1.0 Objective:
The objective of this Standard
Operating Procedure is to lay down the procedure for
operation and calibration of Gas
chromatography (THERMO-TRACE)
2.0 Scope:
This SOP covers
operation and calibration procedure
of Gas chromatography
(THERMO-TRACE).
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 Properly install suitable capillary
column in GC oven injector & selected detector.
4.3
Open the Gas pressure valve
from the distribution panel. (N2 cylinder for N2 set 60 psi for N2).
4.4 Check the backpressure of column.
4.5 Switch on the mains power, stabilizer,
GC power & computer power.
4.6
Double click on the Chrom-card
icon from the desktop, chrom-card window display on
the screen,
double click on Konark icon, select the method from edit G.C parameters
icon, then.
Apply the chromatographic condition and enter ok open run signal. After
completion of
the run, open the instrument in offline, then go to reprocessing menu and
select file and
click on ok button, integrate peak, after integration enter the component
name then click
on ok ,go to edit method than go to in
report parameters and click on ok
to view the
print layout print chromatogram by clicking printer icon.
4.7
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.
4.8
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 Remove the column if attached to the
injector port –1 (to be calibrated).
5.1.3 Connect
soap film flow meter (side arm) and injector 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 Calibration of FPD
5.2.1 Detector precision and consistency of
relative retention time:
5.2.1.1 Preparation of pesticide standard (0.2ppm):
Take 100µl of standard
solution (100ppm) of Methyl parathion in 10 ml volumetric
flask containing 5ml
acetone and dilute to mark with same solvent. Take 2 ml of this
solution in 10 ml
volumetric flask and dilute to mark with acetone.
5.2.1.2 Inject 2.0µl of pesticides standard 5 times
and observe area and RT.
5.2.1.3 ACCEPTANCE CRITERIA:
The deviation of RT ± 0.2 min
The deviation of area ± 10%
5.2.2 Detector Linearity:
5.2.2.1 Prepare the five different concentration
solution as follow to check the detector linearity.
1. Preparation
of pesticide standard 1 (0.025 ppm):
Take 100µl of standard
solution (100 ppm) of Methyl parathion in 10 ml volumetric flask
containing 5ml acetone and
dilute to mark with same solvent. Take 1ml of this solution in
10ml volumetric flask and
dilute to mark with acetone. Take 5ml of this solution in 10ml
volumetric flask and
dilute to mark with acetone. Take further 5ml in 10ml volumetric
flask and dilute to mark
with acetone.
2. Preparation of pesticide standard 2 (0.05 ppm):
Take 100µl of standard
solution (100 ppm) of Methyl parathion in 10 ml volumetric flask
containing 5ml acetone and
dilute to mark with same solvent. Take 1ml of this solution
in 10ml volumetric flask
and dilute to mark with acetone. Take further 5ml in 10ml
volumetric flask and dilute to
mark with acetone.
3.
Preparation of pesticide standard 3 (0.1 ppm):
Take 100µl of standard
solution (100 ppm) of Methyl parathion in 10 ml volumetric flask
containing 5ml acetone and
dilute to mark with same solvent. Take 1ml of this solution
in 10ml volumetric flask
and dilute to mark with acetone.
4. Preparation of pesticide standard 4 (0.2
ppm):
Take 100µl of standard
solution (100 ppm) of Methyl parathion in 10 ml volumetric flask
containing 5ml acetone and
dilute to mark with same solvent. Take 1ml of this solution
in 5ml volumetric flask and dilute to mark
with acetone.
5. Preparation of pesticide standard 5 (0.4
ppm):
Take 100µl of
standard solution (100 ppm) of Methyl parathion in 10 ml volumetric flask
containing 5ml
acetone and dilute to mark with same solvent. Take 2ml of this solution
in 5ml volumetric flask and
dilute to mark with acetone
5.2.2.2 Inject 3 replicates injection of the above
solution and observes area.
5.2.2.3 Plot the graph of linearity and calculate the
correlation coefficient.
Acceptance criteria:
Correlation coefficient minimum 0.99
5.2.2.4 CHROMATOGRAPHIC CONDITION:
Name of the instrument : GC [Thermo Trace
ultra]
Column
: CPSIL-8CB [30m X 0.25 mm ID X 0.25µm]
@ 15°C/min
Oven Temp. :
130°C (3min) 
270°C (10 min)
270°C (10 min)
Injector
Temp.
: 250°C
Detector
Temp.
: 300°C
Carrier Gas
: N2 [2 ml] H2 (115ml) AIR (120ml
Injection
Volume
: 2.0µl
5.3 Calibration of NPD
5.3.1 Detector Precision and consistency of
relative retention time: -
5.3.1.1 Preparation of pesticide
standard (0.2ppm):
Take
100 mL of standard solution (100 ppm) of Chlorpyrifos in 10 ml volumetric
flask containing 5ml acetone and dilute to mark with same solvent. Take 2 ml of
this solution in 10 ml volumetric flask and dilute to mark with acetone.
5.3.1.2 Inject
2.0µL of above pesticide standard 5 times and observe area and RT.
5.3.1.4 ACCEPTANCE CRITERIA:
The
deviation for RT ± 0.2 min and deviation for area ± 10%.
5.3.2 Detector Linearity: -
Prepared
the five different concentrate solution as follow to check the Detector
Linearity
5.3.2.1 Preparation of pesticide standard 1 (0.025ppm): Take 100 mL of standard solution (100 ppm) of
Chlorpyrifos in 10 ml volumetric flask containing 5ml acetone and dilute to
mark with same solvent. Take 1 ml of this solution in 10 ml volumetric flask
and dilute to mark with acetone. Take 5 ml of this solution in 10 ml of
volumetric flask and dilute to mark with hexane. Take further 5 ml in 10 ml
volumetric flask and dilute to mark with acetone.
5.3.2.2 Preparation of pesticide standard 2 (0.050ppm): Take 100 mL of standard solution (100 ppm) of
Chlorpyrifos in 10 ml volumetric flask containing 5ml acetone and dilute to
mark with same solvent. Take 1 ml of this solution in 10 ml volumetric flask
and dilute to mark with acetone. Take further 5 ml in 10 ml of volumetric flask
and dilute to mark with acetone.
5.3.2.3 Preparation of pesticide standard 3 (0.1ppm): Take 100 mL of standard solution (100 ppm) of
Chlorpyrifos in 10 ml volumetric flask containing 5ml acetone and dilute to
mark with same solvent. Take 1 ml of this solution in 10 ml volumetric flask
and dilute to mark with acetone.
5.3.2.4 Preparation of pesticide standard 4 (0.2ppm): Take 100 mL of standard solution (100 ppm) of
Chlorpyrifos in 10 ml volumetric flask containing 5ml acetone and dilute to
mark with same solvent. Take 2 ml of this solution in 10 ml volumetric flask
and dilute to mark with acetone.
5.3.2.5 Preparation of pesticide standard 5 (0.4ppm): Take 100 mL of standard solution (100 ppm) of
Chlorpyrifos in 10 ml volumetric flask containing 5ml acetone and dilute to
mark with same solvent. Take 2 ml of this solution in 5 ml volumetric flask and
dilute to mark with acetone.
5.3.2.6 Inject
3 replicates injection of the above solution and observe Area
5.3.2.7 Plot
the graph of linearity and calculate the correlation-coefficient
5.3.2.8 CHROMATOGRAPHIC CONDITION
Name
of Instrument : GC
[THERMO]
Column :
CPSIL-8CB(30.0Mx0.25 X 0.25mm)
Oven
Temp. :
130°C (3minhold)@5°C/min-->270°C(10min
hold)
Detector :
300 °C(NPD)
Injector :
250 °C
Carrier
Gas :
N2 [2.0ml/min ]
Injection
volume : 2
mL
5.3.3 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.
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