SOP for System Suitability Test in HPLC analysis and HPLC Documentation

OBJECTIVE

To describe the procedure to be followed in HPLC analysis and documentation and to ensure that Good Laboratory Practices are followed in the HPLC analysis. 

SCOPE

This SOP is applicable for the procedure to be followed in HPLC analysis and documentation and to ensure that Good Laboratory Practices are followed in the HPLC analysis. 

RESPONSIBILITY

QC executive is responsible for performing HPLC analysis. 

QC Manager is responsible for ensuring the implementation of SOP in HPLC analysis and its documentation. 

PROCEDURE

• A sequence shall be available for analysis before the instrument is startup.

Mobile Phase:

• The mobile phase shall be prepared as per the composition described in the monograph of respective products.
• The mobile phase container shall have labeling details as follows:

1. Mobile phase :
2. Product Name :
3. Prepared by :
4. Use before:

• Details like the weight of buffer(s), Observed pH, etc. for the mobile phase shall be recorded in the datasheet.
• The mobile phase shall be discarded if any haziness or precipitation is found upon visual examination.

Standard Preparation: 

The standard preparation shall be prepared as per the respective monograph. 

Analysis:

• The injection sequence shall be in the following sequence to be used: Blank → System suitability → Placebo (if required) → Standard → Sample → System suitability (at the end of analysis).
• The Standard shall be injected in five replicates and the sample is required to be injected in duplicate.
• The system suitability (once established) shall be valid for a maximum period of 24 hours.

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• System suitability testing should be performed before the injection of samples. No sample analysis is acceptable unless the requirements of system suitability have been met. Sample analyses obtained while the system fails requirements are unacceptable.
• The RSD of five standard injections and System suitability injections shall be NMT 2%.
• During analysis, the flow rate of the mobile phase shall be kept constant for the entire run after the system suitability is established. If the flow rate of the system flows for more than 200 minutes, fresh system suitability shall be established.
• In case the above-mentioned acceptance criteria are not met, all the data collected during the suspect time period shall be properly identified and reviewed by the supervisor. The system suitability shall be carried by out all over again, before injecting any test samples.

Recording Chromatogram:

• All chromatograms before the establishment of system suitability and up to the entire run shall be recorded and documented.
• In the case where the chromatogram needs disregarding, the analyst shall show it to the supervisor for review and approval.
• The disregarded chromatogram shall be checked and certified by the supervisor.

• The chromatogram, which is disregarded and not considered for calculation, shall be stamped as “DISREGARDED”. The reason for disregarding the chromatogram could be a variation in the area count / inconsistent area, faulty integration, or abnormal drift in the baseline. Ghost peak or any other reason.

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• The analyst performing the analysis shall assign the reason of disregarding a chromatogram on the chromatogram itself. The disregarded chromatogram shall be filled along with the test chromatogram.
• For the system-generated chromatogram, the necessary information shall be printed on each chromatogram.
• The integration parameters such as peak width, peak threshold, minimum peak area, and height shall be recorded, as used for integration of the chromatogram.
• Reprocessing of the chromatogram, if necessary at a letter date /time shall be documented with the reason (s) for reprocessing and certified by the QA Manager.

Calculation 

• The calculation shall be performed as per the respective monograph. All calculations shall be as per the area obtained from the standard injected in the beginning. The area count of the in-between injection of standard (System Suitability) shall not be considered for calculation.

ABBREVIATIONS

SOP – Standard Operating Procedure 

QC – Quality Control 

QA – Quality Assurance 

HPLC – High-Performance Liquid Chromatography

ANNEXURE

Nil

REVISION HISTORY
Nil

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