Experiment 1: Gas Chromatography (GC): Optimization of

Flow Rate and Column Temperature

NAME: NADIRAH BINTI ZAKARIA

ID: 2016586557

PARTNERS’ NAME:

1. NUR SYAZWANI BINTI MUHAMMAD YAZID

(2016596153)
2. SITI ZAHARIAH BINTI ZAHARUDIN (2016552077)
3. WAN NURUL IHSAN BINTI WAN ZAINUDDIN
(2016552007)

DATE EXPERIMENT CONDUCTED: 7 JUNE 2017

DATE EXPERIMENT SUBMITTED: 19 JUNE 2017

LECTURER: MADAM ROHAIZA MOHAMAD

Experiment 1: Gas Chromatography: Optimization of Flow Rate and Column Temperature.

Objective:

To investigate the effect of column temperature and flow rate on theseparation on the compounds.

Introduction:

Gas chromatography, specifically gas-liquid chromatography involves a sample being

vaporized and injected onto the head of the chromatographic column. The sample is transported

through the column by the flow of inert, gaseous mobile phase. The column itself contains a liquid

stationary phase which is adsorbed onto the surface of an inert solid. Efficient separation of

compounds in GC is dependent on the compounds travelling through the column at different rates.

The rate at which a compound travels through a particular GC system depends on several factors

that include volatility of compound, column temperature, carrier gas flow rate and length of the

column.

In volatility of compound, low boiling components will travel faster through the column than

high boiling components. Next is column temperature. In column temperature, raising the column

temperature speeds up all the compounds in a mixture. In carrier gas flow rate, increasing the

carrier gas flow rate increase the speed with which all compounds move through the column. Lastly

is the length of the column. The longer the column, the longer it will take for all compounds to elute.

Longer columns are employed to obtain better separation.

Chemical:

Standard mixture of methyl laurate, methyl myristate, methyl palmitate, methyl stearate and methyl

linoleate.

Apparatus:

Volumetric Flask, micro pipette, vial, dropper and beaker.

Procedure:

1. The standard mixture of methyl laurate, methyl myristate, methyl palmitate, methyl stearate

and methyl linoleate is 0.2 mg mL-1, 0.2 mg mL-1, 1.0 mg mL-1, 0.70 mg mL-1 and 0.35 mg

mL-1 respectively is prepared.

2. Instrument is set up. Split (40:1) is used for injection port. The injection port temperature is

set to 250℃. The column temperature was set to 210℃. The carrier gas flow rate is set at 30

cm sec-1 and the detector temperature is set at 250℃.

3. 0.4 µL standard mixture is injected isothermally at 210℃ at carrier gas flow rate of 30 cm

sec-1. The flow rate is increased to 50 cm sec-1. A few minutes were allowed for the system to

equilibrate and the standard mixture is injected again. The procedure is repeated at flow

rate 70 cm sec-1.

4. 0.4 µL standard mixture is injected isothermally at 170℃, followed by 190℃ , at the optimal

carrier gas flow rate. The effect of column temperature on the separation, resolution and

analysis time is evaluated.

5. At the optimal flow rate, the standard mixture is injected at the optimal carrier gas flow rate

using a linear temperature ramp from 100℃ to 290℃.

6. Each methyl ester individually is injected to identify the various compounds in the standard

mixture using the optimized GC conditions.

Results:
Discussion:

The variation of the mobile phase flow rate will affect the retention time of the compounds in

which slow mobile phase flow rate will give better separation but increased the analysis time. In

contrast, high flow rate will shorten the analysis time but will cause in broadening due to the mass

transfer (C-term) in Van Deemter plot, because the solute does not completely interact with the

stationary phase. To reduce the analysis time and to produce better separation, the optimum gas

flow rate must be used.

In this experiment, the optimum mobile phase flow rate is 70m/s that give good resolution

of 6.33 compared to the others that are far from the ideal resolution value that is 1.5. The column

temperature also affects the separation resolution and the analysis time. High column temperature

will give short analysis time but some of the earlier peaks may be overlapped while low column

temperature produces better separation but will take very long analysis time. The optimum column

temperature must be used in order to separate each compounds adequately.

In this case, 210°C of column temperature is the best temperature to separate each of the

compounds. Based on this experiment, the best condition to separate the methyl ester mixture is by

using 70m/s gas flow rate at 210°C column temperature that will give adequate separation between

compounds and also shorter analysis time.

Optimum flow rate and optimum column temperature will produce better separation, high

efficiency, good resolution and short analysis time for the separation. Since the separation of gas

chromatography is based on the boiling point of the compounds, it can be concluded that methyl

laurate has lowest boiling point followed by methyl myristate and highest boiling point is the methyl

palmitate.
Conclusion:

In this experiment, we investigate the effect of column temperature and flow rate on the

separation on the compounds. The optimized condition for the separation of the methyl esters is

70m/s of gas flow rate and 210°C of column temperature.

References:

1. Analytical separation methods laboratory guide: 2nd edition. Nor’ashikin Saim, Ruziyati

Tajuddin and Mardiana Saaid. (2016)

2. Y. Guillaume and C. Guinchard (January 1997), France, Prediction of Retention Times,

Column Efficiency, and Resolution in Isothermal and Temperature-Programmed Gas

Chromatography: Application for Separation of Four Psoralens, 3/10/2014.