ROYAL COLLEGE OF

PHARMACY AND HEALTH

SCIENCES

TOPIC:
GAS CHROMATOGRAPH

UNDER THE GUIDANCE OF :

MR.SANTOSH PATNAIK
ASST. PROFESSOR.
(M.Pharm.)

SUBMITTED BY:
MINAKHEE KUMARI PANIGRAHI
B.PHARM 7TH SEMESTER.
REGD. NO. : 1603254028.
SUBJECT: PHARMACEUTICAL ANALYSIS-III
BATCH : 2019-20
Gas Chromatograph:-

A Gas Chromatograph is used to detect the components based on the

selective affinity of components towards the adsorbent materials.
The sample is introduced in the liquid/gas form with the help of
GC syringe into the injection port, it gets vaporized at injection port
then passes through column with the help of continuously flowing
carrier stream (mobile phase), mainly H2 (for TCD), and gets
separated/detected at the detection port with suitable
temperature programming. We visualize this on computer in the
form of peaks. Carrier medium can be liquid (e.g. HPLC) or
gas (e.g. GC) for the ease of separation/detection, if it is gas
then called gas chromatography otherwise called liquid
chromatography.

Different chemical constituents of the sample travel through the

column at different rates depending upon,

1. Physical properties

2. Chemical properties

3. Interaction with a specific column filling (stationary phase).

As the chemicals exit the end of the column, they are detected and
identified electronically. The function of the stationary phase in the
column is to separate different components, causing each one to exit
the column at a different time (retention time). Other parameters that
can be used to alter the order or time of retention are the carrier gas
flow rate, and the temperature.
Physical Components involve inlet port, Adsorption column, detector
port, flow controller (to control the flow of carrier gas), etc.
 Two types of columns are used in GC:-

● Packed columns are 1.5 - 10 m in length and have an internal diameter

of 2
- 4 mm. The tubing is usually made of stainless steel or glass
and contains a packing of finely divided, inert, solid support
material (eg. diatomaceous earth) that is coated with a liquid
or solid stationary phase. The nature of the coating material
determines what type of materials will be most strongly
adsorbed.
● Capillary columns have a very small internal diameter, on the
order of a few tenths of millimeters, and lengths between
25-60 meters are common. The inner column walls are
coated with the active materials (WCOT columns).
Some columns are quasi solid filled with many parallel micro pores
(PLOT columns). Most capillary columns are made of fused silica
with a polyimide outer coating. These columns are flexible, so a very
long column can be wound into a small coil.
Temperature dependence of molecular adsorption and of the rate of
progression along the column necessitates a careful control of the
column temperature to within a few tenths of a degree for precise
work. Reducing the temperature produces the greatest level of
separation, but can result in very long elution times.
The choice of carrier gas (mobile phase) is important, with
hydrogen being the most efficient and providing the best
separation. However, helium has a larger range of flow rates that
are comparable to hydrogen in efficiency, with the added
advantage that helium is non-flammable, and works with a greater
number of detectors. Therefore, helium is the most common carrier
gas used Detectors:-
A number of detectors are used in gas chromatography. The most
common are the Flame ionization detector (FID) and the thermal
conductivity detector (TCD). While TCDs are essentially universal and
can be used to detect any component other than the carrier gas (as
long as their thermal conductivities are different than that of the
carrier gas, at detector temperature), FIDs are sensitive primarily to
hydrocarbons, and are more sensitive to them than TCD. Both
detectors are also quite robust. Since TCD is non-destructive, it can
be operated in-series before an FID (destructive), thus providing
complementary detection of the same eluents.
 PROCEDURE:-

● Prepare the samples for calibration with various

compositions. Keep the amount of Isopropyl alcohol fix
equal to 2 grams in each sample.
● Start the apparatus by switching on the hydrogen
supply and set the parameters:
o
Oven temperature = 170 C
o
TCD temperature = 150 C
o
Injector temperature = 200 C
2
Carrier Gas Pressure = 4 . START THE ISOTHERM.
kg/cm

● Before injection of sample wait till the base line of

recorder becomes perfectly horizontal; which indicates
that GC is stabilized or conditioned properly.
● Inject the sample with a micro syringe at the injector port
and START THE RUN
● After all the peaks attained stop the run and get the
integration results. Note the retention time and area
of peak of each of the constituents for each sample.
● Plot the CALIBRATION CURVES and find out the
response factor (slope of the calibration curve) for
each of the constituents with respect to IPA. The
curves are the straight lines passing through origin.
These will be used for analysis of unknown sample.

OBSERVATIONS CALCULATIONS:-

1. Standard Settings
Gas Pressure:
TCD
temperatur
e: Oven
temperatur
e: Injector
temperatur
e:
Amount of Sample injected:
 1. Calibration Table
#
Samp. Wt. Wt. Wt. RT* RT RT AOP AOP AOP
NO. IPA CH3 H2O IPA CH3 H2O CH3 H2O
IPA
(gms) (gms) (gms) (sec) (sec (sec)
)
1.
2.
3.
4.
5.
RT = Retention Time # AOP = Area of Peak
2. Calculation Table

Samp Weight Ratio Area Ratio Weight Area Ratio

CH3/IPA (x1) CH3/IPA (y1) Ratio H2O/IPA (y2)
.
H2O/IPA (x2)
No.
1.
2.
3.
4.
5

DISCUSSION AND RESULTS

● Plot the calibration curves and fit the straight line passing
through the origin.
● Find Response Factor.
● Find out the composition of the unknown sample.