1.0 Objective:
The objective of this Standard
Operating Procedure is to lay down the procedure for
operation and calibration of Gas
chromatography (CHEMITO-7610 ECD)
2.0 Scope:
This SOP covers
operation and calibration procedure of Gas chromatography
(CHEMITO-7610)
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 visibly clean
and free from dust. Wipe all the traces of
solvent/water/moisture by dry
cloth.
4.2
Install properly suitable capillary column
in GC oven injector & selected detector.
4.3
Open the Gas pressure valve
from the distribution panel. Check the backpressure of column.
4.4 Switch on the mains of instrument &
computer.
4.5 Set the column flow as per requirement
from the upper panel by dial gauge
4.6 Setting the parameter:
4.6.1
Press the oven key on the front
panel of the instrument and set the oven temperature.
4.6.2
Then press the temperatures key
for injector and detector temperature and set the required temperature for the
same.
4.7 Software Operation:
4.7.1
Double Click on C2001 1.7 Icon.
Chemitochrom 2000 window display on the screen.
Select
and click login menu, Select GC 1 or GC 2 as per requirement and click.
Add analyst name or Password. My GC
(Work 1 or 2) window display.
4.7.2
Select file and click on New or
Open (for New file or open file), Select File Type display,
Select any one ‘Sequence’,
‘Instrument’, ‘Method’, ‘Project’ etc.
4.7.3
Click on ‘Method’ button, add
file name and click on OK. Now select file menu and click
on open, ‘Open Method File’ window
display. Select require file name and click on OK.
4.7.4
Now select and click on
setting, select method and click, or click on chromatogram with
scale
icon, method window display on the screen Feed require parameters Noise,
threshold etc. and click on OK.
4.7.5 Select and click on setting, select
Instrument and click, or click on column icon,
Instrument window display on the
screen. Feed require parameters column, mobile phase,
Flow
rate, pressure, detection, temperature etc. and click on OK.
4.7.6
Now select and click on
setting, select Analysis and click, or click on injection icon,
analysis window display on the
screen. Feed require parameters Sample ID, File Name
etc. and click on OK.
4.7.7
Select and choose Chromatogram
icon Blue or Red (=>). If choose blue(=>) auto open
the new graph (Running graph) after inject and if choose
red (=>) open the previous
graph after inject. ( Always
choose Red (=>).
4.7.8
Select and choose Chromatogram
with peak report icon Blue or Red
(=>). If choose
blue (=>) auto print out
(Running graph) after completed run and if choose red (=>) no
print out
after completed run. Always choose Red (=>).
4.7.9
Select and click Chromatogram
with peak report icon and click on OK button for print
out.
4.7.10
Now completion of all
parameters and instrument is ready for injection, inject the
solution and immediately click on
‘RUN’ Button.
4.7.11
Select method icon,
chromatogram window display on the screen, select calculation
parameter, open file name, select
require file and click on OK, then again click OK the
chromatogram display with
calibration parameters, then click on printer icon and print
the chromatogram
4.8
Preparation of new print format:
4.8.1
Click on printer icon and click on New button,
add file name and click OK. Now click on
setup button, style set up window
1 or 2 display on the screen.
4.8.2
Select and click on Labor Header, add no of
lines 3, Select line 1st and add on select line.
4.8.3
Select on 1st Page
and border and click OK. Choose Report Header, choose chromatogram
choose portrait and as on screen, then click on OK.
4.8.4
Select Result and click, choose
result table, column performance and click OK. Then
again click OK.
4.9
After completion of the analysis enter the
details in instrument usage log book as per
given.
4.10
After completion of the injection, down the
column temp below 35° C and injector and
detector temperature also cool
down. Now close the gas flow and close the valve of gas
cylinder of Z- air, Hydrogen and
carrier gas ( Nitrogen or Helium).
5.0 Calibration procedure:
5.1 Open the column oven compartment.
5.2
Install the properly capillary
column.
5.3
Connect soap film flow meter to
the detector port outlet with the help of Teflon or rubber tube.
5.4
Fill the pipette bulb partially
with a soap solution and attach to the bottom of the flow meter.
5.5
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
5.6 Check for leaks at each and every point
of attachment using soap solution.
5.7 Open the knob of carrier gas flow
controller & allow carrier gas to flow through
corresponding digital flow
control.
5.8
Adjust the carrier gas flow
rate with flow control (30 ml/min).
5.9
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
5.10
Similarly calibrate the flow
rate after the time interval of 20 min, 40 min, & 60min and find
out the difference between the readings.
5.11
Acceptance criteria: observed
flow rate of the equipment should be within ±2ml/min of set
flow rate
5.12 Detector precision and consistency of
relative retention time:
5.12.1 Preparation of pesticide standard (0.2ppm):
Take 100µl of standard
solution (100ppm) of Gamma-BHC (Lindane) in 10 ml
volumetric flask containing
5ml hexane and dilute to mark with same solvent. Take 2 ml of
this solution in 10 ml
volumetric flask and dilute to mark with hexane.
5.12.2 Inject 2.0µl of pesticides standard 5 times
and observe area and RT.
5.12.3
ACCEPTANCE CRITERIA:
The deviation for RT ± 0.2 min and deviation for area ± 10%
5.13 Detector Linearity:
5.13.1 Prepare the five different concentration
solution as follow to check the detector linearity.
5.13.2 Preparation of pesticide standard 1
(0.025ppm):
Take 100µl of standard
solution (100ppm) of Gamma-BHC (Lindane) in 10 ml volumetric
flask containing 5ml hexane
and dilute to mark with same solvent. Take 1ml of this
solution in 10ml volumetric
flask and dilute to mark with hexane. Take 5ml of this solution
in 10ml volumetric flask
and dilute to mark with hexane. Take further 5ml in 10ml
volumetric flask and dilute
to mark with hexane.
5.13.3 Preparation of pesticide standard 2
(0.05ppm):
Take 100µl of standard solution
(100ppm) of Gamma-BHC (Lindane) in 10 ml volumetric
flask containing 5ml hexane and
dilute to mark with same solvent. Take 1ml of this
solution in 10ml volumetric flask
and dilute to mark with hexane. Take further 5ml in 10ml
volumetric flask and dilute to
mark with hexane.
5.13.4 Preparation of pesticide standard 3
(0.1ppm):
Take 100µl of standard
solution (100ppm) of Gamma-BHC (Lindane) in 10 ml volumetric
flask containing 5ml hexane
and dilute to mark with same solvent. Take 1ml of this
solution in 10ml volumetric
flask and dilute to mark with hexane.
5.13.5
Preparation of pesticide standard 4 (0.2ppm):
Take 100µl of standard
solution (100ppm) of Gamma-BHC (Lindane) in 10 ml volumetric
flask containing 5ml hexane
and dilute to mark with same solvent. Take 1ml of this
solution in 5 ml volumetric
flask and dilute to mark with hexane.
5.13.6
Preparation of pesticide
standard 5 (0.4ppm)
Take 100ml of standard solution (100ppm) of gamma-BHC (lindane) in 10 ml
volumetric
flask containing 5 ml hexane and
dilute to mark with same solvent. Take 2 ml of this
solution in 5 ml volumetric flask and dilute to mark with
hexane.
5.13.7 Inject 3 replicates injection of the
above solution and observes area.
5.13.8 Plot the graph of linearity and calculate
the correlation coefficient.
5.14
CHROMATOGRAPHIC CONDITION:
Name of the instrument : GC [Chemito-7610]
Column
: TR-5 [30m X 0.32 mm ID X 1.0µm]
Oven Temp. :
120°C (1min) @
270°C (3 min)
270°C (3 min)
Injector
Temp.
: 225°C
Detector
Temp.
: 300°C
Carrier Gas
: N2 [1.2 bar]
Injection Volume : 2.0µl
Acceptance
criteria
: Correlation coefficient shall be minimum 0.99
5.15
Calibration of Oven:
Calibration of oven to be done
by calibration department
5.16
Frequency of
calibration: Quarterly
6.0
Routine maintenance:
6.1
Clean all the out side part of
instrument.
6.2
Clean the injector and injector
liner. Change the glass wool and septa if required.
6.3
Check the all connection of
gases for leakage.
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