Journal of Chromatography A, 1255 (2012) 38–55 · Tadeusz Górecki, Ahmed Mostafa, Matthew...
Transcript of Journal of Chromatography A, 1255 (2012) 38–55 · Tadeusz Górecki, Ahmed Mostafa, Matthew...
Tadeusz Górecki, Ahmed Mostafa, Matthew EdwardsDepartment of Chemistry, University of Waterloo (ON)
Journal of Chromatography A, 1255 (2012) 38Journal of Chromatography A, 1255 (2012) 38––5555
• Analysis of volatile and semi-volatile compounds• Max. theoretical peak capacity: ~22 peaks/min
◦ (15m x 0.1mm x 0.1µm column, 50cm/s, n = 150000, 15 min analysis)
Carrier GasColumn
OvenSample
ElectrometerData Recorder
InjectorDetector
3
Segment of a chromatogram of a sediment sample subjected to pyrolysis with TOF-MS detection (AIC)27 peaks visible
4
Overlay of selected masses (magnified 20 - 40 x)95 peaks found through spectral deconvolution.
nc ~ f x N 0.5 f < 2
tfirst tlast
Courtesy of Prof. Pat Sandra 5
6
Column n Peak capacity
25 m x 0.25 mm 100.000 316 – 632
50 m x 0.25 mm 200.000 447 - 894
100 m x 0.25 mm 400.000 632 - 1264
80 m x 0.1 mm 800.000 895 - 1790
Slide courtesy of Prof. Pat Sandra
T.A. Berger, Chromatographia 42 (1996) 63.
450 m x 0.25 mm x 0.25 µm PONA1,300,000 plates
Peak capacity over 1,000
100 min segments
T.A. Berger, Chromatographia 42 (1996) 63.
450 m x 0.25 mm PONA30 m x 0.25 mm PONA
9
… using the statistical theory of peak overlap …
… peak resolution is severely compromised when the number of components present in a sample overrates 1/3 of the peak capacity.J.M. Davis, J.C. Giddings, Anal. Chem. 55 (1983) 418
…in order to resolve 98% of the components, the peak capacity must exceed the number of components by a factor of 100.J.C. Giddings, J. Chromatogr. A 703 (1995) 3
100 analytes peak capacity should be 100 x 100 = 10.000, or N ca. 100.000.000 plates !!!
Slide courtesy of Prof. Pat Sandra
Solvent 1 front
Firs
t dim
ensi
on e
lutio
n
Solvent 1 front
10
Firs
t dim
ensi
on e
lutio
n
Solvent 1 front
First dimension elution11
Sol
vent
1 fr
ont
Sec
ond
dim
ensi
on e
lutio
n
Solvent 2 front
Separated!
Solvent 2 front
First dimension elution12
13
1st dimension retention time
2nd dimension chromatogram
14
1st dimension retention time
2nd dimension chromatogram 2nd
dimension chromatogram
15
1st dimension retention time
2nd dimension chromatograms
1D-GCHeartcut GCGCxGC16
• Interface traps effluent from 1st column and injects into 2nd
column• Times of injections are recorded• Second dimension is fast (0.5 - 10s)
InjectorCarrier Gas
PrimaryColumn
OvenSample
ElectrometerData Recorder
SecondaryColumn
Interface
Detector
1717
1D: 30 m x 0.25 – 0.32 mm x 0.25 – 1 µm, non-polar 2D: 0.5 – 2 m x 0.1 mm x 0.1 µm, polar Modulation period resulting in 2.5 to 3 cuts per peak
All parameters treated somewhat independently
Can it really be that simple?
Directly controllable
Green arrows: parameters whose values increase as the input parameter value increases
Red arrows: parameters whose values decrease as the input parameter value increases
1df 2df
2dc 1dc
1wh
Mass perModulation
Chance of overloading 1D
Chance of overloading 2D
1l 2l
Capacity of 2D
Capacity of 1D
2Δp 1Δp
1u 2u
2D retention
2Rs 1Rs
2wh
Allowed / Required 2D Space
Oven Programming Rate
Te
AnalysisTime
PM
1uopt
Inlet Pressure
1Δp
ΔpT
Thermal modulators◦ heater-based ◦ cooling-based
cryogeniccryogen-free
Flow modulators
Thermal modulators◦ Modulation period◦ Modulation temperature ◦ Stationary phase thicknessFlow modulators◦ Modulation period◦ Carrier gas flow rates
Model 1D separation1st dimension peak width 24s
PM = 6 s (4-5 cuts)
PM = 12 s (2-3 cuts)
0 0.5 1 1.5
5
10
15
20
Retention Time (min)
Sig
nal I
nten
sity
0 0.5 1 1.5
500
1000
1500
2000
Retention Time (min)
Sig
nal I
nten
sity
0 0.5 1 1.5
500
1000
1500
2000
Retention Time (min)S
igna
l Int
ensi
ty
L. Ramos, J. Sanz, in: D. Barcelo (Ed). Comprehensive Analytical Chemistry, Elsevier, Amsterdam, Netherlands, 2009, p. 283.
5 s 6 s 7 s
(1) indeno[1,2,3-cd]pyrene, (2) dibenzo[a,h]anthracene, (3) benzo[ghi]perylene
Liquid-cooled thermal modulatorLibardoni et al., Anal. Chem. 2005, 77, 2786-2794
24
Constant heating voltage
Programmed heating voltage
25
26
C40C16 C20
N-PAHs
120 min
5 s
1D and 2D columns connected through a specially modified segment of a coated stainless steel capillaryCapillary is compressed between two passive coolersCapacitive discharge resistively heats the trapping capillary
2828
2929
30
31
12.80 12.81 12.82 12.83 12.84 12.85 12.86 12.87 12.88 12.89 12.90
150000
200000
250000
300000
350000
400000
450000
500000
e
Signal: 12112204.D\FID1B.CH
25.20 25.21 25.22 25.23 25.24 25.25 25.26 25.27 25.28 25.29 25.30 25.31
120000
130000
140000
150000
160000
170000
180000
190000
200000
210000
220000
230000
240000
250000
260000
270000
280000
290000
300000
310000
e
Signal: 12112204.D\FID1B.CH
LMCSQuad-jet modulatorDelay-loop modulator
T
R
Longitudinally Modulated Cryogenic System (LMCS)
Kinghorn and Marriott (1998-2000)
3535
Temperature difference between the oven and the trap crucialOptimum peak width in both dimensions when ΔT =~70 °C ◦ Smaller ΔT trapping inefficient◦ Larger ΔT release inefficient (peak broadening in
both dimensions)
37
Built by LECO under license from Zoex Corporation
10 °C 20 °C
40 °C 80 °C
39
Carrier gas
40
Carrier gas
41
3.551300
3.5751300
3.61300
3.6251300
3.651300
3.6751300
3.71300
25000
75000
125000
175000
1st Time (s)2nd Time (s)
S2
2.65244
2.7244
2.75244
2.8244
2.85244
2.9244
10000
20000
30000
40000
50000
1st Time (s)2nd Time (s)
S2
100 Hz
60 ms
40 ms
42
1184
3.5184
2188
0.5192
3192
1.5196
0200
2.5200
1204
3.5204
10000
20000
30000
40000
50000
1st Time (s)2nd Time (s)
S2
43
44
1: 2,3-butanediol; 2: n-decane; 3: 1-octanol; 4: 2-ethylhexanoic acid; 5: nonanal; 6: n-undecane; 7: 2,6-dimethylphenol; 8: 2,6-dimethylaniline; 9: methyl decanoate; 10: dicyclohexylamine; 11:
methylundecanoate; 12: methyl decanoate
45
Constant flow Programmed flow
46
Adjustments to the modulation period usually require changes to the loop length or the carrier gas flow◦ When the loop is too short, the analyte band is not
refocused at the second cold spot (breakthrough) ◦ When the loop is too long, multiple injections from
the first cold spot could be present within the loop simultaneously (possible breakthrough, changes in 1D retention times)
47
Journal of Chromatography A, 1218 (2011) 4952– 4959
◦ Analysis of 45 FAME
http://www.chem.agilent.com/cag/prod/GC/2DGC_amj2_05_02_07D1a.pdf
48
Modulation period range limited by the fixed volume of the collecting loop◦ Too long modulation periods lead to loop
overfilling (breakthrough)Typical modulation periods ~2 s
◦ Larger loop volumes result in broader 2D peaksPossible artifacts when high concentration peaks elute
49
(A) Reverse fill/flush (RFF) modulator: flow path of fill cycle. (B) Reverse fill/flush (RFF) modulator: flow path of flush cycle.
Griffith et al., J. Chromatogr. A 1226 (2012) 116– 123
50
Griffith et al., J. Chromatogr. A 1226 (2012) 116– 12351
1D (length m × i.d. µm) 2D (length m ×
i.d. µm) Analyte/sample Reference
Poly(ethylene glycol) (21 m × 250 µm) 100% Polydimethylsilocxane (PDMS) (1 m ×
100 µm)A hydrocarbon mixture and a coal liquids sample [7]
(SolGel + poly(ethylene glycol)) composite phase (SolGel-WAX (30 m ×
250 µm)
5% Phenyl polysilphenylene siloxane (1 m × 100 µm) Roasted coffee bean volatiles [72]
Poly(ethylene glycol) (30 m × 250 µm) 5% Phenyl polysilphenylene siloxane (1 m ×
100 µm)Lipids and roasted coffee bean volatiles [73, 74]
Polyethylene glycol (TPA-treated) (30 m × 250 µm)
35% Phenyl-polysilphenylenesiloxane (1 m × 100 µm) Food analysis [75]
(5%-Phenyl)(1%-Vinyl)- methylpolysiloxane (2 m ×
100 µm)
14% Cyanopropylphenyl) methylpolysiloxane (0.5 m × 100 µm) Test mixtures [35]
100% PDMS (30 m × 250 µm) (SolGel + Poly(ethylene glycol)) composite
phase (SolGel-WAX (1.5 m × 250 µm)Volatile components of Pinotage wines [65]
100% PDMS (50 m × 530 µm) 50% Phenyl-polysilphenylene siloxane (2.2
m × 150 µm)Volatile organic compounds in urban air [76]
100% Cyclodextrin directly bonded to PDMS (10 m ×
100 µm)
(50% Liquid crystal / 50% dimethyl) siloxane column (1 m × 100 µm) PCBs in environmental samples [77]
100% PDMS (1 m × 100 µm) 14% Cyanopropylphenyl)
methylpolysiloxane (2 m × 100 µm) Essential oils [78]
Polyethylene glycol (60 m × 250 µm) (14%-Cyanopropyl-phenyl)-
methylpolysiloxane (3 m × 100 µm) Cigarette smoke condensates [79]
Poly(5%-phenyl–95%-methyl)siloxane phase (40 m ×
100 µm)
1,12-Di (tripropylphosphonium) dodecane bis (trifluoromethanesulfonyl) imide (3 m × 100 µm)
PCBs [80]
Poly(methyltrifluoropropyl siloxane) (30 m ×
250 µm)
Poly(dimethyldiphenylsiloxane) (5 m × 250 µm)
Trace biodiesel in petroleum- based fuel [81]
1D Length of BPX5 column*
Length of BP20 column*
A 20 0B 15 5C 10 10D 5 15E 0 20
D. Ryan, P. Morrison, P. Marriott, J. Chromatogr. A 1071 (2005) 47.
1D Length of BPX5 column*
Length of BP20 column*
A 20 0B 15 5C 10 10D 5 15E 0 20
D. Ryan, P. Morrison, P. Marriott, J. Chromatogr. A 1071 (2005) 47.
Typical 1D columns: 0.25 – 0.32 mm IDTypical 2D columns: 0.1 mm ID
This is not always optimal!
0.1 mm ID
0.25 mm ID
Mixture of pure compounds 1,000 x dilution
(15 m × 0.25 mm i.d) × (1.5 m × 0.1 mm i.d)
(30 m × 0.32 mm i.d) × (1.5 m × 0.18 mm i.d)
J. Beens, H. Janssen, M. Adahchour, U.A.T. Brinkman, J. Chromatogr. A 1086 (2005) 141.
Pin (KPa) 1N 2N 1ū (cm/s) 2ū (cm/s) PRopt
30 m × 0.32 mm + 1.5 m × 0.18 mm
56 86000 9000 16 64 4
88 100000 8000 24 106 5
132 98000 6500 35 140 6
15 m × 0.25 mm + 1.5 m × 0.10 mm
112 44000 15000 10 80 4
224 65000 10000 18 160 7
400 40000 4000 30 280 12
J. Beens in cooperation with www.chromedia.org, Comprehensive Two-Dimensional Gas Chromatography the State-of-Separation-Arts Theory. Part 2, 2010.
No split
35:65 split
P.Q. Tranchida, A. Casilli, P. Dugo, G. Dugo, L. Mondello, Anal. Chem. 79 (2007) 2266.
Primary – IsothermalSecondary - Isothermal
Primary – IsothermalSecondary – Temperature incremented
Primary – Temperature programmedSecondary - Isothermal
Primary – Temperature ProgrammedSecondary – Temperature Incremented
2D peaks very narrow – fast detectors requiredAt least 10 data points across a peak needed for quantitative determinations◦ Data acquisition rate of at least 50 Hz required Detectors must have low internal volume and short time constantsMost popular detectors: FID, TOF-MS
Detector Analyte/sample References
µECD
Pesticides in sediments [116-118]PCBs/OCs/CBz in soils, sediments and sludges [119, 120]Dioxins and dioxin-like PCBs in food and feed [121]Chiral toxaphenes typically found in real-life samples [122]Polybrominated diphenyl ethers [123]Polychlorinated dibenzo-p-dioxins, dibenzofurans and PCBs in food [124, 125]Chiral PCBs in food [125]PCBs in Baltic grey seals [126]Toxaphene [127]Polychlorinated dibenzo-p-dioxins, polychlorinated dibenzofurans and PCAs in a cod liver extract and a standard mixture [99]
NPD
Nitrogen-containing compounds in Brazilian heavy gas oil [128]
Volatile fraction of creosote-treated railway wood sleepers [129]Nanoparticles in roadside atmosphere [130]methoxypyrazines in coffee headspace [131]Fungicide residues in vegetable samples [132]Methoxypyrazines in wine [133]
SCD
Sulfur-containing compounds in straight run diesel oil [134]
Sulfur-containing compounds in heavy petroleum cuts [135]
Sulfur-containing compounds in middle distillates [136]Sulfur-containing compounds in crude oils [137]Sulfur compounds in diesel oils [138]Sulfur-containing compounds in diesel [139]
AED Sulfur-containing compounds in crude oil [140]MPDD Pyrolysis of gasoline (cracked naphtha) and pyrolysis of a polyethylene copolymer [141]
TOF-MS most popular owing to very high data acquisition rates (up to 500 Hz)Quadrupole MS slower, but can be used for qualitative work (quantitative determinations often possible with narrower mass range)HRTOF-MS very promising
Hopanes
Phenanthrenes
TIC
Sample and assistance provided by Jack Cochran and Michelle Misselwitz of Restek Corporation.Data provided by LECO Corporation using a prototype GCxGC-HRT system.
64
Hopanes
Phenanthrenes
Sample and assistance provided by Jack Cochran and Michelle Misselwitz of Restek Corporation.Data provided by LECO Corporation using a prototype GCxGC-HRT system.
65
Hopanes
Phenanthrenes
Sample and assistance provided by Jack Cochran and Michelle Misselwitz of Restek Corporation.Data provided by LECO Corporation using a prototype GCxGC-HRT system.
66
Hopanes
Phenanthrenes
Sample and assistance provided by Jack Cochran and Michelle Misselwitz of Restek Corporation.Data provided by LECO Corporation using a prototype GCxGC-HRT system.
67
HopanesFWHH 0.090 sec
191.179852.2 ppm error
191.179450.12 ppm error
191.17940-0.14 ppm error
191.179530.54 ppm error 191.18078
7.1 ppm error
68
FWHH 0.060 sec
191.085881.8 ppm error
191.085841.6 ppm error
191.085700.91 ppm error
191.08526-1.4 ppm error
Phenanthrenes
69
GCxGC approaches a mature status, but there is still room for improvementCareful optimization is required to reach the full potential of the techniqueGC×GC separation optimization is not as simple as in conventional 1D-GC because of the column coupling◦ any changes to the 1D column, flow rate, modulation,
oven temperature programming rate, etc., affect the separation in the 2D as well
70
71
O. Panic, C. McNeish, T.N. Oldridge, A. Mostafa
◦ NSERC◦ RESTEK◦ LECO◦ SGE◦ DoE◦ Polymicro Technologies
1df 2df
2dc 1dc
1wh
Mass perModulation
Chance of overloading 1D
Chance of overloading 2D
1l 2l
Capacity of 2D
Capacity of 1D
2Δp 1Δp
1u 2u
2D retention
2Rs 1Rs
2wh
Allowed / Required 2D Space
Oven Programming Rate
Te
AnalysisTime
PM
1uopt
Inlet Pressure
1Δp
ΔpT
72