Handling of HPLC and GC Columns

Documentation and Management of Columns

Upon receipt of the new column, check the column for the physical damage, which may have occurred during shipping and column performance certificate.
An in-house column identification number shall be assigned to each HPLC column which shall be in the format of QC/LC/XXX. Where XXX shall indicate serial number starting from 001.
For example: A First HPLC column will be numbered as QC/LC/001
An in-house column identification number shall be assigned to each GC column which shall be in the format of QC/GC/YYY.

Where ,YYY shall indicate serial number starting from 001.

For example: A First GC column will be numbered as QC/GC/001
The in-house column number assigned to the HPLC/GC columns shall be affixed to the column by applying a label of the in-house column identification number to that particular column.
A separate log book shall be maintained for the numbering of new HPLC and GC columns for the identification.
Refer Attachment-1 for Format for Cover Page and Index of Column Log Book, Refer Attachment-2 for Format for HPLC/ GC column ID log
The log book needs to be updated by filling column ID and column description details.
The Column Performance Certificate received from the manufacturer along with the column shall be filed in a file bearing identification as “Column Performance Certificate”.
The in-house column identification number assigned to the column is affixed to the Column Performance Certificate by applying a label of the in-house column identification number with sign/date to respective Column Performance Certificate.
The in-house column identification number shall be recorded in the LNB each time the column is used for analysis as described in Good Documentation Practices (GDP).
HPLC & GC column usage shall be maintained in respective column usage log (Attachment-3).
The columns which can be used under reverse phase or normal phase conditions (i.e. –CN or -NH2), shall be dedicated. For example, if a cyano (-CN) column is issued for the reverse phase use, then it shall not be used for the normal phase.
It is recommended that the manufacturer’s instructions, if any, are to be followed prior to first use of column.
If the column performance is found to be unsatisfactory during the use, then its use shall be discontinued. The identified columns shall be discarded physically and same will be recorded in for Record for Column Discard.

Handling of Reversed phase column:

These columns are generally shipped in Acetonitrile / Water / Methanol.
When a reversed phase column is to be used for the first time, it shall be flushed with the solvent in which it is packed by the manufacturer or with mixture of acetonitrile/ methanol and water (90:10 %v/v), as it can dry out during stocking and shipping.
Flush the column with the flow rate of 0.5 mL/minute for 30 minutes.
After flushing the column with the organic solvent, flush the column with water, at 0.5 mL / minute flow rate for 30 minutes. After washing with water equilibrate the column with mobile phase until baseline is stable.

Handling of Normal phase column:

These columns are generally are shipped in heptane or hexane.
When a Normal phase column is to be used for the first time, it shall be flushed with the organic solvent (Heptane / Hexane or the solvent in which it is packed by the manufacturer) in which it has been shipped as it can dry out during stocking and shipping. Flush the column with the flow rate of 0.5 mL/minute for 30 minutes.
After this, flush the column with methanol, at 0.5 mL / minute flow rate for 30 minutes. After washing with methanol equilibrate the column with mobile phase until baseline is stable.

Post-analysis Column wash

Wash the reversed phase columns with 90:10 %v/v mixture of acetonitrile (ACN)/ methanol and water.
Wash the normal phase columns with Heptane, Hexane or Isopropyl Alcohol (IPA) for at least 30 minutes at a flow rate of 1.0 mL / min.
Disconnect the column from the HPLC system and seal both ends of the column with appropriate plugs/caps for storage.
If the column will be used again for the same analytical methods within 48 hours, the column may be stored in the mobile phase.

Conditioning of GC column

The conditioning of GC column is done by maintaining the column at 10°C below the maximum temperature of the column for 20 minutes.

Regeneration of HPLC/GC Columns

The analytical columns may need to be regenerated when the column efficiency does not comply with the system suitability requirements.
If specified by the manufacturer of the column or reference literature, the procedure for regeneration shall be followed.