1.0
Objective:
The objective of this Standard Operating Procedure is to lay down the
procedure for
operation and calibration of Gas chromatography (HP-6890)
2.0
Scope:
This SOP covers procedure for operation and calibration of Gas
chromatography
(HP-6890)
3.0
Responsibility:
Jr. Research
Officer, Research Officer: Responsible for operation and maintenance of
the
instrument as per procedure.
Head of Department: Responsible for maintenance, timely as
per schedule.
QA Officer/QA Manager: Review the records and
governing the document
4.0
Procedure :
4.1
Ensure that the instrument is clean and free from dust. Wipe all the
traces of
solvent/water/moisture by dry cloth.
4.2
I.
The room temperature should be maintained between 10 to 30°C and
relative humidity
Should be maintained
between 30 to 80 %
II. Avoid direct exposure to air-blow from air
conditioners.
III. Avoid exposure to direct sunlight and
vibrations.
IV. Avoid from equipment which generates
strong magnetic field.
4.3 Install
properly suitable capillary column in GC oven injector & selected detector.
4.4 Open the valve of gas cylinder of Nitrogen,
Hydrogen and z-Air. And checked the
pressure & leakage.
4.5 Open
the Gas pressure valve from the distribution panel.
4.6 Check
the backpressure of column.
4.7 Switch
on the mains power, stabilizer, GC power & computer power.
4.8 Double click on the instrument1 (online)
& instrument1 (offline) icon from the desktop;
create new method from instrument1 (offline) and load method in
instrument1 (online)
from file menu. Apply the chromatographic
condition and enter ok. And view the run
signal window, After completion of the run,
and open the instrument offline select the file
and] integrate the peak using integration
menu, go to calibration task icon and enter the
component name and click on ok, then print the chromatogram by clicking
print option
from file menu.
4.9 Condition the column at 280°C for 10 min.
Enter the programming condition of the sample
through micro syringe, when G.C shows ready signal.
4.10 Cool the G.C after completion of the analysis
& switch it off. Shut down the computer,
mains
power supply. Close the gas supply from the distribution panel & from
cylinder.
5.0 Calibration
procedure:
5.1 Calibration
of Flow rate
5.1.1
Open the column oven compartment.
5.1.2
Install the properly capillary column.
5.1.3
Connect soap film flow meter to the detector
port outlet with the help of Teflon or
rubber tube.
5.1.4
Fill the pipette bulb partially with a soap
solution and attach to the bottom of the flow
meter.
5.1.5
Open the knob of carrier gas (N2) and set up proper pressure (i.e. 500
kpa or 5 Kg/cm2) in
carrier gas pressure controller.
5.1.6
Check for leaks at each and every point of attachment using soap
solution.
5.1.7
Open the knob of carrier gas flow controller & allow carrier gas to
flow through
corresponding digital flow control.
5.1.8
Adjust the carrier gas flow rate with flow control (30 ml/min).
5.1.9
Gentle squeeze the bulb to force a soap film up into the gas stream.
Start the stopwatch as
soon as the film reaches to zero ml mark. Stop the watch when the film
reaches to 30 ml
marking. Note down the time require to reach the film from 0 to 30 ml
mark. Calculate the
flow rate using the following formula.
Flow rate ml/min = 60 X 30 (Dist.
Between two points 30 ml)
Time taken in seconds
5.1.10 Similarly calibrate the flow rate after the
time interval of 20 min, 40 min, & 60min and
find out the difference between the readings.
5.1.11 Acceptance criteria: observed flow rate of the
equipment should be within ±2ml/min of set flow rate
5.2
Detector precision and consistency of relative retention time:
5.2.1
Preparation of calibration solution (2000ppm):
Weigh accurately 100mg of Toluene
and 100mg of 1-Butanol in 50 ml volumetric flask
containing 5 ml methanol and dilute
to mark with same solvent.
5.2.2
Inject 1.0µl of above calibration standard 5 times and observe area and
RT.
5.2.3
ACCEPTANCE CRITERIA:
The RSD of RRT Not More Than 2 %
The RSD of area ratio Not More Than 5%
5.3
Detector Linearity:
5.3.1
Prepare the five different concentration solution as follow to check the
detector linearity.
1.Solution-1 (500 ppm):
Weigh accurately 100mg of Toluene
and 100mg of 1-Butanol in 50 ml volumetric flask
containing 5 ml methanol and
dilute to mark with same solvent. Take 2.5ml of this solution in 10ml
volumetric flask and dilute to mark with same solvent.
2. Solution-2 (750 ppm):
Weigh accurately 100mg of Toluene
and 100mg of 1-Butanol in 50 ml volumetric flask
containing 5 ml methanol and dilute
to mark with same solvent. Take 3.75ml of this
solution in 10ml volumetric flask
and dilute to mark with same solvent.
3. Solution-3 (1000 ppm):
Weigh accurately 100mg of Toluene and 100mg of 1-Butanol in 50 ml
volumetric flask
containing 5 ml methanol and dilute to mark with same solvent. Take 5ml
of this solution
in 10ml volumetric flask and dilute to mark with same solvent.
4. Solution-4 (1250 ppm):
Weigh accurately 100mg of Toluene
and 100mg of 1-Butanol in 50 ml volumetric
flask containing 5 ml methanol and
dilute to mark with same solvent. Take 6.25ml of this
solution in 10ml volumetric flask
and dilute to mark with same solvent.
5. Solution-5 (1500 ppm):
Weigh accurately 100mg of Toluene
and 100mg of 1-Butanol in 50 ml volumetric
flask containing 5 ml methanol and
dilute to mark with same solvent. Take 7.5ml of this
solution in 10ml volumetric flask
and dilute to mark with same solvent.
5.3.2 Inject 3 replicate injections of the above
solutions and observe area.
5.3.3
Plot the graph of linearity and calculate the correlation coefficient.
5.3.4
CHROMATOGRAPHIC CONDITION:
Name of the instrument : GC [HP-6890]
Column
: BP-5 [30m X 0.32 mmID X 1.0µm]
@ 2°C/min
Oven Temp. :
50°C (2min) 70°C (1 min)
Injector Temp. : 250°C
Detector Temp. : 250°C
Carrier Gas
: N2 [1.0 ml], H2-30, AIR-300
Injection Volume : 1.0µl
Split Ratio
: 10:1
Acceptance criteria: Correlation
coefficient minimum 0.99
5.4 Calibration
of Oven:
Calibration of oven to be done by calibration department.
5.5 Frequency of calibration: Quarterly
6.0 Routine
Maintenance:
6.1
Clean the
all out side part of instrument.
6.2
Clean the
injector and injector liner. Change the glass wool of injector liner. Change
the septa of injector port if required.
6.3
Check the
all connection of gases for leakage.
