- To lay down a procedure for the operation and calibration of High Pump for column washing Make: Shimadzu.
2.1 Chemist / Executive – Quality Control – to follow the procedure.
2.2 Head – Quality Control for implementation and compliance.
- 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
- HPLC – High performance liquid chromatography.
- HOD – Head of Department
5. 0 REFERENCES
6. 0 ANNEXURES
- ANNEXURE – 1 – Calibration Record for Wash Pump.
ANNEXURE – 1 – Calibration Record for Wash Pump
|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|
Acceptance Criteria : + 0.02 ml/min of set flow rate
Results : Passes/Fails
Performed By :________ Verified By : _______
Date :________ Date : ________