USP “like” Residual Solvents analysis at Bristol-Myers Squibb Analytical R&D

USP methods • •

•

•

Static Headspace sampling Separate procedures for water soluble/insoluble materials Use of 0.53 and 0.32 mm diameter columns Limit test; Purchase prepared standards

2

Residual Solvent methods at BMS • •

• • •

Primarily Direct Liquid Injections Use of Static Headspace sampling increasing Water is rarely used as a solvent Use of 0.18 mm diameter “fast” columns Quantitation test; prepare fresh standards for every analysis 3

Class 2 Standards mixtures Std A

Std B

Std C

Acetonitrile

Chloroform

N,N-Dimethylacetamide

Chlorobenzene

1,2-Dimethoxyethane

N,N-Dimethylformamide

Cyclohexane

Hexane

2-Ethoxyethanol

1,2-Dichloroethene

Methylbutylketone

Ethyleneglycol

Dichloromethane

Nitromethane

Formamide

1,4-Dioxane

Pyridine

2-Methoxyethanol

Methanol

Tetralin

N-Methylpyrrolidone

Methylcyclohexane

1,1,2-Trichloroethene

Sulfolane

Tetrahydrofuran Toluene Xylene Mixture 4

BMS Approach Direct liquid injection increases sensitivity of high boiling solvents Solvent

BP (°C)

Solvent

BP (°C)

Methylene Chloride

40

2-methoxyethanol

125

cis-1,2-Dichloroethylene

60

Chlorobenzene

132

Chloroform

61

2-Ethoxyethanol

135

Methanol

65

m-Xylene

140

Tetrahydrofuran

66

o-Xylene

140

Hexane

69

p-Xylene

140

Cyclohexane

81

DMF

153

Acetonitrile

82

DMA

166

1,2-Dimethoxyethane

85

Ethylene Glycol

198

Trichloroethylene

87

n-Methylpyrrolidone

202

Methyl butyl Ketone

95

Tetralin

207

Methylcyclohexane

101

Formamide

210

Nitromethane

101

Sulfolane

285

1,4-Dioxane

102

Toluene

111

Pyridine

115

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USP Std C static headspace 20.00 19.00 18.00 17.00 16.00 15.00 14.00 13.00 12.00

mV

11.00 10.00 9.00 8.00 7.00 6.00 5.00 4.00 3.00 2.00 1.00 0.00 0.00

5.00

10.00

15.00

20.00

25.00

30.00 Minutes

35.00

40.00

45.00

50.00

55.00

60.00

6

USP Std C direct liquid 40.00

35.00

30.00

mV

25.00

20.00

15.00

10.00

5.00

0.00 0.00

1.00

2.00

3.00

4.00

5.00

6.00

7.00

8.00

9.00

10.00

11.00

Minutes

7

BMS methods with faster analysis times

USP Headspace

30m x 0.53mm x 3u 30m x 0.32mm x 1.8u

60 min

BMS Headspace

30m x 0.25mm x 1.4u

25 min

Liquid

20m x 0.18mm x 1.0u

12 min

Headspace

20m x 0.18mm x 1.0u

12 min

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Oven Temperature Programs 240° C 20 min USP

10° C/min 40° C 20 min 225° C 2min 40° C/min 75° C

BMS

40° C

0.5 min 10° C/min

2 min

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Chromatograms USP 0.53 mm column Headspace injection 280.00 260.00 240.00

Std A red

220.00

Std B blue

200.00

Std C black

180.00 160.00 mV

140.00 120.00 100.00 80.00 60.00 40.00 20.00 0.00 2.00

4.00

6.00

8.00

10.00

12.00

14.00

16.00

18.00

20.00 22.00 Minutes

24.00

26.00

28.00

30.00

32.00

34.00

36.00

38.00

40.00

10

Chromatograms BMS 0.18 mm column direct liquid injection 40.00 38.00 36.00

Std A black

34.00 32.00

Std B blue

30.00

Std C green

28.00 26.00 24.00

mV

22.00 20.00 18.00 16.00 14.00 12.00 10.00 8.00 6.00 4.00 2.00 0.00 -2.00 0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

4.50

5.00

5.50 Minutes

6.00

6.50

7.00

7.50

8.00

8.50

9.00

9.50

10.00

10.50

11.00

11

BMS Approach Quantitation Test vs. Limit Test Residual solvent levels are used to correct drug substance purity All clinical batches will have specification for Class 2 solvents that may be present

12

BMS Approach Sample concentrations of 100 mg/mL gives adequate sensitivity for all solvents 80 ppm standards had peak area % RSD of 2-5% Nitromethane MQL 4 ppm Methyl Butyl Ketone MQL 11 ppm

Solvent

ppm Limit

Nitromethane

50

2-Methoxyethanol

50

Methyl Butyl Ketone

50

Chloroform

60

Trichloroethylene

80

1,2-Dimethoxyethane

100

Tetralin

100

2-Ethoxyethanol

160

Sulfolane

160

13

Building specific lists

The calibration file is then used to extract mass spectral data for each component from the NIST Mass Spectral Library.

This creates a custom Mass Spectral Library that is a subset of the main library.

Compound List Report Instrument #1 Method Path : C:\msdchem\1\METHODS\ Method File : Fast624highsplit.M Title : USP 467 Class 2 Last Update : Tue Sep 05 11:24:33 2006 Response Via : Initial Calibration Total Cpnds : 27 PK# Compound Name QIon Exp_RT Rel_RT Cal #Qual -------------------------------------------------------------------------1 T Methanol 31 1.349 1.000 L 1 2 T Acetonitrile 41 2.143 1.000 L 3 3 T Methylene Chloride 84 2.232 1.000 L 3 4 T Hexane 57 2.547 1.000 L 3 5 T Nitromethane 61 2.917 1.000 L 2 6 T Furan, tetrahydro42 3.149 1.000 L 3 7 T Trichloromethane 83 3.164 1.000 L 3 8 T Cyclohexane 56 3.338 1.000 L 3 9 T Ethane, 1,2-dimethoxy45 3.494 1.000 L 2 10 T Ethanol, 2-methoxy45 3.519 1.000 L 1 11 T Trichloroethylene 130 3.913 1.000 L 3 12 T Cyclohexane, methyl83 4.026 1.000 L 3 13 T 1,4-Dioxane 88 4.108 1.000 L 3 14 T Ethanol, 2-ethoxy59 4.292 1.000 L 3 15 T Pyridine 79 4.542 1.000 L 3 16 T Toluene 91 4.594 1.000 L 2 17 T 2-Hexanone 43 4.903 1.000 L 3 18 T Formamide, N,N-dimethyl- 73 5.144 1.000 L 3 19 T Benzene, chloro112 5.272 1.000 L 3 20 T o & p-Xylene 91 5.367 1.000 L 3 21 T 1,2-Ethanediol 31 5.373 1.000 L 3 22 T Formamide 45 5.419 1.000 L 2 23 T m-Xylene 91 5.550 1.000 L 3 24 T Acetamide, N,N-dimethyl87 5.752 1.000 L 3 25 T 2-Pyrrolidinone, 1-methyl99 6.690 1.000 L 3 26 T Naphthalene, 1,2,3,4-tetrah... 104 7.026 1.000 L 3 27 T Thiophene, tetrahydro-, 1,1... 41 7.756 1.000 L 3

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Chromatograms 0.18mm column, direct liquid injection

Std A

15

Chromatograms 0.18mm column, direct liquid injection

Std B

16

Chromatograms 0.18mm column, direct liquid injection

Std C

17

AMDIS Deconvolution software

18

BMS USP “like” Method All Class 2 solvents plus some common Class 3 in a validated method „ Dissolution solvent choice DMA, DMSO or NMP „ Choice of injection static headspace or Direct liquid „ Mass spectral detection „ Deconvolution mass spectral method „

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