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.
• 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:             

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