Get the latest updates from us for free
Android App

SOP for Washing of HPLC Columns


Standard operating procedure to wash the columns of High Performance liquid Chromatography used in analysis of Pharmaceutical products.

1.0  OBJECTIVE

       To describe the procedure for washing of HPLC column.

2.0  SCOPE

       This SOP is applicable for washing of HPLC column.

3.0  RESPONSIBILITY

       Officer/ Executive - Quality Control

4.0  ACCOUNTABILITY

       Manager - Quality Control.

5.0  PROCEDURE

5.1  For C18, C8, C6, Phenyl, CN, Amine column:
5.1.1  Change the mobile phase to filtered distilled water.
5.1.2  Allow the water to flow through the column at the rate of 1ml / min. For 30 minutes. Check that the pressure does not exceed 400 BAR.
5.1.3  Change the mobile phase from water to HPLC grade 80% Acetonitrile.
5.1.4  Set the instrument to flow rate 1 ml / minute and wash the column for 30 minutes.
5.1.5  Ensure that the column contains 80% Acetonitrile.
5.1.6  Disconnect the column from the HPLC unit and store in an appropriate place.
5.2  For protein pack column:
5.2.1  Change the mobile phase to 1:10 solution of Isopropyl Alcohol and HPLC water.
5.2.2  Allow mobile phase to flow through the column at the rate of 1 ml / min. for 30 minutes. Check that pressure does not exceed 400 bars.
5.2.3  Ensure that the column contains mobile phase.
5.2.4  Disconnect the column from the HPLC unit and store in an appropriate place.
5.3  For L17 column:
5.3.1  Change the mobile phase to a mixture of 0.01 N H2SO4: ACN (85: 15).
5.3.2  Allow the mixture to flow through the column at the rate of 0.6 ml/min for 2 hours. Flow rate should not exceed 0.6 ml/min.
5.3.3  Change the mobile phase from (H2SO4, ACN) to water.
5.3.4  Set the instrument to flow rate 0.6 ml / min. and wash the column for 2 hours.
5.3.5  Ensure that the column contains water.
5.3.6  Disconnect the column from the HPLC unit and store in an appropriate place.

6.0  ABBREVIATIONS

6.1  SOP - Standard Operating Procedure
6.2  ACN - Acetonitrile
6.3  HPLC - High Performance liquid Chromatography
Ankur Choudhary is India's first professional pharmaceutical blogger, author and founder of Pharmaceutical Guidelines, a widely-read pharmaceutical blog since 2008. Sign-up for the free email updates for your daily dose of pharmaceutical tips.
Email: .moc.enilediugamrahp@ofni Need Help: Ask Question


1 Comment so far : Add yours...

ABDUL KARIM PARMAR said...

Of course there is no doubt that Mr Ankur's efforts are unmatchable and priceless....but I am sure every body will also agree with me that when actually following to get benefit of the material from this site we feel that the CONTENT OF THE SITES SHOULD BE COPYABLE........A huge request to Mr Ankur to allow copying.

Post a Comment