component is accurately quantified without interference.

2. Scope

This SOP applies to the Analytical Method Development and Quality Control departments for assay method development, validation, and routine analysis of multicomponent tablets used in the pharmaceutical industry.

3. Responsibilities

• Analytical Scientist: Develops and validates the assay method for each API in the multicomponent formulation.
• QC Analyst: Performs routine assay testing using the validated method.
• QA Executive: Ensures documentation review, regulatory compliance, and final approval.

4. Accountability

The Head of Analytical Method Development is accountable for ensuring that the assay method is validated as per ICH guidelines and capable of simultaneously quantifying all actives in the formulation.

5. Procedure

5.1 Selection of APIs and Formulation Matrix

• Identify and list all APIs with known potency and excipients used in the formulation.
• Check compatibility and solubility of APIs in the selected diluent.

5.2 Sample Preparation

1. Weigh 10 intact tablets and determine average weight.
2. Crush tablets and weigh equivalent to one dose in a volumetric flask.
3. Add diluent (e.g., 0.1N HCl:Acetonitrile 60:40) and sonicate for 30 minutes.
4. Make up to volume, filter through 0.45 µm filter, and inject.

5.3 Standard Preparation

1. Prepare individual API stock solutions in the same diluent.
2. Combine APIs to prepare mixed standard solutions at 100% concentration.
3. Prepare linearity standards at 50%, 80%, 100%, 120%, and 150% for validation.

5.4 Chromatographic Conditions (Typical)

• Column: C18, 250 mm × 4.6 mm, 5 µm
• Mobile Phase A: Phosphate buffer (pH 3.0)
• Mobile Phase B: Acetonitrile
• Gradient: 0–10 min: 70% A; 10–20 min: 40% A
• Flow Rate: 1.0 mL/min
• Detection: UV at λmax of most sensitive API (e.g., 240–280 nm)

5.5 Validation Parameters

1. Specificity: Peaks of all APIs must be well resolved from each other and excipients.
2. Linearity: r² ≥ 0.999 for each API.
3. Accuracy: Recovery between 98%–102% at three levels (80%, 100%, 120%).
4. Precision: %RSD ≤ 2.0% for six replicates.
5. Robustness: Evaluate flow, pH, and wavelength changes.
6. System Suitability: Resolution ≥ 2.0 between APIs, tailing factor ≤ 2.0, theoretical plates ≥ 2000.

6. Abbreviations

• API: Active Pharmaceutical Ingredient
• HPLC: High Performance Liquid Chromatography
• RSD: Relative Standard Deviation
• QA: Quality Assurance

7. Documents

1. Multicomponent Tablet Assay Log – Annexure-1
2. System Suitability and Chromatogram Set – Annexure-2
3. Validation Summary Report – Annexure-3

8. References

• ICH Q2(R1): Validation of Analytical Procedures
• USP General Chapter <621>: Chromatography
• Ph. Eur. 2.2.29: Liquid Chromatography

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name	Rashmi Kothari	Arvind Narula	Sunita Reddy
Designation	HPLC Analyst	QA Reviewer	Head – AMD
Department	Analytical Method Development	QA	Analytical Method Development

11. Annexures

Annexure-1: Multicomponent Tablet Assay Log

Sample ID	API 1 (%)	API 2 (%)	API 3 (%)	Status
MCT-281-01	99.1	98.7	101.2	Pass

Annexure-2: Chromatogram and Suitability Report

Overlay chromatogram of multicomponent assay showing separation with resolution >2.5. All peaks meet system suitability criteria.

Annexure-3: Validation Summary Report

The HPLC assay method was validated for a combination of paracetamol, caffeine, and phenylephrine. All validation parameters including specificity, linearity, accuracy, precision, and robustness met ICH requirements.

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
21/05/2025	2.0	Updated gradient and validation limits for triple API formulations	Annual Review	Sunita Reddy
20/04/2022	1.0	Initial SOP Release	New SOP	QA Head