Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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Department Bioanalytical and DMPK
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SIGNATURES
Author Reviewed by QAU Approved by TFM
Name Dr.Kanaga Sabapathi S Mr.Sarauya Bhoumi Reddy Dr. T. Kumar
Signature
Date
Change History
Version
No. &
Description of change Reasons for the change
Revision
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Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
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Department Bioanalytical and DMPK
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CONTENTS
1. INTRODUCTION........................................................................................................3
2. scope...............................................................................................................................3
3. Abbreviations, Definitions and Synonyms...............................................3
4.1. Abbreviations...............................................................................................3
4. Responsibility............................................................................................................4
4.2. Definitions......................................................................................................4
4.3. synonyms........................................................................................................4
5. PROCEDURE................................................................................................................4
5.1. Receipt and Numbering Procedure of HPLC & GC Column 4
5.2. Handling and Maintenance of HPLC columns............................5
5.2.1.Usage of HPLC column......................................................................................5
5.3. Maintenance..................................................................................................5
5.4 Disposal of HPLC Column.................................................................................6
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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Department Bioanalytical and DMPK
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1. INTRODUCTION
The purpose of this SOP is to describe procedure for handling of HPLC analytical
columns used for bioanalytical and DMPK studies.
Column: Columns are stationary phase that are filled with adsorbent material leading
to the separation of the sample components.
2. SCOPE
This SOP is applicable to all personnel handling HPLC analytical columns in
bioanalytical and DMPK department.
3. ABBREVIATIONS
µ - micron
o
C - degree Centigrade
% - Percentage
> - greater than
Cm - centimeter
COA - Certificate of Analysis
COL - Column
DMF - Dimethylformamide
DMPK - Drug Metabolism and Pharmacokinetics
DMSO - Dimethyl sulfoxide
EHS - Environment health & safety
GC - Gas Chromatography
GEN - General
He - Helium
HPLC - High Performance Liquid Chromatography
ID - Identification
m - Meter
mm - millimeter
mL/min - milliliter per minute
N2 - Nitrogen
NA - Not Applicable
BA - Bioanalytical
pH - Negative log of base 10 Hydrogen Ion
QAU - Quality Assurance Procedure
SOP - Standard Operating Procedure
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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PBS - Palamur Biosciences
TFM - Test facility Management
V/V - Volume by Volume
4. RESPONSIBILITY
All personnel’s involved in high performance liquid chromatography based activities
update the procedures described in this sop. are responsible for the compliance and
implementation of this SOP.
TFM will ensure the procedure described in this SOP.
4.1. synonyms
NA
5. PROCEDURE
5.1. Receipt and Numbering Procedure of HPLC
An assigned personnel shall be responsible of all HPLC columns and also
responsible to maintain sufficient stocks of new columns.
At the time of receipt of HPLC column, information on stationary phase and
dimension of columns shall be verified against certificate of analysis such as
column name, serial number, dimension, particle size from the column
specification.
If there are any discrepancies observed on the receipt of column the same
shall be intimated to the vendor/supplier for further action.
After Column verification against COA found to be satisfactory, an in-house ID
shall be allotted for each HPLC column received in the following manner:
BA-COL-XXX-For HPLC Column
Where BA stands for Bioanalytical, COL stands for Column and XXX stands for
the serial number of the column, starting from the 001.
A logbook for Column shall be maintained consisting of “Column Index” and
“Column usage” as per Appendix-1 and Appendix-2.
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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“Column Index” shall be used to record the receipt details of Column which
will serve as index/master list and “Column usage” shall be used to record the
usage details of columns. Allot the first few pages of logbook for “Column
Index” (Appendix-1) and the rest of pages for “Column usage” (Appendix-2).
The details of every new column received shall be updated in the “Column
Index” logbook.
The COA of all HPLC columns shall be maintained in a separate file
Label the in house ID as mentioned above on the columns and on their
respective COA.
The user shall be responsible for entering the details of usage in the “Column
usage” as per Annexure-2.
Assign page numbers for each column in the column index log book and
furnish the details as per Appendix-1 of this SOP.
“COLUMN NOT IN USE” and “COLUMN DISPOSED” shall be updated in the
remarks section of the column usage logbook.
Give the reference number of old columns (such as logbook number, page
number) in new logbook, in case of re-entry in new logbook.
5.2. Handling of HPLC columns
5.2.1. Usage of HPLC column
The instruments/equipment will be calibrated in-line with respective operating
procedure.
HPLC column shall be used only in the pH range recommended by the
manufacturer.
After usage of HPLC column, wash it with MS grade water for about 30
minutes.
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
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Followed by water: methanol (50:50 %V/V) or water. Acetonitrile (50:50
%V/V) mixture
For about 10-20 minutes followed by 100 % organic solvent (15-20 minutes)
and shall be stored in designated area. If any specific instruction has been
provided by the manufacture for column maintenance, the same shall be
followed.
HPLC columns shall be regenerated as per manufacturer’s instructions. If
required.
5.3. Maintenance
The following points needs to be taken care during handling of HPLC Columns:
Never let the column dry out.
Always use miscible and compatible solvents for the respective instruments.
Prevent HPLC column from mechanical shock, heating more than 40 0C.
Do not overload or inject dirty samples.
While fitting the column do not apply excessive pressure that will damage the
frit and end capping.
Do not exceed the operating pressure as per manufacture’s recommendation.
While purging and cleaning of HPLC channels, remove the HPLC column and
use a union in place of that.
Always fix the column as per the flow direction unless cleaning is required.
Make sure that end plugs are firmly in place.
5.4 Disposal of HPLC Column
If the Column shows poor chromatography and performance such as high
pressure, broad peak, split peak or poor resolution, ghost peak, peak tailing
or fronting. Inconsistency in results, Column shall be washed and the
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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Department Bioanalytical and DMPK
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respective methods will be verified. Further if the results are not satisfactory
column regeneration procedure shall be followed as per manufacture’s
instructions.
Post Column regeneration if the results are satisfactory column can be further
used, if fails to reproduce the results Column shall be labelled as “COLUMN
NOT IN USE”. The column shall be disposed of as per Waste Management.
“COLUMN NOT IN USE” and “COLUMN DISPOSED” shall be updated in the
remarks section of the column usage logbook.
6. APPENDICES
APPENDIX -1 COLUMN INDEX LOGBOOK
APPENDIX-2 COLUMN USAGE LOGBOOK
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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APPENDIX -1 COLUMN INDEX LOGBOOK
Receive
Date Manufacturer’ Colum Page Remark
Colum Column d by
S.No of s Serial n In- Number s with
n Specificatio (Sign
. Receip Number/ Lot House s Sign &
Name n and
t No. ID Allotted Date
Date)
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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Department Bioanalytical and DMPK
Copy Number Effective Date
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
Title HANDLING OF LC COLUMNS
SOP No. SOP/PBS/BIA/016
Version No. Approval Date
SOP
Revision No. Next Review
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Department Bioanalytical and DMPK
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APPENDIX-2 COLUMN USAGE LOGBOOK
Column In-House Id
Column Name and Specifications:
Issued Checked by
Study no/ Returned by
Date (Sign & Remarks (Sign and
Purpose (Sign & Date)
Date) Date)
Author Reviewed by QAU Approved by TFM
Name Dr. Kanaga Sabapathi S
Signature
Date
