An Improved Method of

Updating Retention Times

for GCMS

Richard Whitney, Ph.D., GC/GCMS Senior Product Specialist, Ronald

D. Snelling, Ph.D., GC/GCMS Senior Product Specialist, Clifford M.
Taylor, GC/GCMS Product Manager, Shimadzu Scientific
Instruments, Columbia, MD, USA
Abstract
One of the common issues in gas chromatography-mass spectrometry
(GCMS) is updating of retention time windows when column
maintenance is performed. Updating 6 to 8 retention times manually is
usually not time-consuming, but many methods have 50 or more
target compounds. Manually updating that many retention times is
very time consuming and, if not done properly, target compounds will
not be determined accurately.

A new technique has been developed to update retention times

automatically in the data processing software. The technique is based
on the use of retention indices, referenced to a hydrocarbon standard
(series of n-alkanes). This is a multipoint correction method, so target
analytes will always be bracketed by the retention index marker
compounds to ensure accuracy of updated retention times for target
compounds.

Examples of this technique illustrate the accuracy of this technique

using a subset of US EPA Method 8270D analytes.

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Method Retention Time Updating
Method retention times (quantitation windows)
(x1,000,000)
1.5

1.0

0.5

10.9 11.0 11.1 11.2 11.3 11.4 11.5 11.6 11.7 11.8 11.9 12.0 12.1 12.2

When a capillary column is cut for maintenance, retention times shift.

(x1,000,000)
1.5

1.0

? ? ? ? ?
0.5

10.9 11.0 11.1 11.2 11.3 11.4 11.5 11.6 11.7 11.8 11.9 12.0 12.1 12.2

Manual updating of retention times can be

time-consuming, especially when the number
of target analytes is large (>50-100).
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Method Retention Time Updating

Two approaches have been employed to deal with

updating of retention times:
Adjust chromatographic conditions to keep retention
times constant after column maintenance.
Use retention time markers (retention indices) to
easily adjust retention times with changing
conditions (column length), keeping other
chromatographic conditions constant.
Based on fundamental chromatographic principles
Maintains optimum chromatographic performance
Easy, inexpensive, accurate
4
What is Retention Index ?
The retention index is information unique to a compound for a given
analytical column phase. It is the retention time of the compound
expressed on a scale based on n-alkane retention times. Kovats
proposed it in 1958.* It is based on a thermodynamic principle and it is
correlated to chemical structure.
*Kovats, E Helv. Chim Acta 41: 1915-32 (1958)

10.253min 14.642min 9.733min 14.040min

Decane Undecane Decane Undecane
(C10) (C11) (C10) (C11)

11.551min 11.017min
Limonene Limonene
Retention Index 1030 Retention Index 1030

Retention index
5
Instrument A
is constant. Instrument B
 Linear Retention Index
LRI (T) = 100n + 100 (RTT - RTCn) / (RTCn+1 - RTCn)
(Temperature Program Condition) Carbon number vs RT

RTCn, RTCn+1 - retention times 2500

of Cn and Cn+1 alkanes 2000

Carbon number x 100

1500

1000
uV(x100,000)
5.5
Chromatogram 500
5.0

4.5 C10 T C12 0

4.0
1 1.5 2 2.5 3 3.5 4 4.5
3.5
RT
3.0

2.5

2.0

1.5 Note: under linear temperature

programming conditions, n-alkanes
1.0

0.5

show a linear relationship between

0.0

1.50 1.75 2.00 2.25 2.50 2.75 min

retention time and carbon number

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 Principle of Retention Index (1)
Method C17 C18 C19
information

1700 1735 1800 1900 R.I.

20 21 22 23 24

Target compound n-alkanes

21.5 min (Method value)

Determining retention indices (done only once for each method)

A series of n-alkane standards are run under the standard
method conditions used for target compounds.
From the method retention times for the target compounds
and the n-alkanes, retention indices are calculated.
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 Principle of Retention Index (2)
Retention indices are used to accurately predict method retention
times for new conditions (done only once for new conditions).
n-alkane standards are run under the revised conditions.
New method retention times are calculated for target compounds
After
column C17 C18 C19
trimming

1735 R.I.
1700 1800 1900
20 21 22 23 24 Retention
time
21.1 min n-alkanes
8
Calculated value
 Principle of Retention Index (3)
Original C17 C18 C19
method
information

1700 1735 1800 1900 R.I.

21.5 min (original method value) n-alkanes
After C19
C17 C18
column
trimming

1700 1735 1800 1900 R.I.

20 21 22 23 24 Retention
time
21.1 min new
9
method value
Advantages of using Retention Index
for Retention Time Updating
Automatic Adjustment of Retention Times (AART)
Retention times of up to 1000 compounds can be
updated simultaneously with one analysis of an n-
alkane
Reduced input by human errors
Easy operation
Less work, regardless of the total number of compounds
Multiple n-alkane standards
Correction is performed at multiple points from low to high retention
times, enabling accurate correction over a wide range of retention
times.
SIM time updating can be performed simultaneously
AART can easily be applied to any existing method
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AART Operation Step 1
Prepare GCMSsolution Method for Target
Compounds (including compound table)

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AART Operation Step 2
Analyze n-alkanes (AART) standard and prepare
AART compound table (for n-alkanes)
Same conditions used for
analysis of target analytes

C15
C30-33

12 2007 SHIMADZU
 AART Operation Step 3
Download retention indices to GCMSsolution method

Run Wizard (modify) Retention indices

under create are calculated for
compound table the target analytes

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 AART Operation Step 4
Run n-alkanes standard under revised conditions and
update retention times for target analytes
Execute AART Target compound
function to update retention times
target compound are updated
retention times

14 2007 SHIMADZU
 AART Results (1)
AART results:
Target compound retention times are accurately predicted for the revised
conditions (with 2-3min retention time change for late-eluters).
All target compounds properly identified; all isomer assignments correct,
indicating accurate updating of quantitation windows.

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 AART Results (2)

AART results:
After executing AART,
GCMSsolution quantitation
properly identified all 95
target analytes.
All isomer assignments
were correct in the
processed data (ie, benzo
(b&k)fluoranthenes).
Retention times predicted
very accurately across the
entire 25min GC run.

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 Library Searching with Retention Index (1)
Many compounds show similar spectra (isomers, homologs, etc.)

Spectra of compounds
with multiple isomers
show library search
results with multiple
hits difficult to
interpret the results (six
hits for this example)
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 Library Searching with Retention Index (2)
Use retention index as library search criterion
(Retention indices are included in many libraries)

Set Retention
Index scale
with data from
n-alkane
standard

Set Retention Index

allowance to limit
library search
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Library Searching with Retention Index (3)
Using retention indices can qualify the library search results

Library search result: Library search result:

Benzo(a)anthracene Chrysene

Note: the library must include accurate, isomer-specific retention

19 indices, corresponding to the column phase used for analyses
AART Summary
AART function is unique to Shimadzu
GCMSsolution
Based on fundamental science retention index
Maintains optimum chromatographic conditions
Easy, fast, inexpensive
Can be applied to any GCMS method with up to
1000 analytes
Extremely accurate - based
on use of multi-point retention
index standards
Retention index is useful in
20 library searching 2007 SHIMADZU