1.0       OBJECTIVE

1.       To lay down a procedure for the operation and calibration of High Pump for column washing Make:  Shimadzu.

2.0 RESPONSIBILITY

2.1       Chemist / Executive – Quality Control – to follow the procedure.

2.2        Head – Quality Control for implementation and compliance.

3.0 PROCEDURE

• 3.1 OPERATION

• Ensure that the working area is clean and check the calibration status of the instrument.  
• Switch on the power to the instrument, wait for initialization.

• Keep the solvent line tube in appropriate solvents.

•  Open the drain valve 2 turns anti clock wise direction.

•  Click on ‘purge’ then pump starts flow.

• Close the drain valve after completion of purge.

• Connect  the column to the column inlet tube

• Set the appropriate flow  rate by pressing function ‘Key’

• Enter the appropriate flow rate and then press ‘Enter’.

• Displays the new set volume of flow rate.

• Press the pump key to start the pumping of set flow rate.

• After completion of washing of the column again click on pump to stop the flow.

• Enter the column washing details as per Annexure – 1 of  SOP QC073.

• Switch off the instrument after usage.

• 3.2  CALIBRATION

• 3.2.1 FLOW ACCURACY TEST

• Fix the C18, 75 x 4.6 mm column to the system.

• Keep degassed methanol (HPLC grade) in solvent line and prime thoroughly, and stabilize for about 30 min. with a flow rate of 1 ml/min.

• Take a 10 ml class ‘A’ calibrated dry volumetric flask.

• When the flow and pressure are stable, insert the outlet tubing into the volumetric flask and immediately start a calibrated stopwatch.

• Stop the stopwatch button when the meniscus reaches the 10 ml mark on the flask.

• Record the elapsed time in seconds.

• Calculate the flow rate using the following equation:   

10 ml * 60

                                        Calculated flow rate =       ————————-

                                                                            Measured Time in seconds

• Record the calculated flow rate in Annexure – 1

•  Similarly test the flow rate accuracy with flow rates of 0.5 and  2.0 ml/min and record the calculated flow rates in Annexure –1.

•  Acceptance criteria: ± 0.02 ml/min of set flow rate.

•  Calibration Schedule: Once three month and after any major maintenance job

4.0        ABBREVIATIONS

• HPLC         –            High performance liquid chromatography.
• HOD           –            Head of Department

5. 0      REFERENCES

Nil

6. 0      ANNEXURES            

• ANNEXURE –  1      –  Calibration Record for Wash Pump.

ANNEXURE –  1 – Calibration Record for Wash Pump

Instrument No.		Make
Calibration date		Next due date
Model		Page no	1of  1

STOP WATCH ID:______________    CALIBRATION DUE DATE OF STOP WATCH:____________

VOLUMETRIC FLASK NO:___________________

1.           FLOW ACCURACY CALIBRATION RECORD

Set flow rate	Observed elapsed Time(sec) to collect 10 ml	Flow Rate         Result    (10*60 /Time in (sec)	Difference of Flow   Rate in ml
0.5   ml/min
1.0  ml/min
2.0 ml/min

Acceptance Criteria : + 0.02 ml/min of set flow rate

Results :                                Passes/Fails

Performed By  :________                                                               Verified By   : _______

Date               :________                                                           Date          : ________

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