Basics of comprehensive GC
•Dr. Frank Michel
sigma-aldrich.com
Overview
•Two-columns-one-MS
•Classical Heart-cut
•Comprehensive GCxGC
•Examples
2
Different Selectivities (37 FAMEs)
Omegawax
SP-2380
SP-2560
3
Two-columns-one-MS
Source:
Shimadzu
•Cost-efficient option
•Saving one MS resp. time for changing
•Two columns simultaneously in MS without compromise in
performance
•Subsequent injections
SPB-5
•Either two subsequent data files or one
data file is recorded
WAX
4
Source:
Shimadzu
Two-columns-one-MS
•Allergenic compounds in cosmetics
•Regulation by EU
Complete Information and Confirmation
•Coelution with matrix
by 2. column:
H. Leijs et al: J. Agric. Food Chem. 53
(2005),6487
2. Injektion
(x100,000,000)
TIC
SPB-5
2.50
WAX
2.25
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
0.00
5.0
10.0
15.0
20.0
25.0
30.0
35.0
40.0
45.0
50.0
55.0
60.0
5
MDGC/GCMS-2010
Source:
Shimadzu
Heart-cut
6
Source:
Shimadzu
Conventional Dean’s Switch
Standby Mode
Switching
pressure
1. Det.
2. Det.
Switching
pressure
1. Det.
Switch Mode
2. Det.
7
Column Switching - Challenges
Source:
Shimadzu
• Switching by valves mechanical system
Problem:
• Void volume
• contact of sample with active surfaces - Absorption
• Impacting heating/cooling
• Dean switching Pressure system
Problem:
• Pressure changes
Standby（1. Det. ）
Cut（1. Det.）
Shift of
retention times
Technical Solution by Shimadzu
Source:
Shimadzu
• Combination of valve (not in anal. line) and Dean
switch
Multi Dean‘s Switch Technik
• Advantages:
• No shift of retention time, one master chromatogram sufficient
for definition of several cuts
• Low void volume because of capillary switching unit
• 100 % Recovery Rate
• Complete inert unit
• Dedicated Software (MDGC Solution)
9
Source:
Shimadzu
Application – Screening of Allergenes
uV(x10,000)
4.25
Chromatogram
Allergen standard
4.00
3.75
1 st Dimension
3.50
Allergen standard cuted
3.25
3.00
2.75
2.50
2.25
2.00
1.75
1.50
1.25
1.00
0.75
0.50
0.25
0.00
-0.25
-0.50
-0.75
-1.00
2.5
5.0
7.5
10.0
12.5
15.0
17.5
20.0
22.5
25.0
min
10
Source:
Shimadzu
Application – Screening of Allergenes
uV
(x10,000)
uV
(x10,000)
Chromatogram
3.00
Chromatogram
2.1
uV(x10,000)
4.25
Chromatogram
2.0
4.00
2.75
3.75
Allergen standard
1.9
3.50
1.8
1 st Dimension
2.50
Allergen standard cuted
3.25
1.7
3.00
2.25
2.75
1.6
2.50
2.00
1.5
2.25
2.00
1.75
1.4
1.75
1.3
1.50
1.2
1.25
1.50
1.00
1.1
0.75
1.25
1.0
0.50
0.25
1.00
0.9
0.00
0.8
-0.25
0.75
0.7
-0.50
-0.75
0.6
0.50
-1.00
2.5
5.0
7.5
10.0
0.5
0.25
12.5
15.0
17.5
20.0
22.5
25.0
min
0.4
0.3
0.00
0.2
-0.25
8.1
0.1
8.2
8.3
8.4
8.5
8.6
8.7
8.8
8.9
9.0
9.1
9.2
9.3
9.4
9.5
9.6
9.7
9.8
min
18.50 18.75 19.00 19.25 19.50 19.75 20.00 20.25 20.50 20.75 21.00 21.25 21.50 21.75 22.00 22.25 22.50
min
11
Application – Screening of Allergenes
Benzyl Alcohol
(x10,000,000)
TIC
5.5
D-Limonene
2 nd Dimension
6.0
5.0
4.5
Source:
Shimadzu
4.0
3.5
3.0
2.5
2.0
1.5
1.0
0.5
6.0
7.0
8.0
9.0
10.0
11.0
12.0
13.0
14.0
15.0
16.0
17.0
18.0
19.0
20.0
21.0
22.0
12
Application – Screening of Allergenes
Benzyl Alcohol
(x10,000,000)
TIC
5.5
D-Limonene
2 nd Dimension
6.0
5.0
4.5
Source:
Shimadzu
4.0
3.5
3.0
%
2.5
125.0
2.0
1.5
68
100.0
1.0
0.5
75.0
93
6.0
7.0
8.0
9.0
10.0
11.0
12.0
13.0
14.0
15.0
16.0
17.0
18.0
19.0
20.0
21.0
22.0
50.0
25.0
0.0
41
53
136
107 121
36
148158 173
200
203
225 237
261
268278 295
30 40 50 60 70 80 90 100110120130140150160170180190200210220230240250260270280290300310
13
Application – Screening of Allergenes
Benzyl Alcohol
(x10,000,000)
TIC
5.5
D-Limonene
2 nd Dimension
6.0
5.0
4.5
Source:
Shimadzu
4.0
3.5
3.0
%
%
2.5
79
100
125.0
2.0
108
90
1.5
80
68
100.0
1.0
70
0.5
60
75.0
93
6.0
7.0
8.0
9.0
10.0
11.0
12.0
13.0 50
14.0
15.0
16.0
17.0
18.0
19.0
20.0
21.0
22.0
40
50.0
30
51
25.0
41
53
136
107 121
20
91
10
0.0
36
148158 173
200
203
225 237
261
268278 295
0
30 40 50 60 70 80 90 100110120130140150160170180190200210220230240250260270280290300310
53
36
121 135 151159 175
194 208 218
236 250 262 278
298
30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 220 230 240 250 260 270 280 290 300 310
14
MDGC - Configurations
Source:
Shimadzu
• GC + GC, FID, NPD, ECD, etc.
• GC + GCMS
• Headspace, Thermal desorption, Pyrolysis, Sniffer,
CryoTrap
• Each of them can be used as single system, too.
19
Comprehensive Chromatography
Source:
Shimadzu
•Comprehensive Chromatography is an “on-line”
multidimensional procedure.
•Perment Heart-Cutting allows a 2-dimensional
separation of the whole sample.
20
Definition
•Giddings Definiton:
• A two-dimensional separation can be called comprehensive if
– 1. Every part of the sample is subjected to two different separations
– 2. Equal percentages (either 100% or lower) of all sample
components pass through both columns and eventually reach the
detector
– 3. The separation (resolution) obtained in the first dimension is
essentially maintained.
•J.C. Giddings, J. High Resol. Chrom. (10) 1987, 319
21
Comprehensive GC
Source:
Shimadzu
•1991 developed Liu and Phillips a new system for transfer:
thermal modulator
•Publications of colored 2D chromatogram of an oil sample*
initiated interest in GCxGC
•
*Z. Y. Liu, J. B. Phillips, J. Chrom. Sci. 29 (1991) 227
22
Comprehensive GCMS
Source:
Shimadzu
Thermal Modulation:
23
Comprehensive GCxGC
Source:
Shimadzu
Shimadzu GCMS
with ZX-1 Modulator
Shimadzu GCMS
with ZX-2 Modulator
24
Source:
Shimadzu
GCxGC - Schematically
Detector
Injector
ZOEX
Loop Modulator
(cold/hot)
nonpolar
30m,
0.25mm,
0.25µ
µm
First column
Second column
polar,
1m,
0.1mm,
0.1µ
µm
25
Source:
Shimadzu
Thermal Modulation
ZOEX LOOP MODULATOR
Capillary column
in a loop
Cold gas stream
Hot gas stream
Release of sample molecules
Molecules from sample are
frozen/focussed
26
Principle – Comprehensive GC
No switching unit close to the columns
Provides extremely narrow peaks
Typical modulated Peak
Loop
Modulator
1st Stage
2nd Stage
Hot Jet
Cold Jet
~ 300 ms
27
Source:
Prof. Oliver Schmitz
Principle – GCxGC
100 % Dimethylpolysiloxane or
5 % Phenyl-95 % Dimethylpolysiloxane
35 – 50 % Phenyl-65 – 50 %
Dimethylpolysiloxane, Polyethyleneglycol
(Carbowax) or Cyanopropyl-phenyldimethylpolysiloxane
hot
nonpolar
cold
polar
28
Source:
Prof. Oliver Schmitz
Principle – GCxGC
hot
nonpolar
cold
polar
29
Source:
Prof. Oliver Schmitz
Principle – GCxGC
cold
hot
nonpolar
cold
polar
30
Source:
Prof. Oliver Schmitz
Principle – GCxGC
hot
cold
nonpolar
hot
polar
31
Source:
Prof. Oliver Schmitz
Principle – GCxGC
cold
hot
nonpolar
cold
polar
32
Source:
Prof. Oliver Schmitz
Principle – GCxGC
hot
4s
cold
nonpolar
hot
polar
33
Source:
Prof. Oliver Schmitz
Principle – GCxGC
cold
4s
hot
nonpolar
cold
polar
34
Source:
Prof. Oliver Schmitz
Principle – GCxGC
hot
4s
4s
cold
nonpolar
hot
polar
35
Source:
Prof. Oliver Schmitz
Principle – GCxGC
cold
4s
4s
hot
nonpolar
cold
polar
36
Source:
Prof. Oliver Schmitz
Principle – GCxGC
hot
4s
4s
cold
nonpolar
4s
hot
polar
37
Source:
Prof. Oliver Schmitz
Principle – GCxGC
cold
4s
4s
hot
nonpolar
4s
4s
cold
polar
38
Source:
Prof. Oliver Schmitz
Principle – GCxGC
4s
4s
4s
4s
39
Principle – GCxGC
Source:
Prof. Oliver Schmitz
40
Principle – GCxGC
Source:
Prof. Oliver Schmitz
4s
41
GCxGC Visulization
Stat. Phase Type 1
Boiling point
54.1 min.
Stat. Phase Type 50
Polarity
Source:
Shimadzu
87.5 min.
42
GCxGC Visulization
Stat. Phase Type 1
Boiling point
54.1 min.
Stat. Phase Type 50
Polarity
Source:
Shimadzu
87.5 min.
43
Source:
Shimadzu
GCxGC Visulization
GC x GC:
Stat. Phase Type 50
Polarity
Stat. Phase Type 1
Boiling point
44
GCxGC Typical Example
Source:
Shimadzu
>500 Peaks !
67 Peaks
45
paraffins & isoparaffins
Source:
Shimadzu
mononapthenes
n-C13
Light Crude Oil
Fraction
diaromatics
binapthenes
n-C12
monoaromatics
n-C11
n-C10
n-C9
n-C8
napthalene
methylenechloride
methylnapthalenes
sulfides
benzothiophenes
46
Source:
Shimadzu
Comprehensive qGCMS of Grapefruit extract
Expansion
1D – 45/50 min
2D – 1.5/3.5 sec
47
Source:
Shimadzu
Comprehensive qGCMS of Grapefruit extract
Expansion
1D – 45/50 min
2D – 1.5/3.5 sec
48
Zeit [s]
Coffee flavor
5
Cooperation with FH Lemgo
Source:
Shimadzu
Detection of >1000 Components
4
3
2
1
49
13
25
38
50
63
75
Zeit [min]
Peak Distribution in second dimension
Source:
Prof. Oliver Schmitz
SLB-5ms + ZB 50
SLB-5ms + SLB-IL 61
51
Comparison: Zebron ZB 50 or SLB-IL61 in second
dimension: Herba Menthae Haplocalyx
Better Separation
1-Phenylpropanone o-/m-/pMethylacetophenone
New Analytes
1-Phenylpropanone
o-/m-/pMethylacetophenone
Menthone isomer
Menthone
Isopulegol ?
α-Terpineol
Isopulegol ?
Citronellal
Source:
Prof. Oliver Schmitz
trans-/ cisIsopulegon
Borneol
Menthol ?
Menthone isomer
Menthone
α-Terpineol
Isopulegol ?
Menthol isomer
Menthol ?
Menthol ?
Octanoic
acid
Zebron ZB 50
Menthol isomer
Citronellal
Lavandulol
Menthol ?
SLB-IL61
Isopulegol ?
52
Source:
Shimadzu
Maximizing Efficiency
(x1,000,000)
TIC
7.0
Classical Gas Chromatography:
6.0
5.0
N = Number of Peaks
4.0
3.0
2.0
1.0
5.0
MDGC/Heartcut:
7.5
10.0
Transfer and separation of single peaks
onto a second column.
N = N1+ N2
(x1,000,000)
12.5
15.0
17.5
(x10,000,000)
1.0 TIC
TIC
7.0
0.9
0.8
6.0
0.7
5.0
0.6
4.0
0.5
3.0
0.4
2.0
0.3
0.2
1.0
0.1
5.0
7.5
10.0
12.5
15.0
17.5
6.0
6.5
7.0
7.5
8.0
8.5
9.0
9.5
10.0
10.5
Comprehensive GCxGC:
Continuous „Injection" –
all components run on both columns,
retention times on both columns are known
N = N1 x N2 x cos β (Orthogonalitätsfaktor)
53
Dziękuję za uwagę!
•Dr. Stephan Schröder (Shimadzu Deutschland GmbH)
•Prof. Oliver Schmitz (University Duisburg-Essen)
54