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SOP for Operation and Calibration of Gas Chromatography (CHEMITO-7610 ECD)

Objective
The objective of this SOP is to lay down the procedure for operation and calibration of Gas chromatography .

Scope
This SOP covers operation and calibration procedure of Gas chromatography.

Responsibility
QC 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 document.

Procedure

OPERATION
  • Ensure that the instrument is visibly clean and free from dust. Wipe all the traces of solvent/water/moisture by dry cloth.
  • Install properly suitable capillary column in GC oven injector & selected detector.
  • Open the Gas pressure valve from the distribution panel. Check the backpressure of column.
  • Switch on the mains of instrument & computer.
  • Set the column flow as per requirement from the upper panel by dial gauge.

Setting the parameter:
  • Press the oven key on the front panel of the instrument and set the oven temperature.
  • Then press the temperatures key for injector and detector temperature and set the required temperature for the same.
Software Operation:
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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.
  • 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 (=>).
  • 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 (=>).
  • Select and click Chromatogram with peak report icon and click on OK button for printout.
  • Now completion of all parameters and instrument is ready for injection, inject the solution and immediately click on ‘RUN’ Button.
  • 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.
Preparation of new print format:
  • 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.
  • Select and click on Labor Header, add no of lines 3, Select line 1st and add on select line.
  • Select on 1st Page and border and click OK. Choose Report Header, choose chromatogram choose portrait and as on screen, then click on OK.
  • Select Result and click, choose result table, column performance and click OK. Then again click OK.
  • After completion of the analysis enter the details in instrument usage log book as per given in annexure – 1.
  • 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).

CALIBRATION
  • Open the column oven compartment.
  • Install the properly capillary column.
  • Connect soap film flow meter to the detector port outlet with the help of Teflon or rubber tube.
  • Fill the pipette bulb partially with a soap solution and attach to the bottom of the flow meter.
  • 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.
  • Check for leaks at each and every point of attachment using soap solution.
  • Open the knob of carrier gas flow controller & allow carrier gas to flow through corresponding digital flow control.
  • Adjust the carrier gas flow rate with flow control (30 ml/min).
  • 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.
  • Similarly calibrate the flow rate after the time interval of 20 min, 40 min, & 60min and find out the difference between the readings.
Acceptance criteria: observed flow rate of the equipment should be within ±2ml/min of set flow rate.

Detector precision and consistency of relative retention time:

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.
  • Inject 2.0µl of pesticides standard 5 times and observe area and RT.
Acceptance criteria:  The deviation for RT ± 0.2 min and deviation for area ± 10%

Detector Linearity:
  • Prepare the five different concentration solution as follow to check the detector linearity.
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.
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.
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.

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.
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.
  • Inject 3 replicates injection of the above solution and observes area.
  • Plot the graph of linearity and calculate the correlation coefficient.
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) @ 5°C/min 270°C (3 min)
Injector Temp. : 225°C
Detector Temp. : 300°C
Carrier Gas : N2 [1.2 bar]
Injection Volume : 2.0µl
Split Ratio : Split less
Acceptance criteria : Correlation coefficient shall be minimum 0.99

Calibration of Oven:
Calibration of oven to be done by calibration department
Frequency of calibration: Quarterly

Routine Maintenance
  • Clean all the out side part of instrument.
  • Clean the injector and injector liner. Change the glass wool and septa if required.
  • Check the all connection of gases for leakage.

Annexure
Annexure – 1 Gas chromatography system usage logbook. 
Annexure –2 Gas chromatography calibration record 
Annexure –3 Gas chromatography calibration record 

 
Revision History
Nil

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