SOP for washing of HPLC columns

1.0 Purpose

To provide an instruction for washing of HPLC columns.

2.0 Objective :
3.0
4.0

To provide a documented procedure for washing of HPLC columns.

Scope
:
Department .

This procedure is applicable for washing of HPLC columns in QC

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 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 :
HPLC column use logbook.
Log Book For HPLC Column

Column ID.
Date

Sample
ID

:
Batch
No.

Product
Mobile
Phase

No.
of
Inj.

Washing
Time
From To

Storage

Pressur
e

Analyst

Remark
s

Format:-F/QC/013