Calibration of Preparative HPLC Pump

Calibration of preparative HPLC pump:


To lay down the procedure for the preparation and calibration of preparative HPLC pump.


This SOP applies to the Quality control department.


AnalystCalibration of RefractometerQuality Control (QC)
Reviewer/ Reviewer Team LeaderReview of reportsQuality Assurance (QA)
Head of DepartmentEnsuring compliance of this SOPQuality Control (QC)
Calibration of preparative HPLC pump


  1. To maintain the flow accuracy degas the solvent before use.
  2. Setting the flow rate
  3. Press the flow key and enter the numerical value for the flow rate in ml/min.
  4. Press enter, and then run. Subsequent changes in the flow rate require only entering the numerical value and pressing enter. changes in flow may also be made by using the up/down scroll key.
  5. Priming the Pump
  6. Open the purge valve 1/2 turn.
  7. Press purge function.
  8. The pump will draw solvent through the inlet lines and expel air out the purge valve to waste. When no more air is present in the intake lines and cannot be seen coming out of the purge valve waste port, then the pump is primed.

Calibration of Pump Flow Rate Accuracy:

  1. Make sure that all flow lines are connected to the pump and put the pump inlet tube in the water.
  2. Make a readily available volumetric flask before starting the procedure.
  3. After connecting all the flow lines of a pump and choose four different flow rates 1ml/min, 2ml/min, 4ml/min and 8ml/min within the allowed range of the pump.
  4. Start the pump and make water flow through the flow lines.
  5. Measure the out water for 1 min in the volumetric flask.
  6. record the details in the given format.

Acceptance Criteria:

The difference between actual measured value and set value should be ± 2%.

Set Flow Rate (ml/min)Allowed deviation (ml/min ± 2%)
1ml± 0.02
2ml± 0.04
4ml± 0.08
8ml± 0.16


Once in a 6 months.

Calibration of Preparative HPLC Column

Calibration of Preparative HPLC Column

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