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.

:

Product

:

Date	Sample ID	Batch No.	Mobile Phase	No. of Inj.	Washing Time		Storage	Pressure	Analyst	Remarks
					From	To
Format:-F/QC/013