the results and approving the batch for release.

5. Procedure

5.1 Sample Collection

1. Collect a representative sample of tablets from the batch, as specified in the batch record.
2. The sample should consist of a minimum of 6 tablets, or as specified in the batch record or pharmacopeial guidelines.
3. Ensure that the tablets are free from defects such as cracks, chips, or contamination.
4. Label the sample appropriately for identification during testing.

5.2 Preparation of Sample and HPLC Equipment

1. Ensure the HPLC equipment is clean, calibrated, and ready for use.
2. Set the HPLC system according to the specified method, ensuring the appropriate mobile phase, column, and detector settings as outlined in the batch record.
3. Weigh the required amount of tablets (typically 10–20 tablets) and crush them to a fine powder.
4. Weigh an appropriate amount of the powdered tablets for dissolution in a suitable solvent, such as methanol or water, depending on the solubility of the drug.
5. Prepare the sample solution, ensuring that the tablet content is completely dissolved and filtered to remove any insoluble particles.

5.3 Calibration of HPLC System

1. Prepare a calibration standard solution by dissolving a known quantity of the API in a solvent to create a standard concentration.
2. Inject the standard solution into the HPLC system and record the chromatogram. Ensure that the retention time of the API matches the expected value for that particular method.
3. Plot a calibration curve using the standard concentrations and their respective peak areas or heights. Ensure that the linearity is acceptable (typically R² > 0.99).

5.4 Performing HPLC Testing

1. Inject the prepared sample solution into the HPLC system.
2. Record the chromatogram and compare the retention time of the sample with that of the standard.
3. Quantify the drug content by calculating the area of the API peak in the chromatogram and comparing it with the calibration curve generated from the standard solution.
4. Ensure the API content falls within the specified range (usually 95%–105% of label claim) as outlined in the batch record or pharmacopeial guidelines.

5.5 Data Recording and Calculation

1. Record the results, including peak area, retention time, and the calculated concentration of the API, in the batch record (Annexure-1).
2. Calculate the percentage of the drug content using the following formula:
• % Drug Content = (Calculated Concentration / Label Claim Concentration) × 100
3. Ensure that the calculated drug content meets the acceptance criteria for uniformity.

5.6 Acceptance Criteria

1. The drug content should be within the acceptable limits specified in the batch record or pharmacopeial guidelines (typically ±5% of the label claim).
2. If the drug content falls outside the acceptable range, investigate and document the findings in the deviation report (Annexure-2).
3. Take corrective actions as necessary and perform re-testing if required.

5.7 Documentation and Record-Keeping

1. Document all test results, including chromatograms, calculations, and observations, in the batch record (Annexure-1).
2. Ensure that all records are signed, dated, and stored according to the company’s record retention policy for future audits and inspections.
3. Maintain all chromatograms, calibration curves, and raw data for future reference and audits.

5.8 Post-Test Cleanup

1. Clean all HPLC equipment, including syringes, injection ports, and sample vials, according to the cleaning SOP to prevent contamination between tests.
2. Ensure the HPLC system is properly maintained and calibrated for future use.

6. Abbreviations

• SOP: Standard Operating Procedure
• GMP: Good Manufacturing Practice
• QC: Quality Control
• QA: Quality Assurance
• API: Active Pharmaceutical Ingredient
• HPLC: High-Performance Liquid Chromatography
• RSD: Relative Standard Deviation

7. Documents

1. Batch Record (Annexure-1)
2. Deviation Report (Annexure-2)

8. References

• USP <621> – Chromatography
• 21 CFR Part 211 – Current Good Manufacturing Practice for Finished Pharmaceuticals (US FDA)
• European Pharmacopoeia (EP) – HPLC Testing

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Batch Record

Batch Number	Tablet Sample	Sample Weight (g)	API Content (%)	Result
Batch 001	Tablet Sample	0.5	99.2%	Pass

Annexure-2: Deviation Report

Deviation Date	Batch Number	Deviation Description	Corrective Action	Responsible Person
15/12/2025	Batch 001	API content out of specification	Reworked formulation and adjusted mixing process	Jane Doe

Revision History:

Revision Date	Revision No.	Revision Details	Reason for Revision	Approved By
01/01/2024	1.0	Initial Version	New SOP Creation	QA Head
01/02/2025	2.0	Updated Testing Procedures	Refined HPLC testing protocols	QA Head