individual monographs.
Method I
Used for samples that are a clear colorless solutions
After addition of a sulfide reagent, the color is compared to both a standard
as well as a sample spiked at the limit.
Method II
Solid organic samples are heated with sulfuric acid and residual carbon is
burned in a muffle furnace.
Metals are then extracted from the residue
Analysis is free from any organic interference
Method III
Solids are first digested with a mixture of sulfuric and nitric acid
Then treated with hydrogen peroxide to ensure complete oxidation

Methods II and III are very aggressive and lead to loss of target analytes

Page 9

USP Webinar
Dec. 08, 2010

Issues with USP<231>

Large sample size required
Sample size for USP<231> is determined by the formula: 2.0/(1000L) L-limit concentration in %

USP<231> is a nondiscriminatory test

The sum of all the sulfide-forming analytes are detected

Reproducibility

Changes to standards/solutions occurs with time

Concerns with reagents use

Use and generation of H2S

Colorimetric test using visual comparison

Visual perception and bias

Significant loss of volatile elements such as Sb and Hg

Page 10

USP Webinar
Dec. 08, 2010

USP<231> Ashing Step Leads to Poor Recovery

Table showing low recovery for several elements due to high temperature
ashing step in USP<231> (600oC ashing leads to almost total loss of
volatile analytes such as Hg and Sb). Issues of low recovery are
eliminated when ICP (acid digestion) sample prep is used.

Pharmacopeial Forum Stimuli Vol. 34(6) [Nov.Dec. 2008]

Page 11

USP Webinar
Dec. 08, 2010

Comparison of USP<231> with ICP-MS

Traditional (wet chemical) method for heavy metals testing in pharma materials
involves an ashing step and so gives low recoveries for all volatile elements!

From: A rapid ICP-MS screen for heavy metals in pharmaceutical compounds; N. Lewen, S. Mathew, M. Schenkenberger and T.
Raglione, Journal of Pharmaceutical and Biomedical Analysis , Volume 35, Issue 4, 29 June 2004, Pages 739-752

Page 12

USP Webinar
Dec. 08, 2010

Recognition of the Problems of USP<231>

USP Meeting dated April 2009

Problem Statement:

We are committed to advancing the current

standards (<231>) so that widely agreed upon
safe limits for key metal impurities are properly
measured, thereby protecting the public health
Clear indication of acceptance that the current method
(USP<231>) does not adequately protect the public health

Page 13

USP Webinar
Dec. 08, 2010

Goals for USP<231> Replacement

Want to set limits for appropriate metal impurities (of known toxicity, that are
sufficiently likely to be present)
Limits based on toxicology data, not method capability
Metal species*
Daily dose and budget fraction
Route of administration
Analytical techniques that measure metal impurities at the set limits must be:
Selective (measure individual metals concentrations, not sum)
Sufficiently sensitive
Robust*
Simple
Instrumentation used for analysis must demonstrate specificity, sensitivity, and
accuracy (performance-based method; any instrument may be used)

Page 14

USP Webinar
Dec. 08, 2010

Proposed USP<231> Replacement

USP<231>

At the same time,

USP developed a new
protocol for dietary
supplements

USP<232>

USP<233>

USP<2232>

Elemental Impurities
Limits

Elemental Impurities
Procedures

Elemental Contaminants
in Dietary Supplements

Page 15

USP Webinar
Dec. 08, 2010

Recent Transformation: <231>

~100 years

July/2010

<232>
October/2010

Proposed Date to Take Effect: September, 2013

Page 16

USP Webinar
Dec. 08, 2010

Impurities Limits
- Grouping class I and II under a single
table
- No change to PDE
- Includes Large volume parenterals
(LVP > 100 mL)
- PDE is now calculated based on the
route of exposure

Page 17

USP Webinar
Dec. 08, 2010

Definitions:
- Elemental impurities are defined as catalysts and environmental contaminants
which may occur naturally, added intentionally, or introduced inadvertently
- Big Four analytes must be tested (As, Cd, Hg, Pb)
- Speciation is required when the concentrations of Hg and As exceed the limit
- Parenterals with daily dose between 10 mL and 100 mL must use the
summation option of limit calculation

Compliance Options:
- Drug product analysis option
Modified daily dose PDE > (measured value) x (Max daily dose)
- Summation Option
Modified daily PDE > (1m (Cm x Wm)) x DD

Page 18

USP Webinar
Dec. 08, 2010

Analytical Procedures USP<233>

Any and all analytical techniques may be used providing they can meet all the validation
requirements
No procedure for speciation is provided, consult USP-NF and published protocols

Definitions
Acids to be used should be ultra pure (HNO3, HCl, H2SO4, HF, Aqua Regia)
Matrix matching should be used
Big Four must be measured for all samples; moreover, those analytes which were used
during production (Catalysts) or may be introduced during manufacturing should be tested
PDE limits are strictly enforced
SRM should be traceable
J

- defined as the w/w concentration of the analyte at the Target Limit

after the necessary dilution

Page 19

USP Webinar
Dec. 08, 2010

Sample Dissolution
Direct aqueous solutions
- Simple solubilization in an aqueous matrix
Direct organic solution
- Simple solubilization in an organic matrix
Closed vessel digestion
- We recommend addition of 0.5% HCl to stabilize Hg
- Dehydrate and predigest 0.5 g sample in 5 mL of strong acid
- Allow to sit loosely covered
- Add 10 mL of strong acid and MW digest
Neat

Page 20

(un-solvated)
- Used in alternative non-ICP-OES / ICP-MS procedures

USP Webinar
Dec. 08, 2010

Analytical Procedures: ICP-MS and ICP-OES

Calibration should be acid matrix matched
- Blank, 0.5J, and 2J
Rinse with dilute acid
Check instrument drift with the 2J standard
solution before and after samples analysis.
Recovery 20%
No indication of ISTD
Add Hg stabilizer to diluted stock solution
Cooled spray chamber is recommended when using ICP-MS
Consult the manufacturer for appropriate operation

Page 21

USP Webinar
Dec. 08, 2010

Validation: Accuracy, Precision, Specificity

Limit Procedure:
- System suitability must be evaluated prior to any sample analysis (at J, accounting for dilution)
- Limit of Detection
- Standard Solution - SRM at the target concentration
- Spike 1 - Actual sample spiked with SRM at the target concentration (J)
- Spike 2 - Actual sample spiked with SRM at 80% of J
- Acceptance: Spike 1 solution standard solution > Spike 2 solution
- Precision
- Six samples of material under test spiked with SRM for the target analytes
- Acceptance: Standard deviation NMT 20%
- Specificity
- Unequivocal demonstration that target analytes may be determined in the presence of
spectral (ICP-OES), isobaric (ICP-MS), and matrix interferences
He mode allows a secondary isotope (also free from polyatomic interferences) to be
used for confirmation. No reaction gas method can do this.

Page 22

USP Webinar
Dec. 08, 2010

Validation - Continued
Quantitative Procedure:
- Accuracy
- Standard Solution - 3 replicate SRM solutions containing 50%-150% J of the target analytes
- Test Samples - 3 replicate samples under test spiked with SRM at 50%-150% J
- Acceptance - 70%-150% spike recovery at each concentration
- Precision
- Test Samples - Six samples spiked with target analytes at J
- Acceptance - Standard deviation, NMT 20%
- Intermediate precision
- Repeatability Test - Repeat analysis (choose one of the following)
- On a different day
- With a different instrument
- With a different analyst
- Acceptance - %RSD, NMT 25% for each target analyte
- Specificity
- False-positive and false-negative check (interference check)

Page 23

USP Webinar
Dec. 08, 2010

USP<2232>: Dietary Supplements ONLY

Limits based on 10 g max daily intake
If total [As] > limit, Inorganic As speciation is required
If total [Hg] > 0.2 g/g, CH3-Hg speciation is required
Three options describing analyte limits:
Drug Product Analysis Option
- Result= g/g contaminant (daily dose) X serving size/day
Individual Component Option
- If each substance in the product meets the component
limit the product does not require further analysis given
the oral daily dose is NMT 10 g.
Summation Option
- For more than 10 g/day dose or for those components that
exceed the Component Limit
- Result = 1n(Ci X Wi)
**If summation option is used, case-by-case validation is required
The following procedure (Distillation, HPLC-AAS) is suggested for determination of inorganic
arsenic, but any validated procedure shown to give equivalent or better results can be used.

Page 24

USP Webinar
Dec. 08, 2010

Organo-Metallic Speciation Analysis

In addition to its common use as a stand-alone metals analyser, ICPMS is eminently suited as a detector for a range of chromatographic
separation methods
Capillary Electrophoresis (CE)
Field-flow Fractionation (FFF)
Ion Chromatography (IC)
Liquid Chromatography (HPLC)
Gas Chromatography (GC)

In this configuration, the separation is performed by the chromatograph,

and the ICP-MS operates as a mass selective detector to measure the
element(s) associated with the compound(s) of interest as they elute
from the chromatograph

Page 25

USP Webinar
Dec. 08, 2010

USP in Federal Law Implementation

USP does not enforce its own standards
FDA enforces USP standards; compliance focus on
manufacturer-approved specs and GMPs
USP standards apply at any time in the life of an article;
when tested, must pass to demonstrate compliance
However, USP does not set frequency of, or specify
circumstances for testing
Repeats, replicates, statistical rejection of outliers, or extrapolation of
results to larger populations, as well as the necessity and appropriate
frequency of batch testing, are neither specified nor proscribed by the
compendia.
Page 26

USP Webinar
Dec. 08, 2010

USP in Federal Law Implementation (continued)

Compliance with USP standards is required at all times.
Frequency of testing and sampling are left to the preferences
of first-party (manufacturer), second-party (buyer) and thirdparty (FDA/regulator) stakeholders, who:
may or may not require additional examination of additional specimens, in
accordance with predetermined guidelines or sampling strategies.
USP General Notices, #3.10

FDA helps promote compliance with applicable USP

compendial standards
Adapted from: Workshop on Metals in Pharmaceuticals and Dietary Supplements, Workshop on Metals in Pharmaceuticals and
Dietary Supplements USP Headquarters Rockville, Maryland April 28-29, 2009

Page 27

USP Webinar
Dec. 08, 2010

Justification for New ICP-MS Method USP<232/233>

Page 28

USP Webinar
Dec. 08, 2010

ICP-MS has been Incorporated into Pharmacopeias

European Pharmacopeia (draft 2008)

Page 29

USP Webinar
Dec. 08, 2010

QC of Traditional Chinese Medicines (TCMs)

Chinese Pharmacopoeia 2005, Appendix IX

Agilent Application Notes:

Determination of Toxic Elements in Traditional Chinese Medicine Using Inductively
Coupled Plasma Mass Spectrometry, P/N 5989-5591EN
Evaluation of Conventional ICP-MS and ORS-ICP-MS for Analysis of Traditional
Chinese Medicines, P/N 5989-2570EN

Page 30

USP Webinar
Dec. 08, 2010

Unmatched Matrix Tolerance and

Unparalleled Interference Removal

Page 31

USP Webinar
Dec. 08, 2010

Unmatched Matrix Tolerance HMI

Sample Introduction
Low-flow (0.15 mL/min
typical)
Temperature stabilized
(Peltier cooled spray
chamber)
Now features HMI (High
Matrix Introduction) Kit as
standard on 7700x model,
allowing auto setup of
plasma conditions and much
higher matrix tolerance

Page 32

USP Webinar
Dec. 08, 2010

Neat Seawater
50 ppb spiked signal stability over 150 samples

Skimmer after
150 sample
introduction

Agilent HMI ultra robust

Page 33

USP Webinar
Dec. 08, 2010

Removing Polyatomic Interferences in ICP-MS

The 7700 Series ORS3 removes interferences using He mode and KED (Kinetic
Energy Discrimination)
Polyatomic
ions
Analyte
ions

Energy distribution
of analyte and
interfering
polyatomic ions
with the same mass

Polyatomic
ions

Analyte
ions

Energy

Energy

At cell entrance,
analyte and polyatomic
ion energies overlap.
Energy spread is
narrow, due to
ShieldTorch System

Energy loss from each

collision with a He atom
is the same for analyte
and polyatomic ion, but
polyatomics are bigger
and so collide more
often

Cell
Entrance

Page 34

Bias voltage
rejects low energy
(polyatomic) ions

Cell
Exit

By cell exit, although

energy spread is
broader, ion energies
have been separated;
polyatomics are
rejected using a bias
voltage step (energy
discrimination)

USP Webinar
Dec. 08, 2010

Polyatomic Interferences in Complex Matrices

Isotope
45
Sc
47

Testing the effectiveness of the He collision

cell design in eliminating ICP-MS polyatomic
interferences involved analyzing extremely
challenging sample matrices. Single
matrices contained:
o 5% HNO3, 5% HCl, 1% IPA, 1% H2SO4,
200 ppm Na, 200 ppm Ca and 500 ppm P
A mixed matrix contained all the above.

o Plasma components (Ar, O, N, H), and

o Matrix components (O, N, H, Cl, S, C, Ca,
Na and P)
The above matrix components are commonly
found in food, agricultural, environmental,
clinical and many other sample matrices.

Page 35

31

Ti
Ti
50
Ti
51
V
52
Cr
53
Cr
54
Fe
55
Mn
56
Fe
57
Fe
58
Ni
59
Co
60
Ni
61
Ni
63
Cu

P16O, 46CaH, 35Cl12C, 32S14NH, 33S14N

P18O, 48CaH, 35Cl14N, 37Cl12C, 32S16OH, 33S16O
34 16
S O, 32S18O, 35Cl14NH, 37Cl12CH
35
Cl16O, 37Cl14N, 34S16OH
36
Ar16O, 40Ar12C, 35Cl16OH, 37Cl14NH, 34S18O
36
Ar16OH, 40Ar13C, 37Cl16O, 35Cl18O, 40Ar12CH
40
Ar14N, 40Ca14N, 23Na31P
37
Cl18O, 23Na32S, 23Na31PH
40
Ar16O, 40Ca16O
40
Ar16OH, 40Ca16OH
40
Ar18O, 40Ca18O, 23Na35Cl
40
Ar18OH, 43Ca16O, 23Na35ClH
44
Ca16O, 23Na37Cl
44
Ca16OH, 38Ar23Na, 23Na37ClH
40
Ar23Na, 12C16O35Cl, 12C14N37Cl, 31P32S, 31P16O2

64

32

65

32

16

66

34

16

67

32

34

68

32

69

32

18

70

34

18

71

34

18

72

40

73

40

74

40

34

75

40

34

77

40

49

Zn

This table shows the main polyatomic ion

interferences arising from:

Principal Interfering Species (mixed matrix)

C16O2, 12C16O2H, 44CaH, 32S12CH, 32S13C, 33S12C

13

Cu
Zn
Zn
Zn
Ga
Zn
Ga
Ge
Ge
Ge
As

Se
78
Se
80
Se

31

S16O2,

32

S2,
32

S O2H,

S S,
S2H,

Ar S,

S2,

48

48

Ca16O

16

Ca OH

18

Ca O

18

N16O37Cl, 16O235Cl

14

S2H,

O237Cl

16

35

Cl2

35

40

Ar31P

Cl2H,

35

37

Cl Cl,

40

Ar16O2

35

Cl37ClH,

40

Ar16O2H

37

Ar SH,
37

48

Ca OH,

Ar32SH, 40Ar33S,
Ar S,

35

N O Cl,

48

S2
34

S O2H,
32

16

34

S O2H,
S O2,

14

33

S SH,
S18O2,

Ar12C16O, 38Ar12C14N,

S2H,
34
33

32

S O2,

36

Cl2

40

Ar 35Cl,

40

40

Ca 35Cl,

37

Cl2H

37

Ar Cl, Ca Cl
40
Ar 38Ar
40
Ar2, 40Ca2, 40Ar40Ca,

S2 16O,

32

32

S16O3

USP Webinar
Dec. 08, 2010

Polyatomic Interferences in No Gas Mode

Color of spectrum indicates which matrix gave each interfering peak

2E5
cps

SO, SOH ClO ArC ClO

CO2

ArO, CaO

SO2, S2,

ArN2H,
SO2H

Ar2, Ca2, ArCa,

S2O, SO3

Unspiked 5% HNO3 + 5% HCl + 1% H2SO4 + 1% IPA Matrix

Unspiked Matrix ALL peaks are due to polyatomic interferences
ArCl

ArN

CO2H

Multiple polyatomic interferences affect almost every

mass Interferences are matrix-dependent

ArC

S2, SO2

Cl2

ArOH,
CaOH

ArS, Cl2
Br,
Ar2H

Ar2
ArCO,
ArCN

Br,
Ar2H

ArCl

CaO,
NaCl
SN

45
No Gas Mode

Page 36

ClO,
NaS

50

55

CaO
CaO,
NaCl

ClN2,
CaOH,
ArNa
NaClH

60 Mass 65

S2, SO2
Cl2H

70

ArS,
Cl2

Ar2

ArS

75

80

USP Webinar
Dec. 08, 2010

Polyatomic Interferences in He Mode

Color of spectrum indicates which matrix gave each interfering peak

2E5
cps

Unspiked 5% HNO3 + 5% HCl + 1% H2SO4 + 1% IPA Matrix

ALL polyatomic interferences are removed in He Mode (same cell conditions)

ALL polyatomic interferences are removed in He Mode

Is sensitivity still OK?

45

50

55

60 Mass 65

70

75

80

He Mode

Page 37

USP Webinar
Dec. 08, 2010

Matrix Mix with Spike (10ppb) in He Mode

Consistent sensitivity and perfect template match for all elements

2E5
cps

10ppb Spike in 5% HNO3 + 5% HCl + 1% H2SO4 + 1% IPA Matrix

Consistent high sensitivity for all isotopes of all elements in He Mode
Good signal for all spike elements in
10 ppb spike. Perfect template fit for
all elements ALL isotopes available
for quantification / confirmation
No residual interferences and no
loss of analyte signal by reaction

45
He Mode

Page 38

50

55

60 Mass 65

70

75

80

USP Webinar
Dec. 08, 2010

Unique Screening Capability of 7700 with He Mode

The 7700 with ORS3 operates effectively in helium mode, allowing all common matrix
interferences (even unknown ones) to be removed from ALL analytes at the same
time (no cell gas mode switching needed).
This provides a unique semiquant screening capability for rapid multi-element
analysis of complex sample matrices. Spectrum below shows a screening analysis
of a commercial antacid sample.
Mg & Al

Ca Fe & Mn
Sc (ISTD)
Ti &
Cr

Rh (ISTD)

Ga &
Ge (ISTD)
Rb & Sr

Tb (ISTD)

Ir (ISTD)

In (ISTD)

Ba & REE
B

Page 39

Zr & Mo

Sn

U
Pb & Bi

USP Webinar
Dec. 08, 2010

Pharmaceutical Sample
Analysis using ICP-MS

Page 40

USP Webinar
Dec. 08, 2010

Gelatin Capsules

PPM

PPB

In-sample spike levels

levels
0.5J
As, Hg
0.75
Cd
0.25
Pb
0.5
Cr-V
5
Os-Ir
1.25

J
1.5
0.5
1
10
2.5

1.5J
2.25
0.75
1.5
15
3.75

250X Dilution
levels
0.5J
As, Hg
3
Cd
1
Pb
2
Cr-V
20
Os-Ir
5

J
6
2
4
40
10

1.5J
9
3
6
60
15

Units

Component
Limit

75 As [He]
111 Cd [NG]
208 Pb [NG]

g/g
g/g
g/g

1.5
0.5
1

0.91
0.011
0.088

201 Hg [NG]

g/g

1.5

0.039

52 Cr [He]
63 Cu [He]
55 Mn [He]
95 Mo [NG]
60 Ni [He]
105 Pd [NG]
195 Pt [He]

g/g
g/g
g/g
g/g
g/g
g/g
g/g

25
250
250
25
25
10
10

0.13
0.43
0.08
0.03
0.16
0.012
0.000061

51 V [He]

g/g

25

0.095

189 Os [NG]
103 Rh [He]
99 Ru [NG]

g/g
g/g
g/g

10

0.064
0.000067
0

193 Ir [NG]

g/g

Gel Caps

0.017

J-Component Limit

Page 41

USP Webinar
Dec. 08, 2010

7700x ICP-MS Calibrations for the Big Four

Page 42

75As

111Cd

201Hg

208Pb

USP Webinar
Dec. 08, 2010

Gelatin Capsules 0.5 J Spike Recovery

120
100
80

% Recovery

60
GelCap 0.5J

40

GelCap 0.5J
20

GelCap 0.5J

Analyte
120
100
80

% Recovery

60
GelCap 0.5J

40

GelCap 0.5J

20

GelCap 0.5J

Analyte

Page 43

USP Webinar
Dec. 08, 2010

Gelatin Capsules 1.0 J Spike Recovery

120
100
80

% Recovery

GelCap 1.0J

60

GelCap 1.0J

40

GelCap 1.0J

20

GelCap 1.0J

GelCap 1.0J
GelCap 1.0J

Analyte
120
100
80

% Recovery

GelCap 1.0J

60

GelCap 1.0J

40

GelCap 1.0J

20

GelCap 1.0J

GelCap 1.0J
GelCap 1.0J

Analyte

Page 44

USP Webinar
Dec. 08, 2010

Gelatin Capsules 1.5 J Spike Recovery

120
100
80

% Recovery

60
GelCap 1.5J

40

GelCap 1.5J
20

GelCap 1.5J

Analyte
120
100
80

% Recovery

60
GelCap 1.5J

40

GelCap 1.5J
20

GelCap 1.5J

Analyte

Page 45

USP Webinar
Dec. 08, 2010

Amoxicillin (API)
Average (n=3)
g/g

g/day

Limit
g/day

Pass/Fail

Arsenic

0.075

0.075

15

Pass

Mercury

0.009

0.009

15

Pass

Lead

0.031

0.031

10

Pass

Cadmium

0.002

0.002

Pass

Avg (n=3) g/g

g/day

Limit (g/day)

Pass/Fail

Chromium

0.194

0.194

250

Pass

Copper

0.032

0.032

2500

Pass

Manganese

0.014

0.014

2500

Pass

Molybdenum

0.013

0.013

250

Pass

Nickel

0.059

0.059

250

Pass

Palladium

0.002

0.002

100

Pass

Platinum

0.009

0.009

100

Pass

Vanadium

0.019

0.019

250

Pass

Rhodium

0.019

0.019

Ruthenium

0.001

0.001

Pass
Pass
Total NMT 100

Iridium

0.005

0.005

Pass

Osmium

0.007

0.007

Pass

Serving size/day-1g

Page 46

Courtesy of Samina Hussain, Exova CA

USP Webinar
Dec. 08, 2010

Amoxicillin Spike Recoveries

Arsenic

Mercury

Lead

Cadmium

Chromium

Copper

Manganese

Molybdenum

Specification (J)
(g/g)

1.5

1.5

0.5

25
J = 10

250
J = 10

250
J = 10

25
J = 10

Avg. %Rec
0.5J (n=3)

81
RSD = 0.86

102
RSD = 2.8

105
RSD = 0.57

98
RSD = 0.82

97
RSD = 0.52

100
RSD = 0.20

103
RSD = 0.29

96
RSD = 0.21

Avg. %Rec
1J (n=6)

86
RSD = 2.4

102
RSD = 0.83

105
RSD = 0.27

99
RSD = 1.7

98
RSD = 0.31

102
RSD = 0.40

103
RSD = 0.31

96
RSD = 0.30

Avg. %Rec
1.5J (n=3)

87
RSD = 0.82

103
RSD = 0.90

106
RSD = 0.33

101
RSD = 0.27

100
RSD = 0.58

111
RSD = 0.64

104
RSD = 0.68

96
RSD = 0.90

Nickel

Palladium

Platinum

Vanadium

Rhodium

Ruthenium

Iridium

Osmium

Specification (J)
(g/g)

25
J = 10

10

10

25
J = 10

J = 2.5

J = 2.5

J = 2.5

J = 2.5

Avg. %Rec
0.5J (n=3)

99
RSD = 0.40

99
RSD = 0.20

100
RSD = 1.4

93
RSD = 0.43

96
RSD = 0.63

98
RSD = 0.41

98
RSD = 0.31

94
RSD = 0.64

Avg. %Rec
1J (n=6)

100
RSD = 0.42

99
RSD = 0.15

101
RSD = 0.30

95
RSD = 0.56

96
RSD = 0.62

98
RSD = 0.49

105
RSD = 0.20

94
RSD = 0.46

Avg. %Rec
1.5J (n=3)

102
RSD = 0.51

105
RSD = 0.19

102
RSD = 0.32

95
RSD = 0.54

102
RSD = 0.22

98
RSD = 0.38

106
RSD = 0.47

95
RSD = 0.30

Courtesy of Samina Hussain, Exova CA

Page 47

USP Webinar
Dec. 08, 2010

Pharmaceutical Sample
Analysis using ICP-OES

Page 48

USP Webinar
Dec. 08, 2010

Agilent ICP-OES
The world's most productive high performance simultaneous ICP-OES
Continuous wavelength coverage
provides extended dynamic range and
reduced interferences, giving you
maximum confidence in your results
Robust plasma ensures reliable and
reproducible resultseven with the most
complex matrices
One view, one step measurement of
major, minor, and trace elements, plus
the fastest warm-up, increases
throughput and productivity
Unique fitted background correction
simplifies method development by
eliminating correction point selection

Page 49

USP Webinar
Dec. 08, 2010

Instrumentation
Agilent 720-ES Simultaneous CCD ICP-OES
Axial configuration
VistaChip custom-designed and
patented CCD detector
Continuous wavelength coverage from
167 to 785 nm
High efficiency 40 MHz RF generator
Cooled-Cone Interface (CCI) displaces
cooler tail of plasma
Increases linear dynamic range
and reduces interferences

Page 50

USP Webinar
Dec. 08, 2010

Agilent 720-ES ICP-OES

- Computer-optimized echelle optical and detector design
Thermostatted to 35oC for stability and fast
start-up
All wavelengths captured in one reading
Fewer optical components
High light throughput
Excellent signal-to-noise

Thermostatted to -35oC for low noise

Adaptive Integration Technology (AIT)
Provides true simultaneous measurement
All wavelengths irrespective of signal intensity
Background signal and internal standards

Page 51

USP Webinar
Dec. 08, 2010

Long Term Stability

Continuing Calibration Verification (CCV)
Ag 328.068
Al 237.312

120

As 188.980

% Recovery of CCV

Ba 585.367
Be 313.042

110

Ca 315.887
Cd 214.439
Co 228.615

100

Cr 267.716
Cu 324.754
Fe 238.204

90

K 769.897
Mg 285.213

80

Mn 257.610
Na 568.821
Ni 231.604

70

Pb 220.353
Sb 217.582
Se 196.026

60

Tl 190.794

Time (Hours)

V 292.401
Zn 206.200
Lower limit
Upper limit

Long-term precision (RSD): 0.98% MAX

Page 52

USP Webinar
Dec. 08, 2010

Specificity
The procedure must be able to unequivocally assess each Target
Element in the presence of components that may be expected to be
present, including other Target Elements and matrix components.

Intensity
331

Intensity
600

As 193.696

Cd 214.439

100 g/L Pt

100 g/L Pt
300
500

5 g/L Cd

15 g/L As

250

400

300

200

200

162
193.664

Page 53

193.680

193.700
Wavelength (nm)
S: 201 B: 173 SBR: 0.163

193.720

193.728

214.405

214.420

214.440
Wavelength (nm)
S: 396 B: 244 SBR: 0.622

214.460

214.474

USP Webinar
Dec. 08, 2010

FACT - Multicomponent Spectral Fitting

European Pharmacopeia 6.0
2.2.57. Inductively coupled plasma-atomic emission spectrometry
Describes the advantage of spectral fitting

Page 54

USP Webinar
Dec. 08, 2010

FACT - Multicomponent Spectral Fitting

Information about position and intensities of potential
interferences is displayed during method development
FACT modelling can be performed and saved before or after
sample measurement. It improves the resolution mathematically
beyond the practical and theoretical spectral resolution
Ru 245.657 nm is typically the better Ru Line for this application
Intensitt
3000

2500

Measured spectrum
Calculated Cr
Calculated Ru

2000

Ru 267.876
Cr 267.879

1500

1000

700
267.833

Page 55

267.860

267.880
Wellenlnge (nm)
S: 1128 B: -1 SUV: -1268.773

267.900

267.921

USP Webinar
Dec. 08, 2010

Impurity Limits
Element

Page 56

Component

Adjusted for 50x Prep Dilution

Limit (g/g)

Limit (g/g)

.5J

1.5J

As

1.5

0.015

0.03

0.045

Cd

0.5

0.005

0.01

0.015

Pb

0.01

0.02

0.03

Hg

1.5

0.015

0.03

0.045

Cr

25

0.25

0.5

0.75

Cu

250

2.5

7.5

Mn

250

2.5

7.5

Mo

25

0.25

0.5

0.75

Ni

25

0.25

0.5

0.75

Pd

10

0.1

0.2

0.3

Pt

10

0.1

0.2

0.3

25

0.25

0.5

0.75

Os,Rh,Ru,Ir

NTE 10 total

0.025

0.05

0.075

USP Webinar
Dec. 08, 2010

Control Sample Data

Page 57

USP Webinar
Dec. 08, 2010

Amoxicillin Sample Spikes

Page 58

USP Webinar
Dec. 08, 2010

Amoxicillin Repeatability

Element
As
Cd
Cr
Cu
Hg
Ir
Mn
Mo
Ni
Os
Pb
Pd
Pt
Rh
Ru
V

Page 59

% RSD
3.73
0.77
0.69
0.48
1.07
0.43
0.44
0.47
0.43
0.96
2.15
0.50
0.40
0.58
0.34
0.43

USP Webinar
Dec. 08, 2010

Sodium Bicarbonate Sample Spikes

Page 60

USP Webinar
Dec. 08, 2010

Sodium Bicarbonate Repeatability

Page 61

Element

% RSD

As

5.99

Cd

2.96

Cr

0.97

Cu

1.34

Hg

1.40

Ir

1.17

Mn

1.00

Mo

0.95

Ni

0.99

Os

1.19

Pb

8.32

Pd

1.39

Pt

1.18

Rh

0.95

Ru

1.24

1.01

USP Webinar
Dec. 08, 2010

Results Confirmation Capability

Page 62

USP Webinar
Dec. 08, 2010

ICP-MS/OES Compliance with 21 CFR Part 11

Agilents complete solution for Compliance ensures:
Data security
Secure central storage of data
Full support of all
requirements mandated
User access limited to authorized individuals
by 21 CFR Part 11
Data integrity
in closed system
Automatic data storage and versioning
Full version control for ALL relevant data including original result reports
Integrated archival and long-term storage in a content management system
Data traceability
Automatic user-independent audit trail
Electronic signatures
Validation (IQ/OQ) is also available on both ICP-MS and ICP-OES

Page 63

USP Webinar
Dec. 08, 2010

ICP-MS and ICP-OES are the new

Gold Standard for metals testing
in pharmaceutical materials.

Page 64

USP Webinar
Dec. 08, 2010

Page 65