1.0 Objective:
6.0 History of Revision:
The objective of this Calibration
procedure is to lay down the procedure for Calibrating the Gas Chromatography.
2.0 Scope:
This procedure covers calibration
of Gas Chromatography.
Jr. Research Officer, Research
Officer: Responsible for calibration of the instrument as per procedure.
Head of Department: Responsible for
calibration, timely as per schedule.
QA Officer/QA Manager: Review the records and governing the document
4.0 Procedure:
4.1 Calibration
with capillary column:
4.1.1 Calibration of Flow rate
Ø Open the column oven compartment.
Ø Install the properly capillary column.
Ø Connect soap film flow meter to the detector port outlet with the
help of Teflon or
rubber tube.
Ø Fill the pipette bulb partially with a soap solution and attach to
the bottom of the flow
meter.
Ø 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.
Ø Check for leaks at each and every point of attachment using soap
solution.
Ø Open the knob of carrier gas flow controller & allow carrier gas
to flow through
Corresponding
digital flow control.to set the flow N2 gas.
Ø 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 3.0 ml
marking. Note down the time require to reach
the film from 0 to 3.0 ml mark.
Calculate the flow rate using the following formula.
Flow rate
ml/min = 60 X 3.0/60 (Dist. Between two
points 3.0 ml)
Time taken in seconds
Ø Similarly calibrate the flow rate after the time interval of 20 Sec,
40 Sec, & 60 Sec and
find out the
difference between the readings.
Ø
Acceptance criteria: observed
flow rate of the equipment should be within ±2ml/min of set flow rate.
Ø Operate the instrument as per respective procedure.
4.1.2 Prepare suitable solvent mixture
and use specified chromatographic conditions as below :
(a) Preparation of Solvents Mixture:
Ø Take 250 mg Ethyl Acetate, 250 mg Toluene and 250 mg Chlorobenzene
in 100 ml volumetric flask and make volume with Methanol. (Solution A).
(b) Chromatographic Status:
Oven Temperature 1 : 40° C
Time 1 : 3.0 minute Rate 1 : 20° C / minute
Oven Temperature 2 : 220° C
Time 2 : 3.0 minute Rate 2 : 0° C / minute
Injector Temperature :
150° C
Detector Temperature : 200° C
Carrier flow : 2 ml/minute
Injection Volume :
1.0 µl
Column : Capillary (VF
624)
Length : 30
meter Diameter : 0.32 mm ID
Film Thickness
: 1.8 µm
(c) Linearity of Detector Calibration:
Ø Prepare different concentration solution of solvent mixture solution
(A). Pipette 1 ml, 2 ml, 3 ml, 4 ml and 5 ml of Solution A in different 10 ml
volumetric flask and make
volume with Methanol. They will be indicated 250 ppm, 500 ppm, 750 ppm,
1000 ppm and 1250 ppm solutions.
Ø Inject 1.0 µl of all different concentration solution, and note down
the response of area.
Ø Calculate the correlation coefficient of the response of all
solvents should not be less than 0.99.
Ø Enter the details of calibration as given in Annexure – 1.
(d) Precision of Detector Calibration:
Ø Inject 1.0 µl five replicate injection of 1250 ppm concentration
solution of solvent mixture.
Ø Note down the response of area and retention time, calculate the RSD
of the area and retention time should not be more than 3.0 %.
Ø Enter the details of calibration as given in Annexure – 1.
4.2 Calibration
Frequency:
The instrument will be calibrated
once in three months (Quarterly).
4.3 Column
Calibration:
Ø Preparation of Solvent mixture solutions and use chromatography
conditions as given in Gas Chromatography system calibration foe respective column.
Ø Calculate the area/ area ratio and the relative retention time (RRT)
of first two peaks by considering the last peak as internal standard.
Ø Calculate the RSD of ratios, and relative retention times. The
obtained RSD value should not be more than 3.0 %.
Ø Calculate the resolution and number of theoretical plates. The Resolution
between two peaks should not be less than 2.0 and theoretical plate should not
be less than 35 % of Initial Value.
Ø Record the details in the column calibration record, as given in
Annexure - 2.
Calibration
Frequency:
Quarterly
5.0
Documentation:
5.1 Annexure - 1 Gas
Chromatography Calibration Record XXX/FRM/000
5.2 Annexure - 2 Gas
Chromatography Column Performance Record XXX/FRM/000
Revision No.
|
Effective
Date
|
Revision
details
|
Reason for
revision
|
Annexure – 1
Calibration of Gas Chromatography
|
Page
no. :
01 of 03
|
|||||||||||||||||||||||||
1.0 Calibration of carrier gas flow rate
|
||||||||||||||||||||||||||
Name of Equipment : Gas Chromatography Model No:
Instrument ID
No. :
Make
:
|
||||||||||||||||||||||||||
Required Gas flow: 3.0ml/min
Remark
: Complies / Does not Complies
Calibration on
:
Calibration Due Date:
Calibrated By
:
Checked By :
|
||||||||||||||||||||||||||
Format No. : XXX/FRM/000
|
||||||||||||||||||||||||||
CALIBRATION OF GAS
CHROMATOGRAPHY
|
Page
no. :
02 of 03
|
(1) Detector Calibration Linearity :
|
|
Date of
Calibration : Due date for next
Calibration :
Instrument ID
No. : Model
No. :
Make :
|
|
(1) : Make :
(Solvent a) Batch No. :
(2) : Make :
(Solvent b) Batch No. :
(3) : Make :
(Solvent c) Batch No. :
Preparation of Solvents Mixture :
Take (250 mg)
mg Ethyl Acetate, (250 mg)
mg Toluene and (250 mg)
mg Chlorobenzene in 100 ml volumetric flask and make volume with
Methanol. (Solution A). Pipette 1 ml, 2 ml, 3 ml, 4 ml and 5 ml of Solution A
in different 10 ml volumetric flask and make volume with methanol. They will
be observed 250 ppm, 500 ppm, 750 ppm, 1000 ppm and 1250 ppm solutions.
CHROMATOGRAPHIC STATUS :
Oven
Temperature 1 :
° C Time 1 :
- Rate 1 : ° C / minute
Oven
Temperature 2 :
° C Time 2 : minute Rate 2 : ° C / minute
Injector
Temperature : ° C Detector Temperature : ° C
Carrier
flow : ml/minute Injection Volume :
Column ID No. :
Column :
Manufacturer :
Length : Diameter : Film
Thickness :
|
|
Calibrated by : Checked by :
|
|
CALIBRATION OF GAS
CHROMATOGRAPHY
|
Page
no. :
03 of 03
|
||||||||||||
(1) Detector Calibration Linearity
|
|||||||||||||
Date of
Calibration : Due date for next
Calibration :
|
|||||||||||||
Sr. No.
|
Concentration
|
Area
|
Remarks
|
||||||||||
Ethyl
Acetate
|
Toluene
|
Chlorobenzene
|
|||||||||||
1
|
250 ppm
|
Satisfactory / Not Satisfactory
|
|||||||||||
2
|
500 ppm
|
||||||||||||
3
|
750 ppm
|
||||||||||||
4
|
1000 ppm
|
||||||||||||
5
|
1250 ppm
|
||||||||||||
Correlation
Coefficient
(Limit : NLT 0.99)
|
|||||||||||||
(2)
Precision of Detector Calibration :
|
|||||||||||||
Preparation of Solvents Mixture :
Pipette
5 ml Solution A in a 10 ml volumetric flask and make volume with methanol
(1250 ppm). Inject 5 replicate injections.
|
|||||||||||||
Sr. No.
|
Area
|
Retention Time
|
|||||||||||
Ethyl Acetate
|
Toluene
|
Chlorobenzene
|
Ethyl Acetate
|
Toluene
|
Chlorobenzene
|
||||||||
1
|
|||||||||||||
2
|
|||||||||||||
3
|
|||||||||||||
4
|
|||||||||||||
5
|
|||||||||||||
Average
|
|||||||||||||
RSD
Limit
|
|||||||||||||
NMT 3.0 %
|
NMT 3.0 %
|
||||||||||||
Remarks :
Satisfactory / Not Satisfactory
|
|||||||||||||
Calibrated by : Checked
by :
|
|||||||||||||
Annexure
– 2
Gas
Chromatography Column Performance Record
|
Page No. :
01 of 02
|
Date of
Calibration : Due date for next
Calibration :
Column No. :
Column :
Manufacturer :
Length : Diameter : Film
Thickness :
|
|
(1) : Make :
(Solvent a) Batch No. :
(2) : Make :
(Solvent b) Batch No. :
(3) : Make :
(Solvent c) Batch No. :
Preparation of Solvents Mixture :
Take (250 mg)
mg Ethyl Acetate, (250 mg)
mg Toluene and (250 mg)
mg Chlorobenzene in 100 ml
volumetric flask and make volume with Methanol. (Solution A). Pipette 5 ml of
Solution A in different 10 ml volumetric flask and make volume with methanol.
They will be observed 1250 ppm solutions.
CHROMATOGRAPHIC STATUS :
Oven
Temperature 1 : ° C Time 1 : Rate 1
: ° C / minute
Oven
Temperature 2 : ° C Time 2 : minute Rate 2
: ° C / minute
Injector
Temperature : ° C Detector Temperature : ° C
Carrier
flow : ml/minute Injection Volume :
Column :
|
|
Calibrated by : Checked by :
|
|
Format
No. : XXX/FRM/000
| |
Gas
Chromatography Column Performance Record
|
Page No. :
02 of 02
|
||||||||
Date of
Calibration : Due date for next
Calibration :
Column
No. :
Length : Diameter : Film Thickness :
|
|||||||||
Sr. No.
|
Relative Ratio
|
Relative Retention Time
|
|||||||
Ethyl Acetate
|
Toluene
|
Ethyl Acetate
|
Toluene
|
||||||
1
|
|||||||||
2
|
|||||||||
3
|
|||||||||
4
|
|||||||||
5
|
|||||||||
Average
|
|||||||||
RSD
Limit
|
|||||||||
NMT 3.0 %
|
NMT 3.0 %
|
||||||||
Sr. No.
|
Theoretical Plate
|
Resolution
|
|||||||
Ethyl Acetate
|
Toluene
|
Chlorobenzene
|
Ethyl Acetate
|
Toluene
|
|||||
1
|
|||||||||
2
|
|||||||||
3
|
|||||||||
4
|
|||||||||
5
|
|||||||||
Average
|
|||||||||
Limits
|
NLT 2500
|
NLT 12000
|
NLT 25000
|
NLT 2.0
|
NLT 2.0
|
||||
Remarks : Satisfactory / Not Satisfactory
|
|||||||||
Calibrated by : Checked by :
|
|||||||||
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