HPLC COLUMN MAINTENANCE

1.0 OBJECTIVE 

          To lay down a procedure for maintenance of HPLC column. 

2.0 SCOPE

This SOP shall be applicable for receipt, performance, storage, regeneration and withdrawal of all HPLC column received in Quality Control Laboratory.

3.0RESPONSIBILITY

3.1       Execution – Tr. Executive & above – QC Department 

3.2       Checking – Asst. Manager & above – QC Department

ACCOUNTABILITY

Head of the QC Department  

PROCEDURE

• Receipt Of New Column:

1. 1. On receipt of new column check the column as per indent.
   2. Check the column for any physical damage which may occur during its transportation. 
   3. Check the column with respect to the certificate of analysis received from the supplier for its make, type, Serial number, dimension etc.
   4. Verify that the column and the instrument fittings are compatible.
   5. In case of any non compliance with the user requirement, the column should be returned to the supplier.
   6. If case of compliance of user requirements, make the entry of the column.

Issuance and Performance of Column.

1. 1. The column will be issued according to the requirement as mentioned in the standard test procedure of that particular product. 
   2. Upon issuance of the column, the column will be tested to meet the system suitability criteria of the product for which it has been issued. 
   3. If the column meets the system suitability criteria, attach the printout of the chromatogram showing the system suitability criteria along with the certificate of analysis received form the supplier and maintain the record.
   4. In case the column does not meet the system suitability criteria then the column is to be returned back to the concerned person and should be tested for its performance on any other product. 
   5. If the column fails to meet the system suitability parameter repetitively, then it should be returned to the supplier.
   6. Assign column ID to column successfully meeting the system suitability criteria, as per the procedure described in point 5.3 and mark its entry.
   7. Once the column is in use, maintain column history card of its usage.
2. Column ID:
   1. Each column will be assigned a unique ID. The ID will be in the format:  

                             C-XXX/YY

XXX indicates serial no. in series starting from 001.

                                      YY  indicates last two digit of current year.

• Column Regeneration for Reverse Phase Column

1. Flush the column using warm water (55oC) at flow rate of 0.7 ml/min. for 10 minutes .Maintain      the column at (55oC) in column oven during flushing.     
2. Inject 100 µl of DMSO four times at interval of 2 minutes.
3. Flush it with water for at least 30 min. 
4. Flush it with methanol at flow rate of 0.7 ml/min in reverse direction for 60 minutes.
5. After flushing with methanol, flush the column with chloroform for 60 minutes at flow rate of 0.7 ml /min in reverse direction. 
6. At the end, wash the column in methanol for 60 minutes at flow rate of 0.7 ml/min. in reverse direction.  
7. After regeneration of column, perform the system suitability test as per the standard test procedure of the concerned product.  
8. Store the column in methanol, after flushing with methanol for 20 minutes at flow rate of 1.0 ml/min. in normal direction.
9. Make the entry of regeneration in the column history card. 

• Column Regeneration for Normal Phase Column    

1. Flush   the column with Hexane for 60 minutes at flow rate of 2.0 ml/min.
2. After flushing with Hexane, wash the column with Methylene chloride for 60 min at flow rate of 2.0 ml/min.       
3. After flushing with methylene chloride, wash the column with Isopropyl alcohol for 60 minutes at flow rate of 2.0 ml/min. 
4. At the end, flush the column with methanol for 60 minutes at flow rate of 2.0 ml/min. 
5. After regeneration of column, perform the system suitability test as per the standard test procedure of the concerned product.  
6. Store the column in Hexane, after flushing with Hexane for 20 minutes at flow rate of   2.0   ml/min in normal direction.
7. Make the entry of regeneration in the column history card. 

• Storage condition for HPLC columns

Withdrawal of Column:

      Withdrawal of column from further analysis is done in case:

1. Any physical damage.
2. Failure to comply the system suitability requirement as per specification after regeneration.
3. In case of dedicated column, upon regeneration if the column is not complying the system suitability criteria of that product but can pass system suitability criteria of any other product than continue using the column for that vary product with the approval of Head of Department. 
4. Indicate the withdrawal of the column in the remarks column.

5.  After withdrawal of column bent it and paste the label of Discarded, keep this column in specified place for discarded column.

About the Author

Author

Administrator

"A seasoned professional with a Master's in Chemistry, the author brings more than two decades of expertise in Quality Control, Analytical Method Validation, and advanced QMS investigation. Their experience is globally validated by successful management of major regulatory audits, including those conducted by the US, EU, Brazil, and Russian authorities."

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