Transcript ICP-MS in Pharma Analysis

ICP – Mass Spectroscopy
Applications
in
Pharma and Bio-Medical Sciences
Prasenjit Kar
Product Manager - SPSD
1
7/16/2015
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 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
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problems be fully investigated in a timely fashion
July 16, 2015
Page 2
Issues with USP<231> Part 1
Large sample size required
• Sample size for USP<231> is determined by the formula: 2.0/(1000L)
– where L is the limit concentration in percent
• For a limit of 10 ppm (0.001%) at least 2 g of material is required
– This may be impractical for many substances
• For instance proteins or peptides
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July 16, 2015
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Issues with USP<231> Part 2
Quantification issues with USP<231>
• USP<231> is a compendial test
– Individual elements cannot be quantified, only the sum of all the elements
in the group (sulfide-forming elements)
– Only 10 analytes are determined using USP<231> (Ag, As, Bi, Cd, Cu, Hg,
Mo, Pb, Sb, Sn). Does not cover many potentially toxic elements (such as
Cr, but also including many catalyst metals, such as Pd, Rh, etc)
• The aggressive sample preparation causes significant loss of volatile
elements such as Sb and Hg
– Leads to false negatives
Note:
While False Positives mean verification
reruns and wasted
time,
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False Negatives could mean potentially
harmful material entering the market
July 16, 2015
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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,
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and
accuracy (performance-based method; any instrument may be used)
July 16, 2015
Page 5
Comparison Between Modern Instrumentation and
USP <231> Method
Average Recoveries
of Elements (%)
120
100
80
ICP-MS
60
USP <231>
40
20
0
Pb As Se Sn Sb Cd Pd Pt Ag Bi Mo Ru In Hg
Elements
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Ref.:Lewen, N et al, J.Pharm & Biomed.Anal. 35 (2004) 739-752
Reprinted by Labcompliance with permission from Elsevier Limited.
Slide 6
Elemental Impurities for Drug Products
Daily
Dose
(µg/day)
LVP
Limit
(µg/g)
Daily
Dose
(µg/day)
LVP
Limit
(µg/g)
Cadmium
5
0.05
Rhodium
100
1.0
Lead
10
0.1
Ruthenium
100
1.0
Inorg. Arsenic
15
0.15
Chromium
250
2.5
Inorg. Mercury
15
0.15
Molybdenum
250
2.5
Iridium
100
1.0
Nickel
250
2.5
Osmium
100
1.0
Vanadium
250
2.5
Palladium
100
1.0
Copper
2500
2.5
Platinum
100
1.0
Manganese
2500
2.5
Element
Element
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PDE = Permissible Daily Exposure on a 50 kg person
LVP = Large Volume Parenteral (daily dose greater than 100 mL)
Modified daily dose (m)PDE = Exposure factor x Daily Dose PDE
Ludwig Huber
Slide 7
Proposed USP<231> Replacement
USP<231>
At the same time,
USP is developing a
new test for dietary
supplements
USP<232>
USP<233>
USP<2232>
Elemental Impurities
Limits
Elemental Impurities
Procedures
Elemental Contaminants
in Dietary Supplements
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July 16, 2015
Page 8
USP Implementation Plan
Stage 1: Feedback to PF Stimuli Papers, expired in July 31,
2011, update expected in 2012 (draft or final)
Stage 2: Adoption of General Notice to be published in PF
Official release date should coincide with the official EMA (Sept
2013)
Stage 3: All references to <231> removed and can not be used
any more. Timing coincides with Approval of General Notices
For ongoing updates check www.usp.org/hottopics/metals.html
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PF
= Pharmacopeial Forum
EMA = European Medicines Agency
Slide 9
Analytical Procedures: ICP-MS and ICP-OES
USP<232> permits the use of either ICP-MS or
ICP-OES (or any other “suitable” technique)
Decision will typically come down to customer’s
budget and DL requirements:
When to offer ICP-MS:
• When low DLs are needed (for example, large volume
parenteral dose component limit for Cd is 50ppb in the
sample. After 100x dilution (recommend 0.5g  50mL),
means quantification at 0.5ppb in solution.
• When speciation is required
• When measuring traces after prep in organic solvent
• When research applications will be done
When to offer ICP-OES:
When samples don’t need lowest DLs
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Atomic Spectroscopy for USP Webinar
Page 10
Polyatomic Interferences in Complex Matrices
Table shows Main Polyatomic Ion
interferences arising from:
Plasma Components (Ar, O, N, H), plus
Matrix Elements (N, Cl, S, C, Ca, Na & P)
Every isotope of every element in this mass
range suffers from multiple polyatomic
overlaps
Cell Mode test using several common
matrices. Single matrices containing:
5% HNO3, 5% HCl, 1% H2SO4, and 1%
IPA
Isotope
45
Sc
Principal Interfering Species (mixed matrix)
13 16
C O2, 12C16O2H, 44CaH, 32S12CH, 32S 13C, 33S12C
47
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
31
49
31
64
Zn
32
S16O2,
65
Cu
32
S16O2H,
Zn
34
Zn
32
Zn
32
Ga
32
Zn
34
66
67
68
Plus a Mixed Matrix containing all the
above
69
70
71
Commonly found matrix components in
environmental, food, agricultural, Pharma,
clinical and many other sample matrices
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, 31P 32S, 31P 16O2
32
S2, 36Ar12C16O, 38Ar12C14N,
16
S O2,
33
S SH,
S O2,
33
S2H,
48
S 2,
S2H,
Ge
40
Ar S,
73
Ge
40
Ar32SH, 40Ar33S,
Ge
40
Ar S,
As
40
34
Se
78
Se
80
Se
40
77
Ca O
18
16
Ga
75
Ca16OH
14
N16O37Cl, 16O235Cl
O237Cl
Cl2
18
35
S O2H,
34
48
18
35
72
74
48
Ca OH,
34
32
N16O35Cl,
S2
34
S O2H,
S O2,
34
Ca16O
34
18
18
14
S2H,
S S,
34
18
32
32
48
35
37
Ar SH,
40
Cl2H,
37
Cl Cl,
40
Ar31P
Ar16O2
35
Cl37ClH,
Cl2
40
Ar 35Cl,
40
40
Ar16O2H
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37
Ca 35Cl,
Ar 37Cl, 40Ca 37Cl
40
Ar 38Ar
40
Ar2, 40Ca2, 40Ar40Ca,
32
S2 16O,
Cl2H
32
S16O3
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Page 11
Interference Management : Principle of Helium Collision
Mode and Kinetic Energy Discrimination (KED)
Polyatomic
ions
Analyte
ions
Energy distribution
of analyte and
interfering
polyatomic ions
with the same mass
Polyatomic
ions
Bias voltage
rejects low energy
(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
Cell
Exit
By cell exit, although
energy spread is
broader, ion energies
have been separated;
polyatomics are
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rejected
using a bias
voltage “step” (energy
discrimination)
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Page 12
Blank Acid Matrices and IPA 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
ClO,
NaS
50
55
CaO
CaO,
NaCl
ClN2,
CaOH,
ArNa
NaClH
60 Mass 65
S2, SO2
Cl2H
70
ArS,
Cl2
Ar2
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ArS
75
80
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Blank Acid Matrices and IPA 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?
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45
He Mode
50
55
60 Mass 65
70
75
80
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Reliable Results with ORS3 in He Mode (1)
He mode removes ALL polyatomics at each analyte mass, not just reactive ones – reaction
gases only remove reactive interferences
Plot of Background Equivalent Concentration (BEC) for
51V
ClO interference in NoGas mode is not
completely removed in H2 mode – ClO
is not very reactive with H2 cell gas, so
residual interference remains
51V
in various matrix blanks
Consistent low blanks
obtained in He mode for 51V
in all matrices and mix
51V
in NoGas mode suffers
interference from ClO.
ClO is not completely
removed in H2 mode (ClO is
not very reactive with H2).
ClO
SOH
User has to identify
interferences before
choosing reaction gas (i.e.
must know matrix in
advance)
He mode
removes all
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polyatomics – reaction
gases don’t
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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]
µg/g
µg/g
µg/g
1.5
0.5
1
0.91
0.011
0.088
201 Hg [NG]
µg/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]
µg/g
µg/g
µg/g
µg/g
µg/g
µg/g
µg/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]
µg/g
25
0.095
189 Os [NG]
103 Rh [He]
99 Ru [NG]
µg/g
µg/g
µg/g
10
193 Ir [NG]
µg/g
Gel Caps
0.064
0.000067
0
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J-Component Limit
Atomic Spectroscopy for USP Webinar
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Gadolinium estimation in Bone sample: Clinical
Estimation of Gadolinium
Building calibration of Gd ( mass: 157)
Building calibration of Na, K, Ca,Fe,Zn
Digested standards of Gd comparison with aqueous standards
Optimization of Digestion procedure of Bone samples (Hip and Knee)
Quantification and recovery of Gd on digested samples
Quantification of Other elements like
Calcium , Sodium, Potassium, Iron & Zinc
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Calibration standards for Gd
Standard Calibration Curve : Gadolinium
Based on the sample weight of 0.3g the standard concentration was calculated.
Aqueous standard are made up in 5% HNO3
LEVEL
Gadolinium
Conc.(µg/G
bone)
STD-1
0.1
0.03
30
0.6
STD-2
0.2
0.06
60
1.2
STD-3
1
0.3
300
6
STD-4
2
0.6
600
12
STD-5
4
1.2
1200
24
STD-6
8
2.4
2400
48
STD-7
12
3.6
3600
72
STD-8
16
4.8
4800
96
STD-9
20
6
6000
120
(µg/0.3G
ng/0.3gbone ppb( 50mL)
bone)
Calibration curve built on the
Instrument.
0.3g sample made up to 50mL, implies
Standards when diluted to the same volume
result in this concentration hence
Instrument was calibrated
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Gadolinium content in HIP BONE
(ppb: ng/g)
Element
157 Gd [ 2 ]
Sample-1
1.060
Sample-2
0.854
Sample-3
1.247
Sample-4
2.181
Sample-5
1.650
Sample-6
1.122
Sample-7
2.387
Avg(ppb)
1.641
Std.dev
0.677
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Gadolinium mass spectrum in HIP bone sample
Analyte of interest
Internal standard used
In method
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Advanced Applications: Hyphenated ICP-MS
Determination of Clinical Cancer Treatment Biomarkers by LC-ICP-MS.
Platinum containing drugs like Cisplatin are widely used to treat cancer.
Using an LC-ICP-MS its possible to carry out determination of Pt based
markers.
Speciation Studies:
• As (III) Vs. As (V)
•Cr (III) Vs. Cr (VI)……
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Arsenic Speciation with LC-ICP-MS
Mobile Phase (Basic):
2 mM phosphate buffer solution (PBS)
pH 11.0 adjusted with NaOH
0.2 mM EDTA
10 mM, CH3COONa
3.0 mM NaNO3
1% ethanol
* Tetsushi Sakai and Steven
Wilbur, Agilent Technologies
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*Adjusted to pH 11 to separate AsB. Ionic strength of mobile phase shortens elution of As V
10ug/L each As species
Page 23
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