Saturday, November 27, 2010

Instruction for washing of HPLC columns

1.0     Purpose     :      To provide an instruction for washing of HPLC columns.

2.0     Objective   :      To provide a documented procedure for washing of HPLC columns.

3.0              Scope      :     This procedure is applicable for washing of HPLC columns in QC Department ---Pharmaceuticals Ltd, Ankleshwar.

4.0      Responsibility   :
·        Primary  :    QC Chemist / QC-Officer
·        Secondary  :  Overall QC – In charge

5.0       Procedure  :
·        After completion of analysis stop the pump, keep the reservoir in the distilled & filtered water.
·        Open the purging valve of the pump; purge the system for 3 to 5 minutes to remove air bubbles. Close the purging valve by decreasing the flow rate up to zero.
·        Wash the HPLC column initial with HPLC grade water for about 15-20 minutes.
·        Then wash the HPLC column with respective solvent mixtures (e.g. 50:50 Methanol: Water or Acetonitrile: Water) for minimum ½ hr to 1 hr.
·        Finally wash the HPLC column with respective solvent (Acetonitrile / Methanol) for about 15-30 minutes.
·        Keep flow rate 0.5 ml / min or 1.0 ml / min. increase flow rate gradually.
·        Check the pH of eluent on pH paper (it should be neutral) 
·        Store the HPLC column in solvent (acetonitrile or methanol).
·        Disconnect the HPLC column and keep it at respective column storage box.
·        Maintain the cleaning record in HPLC column logbook.

6.0      Reference Document  :
v      HPLC column use logbook.

Log Book For HPLC Column

Column ID.

Sample ID
Batch No.
Mobile Phase

No. of Inj.
Washing Time


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