Developments in
2D GC with Heartcutting
Kevin Mac Namara,a Riccardo Leardib and Andreas Hoffmannc
aIrish Distillers Ltd, Midleton, Co. Cork, Ireland,
bDipartimento di Chimica e Tecnologie Farmaceutiche e Alimentari, Genova, Italy,
cGerstel Gmbh & Co. KG, Mülheim an der Ruhr, Germany.
Introduction
Modern capillary gas chromatography is a mature technique
offering high separation efficiency complemented by state-of-
the-art hardware and detectors. However, real-world samples
(e.g., natural flavour extracts and concentrates, gasoline streams
and environmental samples) are often too complex for
sufficient resolution on any one chromatographic phase. In
natural products, such as alcoholic beverages, the problems are
exacerbated in that compounds present at trace amounts can
often be the species contributing most to the aroma and taste
because of their much lower natural sensory thresholds.
Resulting coelution with larger amounts of matrix compounds
of much less sensory interest pose real problems for detection
and quantification. A modern capillary column contains in
excess of 100 000 plates, but Giddings calculates that 500
million theoretical plates would be required for 0.99 separation
probability of a 100 component mixture.1 Viewed in a different
way Smits estimates the total number of peaks that can be
separated on a standard capillary column is only 0.1% of all
known volatile compounds.2 Use of selective element-specific
detectors can help but the problem of obtaining a clean mass
spectrum for positive compound identification remains.
Therefore, the idea of using the consecutive separation
power of two different chromatographic phases has always been
an attractive proposition for such difficult problems. Resolution
in chromatography is described by
N
R 
4 ( 1
 (( k
1k ( [1]
where N is the number of theoretical plates,  is the
separation factor of a given solute pair describing the selectivity
and k is the capacity factor of the more retained solute. As R
depends on the square root of N, a substantial increase in
resolution can only be achieved by a very large increase in
column length with a corresponding increase in retention time.
14 RECENT APPLICATIONS IN MULTIDIMENSIONAL CHROMATOGRAPHY December 2003
A more effective approach is to increase the selectivity factor ,
as a small increase in this parameter will give a major increase in
resolution. This can be achieved through multidimensional GC
on dissimilar phases.
A multidimensional separation means that the sample is
dispersed in different time dimensions.3 The concept of peak
capacity, or the maximum number of compounds in a mixture
which a chromatographic system can resolve, can be applied
here. When a sample is separated using two dissimilar columns
the total peak capacity will be the product of the individual
column peak capacities. This represents a huge gain in
separation space. For maximum resolution gain separation in
the individual time dimensions must be totally uncorrelated or
orthogonal. This is rarely achieved in practice as
chromatographic phases have complex mixed separation modes
and so will have at least some similarities. The resolution gain
will still be very significant even without total orthogonality.
The difference between two-dimensional (2D) GC with
heartcutting (GC–GC) and comprehensive 2D GC (GCGC)
is now well defined.3,4 In the classic heartcut approach a
fraction from the first retention axis is transferred for separation
on the second retention axis. By contrast, in the comprehensive
approach the entire sample, after a single introduction to the
first column, is subjected to the two different separations. This
review is only concerned with modern heartcutting 2D GC.
A Short History of GC–GC
The earliest work in 2D GC employed valve systems to connect
the two columns and for fractionation of the first column
eluent. The valves were metal multiport units and suffered
from catalytic activity on the metal surface. In addition,
unswept volumes and cold trapping effects reduced efficiency
and introduced undesirable band broadening.
In 1968 Deans introduced a valveless switching system or
stream switching using pneumatic pressure balancing.5 The
principle of operation has been extensively reviewed by

Bertsch.6 By using controlled carrier gas flows to the injection
port and the column intersection point, fractions could be
diverted to the second column or vented to waste. This design
obviated the disadvantages of a valve connection system but
introduced the challenge of stabilizing carrier-gas pressure in
the injector and at the midpoint connector.7 Several
commercial instruments were subsequently introduced to the
market based on the Deans principle. Some were add-on
pneumatic modules for existing GC units while others were
complete dedicated systems using double-oven configurations
for independent temperature control of the two different
columns. Gordon and colleagues reviewed several of these
systems in 1986 for both activity and ability to analyse complex
samples.8 All the systems functioned adequately for
heartcutting from complex samples, but the operations
required were still rather complex and manual, and as such
these instruments never integrated fully into the routine
general chromatographic community. Practically all of these
systems have ceased to be available commercially. The problem
principally centred on the considerable technical difficulty of
achieving the necessary pressure equilibration for reproducibility
of cutting and retention-time stability.
This basic problem can be solved by use of modern
electronic mass flow controllers and electronic proportional
backpressure regulators with pressure sensing. The following
sections of this review will attempt to describe such a unit
incorporated into Agilent GC and GC–MS (Agilent
Technologies, Waldbronn, Germany) equipment to produce a
system that allows rapid access on demand to 2D capability.
The only intrusive addition to the first GC is a single miniature
device at the intersection of the two columns that can remain
in situ in this GC whether or not the system is in 2D mode.
Modern electronic pneumatic control (EPC) technology is
used for the necessary heartcutting pressure equilibrations.
When a programmed temperature vaporization injector (PTV)
is used for sample introduction powerful techniques such as
stir-bar sorptive extraction (Twister) and large-volume injection
with solvent venting can be used to productive effect. Fully
automated sequences can be programmed for repetitive
injection to the first column, with single or multiple
Figure 1: Schematic of the GC–GC system. 1  injector for
either oven; 2  PTV inlet on instrument 1; 3  PTV inlet on
instrument 2 or 3; 4  monitor FID on instrument 1;
5  sulphur chemiluminescent detector; 6  nitrogen
chemiluminescent detector; 7  pneumatics unit for column
switching device.
1 the precolumn.
6 5 7 2 4
3 Table 1: Description of the instruments used to establish the
Chemstation MSD Instrument 2 or 3 Coolant Instrument 1
www.lcgceurope.com
Mac Namara, Leardi and Hoffmann
Modern electronic pneumatic control (EPC)
technology is used for the necessary
heartcutting pressure equilibrations.
consecutive cuts to the second column. With software control
of all aspects of the 2D analysis the entire process becomes as
straightforward as a routine sequence to a single column.
Modern GC–GC
Build-up of a 2D system: The system described in this section
has been in use in our laboratory for some years. Initial work
on flavour compounds in distilled spirits revealed the obvious
limitations of single compound capillary GC for identification
of important trace nitrogen and sulphur. Interesting patterns
were available from selective detectors but mass spectrometric
identification was very difficult because of matrix interference
and relative detector sensitivity. In a first application using 2D
heartcutting we were able to identify several new medium
boiling sulphur compounds in whiskey.9 Rather than say we
logically pre-planned the design of our particular GC–GC
system it is more realistic to say that our current system has
evolved to its present format from a learning process based on
practical applications and taking advantage of continuing
advances in GC design and operation.
Figure 1 is a schematic of a system offering total flexibility
for both routine GC and GC–GC. The main oven with MSD
and selective detectors already existed and the monitor GC was
added and installed beside the GC–MS. These two systems are
then connected by the cryotrap system (CTS). The column-
switching device (CSD) is installed in the first oven and linked
to the external pneumatics box. For optimum instrumental
flexibility the entire configuration is then run as three separate
systems from the Agilent Chemstation. These are described in
Table 1.
The Master software (Gerstel GmbH, Mülheim an der Ruhr,
Germany) for sample introduction, cutting and transfer
operations, cryotrapping and remote start of monitor GC and
GC–MS is also run from the Chemstation.
When GC–GC is required the main GC column is
disconnected from its injector and threaded through the
cryotrap for connection to the exit of the column-switching
device. The outlet of the precolumn is similarly connected to
the inlet of the column-switching device. Additional permanent
inlet and outlet lines connect the column-switching device to
the mass flow controller and pressure regulator with sensor
in the pneumatics box. This will be described in more detail
later. A final deactivated 0.05 mm i.d. fused-silica line takes a
small split from the column switching device to a flame
ionization detector (FID) for monitoring the eluent from
different two dimensional configurations.
Instrument 1 Agilent 6890 GC with FID and Gerstel PTV injector.
Instrument 2 Agilent 6890 GC with 5973 mass selective
detector and Gerstel PTV injector.
Instrument 3 Agilent 6890 GC with chemiluminescent nitrogen
and sulphur detectors (Sievers Ionics Inc.) and
Gerstel PTV injector.
15
 Mac Namara, Leardi and Hoffmann

The advantages from this instrumental design are: the pressure at the column switching device will be the head
• The entire configuration does not constitute a dedicated pressure on the main column after the selected compounds
GC–GC system. Each instrument is self-contained and can be are transferred. In addition, the commencement of ballistic
run entirely separately for specific applications when heartcutting heating of the cryotrap is the start signal for the chosen
operations are not required. For heartcutting the Master analysis on the main column. Therefore, this combination of
software links all individual modules in an orderly sequence. “column head pressure” (pressure at the column switching
• The only connections required are those of the pre- and device) and “injection” (heating of the cold trap) can be used
main columns to the column switching device. The to establish a retention-time locked analysis for heartcuts on the
connections are made with Graphpack technology ensuring a main column. We applied this procedure to organophosphorus
leak-free coupling. (The most recent commercial addition to pesticides in a complex lemon oil and the modus operandi will
the GC–GC market is an add-on Deans unit but this requires be explained in detail in the applications section.
connection to three micro-volume tees). • Total automation is possible with all aspects of the 2D
• The cryotrap can also be ballistically heated after trapping analysis. The selected cut time windows together with the
selected compounds and therefore acts as a proxy injector for relevant cryotrap cool and heat times are programmed to the
the transfer of selected compounds to the main column. Master software. A sequence with data files is established for
• The EPC principle used for cutting and venting means that each of the chosen instrument pairs and the injection
sequence starts all other sequences.
• Sequences can be multiple injections of different samples
Figure 2: (a) Pneumatics and the column switching device in with similar cuts for each sample or multiple injections of the
heartcutting condition, countercurrent flow-off; same sample with different consecutive cuts transferred to
(b) Pneumatics and the column switching device in venting the main column. In this way a complex sample can be
condition, countercurrent flow-on. totally profiled by automated heartcutting. An application
example to a whiskey extract will be shown.
Pneumatics and the column-switching device: Figures 2(a) and
(a)
2(b) show the principle and the simplicity of the column-
PTV-inlet switching device. The external pneumatics unit contains the
(CIS 4) mass flow controller and proportional valve plus sensor, with
corresponding connections in and out of the column-switching
Precolumn device. Figure 2(a) can be defined as the zero condition. The
mass flow controller is off and the carrier gas flow to both
FID columns is supplied through the injector. A digital pressure
reading at the column-switching device is available at the
kPa Chemstation. This zero condition applies when a cut is to be
Countercurrent transferred to the main column. For venting of all unwanted
Countercurrent mL flow-in
flow-out eluent from the precolumn, a countercurrent flow greater than
Cooled TRAP
the column flows is established across the column switching
(CTS)
device by the mass flow controller. This condition is depicted in
Main column Figure 2(b) (for capillary operation this can be 10 mL/min).
The outlet proportional valve then acts as a programmable
restrictor and quickly re-establishes the zero condition pressure
at the column-switching device through the software.
This combination of mass flow control and programmable
restriction guarantees extremely rapid pressure re-equilibration
at the mid-point connector after transfer of cuts. The entire
(b)
operation is software driven and user friendly. After all compounds
PTV-inlet of interest have been transferred to the cold trap the switching
(CIS 4) pneumatics will be in the non-zero condition The mass flow
controller supplies both the countercurrent venting flow for
Precolumn precolumn eluent and the main carrier gas flow to the column
switching device. The Master software now allows a suitable
FID pressure to be established at the switching device for
subsequent application of retention-time locking technology.
kPa
Countercurrent Applications
Countercurrent mL flow-in Organophosphorus pesticides in lemon oil: Natural essential oils
flow-out Cooled TRAP represent a very complex matrix and offer very difficult
(CTS) challenges for detection of possible pesticide residues. Element-
Main column
specific detectors are useful for indicating possible positives but
spectral confirmation on any single column is severely
compromised by extensive matrix coelution.
The following discussion will show in detail the elaboration
(essentially software operations) of a 2D procedure for

16 RECENT APPLICATIONS IN MULTIDIMENSIONAL CHROMATOGRAPHY December 2003

 Mac Namara, Leardi and Hoffmann

identification of several organophosphorus (OP) pesticides in
lemon essential oil. Six compounds were cut after a 1 µL
splitless injection to a DB-17MS precolumn and then profiled
under retention-time locking GC–MS after transfer to a HP5-MS
main column. Detailed information on retention-time locking
concepts and techniques can be found in relevant Agilent
publications.10–12 The following are the initial operations
required to set up this procedure in GC–GC mode. The
sequence of operations is quite logical and constitutes a
general-purpose template, which is applicable whether single or
multiple cuts are desired.
• With the HP5-MS column installed in the main oven only
(Instrument 2, see Figure 1), run the five injections required
for retention-time locking set-up. In this instance, this
calculated a locked inlet pressure of 160.4 Kpa for elution of
chlorpyrifos methyl (locking compound) at 16.593 min in
constant pressure mode.
• Disconnect the HP5-MS from the inlet in the main oven.
Thread the column through the cryotrap and reconnect to
the exit of the column-switching device in the pre-oven. The
DB-17MS precolumn is installed from the pre-oven inlet to
the entry of the column switching. The lines to and from the
pneumatics box, together with the line to the monitor FID
are always in place and so the two columns are the only
connections required for full 2D operation.
Table 2 : Total parameter set for GC–GC–MS analysis of
pesticides in lemon oil.
A. Sample Lemon oil spiked with 1 and 10 ppm
(MS Scan) or 50 ppb (MS-Sim)
Organophosphorus pesticides mix A
(Sigma-Aldrich)
B. Injection 1 µL splitless to PTV injector (CIS 4 –
Gerstel GmbH) 60 °C, 10 °C/s to 300 °C
10 min. 1 min splitless time.
Carrier gas: helium.
• In theory, 160.4 Kpa will be the pressure required at the
column switching device when retention-time locked analysis
is required for compounds transferred to the main column.
However, this pressure should now be retuned as the
characteristics of the “ new injector ” (column switching
device and cold trap) will be slightly different. The compounds
are already focused in the “liner”, as represented by the first
length of the second column.
• Cap the injector in the main oven (Instrument 2) and
establish 160.4 Kpa as a blank head pressure in a GC–MS
method. This instrument is then ready for a retention-time
locked run but with injection from the column switching
device and the cold trap.
• Run the locking compound on the DB17-MS with transfer
to the HP5-MS. Relock in the software and read the new
headpressure required for the expected locking compound
time. For this column combination 140.7 Kpa was indicated
and this is now the required locking head pressure at the
column-switching device. When heartcuts are analysed on
the main column under these conditions they can be
Figure 3: (a) Scan TIC after injection of lemon oil with
10 mg/L OP pesticides to HP5-MS column only; (b) Scan TIC
after injection of lemon oil with 1 and 10 mg/L OP pesticides
to DB-17MS precolumn with three heartcut transfers to
HP5-MS column.
(a)
2.5107
2107
1.5107
1107
5106

C. Instrument 1 DB-17MS precolumn (J&W Scientific) (b) 10 15 20 25 30 35

Oven program, 60 °C, 2 min;
5 °C/min to 250 °C for 15 min; pressure 5106
program; 275 kpa, 2 min; 4106
2.7 kpa/min to 377 kpa for 15 min;
Abundance

detector: FID 3106

D. GC–GC Parameters Pneumatics – mass flow-controlled 2106 10 ppm

countercurrent flow @ 10 mL/min. 1 ppm
1106
Time (min) Cut Pressure (kpa)
Cut times – initial 0.00 on 140.7 5 10 15 20 25 30 35
31.80 off –
32.10 on 140.7 (c)
32.50 off – 3
2
32.80 on 140.7 3106
33.10 off –
final 33.40 on 140.7
2106 1 10 ppm
Cold Trap Initial temperature 260 °C 1 ppm
Initial time 30 min 1106
Rate 1 20 °C/min to 50 °C for 5 min
Rate 2 20 °C/min to 260 °C

E. Instrument 2 HP5-MS main column (Agilent 16 17 18 19

Technologies); oven program: 70 °C Time (min)
1.5 min, 25 °C/min to 150 °C, 3 °C/min
to 200 °C, 8 °C/min to 280 °C, 10 min;
Detector: 5973 MSD in Scan or Sim. Peaks: 1  methyl parathion, 2  fenchlorphos, 3  chlorpyrifos.

www.lcgceurope.com 17
 Mac Namara, Leardi and Hoffmann

automatically screened using the Agilent 567-compound
retention time locked pesticide mass spectral library.12
• A final advantageous step can now be incorporated to
compensate for pressure changes resulting from the
temperature program in the first oven. With the column
switching pneumatics in zero condition (mass flow controller
off) set the first oven at its starting temperature and adjust
the inlet head pressure until 140.7 Kpa is achieved at the
column switching device. Set the first oven to its final
programmed temperature and again adjust the inlet pressure
to the new higher value to achieve 140.7 Kpa at the
switching device. Using these initial and final head pressure
values include a pressure program in the GC method with a
rate, and initial and final times, to match the corresponding
oven temperature program. In this way the column-switching
device will always be at 140.7 Kpa during the precolumn
run and for chosen heartcuts at any point in this run. After
cuts the EPC pneumatics instantly re-establish 140.7 Kpa
and the end result is very precise retention time
reproducibility on the first column and absence of pressure
disturbance during heartcutting.
These parameters are summarized in Table 2, together with
Master software entries for injection, pneumatic options,
heartcutting and cold trap refocusing.
Figure 4: Mass spectra of (a) methyl parathion,
(b) fenchlorphos and (c) chlorpyrifos at 10 mg/L after
heartcutting from lemon oil.
(a)
109
1.2105
O O
125
1105
O S
8104
6104
4104 79 N
63 93
2104 47 O O
The heartcut entries are for the three compounds methyl
parathion, fenchlorphos and chlorpyrifos after their retention
times were established on the first column. The reproducible
narrow heartcut windows (0.3 min) are possible because of the
retention time stability on the precolumn.
Figure 3 compares the scan TIC traces for direct injection to
the HP5 column only (no heartcutting) of the lemon oil spiked
with 10 mg/L (ppm) of the OP mix, with a similar TIC trace
but after three heartcuts from the DB17 precolumn.
The 2D procedure was at both 10 and 1 mg/L spiking
levels. The analysis parameters are as outlined in Table 2. What
is immediately apparent is the degree of matrix removal with
heartcutting, which allows easy detection of the three
compounds in the MS. Both the direct analysis and the
heartcut analysis to the HP5 column were run under retention-
time locking conditions, but the direct analysis only reported
fenchlorphos when the data file was screened. At the 1 mg/L
level no compounds were found without initial heartcutting.
Figure 4 shows the mass spectra of these compounds from the
10 mg/L 2D analysis.
Lower detection limits using retention-time locking can be
achieved by setting up a GC/MS-SIM for the compounds of
interest.13 The same parameters as in Table 2 are used but now
the cutting is extended to six compounds — six individual 18 s
cuts from the precolumn. Figure 5 shows the main column
SIM trace after heartcutting for the lemon oil as is and the
lemon oil spiked with 50 µg/L (ppb) of the OP mix. The
lemon oil itself contains low µg/L amounts of disulphoton,
methyl parathion and chlorpyrifos.
Table 3 shows the retention-time locked screener report for
the six compounds in the 50 µg/L spike sample.
P Figure 5: (a) SIM trace for six pesticides after heartcutting
263
from lemon oil; (b) SIM trace for six pesticides spiked at
50 µg/L after heartcutting from lemon oil.
(a)
200 233

60 100 140 180 220 260 300 8000

(b) 285 2
6000
O O 5
P 4000 3
2105 O S
Abundance

Cl
2000
125
Cl
Abundance

1105 Cl
109 12 14 16 18 20 22 24
79 93
47 63 167 270 (b)

60 100 140 180 220 260 300

8000 2
(c) 3 5
197
97 Cl
O 6000 4
10105 O 6
P 1
8104 N O 4000
Cl S
Cl 314
6104 2000
258
4104 125
268 12 14 16 18 20 22 24
65 213
2104 47
169 244
349 Time (min)

60 100 140 180 220 260 300 340

m/z Peaks: 1  ethoprophos, 2  disulphoton, 3  methyl parathion,
4  fenchlorphos, 5  chlorpyrifos, 6  prothiofos.

18 RECENT APPLICATIONS IN MULTIDIMENSIONAL CHROMATOGRAPHY December 2003

 Mac Namara, Leardi and Hoffmann

Table 3 : GC–MS retention-time locked screener report for lemon oil spiked with six pesticides at 50 µg/L each after direct
injection to a DB-17MS precolumn and heartcutting to a HP5-MS main column.
Target Qualifiers
Compound Status ExpRT Delta m/z Resp. Out of Range XCR
86 Ethoprophos ✕ 10.742 0.058 158 28076 0.82
170 Disulphoton ? 14.551 0.045 88 53845 89,97,142 0.47
217 Methyl parathion ? 16.594 0.059 263 40648 109,125 0.37
236 Fenchlorphos ✕ 17.330 0.045 285 60596 0.95
278 Chlorpyrifos ? 19.234 0.007 197 92456 97 0.70
383 Prothiofos ? 23.752 0.003 309 26145 162,113 0.77

3-Alkyl-2-methoxypyrazines in wine: 3-isopropyl-2 but at the expense of considerable individual manipulation. In

methoxypyrazine (IPMP) and 3-isobutyl-2-methoxypyrazine
(IBMP) are known to contribute to the bell pepper character of
Sauvignon Blanc wines.14,15 These compounds have olfactory
detection thresholds at the ng/L (ppt) level and various
approaches have been employed for their detection and
quantification. Harris and colleagues used both distillative and
dynamic sparging enrichment followed by trapping on an ion-
exchange resin to isolate the pyrazine bases from wine. This
was followed by selected ion monitoring on two different
capillary columns and using mass spectrometry in both electron
ionization and positive chemical ionization modes.15 The
pyrazines are volatile compounds with relatively simple mass
spectra (Figure 6) and a major concern must be the possibility
of matrix interference in any single dimensional separation.
Another approach used a similar distillative process for ethanol
removal and compound enrichment followed by a four-hour
headspace solid-phase microextraction (SPME) procedure with
injection to a nitrogen phosphorus detector.16 In both of these
applications single figure ng/L detection limits were achieved
Figure 6: Mass spectra of alkyl methoxypyrazines: (a) IPMP;
(b) IBMP.
(a)
8105
6105 O
N
N 1
4105
the following application a combination of stir-bar sorptive
extraction (SBSE) followed by GC–GC with either selected ion
monitoring or specific nitrogen chemiluminescence detection
allows detection of the pyrazines at the required trace level.
SBSE enriches the compounds directly from the wine and GC–GC
removes interfering matrix before analysis on the main column.
SBSE (Twister, Gerstel GmbH) was introduced in 199717
and is similar to SPME in that analytes are partitioned between
the sample and a PDMS phase. In SBSE the amount of PDMS
used is approximately 100 times greater than in SPME, and
because analyte recovery for both techniques is a function of
the relative volumes of PDMS and sample (phase ratio-), it
follows that for equal sample volumes recovery and enrichment
in SBSE will be much greater. This recovery can be correlated
with published octanol/water partitioning coefficients, which
reflect the hydrophobicity of a compound. Therefore, use of
Figure 7: Second column SIM trace for methoxy pyrazines in
(a) wine and (b) 10 ng/L standard after stir-bar sorptive
extraction and heartcutting.
(a)
2
137 10000
8000
6000
4000
152 2000
Abundance

124
2105 9 10 11
68 95 105
Abundance

(b)
40 60 80 100 120 140
(b) 10000
124

1.6106 8000
1.4106 6000
1.2106
1106 O 4000 2
N
8105 1
N 2000
6105
94 151
4105 81 9 10 11
2105 166
Time (min)
40 60 80 100 120 140 160
m/z Peaks: 1  IPMP, 2  IBMP.

www.lcgceurope.com 19
 Mac Namara, Leardi and Hoffmann

these values can predict the extraction efficiency for a compound
from an aqueous matrix to a PDMS-coated device. For the
pyrazines this was calculated as 60% recovery from a 10 cc
sample for a standard 10 mm Twister, making them suitable
candidates for this technique.
For this analysis, the same strategy in terms of pressure and
flow programming was used as for the pesticide application,
except in this instance a more polar column (Supelcowax-10,
Supelco Inc. Bellefonte, Pennsylvania, USA) could be used as
precolumn to maximize the selectivity difference for the
compounds of interest. This integration of SBSE and GC–GC
now means that that the sample preparation step is the
combination of the Twister enrichment and the first dimension
separation. In SBSE the glass-lined and PDMS-coated
magnetic stir bar is simply dropped into 10 cc of sample and
stirred for 1 h. 50 to 100 samples can be treated simultaneously
and all stir bars can then be analysed in a sequence using a
modified PAL autosampler (Gerstel GmbH).
For each analysis all aspects of the subsequent GC–GC
separation will also be sequence automated, thus the entire
procedure is a good example of the applicability of modern
system development for complex routine automated analysis.
Use of intermediate cold trapping for cuts transferred to the
main column allows the use of standard capillary columns.
Cold trapping may not be necessary in all instances, but if it is
needed and not available then the analyst must rely on phase-ratio
refocusing (thick film columns) or use of PLOT columns.
These introduce other complications such as much longer
elution times for semivolatile compounds and are best used
only if a dedicated analysis with one cut is required. With cold
trapping available all eventualities are catered for and maximum
flexibility is attained.
Figure 7 shows the second column SIM trace for a wine and
a 10 ng/L standard of the pyrazines after SBSE and subsequent
transfer of two cuts from the polar precolumn to the HP5-MS
main column. This analysis used the combination of
Instrument 1 and Instrument 2 (Table 1).
Figure 8 shows the same analysis for the wine only but now
using the combination of Instrument 1 and Instrument 3,
Figure 9: (a) Monitor FID trace after injection of flavour
extract to precolumn. Cut region at 16–18 min for transfer to
second column indicated; (b,c,d) Corresponding MS, N & S
traces for heartcut run on second column.
(a)
cut
8105

Figure 8: Second column nitrogen trace for methoxy pyrazines 6105 Monitor FID
in wine after stir-bar sorptive extraction and heartcutting. 4105

2 2105
Abundance

1.5106 5 10 15 20 25 30 35
(b)
Response

8106
6106 MS trace
1106
4106
1
2106

9 10 11 10 20
Time (min) (c)
1.4107
Peak identification: 1  IPMP, 2  IBMP. 1.2107
1107 Nitrogen trace
8106
6106
Table 4 : Description of the different sequences needed for 4106
total profiling of a sample by GC–GC. 2106
Response

Gerstel Master: Establish a sequence with the selected

injection mode and specify cold trap and 10 20
(d)
cut times for each repetitive injection to the
precolumn.
2.5107
Instrument 1: Establish a sequence with as many data files
(Precolumn) as injections. Monitor FID traces will all 2107
Sulphur trace
look similar. The injection process will start 1.5107
this instrument.
1107
Instrument 2 or 3: Establish a sequence with as many data files
(Main column) as heartcuts. Each MS and selective 5106
detector trace will be unique for each cut.
Heating of the cold trap (“re-injection”) will 10 20
start this instrument. Time (min)

20 RECENT APPLICATIONS IN MULTIDIMENSIONAL CHROMATOGRAPHY December 2003

 Mac Namara, Leardi and Hoffmann

i.e., the main column exits to the nitrogen detector. Both

Figure 10: See text for details. traces agree and indicate about 10 ng/L of IPMP and 50 ng/L
of IBMP in the wine.
For any number of specific compounds of interest a custom
6106 screener library can be established for cuts transferred to the
main column. This can be useful when multiple compounds in
5106 multiple samples must be automatically screened.
Abundance

Total profiling of a complex extract: In this example the same

4106
column pair as for the methoxy pyrazines was used. However,
3106 the strategy now will be to totally deconvolute a complex
flavour extract by repetitive injection to the precolumn with
2106
different consecutive cuts being transferred to the main
1106 column. This total profiling was applied recently to garlic
flavour volatiles using headspace solid-phase microextraction
8 9 10 11 12
(HS-SPME) and the newer technique of GCGC. Time-of-
flight mass spectrometry is mandatory for spectral data using
Time (min)
this approach because of the speed of analysis in the second
column.18 Using the automated heartcutting approach the
sample is first heartcut to simultaneous nitrogen and sulphur
detectors (Instruments 1 and 3), and then repeated to the
Figure 11: See text for details. MSD (Instrument I and 2). Each sequence of 14 injections
with heartcuts was run overnight and at completion 3  14
sets of data files (MS, N & S) were assembled for processing.
3107
Table 4 describes the sequence set-up for automated running.
2.5107 (a) Figure 9 shows the monitor trace and the related
2107 combination of MS, S & N traces on the main column for the
1.5107 cut 16–18 min from the precolumn. Extensive detail is now
1107 MS traces available from these correlated cut traces and Figure 10 shows
510 6 an overlay of the MS and nitrogen traces for the time segment
0 7 to 13 min. The entire data set for all cuts for each detector
26 can also be assembled and portrayed in 3D format; that is, cut
22
18
30
time from the precolumn, retention time on the main column
14 25
Precolumn 10 20
cut time (min) 6 15
10
5 Analytical column
retention time (min) Figure 12: See text for details.

1.2108
(a)
1108 (b)
8107 8106
Abundance

6107
6106
4107 Sulphur traces
2107 4106
0
2106
26
22 15
18
Precolumn
14 10 7.0 8.0
10 5
cut time (min) 6 Time (min)
0
Analytical column
retention time (min) (b)
2.2107

(c) 100
2107

1.5107

1107 Nitrogen traces 80

m/z

5106

0
60
12
10 10
8 8
6 6
4 4
Precolumn 2 2 40
cut time (min) 0
Analytical column 200 400
retention time (min)
Scan number

www.lcgceurope.com 21
 Mac Namara, Leardi and Hoffmann

and the corresponding peak areas. Figure 11 shows this format

for the three detector responses. An overview of mass peaks in
the MS heartcut traces can also be represented as a contour
plot. Figure 12 shows this format for a time segment of the MS
trace in Figure 9. All 3D and contour plots were constructed
using Matlab Version 6 (The MathWorks Inc., Natick,
Massachusetts, USA).

Conclusions
Automated 2D gas chromatography with heartcutting is
available as a powerful routine tool for complex sample analysis.
Most of the modules required to construct a system are
probably available in candidate laboratories. Modern
instrument control networking ensures that the system
components are always available for individual one-dimensional
analysis. In fact, successful GC–GC application is simply a
sequence extension of the individual operations.
The all-important pressure control and manipulation at the
column intersection is achieved with modern EPC pneumatics.
Integration of cryofocusing into the system allows maximum
flexibility and will cater for the entire volatility range of
compounds. The same cryofocusing unit can be used for sample
introduction with stir-bar sorptive extraction or large-volume
injection with solvent venting. Combination of these low
sample preparation techniques and the automated features of
modern GC–GC allow the build-up of very powerful systems
capable of processing large numbers of samples. A further
developmental stage is the use of short 0.1 mm i.d. capillary
columns for lower total analysis time.

References
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13. H. Prest and D. Peterson, Agilent Technologies, Applications Brief
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Kevin Mac Namara is Head of Technical Development at Irish

Distillers Ltd, a major subsidiary of Groupe Pernod-Ricard.
Riccardo Leardi is a Research Chemometrician at the University of
Genoa. He is a member of the Editorial Board of the Journal of
Chemometrics and is also active in chemometric consultancy.
Andreas Hoffmann is Applications Manager at Gerstel GmbH.

22 RECENT APPLICATIONS IN MULTIDIMENSIONAL CHROMATOGRAPHY December 2003