headspace analysis of residual solvents in … using flow-modulation chromatography. ... 3 3...

CHRISTOPHER M. ENGLISH, Rebecca E. CHRISTOPHER M. ENGLISH, Rebecca E. Wittrig, Frank L. Dorman.Wittrig, Frank L. Dorman.

Headspace Analysis of Residual Solvents in Pharmaceuticals, Using Flow-Modulation

Chromatography.

Application: Residual SolventsApplication: Residual SolventsPharmaceutical FormulationsPharmaceutical FormulationsGuidelines for TestingGuidelines for Testing

International Conference on HarmonizationInternational Conference on HarmonizationEuropean PharmacopoeiaEuropean Pharmacopoeia

Compound Lists VaryCompound Lists VaryOver 60 compounds of regulatory interestOver 60 compounds of regulatory interestClasses based on toxicitiesClasses based on toxicitiesResolution of large lists on a single stationary phase can be Resolution of large lists on a single stationary phase can be extremely difficultextremely difficult

The Three Approaches: The Three Approaches:

Evaluate Existing Phases Evaluate Existing Phases Develop a New Stationary PhaseDevelop a New Stationary PhaseStopStop--Flow GC TechnologyFlow GC Technology

Using Existing RT DataUsing Existing RT DataApplying RT Data for StopApplying RT Data for Stop--FlowFlow

Existing Phases Evaluated Existing Phases Evaluated

Change in selectivity Change in selectivity Low bleedLow bleedCritical resolutionCritical resolutionFID or MS detectionFID or MS detection

Column Design. Column Design.

n-propanolmethylene chloride

1,1-dichloroethene diethyl ether

acetonitrile/ aceteone/ isopropanolethanol

ethylene oxide methanol

MEK / C-6

benzene/C-tet/n-butanol

THF

1,1,1-TCA

ethyl acechloroform

1,2-DCA

C7

pyridine

1,4-dioxane

TCE

toluene

Rtx®-G27USP <467> Common Solvents

20 minutes

The RtxThe Rtx--G27 Unresolved G27 Unresolved

acetonitrile (acetonitrile (II),acetoneII),acetone (III), IPA (III)(III), IPA (III)MEK (III), C6 (II)MEK (III), C6 (II)benzene (I), carbon benzene (I), carbon tettet. (I), . (I), nn--butanolbutanol (III)(III)C7 (III), TCE (II)C7 (III), TCE (II)

RtxRtx®®--VGCVGCUSP <467> Common SolventsUSP <467> Common Solvents

1,1-dichloroethenediethyl ether

ethylene oxidemethanol

5

2 4 6 8 10 12 14

acetone / hexane2-propanol

methylene chloride

1-propanolacetonitrile

e.ace/111TCA/ THFcarbon tet.

chloroform

toluene

pyridine

1,4-dioxane1-butanolTCE

MEKbenzene/C7

1,2-DCA

The The RtxRtx--VGC Unresolved VGC Unresolved

acetone (III), C6 (II)acetone (III), C6 (II)E.aceE.ace (III),111TCA (I), THF (III)(III),111TCA (I), THF (III)benzene (I), C7 (III)benzene (I), C7 (III)

Bonded Polymer Examined for GC Bonded Polymer Examined for GC Applications.Applications.

Si O

C

Si O

(CH )2 2

Si O

(CH )2 3

C

Si O

(CH )2 3

CH3

CH3

CH3CH3

F

F F

FF

F

F

F

CH3

F

F

F

F

F F

FF

F

F

C

C

CC

CFF

F

Experimental Fluorinated Phase

Modeling for a New Bonded PhaseModeling for a New Bonded Phase

0.10n-

prop

ylbe

nzen

e

2-ch

loro

tolu

ene

4-ch

loro

tolu

ene

1,3,

5-tri

met

hylb

enze

ne

tert-

buty

lben

zene

1,2,

4-tri

met

hylb

enzn

e

sec-

buyl

benz

ene

1,3-

dich

loro

benz

ne

1,4-

dich

loro

benz

ne

p-is

opro

pylto

luen

e

1,2-

dich

loro

benz

ene

n-bu

tylb

enze

ne

0.10

0.18

0.18

0.15

0.15

0.11

0.45

0.18

0.44

0.15

0.17

0.38

0.18

0.17

0.11

0.45

0.15

0.160.13

0.06

0.07

Stationary Phases Used for ModelingStationary Phases Used for Modeling

CF

Si O

C H2 4

CH3

3

trifluoropropylmethylpolysiloxane

CH

Si

CH

O

3

3

dimethylpolysiloxane

Si O

diphenylpolysiloxane C N

Si O

(CH )2 3

cyanopropylphenylpolysiloxane

Current Issues with the ICH Compounds.

SeparationConcentration

Headspace & Stop Flow

Stop Flow GC System: Sacks, et. al.*

C1 C2FID#1

Inlet

V

FID #2(or MS)

BC

EPC*Richard Sacks, University of Michigan

Summary of StopSummary of Stop--Flow GCFlow GC

Ability to “Tune” the SelectivityAbility to “Tune” the SelectivityFlexibilityFlexibility

Standard dimension columnsStandard dimension columnsCan vary the pulse sequencesCan vary the pulse sequences

Significant Improvements in Analysis Times PossibleSignificant Improvements in Analysis Times PossibleFast oven programs, high flow ratesFast oven programs, high flow rates

Class I & II Residual SolventsClass I & II Residual Solvents

sulfolone361,4-dioxane18

formamide35toluene17

1-methyl-2-pyrrolidinone (1-MP)34chloroform16

ethylene glycol (EG)33acetonitrile (MeCN)15

1,2,3,4-tetrahydronaphthalene (THN)32trichloroethene (TCE)14

N,N-dimethylacetamide (DMA)31cis-1,2-dichloroethene13

dimethyl formamide (DMF)30benzene12

1,1,2-trichloroethane (1,1,2-TCA)29methylene chloride (CH2Cl2)11

2-ethoxyethanol281,2-dimethoxyethane10

chlorobenzene27methanol9

o-xylene261,1,1-trichloroethane (1,1,1-TCA)8

pyridine25carbon tetrachloride (CCl4)7

2-methoxyethanol24trans-1,2-dichloroethene6

nitromethane23methyl cyclohexane5

m-xylene221,1-dichloroethene (1,1-DCE)4

p-xylene21methyl cyclopentane3

2-hexanone (MBK)20hexane2

1,2-dichloroethane (1,2-DCA)192-methylpentane1CompoundCompoundPeak #Peak #CompoundCompoundPeak #Peak #

CoelutionsCoelutions by Phase by Phase Class I & IIClass I & II

Rtx-11,2-dichloroethane / methyl cyclopentane1,4-dioxane / trichloroethenep-xylene / m-xylene

Rtx-200hexane / methylene chloride1,2-dichloroethane / trichloroethene2-ethoxyethanol / toluene

Stabilwaxcarbon tetrachloride / 1,1,1-trichloroethanecis-1,2-dichloroethene / trichloroethene

Rtx-VMSCarbon tetrachloride / 1,1,1-trichloroethane (close pair)Trichloroethene / methyl cyclohexanep-xylene / m-xylene

Class I & II OVIs: Total of 9 PulsesAt the End Detector – all 36 resolved

1 2 43

9

6

57 11

8 12 10

13

16

14

15 19

17

18

23

24

20

21

22

25 28

26

27

29

30

31

32

33

35

34

36

Residual Solvents: Run ConditionsResidual Solvents: Run Conditions

Dual FIDs @ 250°CDual FIDs @ 250°CDetectors

0.2 µL HS, 200:1 split0.2 µL HS, 200:1 splitInjection

230°C230°CInjector

2.5 mL/min. to 9.5 min.3.5 mL/min. at 10 min.

1.5 mL/min. constant flow

Column Flow

40°C (1 min. hold)to 65°C at 6°C/min.,

to 100°C at 12°C/min.,to 250°C at 70°C/min.,

1.8 min. hold

40°C (6 min. hold)to 100°C at 4°C/min.,

to 220°C at 15°C/min.,5 min. hold

Oven Program

Stabilwax 15m x 0.25mm, 0.5µm

Rtx-20030m x 0.25mm, 1µm

Stabilwax 15m x 0.25mm, 0.5µm

Rtx-20030m x 0.25mm, 1µm

Analytical Columns

Fast ProcedureStandard Procedure

Class I & II OVIs: Total of 9 PulsesAt the Junction

Fid#1Inlet

FID #2

31psi

StabilwaxStabilwax RtxRtx--200200

Flow Modulation Pulses

2.3ml/min 2.0 ml/min

0.3 ml/min

1.7 ml/min

0.6 ml/min

3.9 ml/min

0.4m 0.05mm tubing

1.0m 0.10 mm tubing

min2 4 6 8 10 12 14 16

pA

0

500

1000

1500

2000

2500

3000

3500

4000

FID1 A, (C:\HPCHEM\1\DATA\HT3\Q5022A02.D)

Stabilwax 15m x 0.25mm x 0.5dfStabilwax 15m x 0.25mm x 0.5dfRtxRtx--200 30m x 0.25mm x 1.0df200 30m x 0.25mm x 1.0df

Analysis without Pulses 2Analysis without Pulses 2ndnd FIDFID

m4.8 5 5.2 5.4 5.6 5.8 6

pA

0

200

400

600

800

1000

1200

1400

1600


m7.1 7.15 7.2 7.25 7.3 7.35

pA

0

100

200

300

400

500


Cl3CHMeCN/TCE

CCl4/MCC6CH2Cl2111TCA

min9 9.25 9.5 9.75 10 10.25 10.5 10.75

pA

0

100

200

300

400

500


min3 4 5 6 7

pA

0

10

20

30

40

50

FID1 A, (C:\HPCHEM\1\DATA\HT3\Q4070626.D)

min3 4 5 6 7

pA

0

200

400

600

800

1000

FID2 B, (C:\HPCHEM\1\DATA\HT3\Q4070626.D)

Three Flow Modulation PulsesThree Flow Modulation Pulses

End FID

Junction FID

MeC

N

TCE/

C12

DC

E

Cl3

CH

111T

CA

/CC

l4

MeC

l2

MC

C6

RT=

1.56

MeCl2CCl4

MCC6

111TCAmin4.8 5 5.2 5.4 5.6 5.8 6

pA

0

200

400

600

800

1000

1200

1400

1600


CC

l4/M

CC

6

CH

2Cl2

111T

CA

15°C/min

25°C/min

Teledyne Tekmar HT7000Platen Temperature: 140°CSample Equilibration Time: 5 minutesMixing Time: 10 minutesMixing Power: 2Mixture Stabilization Time: 1Pressure Time: 0.2 minutesPressure Equilibrium: 0.3 minutesLoop Fill Time: 0.1 minutesLoop Equilibration Time: 0.05 minutesInjection Time: 1.0 minutesLoop/Line Temperature: 250°CVial Size: 22ml high temperature vialsSample Loop Size: Silcosteel 1ml Standard Size.Static Vial Pressure: 3.5 PSI HeliumVial Pressurization: 8 PSI HeliumVariable Injection Pressure (VIPR) 5 PSI HeliumConcentration: 200ppm each component.Solvent: 1,3-dimethyl-2-imidazolidinone (DMI)Interface: plumbed through injection port 1:20 split

GC Agilent 6890 Injection Port Temperature: 250°CCarrier gas: helium, constant flow Flow rate: 2.3 ml/min, 25.6psi @ 40°C. Oven temperature: 40°C (hold 2 min.) to 55°C @ 4°C/min. (no hold)

to 110°C @ 25°C/min. (hold 2 min.) to 250°C @25°C/min (hold 5 minutes)

Total Runtime: 20.55 minutesAgilent Dual Flame Ionization Detectors

Restek’s Stop-Flow Unit Pulse Settings:Injection Port Connection: cool-on-column injector.Pressure: 31.0 PSI constant pressurePulses: Valve Closed 0.00 minutes.

Valve Open 3.00 minutesValve Closed 3.15 minutesValve Open 4.65 minutesValve Closed 5.02 minutesValve Open 5.10 minutesValve Closed 5.40 minutes

min4 6 8 10 12 14 16 18

pA

0

100

200

300

400

500

600

700

t12D

CE11

DC

E MEO

H

MC

C6

CC

l4M

eCl2

Ben

zene

111T

CA 12

DM

etha

neTC

Ec1

2DC

EC

l3C

HM

eCN

12D

CA 1,4-

Dio

xane

Tolu

ene

NN

DM

A

1MP

Form

amid

e

Sulfo

lane

** **

*

*

* DMI Contaminant

SF-HS7000/GC/FID for ICH Class I & II200 ppm 5ml DMIC

6

N-m

e

2MC

5

2mEt

ohM

BK

P-xy

lene

M-x

ylen

epy

ridin

e2e

Etoh

Cl-B

O-x

ylen

e11

2TC

A

MC

C5

DM

F

Solvent:1,3-dimethyl-2-imidazolidinone

(DMI)

Teledyne Teledyne TekmarTekmar HT3 HS/HSHT3 HS/HS--TrapTrap

Instrument SetupInstrument Setup

HT3 Conditions Using DMIHT3 Conditions Using DMIConstant Heat Time ONG.C. Cycle Time 23.00 minValve Oven Temp 150 CTransfer Line Temp 150 CStandby Flow Rate 50mL/minPlaten / Sample Temp 150 CPlaten Temp Equil. Time 0.10 minSample Equil. Time 1.00 minMixer ONMixing Time 10.00 minMixing Level Level 5Mixer Stabilize Time 0.50 minPressure 8 PSIGPressure Time 2.00 minPressure Equil. Time 0.50 minLoop Fill Pressure 5 PSIGLoop Fill Time 2.00 minLoop Fill Equil. Time 0.50 minInject Time 1.00 min

Class I & II Residual SolventsClass I & II Residual Solvents

sulfolone361,4-dioxane18

formamide35toluene17

1-methyl-2-pyrrolidinone (1-MP)34chloroform16

ethylene glycol (EG)33acetonitrile (MeCN)15

1,2,3,4-tetrahydronaphthalene (THN)32trichloroethene (TCE)14

N,N-dimethylacetamide (DMA)31cis-1,2-dichloroethene13

dimethyl formamide (DMF)30benzene12

1,1,2-trichloroethane (1,1,2-TCA)29methylene chloride (CH2Cl2)11

2-ethoxyethanol281,2-dimethoxyethane10

chlorobenzene27methanol9

o-xylene261,1,1-trichloroethane (1,1,1-TCA)8

pyridine25carbon tetrachloride (CCl4)7

2-methoxyethanol24trans-1,2-dichloroethene6

nitromethane23methyl cyclohexane5

m-xylene221,1-dichloroethene (1,1-DCE)4

p-xylene21methyl cyclopentane3

2-hexanone (MBK)20hexane2

1,2-dichloroethane (1,2-DCA)192-methylpentane1CompoundCompoundPeak #Peak #CompoundCompoundPeak #Peak #

Class III Added Class III Added Residual SolventsResidual Solvents

compoundcompound classclass

pentanepentane 33

methylalmethylal 44

pentanepentane 33

nn--heptaneheptane (C7)(C7) 33

methyl acetatemethyl acetate 33

acetoneacetone 33

22--propanolpropanol 33

tetrahydrofurantetrahydrofuran 33

ethyl acetateethyl acetate 33

isopropyl acetateisopropyl acetate 33

22--butanolbutanol 33

11--propanolpropanol 33

1,11,1--diethoxypropanediethoxypropane 44

hexanonehexanone (MIBK)(MIBK) 33

11--butanolbutanol 33

isoamylisoamyl alcoholalcohol 33

isoamylisoamyl acetateacetate ss

11--pentanol pentanol 33

anisoleanisole 33

dimethyldimethyl sulfoxidesulfoxide 33

min2 2.5 3 3.5 4 4.5

pA

0

1000

2000

3000

4000


min2 4 6 8 10 12 14 16 18

pA

0

1000

2000

3000

4000


Pent

ane

2-m

ethy

lpen

tane

hexa

ne

1,1-

dich

loro

ethe

ne

met

hyl c

yclo

pent

ane

met

hano

l n-he

ptan

et-1

,2-d

ichl

oroe

then

e

m. a

ceta

teac

eton

e

acet

alde

hyde

Class I,II,III 2Class I,II,III 2ndnd FID by HSFID by HS

met

hyla

l

min5 5.5 6 6.5 7 7.5

pA

0

250

500

750

1000

1250

1500

1750

2000


min2 4 6 8 10 12 14 16 18

pA

0

1000

2000

3000

4000


m. c

yclo

hexa

ne

c. te

t2-

prop

anol

met

hyle

nech

lorid

e

1,1,

1-tr

ichl

oroe

than

e

ethy

l ace

tate

THF

benz

ene

1,2-

dim

etho

xyet

hane

C-1

,2-D

ichl

oroe

then

eI.P

.ace

tate

chlo

rofo

rm

acet

onitr

iletr

ichl

oroe

then

e

1-pr

opan

ol2-

buta

nol

1,1-

diet

hoxy

prop

ane

ND


min8.5 9 9.5 10 10.5

pA

0

200

400

600

800

1000


min2 4 6 8 10 12 14 16 18

pA

0

1000

2000

3000

4000


1,2-

dich

loro

etha

ne

1,4-

diox

ane

tolu

ene

Hex

anon

e(M

IBK

)

1-bu

tano

l

nitr

omet

hane

2-m

etho

xyet

hano

l

2-he

xano

ne (M

BK

)

p-xy

lene

m-x

ylen

eis

oam

ylal

coho

l

2-et

hoxy

etha

nol

isoa

myl

acet

ate

1-pe

ntan

ol

1,1,

2-tr

ichl

oroe

than

e

o-xy

lene

chlo

robe

nzen

e

pyrid

iine


min11 12 13 14 15 16 17 18 19

pA

0

50

100

150

200

250

300

350

400


min2 4 6 8 10 12 14 16 18

pA

0

1000

2000

3000

4000


DM

F

1-m

ethy

l-2-p

yrili

done

DMI

anis

ole

Sulfolane ND

Formamide ND

Class I,II,III 2Class I,II,III 2ndnd FID by HS (spiked @ 400ppm)FID by HS (spiked @ 400ppm)

DM

SO

N,N

-dim

ethy

lace

tam

ide

EG

StabilwaxStabilwax / Rtx/ Rtx--200 Unresolved200 UnresolvedUnder Current ConditionsUnder Current Conditions

methylalmethylal (s) / methanol (II)(s) / methanol (II)nn--heptaneheptane (III) / isooctane (s)(III) / isooctane (s)tt--1,21,2--DCE (II) / ethyl DCE (II) / ethyl formateformate (III)(III)ethanol (III) / methyl ethanol (III) / methyl cyclohexanecyclohexane (II)(II)22--butanone (III) / 1,2butanone (III) / 1,2--dimethoxyethane (II)dimethoxyethane (II)isobutyl acetate (III) / isobutyl acetate (III) / hexanonehexanone (III)(III)11--pentanol (III) / pentanol (III) / isopropylbenzeneisopropylbenzene (III)(III)cc--1,21,2--dichloroethene (II) / isopropyl acetate (III)dichloroethene (II) / isopropyl acetate (III)

Summary of StopSummary of Stop--Flow GCFlow GC

Ability to “Tune” the SelectivityAbility to “Tune” the SelectivityFlexibilityFlexibility

Standard dimension columnsStandard dimension columnsCan vary the pulse sequencesCan vary the pulse sequences

Significant Improvements in Analysis Times PossibleSignificant Improvements in Analysis Times PossibleFast oven programs, high flow ratesFast oven programs, high flow rates

ConclusionsConclusions

Continue evaluation of current phases Continue evaluation of current phases Continue work on a new stationary phasesContinue work on a new stationary phases

Using computer modelingUsing computer modelingGoal: resolve Class I, II, III, other solvents. Goal: resolve Class I, II, III, other solvents.

Continue with StopContinue with Stop--Flow technology Flow technology

Special Thanks to Ed Special Thanks to Ed Price, Eric Price, Eric HeggsHeggs, , Brian Wallace & Terry Brian Wallace & Terry Jeffers of Teledyne Jeffers of Teledyne TekmarTekmar for their for their assistance in setup assistance in setup and operation of the and operation of the HT3 Headspace unit. HT3 Headspace unit.

headspace analysis of residual solvents in … using flow-modulation chromatography. ... 3 3...

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