gc gc in analysis of enantiomers - a leading global university · gc×gc principle 187 167 185 201...

GC×GC in analysis of enantiomers

Peter Korytár

The Netherlands Institute for Fisheries Research

GC×GC principle

injector detector

modulator

1st columnconventional

(30 m × 0.25 mm × 0.25 µm)

2nd columnfast

(1 m × 0.1 mm × 0.1 µm)

2nd dimension retention time

2nd dimension retention time1st dimension retention time

1st dimension retention time

GC×GC principle

a

aa

ab

bb

b

b

Raw GCxGCchromatogram

GC×GC principle

a b

1D-GC chromatogram

GC×GC principle

187167

185

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ec)

1st dimension retention time (min)

201 180

198

187 183

128167

185

171

156 157

180

198173

201202

187187183183

128128

167167 185185171171

156156 157157180180

198198

173173

201201

202202

Contour plot

GC×GC principle

187167

185 201 180

198

187187183183

128128

167167 185185171171

156156 157157180180

198198

173173

201201

202202

2nddi

men

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ec)


187 183

128167

185

171

156 157

180

198173

201202

APEX plot

Enantioseparation by GC×GC

• GC × enantio-GC

• enantio-GC × GC

GC × enantio-GC: (normal) set-up

Non-polar phase


(30 m × 0.25 mm × 0.25 µm)

Enantio selective phase

2nd columnfast

(1 m × 0.1 mm × 0.1 µm)

injector detector

modulator

Fast enantioseparationColumn:EtTBS-βCD phase1m × 0.1mm × 0.1µm

Isothermal:70°C

Linear carrier gas velocity:60 cm/s = ūopt(2ūopt or 3ūopt ⇒ 58 s or 37 s)

limon

ene

cam

phor

linal

ool

linal

ylac

etat

e

retention time (s)

R. Shellie, Ph.J. Marriott, Anal. Chem. 74 (2002) 5426.

GC × enantio-GC: (alternative) set-up

Non-polar phase

1st columnconventional DB-5

(10 m × 0.10 mm × 0.10 µm)


2nd columnfast EtTBS-βCD

(1 m × 0.25 mm × 0.25 µm)

MS detector(vacuum)injector

modulator

GC × enantio-GC: (alternative) set-up

(-) linalool

(+) limonene

(-) limonene

(+) linalool

(-) linalylacetat

(+) linalylacetat1,

8-ci

neol

e(±

)α-p

inen

e

1,8-cineole

(-)limonene

(-) limonene

(+) limonene

1,8-cineole(+) limonene

(-) limonene (+) limonene

6136 14028

30156894= =

Rs (limonene) = 1.4(linalool) = 1.6(linalyl acetat) = 0.8

= 0.437

2tR ((+) linalyl acetat, MS) = 38 s2tR ((+) linalyl acetat, FID) = 154 s

R. Shellie, Ph.J. Marriott, Anal. Chem. 74 (2002) 5426.

GC × enantio-GC: conclusions

• Enantiomeric separation is worse compared to heart-cut MDGC

• Separation of enantiomeric pair from other co-elutants is questionable

• Suitable only for well resolved pairs of enantiomers

• Vacuum set-up is rather difficult to operate

• Not really a viable option ?

GC × enantio-GC: SolGel × Cyclodex-B

traditional Chinese medicine• more than 800 peak detected• 394 compounds positively identified• some of those compounds are enantiomers

J. Wu, X. Lu, W. Tang, H. Kong, S. Zhou, G. Xu, J. Chromatogr. A 1034 (2004) 199.

enantio-GC × GC



(30 m × 0.25 mm × 0.25 µm)

Polar or shape selective phase

2nd columnfast

(1 m × 0.1 mm × 0.1 µm)

injector detector

modulator

EtTBS-β-CD × BP-20

R. Shellie, Ph. Mariott, C. Cornwell, J. Sep. Sci. 24 (2001) 823.

(EtTBS-β-CD+CycloSil B) × BPX-50Red wine volatiles

Y. Shao, Ph. Mariott, Anal. Bioanal. Chem. 375 (2003) 635.

(EtTBS-β-CD+CycloSil B) × BPX-50Strawberry volatiles

linalool

2,5-dimethyl-4-hydroxy-(2H)-furan-3-one(DMHF)

A. Williams, D. Ryan, A.O. Guasca, Ph. Mariott, E. Pang, J. Chromatogr. B 817 (2005) 97.

Xenobiotics: PCBs on Chirasil-Dex

M. Harju, A. Bergman, M. Olsson, A. Roos, P. Haglund, J. Chromatogr. A 1019 (2003) 127.

Chirasil-Dex × LC-50

Standard of the 144 Aroclor CBs

Grey seal blubber sample


Chirasil-Dex × VF-23ms

Standard of the 144 Aroclor CBs

Grey seal blubber sample


Quantitative parameters


(-)

(-)(-)

(-)(+)

(+)(+)

(+)

(+)

enantio-GC × GC: quantification

(-) (+)

enantio-GC × GC: conclusions

• Enantiomeric separation is not improved but 1D-GC separation is maintained

• Excellent and fast separation of the enantiomersfrom other sample constituents

• GCxGC – excellent tool for determination of isomers well separated in the 1D-GC

• Heart-cut MDGC still the method of choice if isomers are eluting very closely

• Method development • select chiral column • select second column

DB-1×007-65HT: overlay

20 30 40 50 60 700

1

2

3

4

5

6

7

8

10

9

10 PBDEsPCBsPCNsPCDTsPCDEsPBBsPCDD/FsToxaphene

2nddi

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OCPs

DB-1×007-65HT: PCAs Clx

1

2

3

4

5

6

010 20 30 40 50 60

2nd

dim

ensi

on re

tent

ion

time

(s)


C10–C13 with 52, 56 and 63 wt.% Cl content

C14–C17 with 42 and 57 wt.% Cl content

C18–C20 with 36 wt.% Cl content

1920

21

14

15

1617

1819

2021 22

23

2322

18

17

24

21

2223

24

25

26

C18Cl5

C18Cl6C19Cl5C14Cl5 C14Cl6

C15Cl5

C11Cl8C12Cl7C13Cl6

DB-1×007-65HT: PCAs Clx

A – Slovak dust

B – Spanish dust

2nddi

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)


shortmedium long

shortmedium long

O

Brx BryDB-1×007-65HT: PBDEs

0

1

2

3

4

5

6

7

8

10 15 20 25 30 35 40

2nd

dim

ensi

on re

tent

ion

time

(s)


0

1

2

3

4

5

6

7

8

45 50 55 60 65 70

155

105

126

85

114106

127

116

124

99104

103121

102100

101120

125

88109

98,119

40,77

81

55

78

42,66

58,79

74

47

67

80

49

68

7146,48

726F-47

62

51

53

69,73

50

22,37

20,35

38

30

27 19

18

34

32

2629

36

17

39

16,33

28

10

4

9

14

12,13

1511

8

23

115

76

7

6

25,31

97,118

108,123

8687

4MeO-49

4OH-42

3F-100

6F-664’F-69

2OH-28

BB52

BB49

4’F-27BB15

1 4’F-253’F-28

BB101

6OH-47

4OH-49

154

me-TBBP-A

BB153

6OH-99

144161

168

153TBBP-A

139

131,158,140

141

160142

159 167

HBCD

138

166

156184

128

BB169183

182185

192

191

181

173,190204

4’,6F-1992,4’F-198

198

203205

208207 206

4’F-208

209

80

4’F-160

3F-119

6MeO-47

203

198

183 181

PCAs

HBCD138

139

153

TBBP-A

144154

85116

99101

100

DB-1×007-65HT: dust extract

203

183181

138

139

153HBCD

144

154

85116

99101

10042 or 66

47

46 or 48

28

7

PCAs

TBBP-A 198

DB-1×007-65HT: PCTs, toxaphene, PCAs

0

1

2

3

4

5

6

7

8

15 25 35 45 55 65 75 85

PCAs

Toxaphene

PCTs

2nddi

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DB-1xLC-50: overlay

20 30 40 50 60 700

1

2

3

4

5

6

7

8

10

9

10

11PCDD/FsPCBs

81

77 126 169

123,118114

105

167

156157

189

2nddi

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PBBsPBBs

15

5249

169

101 153

PBDEsPCDEsOCPsToxaphenesPCNsPCDTs

DB-XLBxLC-50

20 30 40 50 60 700

1

2

3

4

5

6

7

8Sediment sample – dioxin fraction

2nddi

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4F1 4D1

5F1

5F2

5D1

6F1

6F2

6F3

6D16D2

6D3

7F1

6F4

7D1

7F2

OCDD

OCDF

DB-1xVF-23: overlay2nd

dim

ensi

on re

tent

ion

time

(sec

)

1st dimension retention time (min)20 30 40 50 60 70

0

1

2

3

4

5

6

7

8

10 80

OCPsPBDEsPCBsPCNsPCDTsPCDEsPCDD/FsPBBs

DB-1xVF-23: toxaphene2nd

dim

ensi

on re

tent

ion

time

(sec

)


0

1

2

3

4

5

6

7

8

10 80

DB-1xVF-23: PCA-602nd

dim

ensi

on re

tent

ion

time

(sec

)


0

1

2

3

4

5

6

7

8

10 80

gc gc in analysis of enantiomers - a leading global university · gc×gc principle 187 167 185 201...

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