Running head: LAB REPORT ON DETERMINING THE CONCENTRATION BY

1
USING GC-MS

Lab Report On Determine the Concentration by Using GC-MS

Firstname Lastname

Name of Institution
LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 2

Abstract
Gas chromatography mass spectrometry is a method used to separate volatile mixtures, which
have low molecular mass found in environmental material. GC-MS analysis requires a thermally
stable molecule to avoid effects of adsorption. The trial is added into the Gas Chromatograph
(GC) for vaporization. The sample undergoes stationary and mobile phases with the columns
where it is converted into ions. The GC-MS column types ranges from ionic and nonionic, polar
and non-polar. The specific types of GC-MS column type are Polar GC columns, Mid-Polar GC
Columns, Low-Polar GC Columns, Non-Polar GC Columns, Ultrafast GC Columns, and PLOT
Columns

The ion production methods include electron ionization and chemical ionization. EI involves
results into loss of an electron while the alternative CI is preferred for its less energy requirement
during ionization but its first step is ionization of methane. The subsequent section is the filter
mostly referred to as a mass analyzer that filters out positive charges. The filters are varieties
such as ion traps, radio frequency, quadrupoles, and many others. After ion detection, they move
to detector where signal is improved. The data is received as visual after conversion from
electrical impulses. The values are displayed as ion peaks.

Liquid chromatography–mass spectrometry (LC–MS) used in determination of concentration has

more benefits than gas chromatography–mass spectrometry (GC–MS). It can be pointed out that
LC–MS can identify and give measurement of variable compounds and requires less time for
preparation offers the advantage of identifying and measuring a broad range of compounds
needing comparatively minimal preparation. Having known concentration by using GC analysis,
the area of peak is determined by integration followed by calculation of response factor for
individual compound.

LC–MS technology is therefore a suitable alternative confirmation technology to test for

concentration. (Roman, Knapp, Horn, Stillman, & Evans, 2016)

Introduction
This lab report entail the activities that were undertaken to determine the concentration of
standard solutions using GC-MS after which comparison of their corresponding ionization
difference is determined using the data obtained using LC-MS.
LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 3

Calculating concentration requires comparing the peak area of the analyte in the sample with a
known concentration based on its peak area of standard. If internal standard is used, then the
analyte’s ratio to the internal standard should be used. The assumption is that one need to use at
least one standard point of known concentration. The setting of spectrometer will determine how
you find the correct area of the correct peak. You need to filter out the mass charge ratios are
filtered in case of too many peaks.
Having pure internal standard, one will only get a single peak readily in the chromatogram.
Using EI souse may not give you molecular ion peak. Next, go on the selected ion chromatogram
peak option where you need to enter the m/z that corresponds to your IS. In the option of peak
labeling, particular peak area is asked. Concentration is computed using the peak area ratio and
then linearity curve plotted.

Objective of the experiment

The objective of the experiment was to determine the concentration by using GC-MS then
comparing the ionization difference with the data obtained using LC-MS

Experimental
The apparatus used to perform the experiment included volumetric flask, pipette, Machines i.e
GC machine and stopwatch
The Chemicals and Reagents
Standard solution:
Methyl heptanoate C8H16O2
Methyl octanoate C9H18O2
Methyl nonanoate C10H20O2 ------------- internal standard
Methyl decanoate C11H22O2
Solvent: Chloroform
FAMEs was made: 1x10-5 mg/microliter
Chemical Solutions Preparation:
Sample:
Stock solution 10mg/ml
LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 4

2ml volumetric flask was used to transfer solution to a 20 microliter. CHCl3 solvent was added
to the solution up to the mark. After preparation of the solution, using GC-MS vials, 50
microliter of the sample was pipetted together with 450 microliter of CHCl3.
Unknown solution:
GC-MS vials and 2ml volumetric flask was used in pipetting 20 microliter of internal standard
C10H20O2 solution, give the instructor to get unknown, and diluting to the mark with solvent
CHCl3. using a GC-MS vials, pipetting 50 microliter of the sample and continued to pipette 450
microliter of CHCl3.

Results and Discussion

Lab Results
Standard solution data obtained by GC machine
Solution Retention time (min) Area (mm)
C9H18O2 11.346 3628.352
C10H20O2 13.319 2546.082
C11H22O2 15.139 3200.583
C13H26O2 18.447 6067.360

Unknown solution data obtained by GC machine

Solution Retention time (min) Area (mm)
C9H18O2 11.346 25352.934
C11H22O2 15.133 23555.361
C13H26O2 18.441 41814.323

From the results obtained from the GC machine, it can be simply pointed out that retention time
and area increases with increase in the size of methyl molecules whereby C13H26O2 has the
highest time and area of 18.447 minutes and 6067.360 mm2 respectively, While C9H18O2 has the
lowest time and area as it can be seen on the table. This can be attributed by the fact that methyl
nonanoate has simple molecular structure and hence requires shorter time to be ionized. The
unknown solutions also behaves that way with C9H18O2 having lower retention time and area.
There was no data for C10H20O2 for unknown solution.
LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 5

Graphical output for Standard Solution

Graphical Output for Unknown solution

LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 6

There was a similar trend between the graph for standard solution and that for unknown solution
still there was no output for C10H20O2 for unknown solution.

Calculation:
Standard solution response factor calculation:
Response factor F = Ax /As
Ax: Area of analyte
As: Area of Internal Standard
Solution F
C9H18O2 1.1337
C10H20O2 0.79551
C11H22O2 1
C13H26O2 1.89570

Computing concentrations of unknown solution

LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 7

Convert Internal Standard C11H22O2 solution 1x10-5mg/uL

𝑚𝑔 1𝜇𝐿 1𝑔 1𝑚𝑜𝑙 𝑚𝑜𝑙

1𝑥10−5 ∗ ∗ ∗ = 5.3678 𝑥10 −5
𝜇𝐿 1𝑥10−6 𝐿 1000𝑚𝑔 186.295𝑔 𝐿
The formula used to compute unknown concentration is :
𝐀 𝐱 ∗ 𝐂𝐨𝐧𝐜𝐞𝐧𝐭𝐫𝐚𝐭𝐢𝐨𝐧 𝐨𝐟 𝐈𝐒
𝐔𝐧𝐤𝐧𝐨𝐰𝐧 𝐜𝐨𝐧𝐜𝐞𝐧𝐭𝐫𝐚𝐭𝐢𝐨𝐧 =
𝐀 𝐬 ∗ 𝐑𝐞𝐬𝐩𝐨𝐧𝐬𝐞 𝐟𝐚𝐜𝐭𝐨𝐫 𝐅
Ax: Area of analyte
As : Area of Internal Standard
Applying the formula to the rest of the analyte, we found the following values of the
concentration
Solution Concentration (M)
C9H18O2 5.0961 x10-5
C11H22O2 5.3678x10-5
C13H26O2 5.0265x10-5
The results for the concentrations shows that the higher the response factor F and area of internal
standard, As the lower the concentration and vice versa for other analyte .

Method Improvement Discussion

Integration was performed by use of agilent systems which integrated function located on the top
of the tool bar under the chromatogram tab. Through pressing the button of integrate the software
automatically lead to integrating of the all the peaks in chromatogram above a set area. The
minimum peak area can changed by typing integration etc. from that same tab. Clicking on
integration results brings results in the form of a table that can be exported to excel through
various ways such as copy pasting. (Lian & Yon, 2013). Mass spectra charges the molecules of
the specimen electrically, accelerates the molecules and breaks it using magnetic field strength.
Breaking molecules by magnetic fields produces specific fragments for GC-MS and HPLC-MS.
It was found that the number needed is 3-5 replicate injections each at five concentrations. One
should be near and another below the limit of quantification. This must be performed to
determine confidently the uncertainty. LOD qualifies as the lowest analyte concentration that can
be distinguished reliably from the LOB with feasible detection. To determine LOD, both the
LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 8

measured LOB test replicates of a known sample containing low concentration of analyte are
utilized.

LOD = LOB + 1.645(SD low concentration sample).

The lowest concentration where analyte cannot be detected reliably but where still some
predefined goals meant for bias imprecision are met is referred to as LOQ. Sometimes, LOQ
could be equivalent to LOD or its concentration can be much higher. Excel sheet can also be an
alternative way of performing the experiment. GC-MS software serves as an alternative

Conclusion
In conclusion, the experiment verified how simple and accurate the use of GC machines is in
determining the concentration of unknown solution from the standard ones. The Lab experiment
also confirmed that standard or calibration curve requires one to conduct it more often since the
response from the detectors may vary.
Lastly using GC-MS should make it easier for one to identify the internal standards from the
mass spectrum. The challenge of this may arise if the compound being used as internal standard
is equally present in your sample. The peak may be a combination of both the sample and
internal standard leading to inaccurate quantification thus becoming a problem.
LAB REPORT ON DETERMINING THE CONCENTRATION BY USING GC-MS 9

References
GAS CHROMATOGRAPHIC (GC) ASSAY OF FLAVOUR CHEMICALS. (2017, February 12).
Retrieved from fao.org: http://www.fao.org/docrep/x3860e/X3860E44.htm
Lian, Y. Y., & Yon, C. (2013). Gas chromatography-mass spectrometry. Gas
Chromatography/Mass Spectrometry-Based Metabonomics, 2-15.
Roman, E., Knapp, J., Horn, C., Stillman, S., & Evans, J. (2016). Comparison of LC–MS-MS
and GC–MS Analysis of Benzodiazepine Compounds Included in the Drug Demand
Reduction Urinalysis Program. Jounal of Analytical Toxicology, 1-20.
Armbruster, D. A., Tillman, M. D., & Hubbs, L. M. (1994). Limit of detection (LQD)/limit of
quantitation (LOQ): comparison of the empirical and the statistical methods exemplified with
GC-MS assays of abused drugs. Clinical chemistry, 40(7), 1233-1238.