to Quality Control personnel performing assay testing of all sterile injectable formulations produced in the facility using validated analytical methods such as HPLC, UV, or titration.

3. Responsibilities

• QC Analyst: Responsible for executing assay testing and recording data accurately.
• QC Reviewer: Responsible for verifying calculations, chromatograms, and final reports.
• QA Officer: Responsible for batch release approval based on results.

4. Accountability

The Head of Quality Control is accountable for ensuring that assay testing is carried out in accordance with this SOP and applicable regulatory standards.

5. Procedure

5.1 General Guidelines

1. Perform assay testing as per the approved specification and validated test method.
2. Ensure all reagents and instruments are within calibration and validity dates.
3. Follow ALCOA+ principles for data integrity and GMP compliance.

5.2 Sample Preparation

1. Receive samples from production or stability in double-sealed condition.
2. Prepare sample solution as per the defined method—dilution, filtration, or extraction.
3. Use Class A volumetric glassware and calibrated balances.

5.3 HPLC Method (if applicable)

1. Set the chromatographic conditions as per the method (e.g., mobile phase, flow rate, wavelength).
2. Inject system suitability standard and confirm parameters (theoretical plates, tailing factor, RSD).
3. Inject sample and standard preparations in duplicate or triplicate.
4. Calculate the assay by comparing the peak area of sample vs. standard.

5.4 UV Spectrophotometry Method (if applicable)

1. Zero the instrument using blank solution.
2. Measure absorbance of standard and sample solutions at specified wavelength.
3. Calculate concentration based on standard curve or absorbance ratio.

5.5 Titration Method (if applicable)

1. Prepare the sample and titrant as per the method SOP.
2. Titrate using indicator or potentiometric endpoint.
3. Calculate API content based on titrant volume and equivalence factor.

5.6 Documentation and Calculations

1. Record raw data, instrument printouts, chromatograms, and calculations.
2. Fill the Assay Testing Report (Annexure-1).
3. Attach Certificate of Analysis (Annexure-2) for batch review.

5.7 Acceptance Criteria

API content must be within ±5% of the labeled claim unless otherwise specified in pharmacopoeial monograph or regulatory filing.

6. Abbreviations

• API: Active Pharmaceutical Ingredient
• HPLC: High Performance Liquid Chromatography
• UV: Ultraviolet Spectrophotometry
• ALCOA: Attributable, Legible, Contemporaneous, Original, Accurate
• SOP: Standard Operating Procedure

7. Documents

1. Assay Testing Report – Annexure-1
2. Certificate of Analysis (CoA) – Annexure-2
3. Instrument Calibration Log – Annexure-3

8. References

• USP General Chapter <621>, <731>, <851>
• ICH Q2(R2): Validation of Analytical Procedures
• 21 CFR Part 211 – Current Good Manufacturing Practice

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation	QC Analyst	QC Reviewer	Head QA
Department	Quality Control	Quality Control	Quality Assurance

11. Annexures

Annexure-1: Assay Testing Report

Batch No.	Product	Test Method	Result (%)	Tested By
AMP-2025	Ampicillin Injection	HPLC	99.4	Neha Reddy

Annexure-2: Certificate of Analysis (CoA)

Product	Batch No.	API Assay	Complies	Released By
Meropenem Injection	MER-025	101.2%	Yes	Sunita Patil

Annexure-3: Instrument Calibration Log

Date	Instrument ID	Type	Calibrated By	Status
17/06/2025	HPLC-03	HPLC	Rajesh Kumar	Pass

Revision History

Revision Date	Revision No.	Details	Reason	Approved By
05/01/2022	1.0	Initial version	New SOP	QA Head
18/06/2025	2.0	Revised limits and included UV method	Periodic Review	QA Head