WASHING OF HPLC COLUMNS

• OBJECTIVE:

• To lay down the procedure for washing of HPLC columns.

• SCOPE:

• This SOP is applicable to the procedure for the washing of HPLC columns at {Company Name} {Location}.

• RESPONSIBILITY:
• Quality Control Officer/Executive: is responsible to perform the activity as per the SOP
• Head Quality Control: is responsible to ensure the compliance of this SOP
• ACCOUNTABILITY:

             QA Head shall be Accountable for implementation of SOP.

• PROCEDURE:
• For C18, C8, C6, Phenyl, CN, amine columns:
• Change the mobile phase to Milli Q water.Allow the water to flow through the column with flow rate of 0.5 ml increment at the rate of 1ml / min. for minimum 30 minutes.
• Change the mobile phase from water to HPLC Methanol or Acetonitrile.
• Set the instrument to flow rate 1 ml/min. and wash the column for 30 minutes.
• In case of Reverse phase columns, ensure that the column contains methanol.
• In case of normal phase columns, ensure that columns shall be washed and stored in Hexane.
• Disconnect the column from the HPLC system and store in an appropriate place
• For protein pack column:
• Change the mobile phase to 10% Isopropyl Alcohol.
• Allow 10% Isopropyl Alcohol to flow through the column with flow rate of 0.5 ml increment at the rate of 1ml/min. for minimum 30 minutes.
• Ensure that the column contains 10% Isopropyl Alcohol.
• Disconnect the column from the HPLC unit and store in an appropriate place.
• For L17 column:
• Change the mobile phase to a mixture of 0.01N H2SO4: ACN (85: 15).
• Allow the mixture to flow through the column at the rate of 0.6ml/min for 2 hours.
• Flow rate should not exceed 0.6ml/min.
• Change the mobile phase from (H2SO4, ACN) to water.
• Set the instrument to flow rate 0.5ml/min. and wash the column for 2 hours.
• Ensure that the column contains water.
• Disconnect the column from the HPLC unit and store it in an appropriate place.

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• REFERENCES:

Not Applicable

• ANNEXURES: 

Not Applicable

        ENCLOSURES: SOP Training Record.

•  DISTRIBUTION:
• Controlled Copy No. 01  :          Head Quality Assurance
• Controlled Copy No. 02  :         Head Quality Control
• Master Copy                    :          Quality Assurance Department
• ABBREVIATIONS:

• REVISION HISTORY:             

CHANGE HISTORY LOG

Click the link for download word file copy of this document:
https://pharmaguidehub.com/product/washing-of-hplc-columns/