CHM 510 LABORATORY REPORT

EXPERIMENT 1
GAS CHROMATOGRAPHY(GC):
OPTIMIZATION OF FLOW RATE AND
COLUMN TEMPERATURE

NAME :
MATRIC NO :
GROUP :3
DATE OF EXPERIMENT:
LECTURER’S NAME

INTRODUCTION

Gas chromatography is a method for separating components of mixtures of

volatile compounds. This is because in the gas chromatography the mobile phase is
gas which is called as carrier gas. The stationary phase is in liquid which has a high
boiling point since the liquids will be used to modify the stationary phase and cannot
be volatile. In most applications, the separations in gas chromatography are made to
identify and determine the quantity of each component of a sample of the mixture.
Separation is achieved by partition of the component of the sample between the
phases. There are some factors that will affect Gas Chromatography separations,
such as volatility of compound, polarity of compounds, column temperature, column
polarity, flow rate of gas and length of column. In this experiment two factors will be
investigated which is the effect of column temperature and flow rate of gas. The
effects of the two factors on the separation of methylester were studied to determine
the optimized condition to separate the compound in methylester. In order to
determine the optimized condition the resolution was calculated. Resolution Rs is
used to describe how well the species in the mixture are separated. Ideal resolution is
1.5 that indicates adequate separation between species which means the compounds
are well separated with each other. Lower than 1.5 resolutions is when the two species
is not completely separated while too high resolution means the separation is not
efficient and having a long analysis time. This is is to explore gas chromatography;
including the concepts of retention time and resolution using a mixture of methyl
esters; methyl laurate, methyl myristate, methyl palmitate, methyl stearate and methyl
linoleate. The effects of column temperature and flow rate on the separation of these
compounds will be investigated

METHADOLOGY

Instrument set-up :
Injection port : Split (40:1)
Injection port temperature: 250°C
Column temperature : 210°C
Carrier gas flow rate : 30 cm sec-1
Detector temperature :250°C

Effect of carrier gas flow rate on isothermal GC separation of methyl esters.

0.4 µL standard mixture was injected isothermally at 210°C at carrier gas flow
rate of 30 cm sec-1. The flow rate then increased to 50 cm sec-1. The system was
allowed to equilibrate for few minutes and the standard mixture was injected again.
The same procedure was repeated at flow rate of 70cm sec-1. The most optimized flow
rate was determined.

Effect of column temperature on the isothermal GC separation of methyl esters.

 0.4 µL standard mixture was injected isothermally at 170°C, followed by 190°C
at the optimal carrier gas flow rate. The effect of column temperature on the
separation, resolution and analysis time was evaluated.

Separation of methyl esters using column temperature programming

The standard mixture was injected at the optimal carrier gas flow rate using a
lineae temperature ramp from 100°C to 290°C at optimal rate. The separation of
compound was commented.

Identification of components in methyl esters mixture

Each methyl ester was injected individually to identify the various compounds in
the standard mixture by using the optimized GC conditions

RESULTS AND DISCUSSION

A.30 cm/s, 210 °C First Injection

A.30 cm/s, 210 °C Second Injection
A.50 cm/s, 210 °C First Injection
A.50 cm/s, 210 °C Second Injection
A.70 cm/s, 210 °C First Injection
A.70 cm/s, 210 °C Second Injection
B.70 cm/s, 170 °C First Injection
B.70 cm/s, 190 °C First Injection
B.70 cm/s, 190 °C Second Injection
B.70 cm/s, 210 °C First Injection
B.70 cm/s, 210 °C Second Injection
C. Optimum Condition First Injection
C. Optimum Condition Second Injection
Methyl laurate
Methyl myristate
Methyl palmitate
Methyl linoleate
Methyl stearate

A. Effect of the variation of gas flow on the resolution.

 Retention time Peak width of
Average
Condition Injection of peak 2 and 3 peak 2 and 3 Resolution
resolution
(min) (min)
30 cm/s, 210 1 4.440, 6.178 0.0491, 0.0736 28.3293
31.9199
°C 2 4.484, 6.240 0.0428, 0.0561 35.5106
50 cm/s, 210 1 2.618, 3.655 0.0301, 0.0305 31.8587
31.1270
°C 2 2.651, 3.689 0.0300, 0.0383 30.3953
70 cm/s, 210 1 1.887, 2.632 0.0224, 0.0339 26.4654
24.9918
°C 2 1.865, 2.607 0.0306, 0.0325 23.5182

Resolution, Rs = tR3-tR2
1/2 (W2+W3)

B. Effects of the variation of column temperature at optimized column temperature on

resolution.
Retention time Peak width of
Average
Condition Injection of peak 2 and peak 2 and 3 Resolution
resolution
3 (min) (min)
70 cm/s, 1 3.634, 7.035 0.0361, 0.0602 70.6334
70.6334
170°C 2
70 cm/s, 1 2.488, 4.023 0.0294, 0.0377 45.7526
45.1018
190°C 2 2.489, 4.027 0.0297, 0.0395 44.4509
70 cm/s, 1 1.929, 2.682 0.0259, 0.0277 28.0970
27.0815
210°C 2 1.932, 2.684 0.0273, 0.0304 26.0659

C. Retention time of standard compound of the methyl ester at optimum condition.

Average
Standard compound Injection Retention time
retention time
1 1.929
Methyl laurate 1.930
2 1.931
1 2.685
Methyl myristate 2.6855
2 2.686
1 4.213
Methyl palmitate 4.2135
2 4.214
1 6.698
Methyl linoleate 6.6985
2 6.699
1 7.258
Methyl stearate 7.259
2 7.260

In the experiment, the effect of carrier gas flow rate on isothermal GC separation
of methyl esters and the effect of column temperature on the isothermal GC
separation was studied to determine the optimun condition to separate standard
compound in methyl esters. For the effect of carrier gas flow rate on isothermal
separation, the methyl esters was separated at same temperature which is 210 °C and
the manipulated variable is the carrier gas which the separation was evaluated at
different flow rate. The flow rate studied was 30 cm/s, 50 cm/s and 70 cm/s. From the
observation the retention time of peak 3 and peak 3 with their width was used to
calculate the resolution. At flow rate 30 cm/s the resolution gives 31.9199, at flow
rate 50 cm/s the resolution gives 31.1270 and at 70 cm/s the resolution gives 24.9918.
From the resolution it shows that peak 2 and peak 3 achieved base line separation
since the resolution > 1.5. but the optimum flow rate is 70 cm/s since the higher the
resolution results in longer retention time so the separation was not efficient. Since the
retention time for separation at flow rate 70 cm/s is the shortest so it is the optimum
flow rate.

Next, the effect of the column temperature on the isothermal GC separation of

methyl esters was observed. To observe this the optimum flow rate from the
observation earlier was used as constant variable. For every separation it will use 70
cm/s for the carrier gas flow rate. The column temperature observed is 170 °C , 190
°C and 210 °C. Isothermal means for each separation the column temperature will be
same from beginning to the end of separation. From the results the resolution of peak
2 and peak 3 was calculated, at temperature 170 °C, the average resolution is 70.6334,
at temperature 190°C the average resolution is 45.1018 and at temperature 210 °C the
resolution is 27.0815. From the resolution it shows that peak 2 and peak 3 achieved
base line separation at every temperature since the resolution > 1.5. but the optimum
temperature is 210 °C since the higher the resolution results in longer retention time
so the separation was not efficient. Since the retention time for separation at flow rate
210°C is the shortest and it achieved base line separation so it is the optimum
temperature.
Using the optimum carrier gas flow rate and the optimum column temperature the
components in methyl esters mixture was identified. The components of the methyl
esters mixture consits of Methyl laurate , Methyl myristate, Methyl palmitate, Methyl
linoleate and Methyl stearate. The average retention time for the compounds to elute
are 1.930, 2.6855, 4.2135, 6.6985, and 7.259 respectively. Since the column used in
the GC is HP-5 capillary column so it means the column only have 5% phenyl group
which shows it is non-polar column. The reason for using the column because in order
to retain standard methyl ester mixture which is non-polar a same polarity for the
stationary phase was used. From the results methyl laurate was elute first this is
because it is the most volatile and have the lowest boiling point.

CONCLUSION

In conclusion, the optimum condition for the separation of methyl esters is 70

cm/s and 210 °C. With the optimum condition, the identification of components in
methyl esters mixture was done with methyl laurate eluted first and methyl stearate
eluted last. This is because methyl laurate is the most volatile and have the lowest
boiling point.

REFERENCES

1. BalkisHazmi (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

2. Amirul Azhar (2014) GAS CHROMATOGRAPHY (GC), OPTIMIZATION OF

FLOW RATE AND COLUMN TEMPERATURE. Retrieved from
https://www.scribd.com/document/249342167/GAS-CHROMATOGRAPHY-GC-
OPTIMIZATION-OF-FLOW-RATE-AND-COLUMN-TEMPERATURE

3. News Medical Life Science (n.d.) Capillary HP-5ms GC/MS Columns from
Agilent. Retrieved from https://www.news-medical.net/Capillary-HP-5ms-GCMS-
Columns-from-Agilent

4. PubChem (n.d.) Methyl Laurate. Retrieved from

https://pubchem.ncbi.nlm.nih.gov/compound/Methyl_laurate