CHM510 LABORATORY REPORT

EXPERIMENT 1
GAS CHROMATOGRAPHY (GC): METHOD DEVELOPMENT

NAME: NUR AFIQAH SYAURAH BT MOHD HIFNI

MATRIC NO: 2018235124
GROUP: AS2453M2
PARTNER’S NAME: I)NADA NATASHA BINTI ABU BAKAR
II) SITI NURHAMIMI BINTI AZIZAN
DATE OF EXPERIMENT: 23rd OCTOBER 2019
LECTURER’S NAME: PN. HALIZA BINTI KASSIM
1.0 ABSTRACT:
Gas chromatography is a sensitive, reproducible, accurate, quantitative and versatile tool well
adapted for the analysis of complex mixtures. The steps plays an important role in analysis of
drugs and pharmaceutical products. However the use of GC is limited to volatile thermally
stable compounds or the molecules or the molecules that may undergo derivation reactions to
thermally stable products. Method development and validation plays an important role in the
discovery, development and manufacture of pharmaceuticals. The objective of this experiment
is to obtain the optimal flow rate and optimal temperature in order to achieve optimum
condition.
2.0 METHODOLOGY:
Standard mixture 1 were injected isothermally starting at 210˚C at flow rate 70 for at
least two times reproducible results. Then, the experiment were proceed to the another two
flow rate (50 and 30) at the same temperature with another two reproducible results. After
that, the most suitable flow rate for that separation were obtained by calculating the resolution
(Rs) considered as optimal flow rate. Then, the most suitable temperature were obtained by
calculating the resolution (Rs) again considered as optimal temperature. Once the optimize
condition (optimal temperature, optimal flow rate) were obtained, the best resolution and the
best separation would be obtained.
3.0 RESULTS
a) Effect of carrier gas flow rate
Condition: 30 cm sec⁻¹ , 210˚C
Injection Peak Retention Base peak Resolution Average
time (min) width (min) (Rs) resolution
(Rs)
1 1 and 2 4.888 0.0482 34.92 42.75
7.067 0.0766
2 2.838 0.0319 50.58
4.889 0.0492

Condition: 50 cm sec⁻¹ , 210˚C

Injection Peak Retention Base peak Resolution Average
time (min) width (min) (Rs) resolution
(Rs)
1 1 and 2 1.698 0.0257 39.39 39.50
2.925 0.0366
2 1.698 0.0257 39.61
2.924 0.0362
Conditon: 70 cm sec⁻¹ , 210˚C

Injection Peak Retention Base peak Resolution Average

time (min) width (min) (Rs) resolution
(Rs)
1 1 and 2 2.104 0.0231 30.36 40.23
3.042 0.0387
2 1.220 0.0117 50.09
2.099 0.0234

b) Effect of variation of column temperature at flow rate on the resolution

Condition Injection Retention Peak width Resolution Average
time of peak of peak 1 (Rs) resolution
1 and 2 and 2 (min) (Rs)
(min)
70 cm sec ⁻¹ 1 2.830 0.0310 42.87 59.66
190˚C 4.787 0.0603
2 1.226 0.0116 76.44
2.835 0.0305
70 cm sec ⁻¹ 1 1.197 0.0114 100.72 101.47
170˚C 4.294 0.0501
2 1.196 0.0114 102.21
4.293 0.0492
70 cm sec⁻¹ 1 2.104 0.0231 30.36 40.23
210˚C 3.042 0.0387
2 1.220 0.0117 50.09
2.099 0.0234

c) Retention Time of Standard Compound of Methyl Esters

Standard Compound Retention Time (min)

Methyl laurate 1.91

Methyl myristate 2.65
Methyl palmitate 4.17
Methyl stearate 7.21
Methyl linolate 6.74
d) Simple Calculation
Condition: 70 cm sec⁻¹, 210˚C
1st injection
Rs peak 1 and 2
2(3.042−2.104)
Rs=
0.0387+ 0.0231
=30.36

2nd injection
Rs peak 1 and 2
2(2.099−1.220)
Rs=
0.0234+ 0.0117
=50.09
Average resolution of peak 1 and 2
=40.23

4.0 DISCUSSION
In this experiment, there were three conditions used which are 30 cm sec⁻¹, 50 cm sec⁻¹
and 70 cm sec ⁻¹. It was found that 70 cm sec⁻¹ was the highest flow rate which will result in the
lowest value of average resolution compared to the other two conditions. The most ideal
separation is when the resolution is 1.5. High flow rate will elute the analyte faster in lower
analysis time. However, band broadening will occur because of increase in mass transfer. To
restrain, optimum gas flow rate must be obtained and used to gain better separation. In this
experiment, flow rate of 70 cm sec⁻¹ is ideal.
Next, the flow rate of the carrier gas was kept constant while the temperatures were
manipulated into three values which are 170˚C, 190˚C and 210˚C. The injection with the
temperature of 210˚ has the lowest retention time for the first and second injection compared to
the other injections with the temperature 170˚C and 190˚C. High temperature will lead to column
bleeding. Column bleeding is when the stationary phase decompose due to high temperature. The
optimum temperature in this experiment is 210˚C.
 There were five standard compound used. From the experiment, the least non-polar
compound is methyl linolate with the retention time of 6.74 minutes because it elutes last. It is
the most retained compound by the stationary phase. Methyl laurate is the most non-polar as it is
the first one to be eluted with the retention time of 1.91 minutes.
5.0 CONCLUSION
The optimized condition for separation of methyl ester is with the flow rate is 70 cm
sec⁻¹ and temperature 210˚C. The first peak of chromatogram is methyl laurate while the last
peak is methyl linolate with retention time of 1.91 and 6.74 minutes respectively.

6.0 REFERENCES

1. Analytical Separation Methods Laboratory Guide 2nd edition

2. Hazmi, B. (n.d.). Experiment 1: Gas Chromatography (GC)-Optimizing of Flow Rate and
Column Temperature. Retrieved from
https://www.academia.edu/27450633/Experiment_1_Gas_Chromatography_GC_-
Optimizing_of_Flow_Rate_and_Column_Temperature
3. Libretexts. (2018, November 26). Gas Chromatography. Retrieved from
https://chem.libretexts.org/Textbook_Maps/Analytical_Chemistry/Supplemental_Modules_
(Analytical_Chemistry)/Instrumental_Analysis/Chromatography/Gas_Chromatography
4. Knowledge Base. (n.d.). Retrieved from https://kinesis-usa.com/knowledgebase/achieving-
low-levels-of-gc-column-bleed
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https://www.chromatographytoday.com/news/gc-mdgc/32/breaking-news/what-is-a-
response-factor/31169