HPLC Column Management; Purpose

The tenacity of this SOP is to define proper controlling of all HPLC columns during receive, use and storage period to confirm that the HPLC column performs satisfactorily and gives reproducible results.

HPLC Column Management; Scope

This process is applicable for stock usage of HPLC columns required for Materials and Products analysis in quality control laboratory of general block of XX Pharmaceuticals Limited.

Definitions / Abbreviation:

[] QC: Quality Control.

[] HPLC: High performance liquid chromatography.

[] CoA: Certificate of analysis.

Responsibilities:

The roles and responsibility is as follows

Officer/Executive/Senior Executive, Quality Control

• To confirm that this procedure is followed.
• To maintain the records properly as per SOP.

Assistant Manager, Quality Control

• To confirm that this procedure is kept up to date.
• To ensure that the SOP is technically sound & reflects the required working practices.
• Organize training on the SOP to all relevant personnel.
• To confirm implementation of the SOP after training.
• Schedule calibration of the instrument at the defined intervals.

Head of Quality Assurance

• Initiative to take approval of the SOP.
• To confirm overall implementation of this SOP.

Procedure:

• Preserve column with appropriate end fitting during storage.
• Prevent the column from any type of mechanical shocks.
• Always use the column in the indicated direction.
• Clean solvent reservoir filter in the mobile phase by sonication in the Methanol for 30 minutes, when required.
• Test sample should be free from any type of particulate matter.
• Stock review and raise requisition

Executive, QC will raise a purchase requisition considering the following elements:

• Product & Method wise dedicated column.
• New method related to Product Development which is in product launch pipeline for commercialization.
• New Pharmacopoeia (BP, USP & EP) requirements.
• Underprivileged system suitability performance of a column that has been triggered from HPLC column usage register and included in retired HPLC column register.
• Executive, QC will make a list for purchasing of new columns on the basis of purchase requisitions to ensure uninterrupted HPLC analysis.
• In-Charge, QC will finally review the list for purchasing of new columns by checking the HPLC column stock and issue and retired HPLC column register status or considering column requirement for new product.

Column receiving

• Executive, QC will receive new column with CoA.
• After receiving a new HPLC column, QC Executive will check the column CoA, specification & physical condition and compare with Purchase Order and Invoice description.
• If complies with ordered description, then assign an individual column serial number (e.g. GQC-H001, where H represents HPLC column and 001 is the serial number) and record necessary information [Column receiving date, Column description (Name, brand, particle size and dimension etc.), Part number/ Catalogue, Manufacturer name, Supplier name, Column Sl. No., Column dedicated for (Product /Material), Column issue date etc.] in the HPLC column stock and issue register.

Column performance check for new column: HPLC Column Management

• Check availability of reference reagents /standards required for performance test. If unavailable, then promptly raise requisition.
• Carry out the column performance test for new HPLC column, as per manufacturer’s instructions. (If particular chemicals as per Manufacturer’s CoA are available.)
• Check column with respect to Manufacturer’s CoA for retention time, theoretical plates using the material provided with the column & method specified by manufacturer.
• In case of emergency carry out performance check only with the available reference standards/reagents.
• Connect the column to HPLC system.
• Flush (condition) specific column with shipping solvent (the details available with manufacturer’s CoA) or methanol for at least 30 minutes with a flow rate of 0.5 ml/min.
• Flush column with mobile phase as mentioned in the technique for respective columns.
• Prepare test solution in mobile phase as recommended in the procedure.
• After column is conditioned, inject 20 µl or as per manufacturer’s CoA & record chromatograph of test solution.
• Calculate tangent (theoretical plates) as per the USP method. The result should meet system suitability parameters.

Theoretical plate: (N) = 16 (t/W)2

Asymmetry (10%) = w/2f

Where, t = Retention time

W = Width of peak measured by extrapolating the relatively straight sides to the baseline

W = Width of peak at 10% of peak height

f = Distance between perpendicular dropped from apex of the peak and leading edge of peak at 10% of peak height

• Keep records of the performance check on the HPLC Column Performance Check Record (as per Annexure-IV).
• Accept column for use if observed values are within limit specified in manufacturer’s CoA.
• Inform manufacturer through Purchase Department, if test result doesn’t comply with the accepted limit mentioned in the manufacturer’s CoA.
• If problem is not resolved, reject same & information Purchase Department regarding rejection.

Material/Product Dedicated Column

• In-Charge, QC will ensure method and/or product wise dedicated columns are available for HPLC analysis.
• Same column can be used for analysis of two or more products in different dosage form where product formulation & active ingredients are same. But prior to use, method shall be validated using same column.
• If dedicated column is not available, instantly discuss with In-Charge, Quality Control.
• Column other than material and/or product dedicated column can be used in case of emergency with the concern of In-Charge, Quality Control.

Column Issue

• Prior to issue a new column, review previous column history & record all information [Initial theoretical plates (Before issue), Column issue date, Issued by, Remarks] in the HPLC column stock and issue register.
• If dedicated column is not available for a new material/product analysis, discuss the matter with In-Charge, QC for alternate decision.
• All decisions shall be recorded in HPLC column stock and issue register (Annexure-I).
• After issue, record the column information in a new page in HPLC Column usage register. (Annexure-II).

Column Care

• Confirm that the integrity of the column packing is not be disturbed by needless opening of the column ends.
• Handle HPLC columns as per manufacturers’ instructions, e.g. avoidance of physical shock, vibration and storage at high temperature.
• For silica-based columns, avoid eluents outside the pH range of 3-8 where possible, except when column manufacturer specifications acclaim otherwise.
• Wash column as per HPLC column washing procedure.
• After use, wash the column to make it free from buffer salts with a suitable solvent (typically 1:1 aqueous/organic solvent for reversed phase columns) about 45 to 60 minutes maintaining a flow rate of 1.5 to 2.0 ml per minute.
• After washing cap or seal column for storage in designated place.
• In case of poor system suitability performance or column blockage caused by precipitation of buffer salts, always incorporate a flush with a solvent/water mixture before switching to pure solvent as per HPLC analysis and system suitability check procedure SOP No.: (SOP/QC/009).
• Record the HPLC column flushing time in HPLC log book. Maintain HPLC column cleaning and storage as per manufacturer’s instructions.

Regeneration of a column

• If the system suitability fails only then do the column regeneration.
• For columns C6, C4, C18, C8, Phenyl, Amino, Polymeric follow the sequence given below with a 1.0 ml flow per minute flow rate.
• Flush with filtered and de-gassed water for 30 minutes. Inject 20 µL of 1.0% of Acetic acid in purified water.
• Flush with Methanol for 30 minutes, inject 20 µL of 1.0% Dimethyl Sulphonic Oxide (spectroscopic grade) in Methanol.
• Flush with

1. Acetonitrile for 15 minutes;
2. Chloroform for 10 minutes;
3. Methanol for 30 minutes sequentially;
4. Isopropanol for 15 minutes;
5. Purified water for 15 minutes

• Check pH of water at inlet and outlet, which should be identical.
• Mobile phase for the compound to be analyzed.

 

Retired HPLC Column

• After washing, if the HPLC column performance is not improved then In-Charge, QC will review column washing process and column history prior to discarding/retiring a column.
• The column retiring information must be recorded in the respective page of HPLC column usage log sheet (Annexure-II) and retired column register (Annexure-III).
• Keep all retired columns under lock & key in a separate locker in the lab area. Before keeping analyst must ensure that the retired column has been separated from usage stock and de-shaped.
• Periodically the HPLC column usage log sheet and retired column register shall be reviewed and retired column shall be destroyed as per column destruction process and manufacturers’ instructions.
• Retired column storage period should not be less than last test product shelf-life plus one year.

Download All Annexure: HPLC Column Management

Annexure I HPLC Column Stock and Issue Register

Annexure II HPLC Column Usage Register

Annexure III Retired HPLC Column Register

Annexure IV HPLC Column Performance Check Record