SOP Title-Calibration of Gas Chromatography

SOP Title-Calibration of Gas Chromatography

 

 

• OBJECTIVE

To lay down the procedure for Calibration of Gas Chromatography.

• SCOPE

This SOP is applicable to Calibration of Gas Chromatography in Quality Control laboratory.

• RESPONSIBILITY
• QC personnel shall be responsible for Implementation of the procedure.
• QC Head /designee shall be responsible for compliance of this procedure.

 

• CALIBRATION PROCEDURE
  • Calibration Frequency:
    • Every six months and after critical parts replacement.
  • Chromatographic condition:

Column                       :           ZB 624, Length 30m, Inner Diameter                                          0.53mm, Film Thickness 3.0µm (or equivalent)

Detector                      :           Flame Ionization Detector

Carrier Gas                  :           Nitrogen gas

Flow rate                     :           2 ml/min

Injection temperature  :           150°C

Oven temperature       :           100°C

FID temperature         :           200°C

Injection volume         :           1 µl

Split ratio                    :           1:20

Run time                     :           10 min.

• Head space sampler parameter:

                        Incubation /vial temperature   :           80°C

Syringe temperature                :           85°C

Injection volume                     :           1 ml

Incubation time                       :           15 min.

• Standard stock solution:
  • • 5 % v/v Acetone solution in water:-

                 5ml of Acetone dissolve in 100ml of water.

    • 5 % v/v Tetrahydrofuran (THF) in water:-

                5ml of THF dissolve in 100ml of water.

  Standard solution A:

Take 5 ml Acetone standard stock solution and 20 ml Tetrahydrofuran standard stock solution in 200 ml with water.

              Standard solution B:

    Take 5 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

Standard solution C :

           Take 10 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

Standard solution D:

       Take 15 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

Standard solution E:

Take 20 ml Acetone standard stock solution and 10 ml Tetrahydrofuran standard stock solution in 100 ml with water.

• Precision:
  • Pipette out accurately 1 ml of standard solution C into six separate 20 ml HS vial

5.5.2     Seal all the vials properly using septa.

5.5.3  Procedure:   Inject the blank vial (system blank) and six standard vials and again blank vial (for carry over) as per the procedure and calculate the RSD for Retention times and peak areas response ratio of individual peaks.

5.5.4                The relative standard deviation of area response ratio for acetone with respect to tetrahydrofuran for six replicate injections should not be greater than 5.0 %.

5.5.5   The relative standard deviation of the retention time for acetone with respect to tetrahydrofuran for six replicate injections should not be greater than 1.0 %.

5.5.6   Carry over: NMT 0.1 % (Blank after six injection/ Average peak area)

 

5.6 Linearity:

5.6.1   Pipette out accurately 1 ml of standard solution A into two separate 20 ml HS vial and seal the vials properly using septa. Similarly prepare two vial standard solution B, two vial standard solution C, two vial standard solution D and two vial standard solution E.

5.6.2    Inject all the vials, one by one.

5.6.3    Record the chromatogram of each injection and calculate the average area response ratio for acetone with respect to tetrahydrofuran.

• Draw a graph with injection volume on X-axis and average area response ratio on Y-axis.
• Curve should be linear and correlation co-efficient should not be less than 0.999.

5.7  Calibration of Column Oven Temperature Test

5.7.1   Take the first temperature reading from the thermometer and record the temperature.

5.7.2     Set the oven temperature to 100°C, 150°C and 230ºC and wait for at least 5 minutes after the oven goes ready for temperature stabilization

• Take the first temperature reading from the thermometer and record the temperature.

5.7.4                Acceptance Criteria :-

Difference at 100°C should be not more than 3°C.

Difference at 150°C should be not more than 3°C

Difference at 230°C should be not  more than 3°C.

• Software Verification

• Go to the tools bar and select “Tools”, than select “Check program files”, a window “Check the program files” will display.
• Click on execute and wait for few seconds.
• Now click on Browse & than take the report print.

Acceptance criteria: It should “Pass”

• Calibration Report of Gas Chromatography shall be prepared as per Format.
• After the successful completion of calibration. Calibration label shall be affix on the
• If the Calibration parameters are out of limit affix the Out of Calibration label and Inform to QC Manager/Designee.
• Record shall be maintained in instrument usage logbook.

 

• ABBREVIATION(s)

Abbreviation	Full Description
QC	Quality Control
QA	Quality Assurance
SOP	Standard Operating Procedure.
µl	Micro liter
µm	Micrometer
%	Percentage
°C	Degree Centigrade
RSD	Relative standard Deviation
GC	Gas chromatography
No.	Number
RT	Retention time

 

*Note – Ready to use SOP available in “DOWNLOAD” Section.