SOP For HPLC Column Maintenance and Washing


SOP For HPLC Column Maintenance and Washing



1.1 To describe the procedure for the Maintenance, Column Washing and Usage Record of HPLC Column.


2.1 This SOP is applicable for the Maintenance, Column Washing and Usage Record of HPLC Column.


3.1 QC Officer or QC Executive


4.1 QC Manager

5.0 Procedure

5.1 The Column has to be taken serial number has to be seen.
5.2 Check the column there should be no damage.
5.3 Check the Column according to Certificate of Analysis, make, serial number, supplier name, etc.
5.4 Change the mobile phase and column washing with HPLC grade water.

5.5 If everything complies of user requirement, make the entry of the column in as per Annexure also to be write the column ID.

5.6 The Column will be given according to the requirement as mentioned in the standard test procedure at that particular product.

5.7 If the Column Complies the system suitability criteria use the column for HPLC Instrument.

5.8 In case the Column does not complies system suitability Criteria than the column cannot even use an instrument.

HPLC should be tested for column specification.

5.9 Column Fail for system suitability parameter repetitively than it should be returned to the supplier.

5.10 Flush the column using warm water at 55°C at flow rate 0.5 ml/min for 15 minutes. Maintain the

Column temp. at 55°C in the column oven.

5.11 Flush the Methanol : Water (50 : 50) flow rate 0.5 ml/min in 60 minutes.

5.12 Change the Mobile Phase from water to HPLC Grade 80 % Acetonitrile.

5.13 Flush the column using HPLC Water 0.5 ml/min for atleast one hour.

5.14 Disconnect the column from the HPLC instrument and store in an appropriate place.

5.15 Do not use the mobile phase having pH out of the range given in the column.

5.16 Do not over tight the ferrules of the column.

5.17 Wash the column properly minimum for 60 min, the water, methanol & Acetonitrile or specified

solvent given for the column washing.


Sr. No. Abbreviation used Extended Form
1.0 SOP Standard Operating Procedure
2.0 No Number
3.0 QC Quality Control


Annexure-1 : HPLC Column Usage Entry log Book.


Sr. No. Reference Title
01. In-House


Annexure-1 : HPLC Column Usage Entry log Book


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