suspension using UV-Visible Spectrophotometry or High Performance Liquid Chromatography (HPLC).

2. Scope

This SOP is applicable to the Analytical Method Development and Quality Control departments responsible for the development, validation, and routine application of assay methods for oral granules to ensure dose uniformity after reconstitution.

3. Responsibilities

• Analytical Scientist: Develops and validates the assay method, selects appropriate diluents and instrumentation.
• QC Analyst: Executes routine testing using validated methods.
• QA Executive: Reviews validation documentation and ensures method compliance with regulatory guidelines.

4. Accountability

The Head of Analytical Method Development is accountable for ensuring that assay methods provide accurate, precise, and specific quantification of APIs in granules for suspension, both pre- and post-reconstitution.

5. Procedure

5.1 Sample Preparation

1. Weigh an amount of granules equivalent to one dose (as per label claim).
2. Disperse in a suitable diluent (e.g., purified water, buffer) and sonicate for 15–30 minutes.
3. Filter the suspension through 0.45 µm membrane filter to remove insoluble excipients.
4. Dilute the filtrate to a known volume and analyze.

5.2 Standard Preparation

1. Prepare a stock solution of the API reference standard using the same diluent.
2. Prepare a working standard solution at target concentration matching sample strength.

5.3 Method Selection and Optimization

• HPLC (Preferred): Suitable for APIs with low UV absorbance or potential degradation.
• UV-Vis Spectrophotometry: Acceptable for stable APIs with distinct λmax values.

5.4 Typical HPLC Conditions

• Column: C18, 250 mm × 4.6 mm, 5 µm
• Mobile Phase: Phosphate buffer: Acetonitrile (60:40) or as optimized
• Flow Rate: 1.0 mL/min
• Detection: UV at suitable wavelength (e.g., 230–280 nm)
• Injection Volume: 20 µL

5.5 Validation Parameters

1. Specificity: No interference from excipients or granule matrix.
2. Linearity: r² ≥ 0.999 over a range of 50% to 150% of label claim.
3. Accuracy: Recovery between 98%–102% at three concentration levels.
4. Precision: RSD ≤ 2.0% for assay repeatability and intermediate precision.
5. Robustness: Evaluate impact of small changes in wavelength, flow rate, or pH.

5.6 Acceptance Criteria

• The % assay of the API should fall within 90%–110% of the label claim.
• Recovery of spiked samples should be between 98%–102%.

6. Abbreviations

• HPLC: High Performance Liquid Chromatography
• UV: Ultraviolet
• RSD: Relative Standard Deviation
• API: Active Pharmaceutical Ingredient

7. Documents

1. Granule Assay Raw Data Log – Annexure-1
2. Chromatograms or Spectra Summary – Annexure-2
3. Validation Summary Report – Annexure-3

8. References

• ICH Q2(R1): Validation of Analytical Procedures
• USP <621>: Chromatography
• USP <851>: Spectrophotometry and Light Scattering

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name	Anjali Sharma	Rahul Pillai	Sunita Reddy
Designation	Method Developer	QA Reviewer	Head – AMD
Department	Analytical Method Development	QA	Analytical Method Development

11. Annexures

Annexure-1: Granule Assay Raw Data Log

Sample ID	% Assay	RSD (%)	Status
GFS-286-01	99.2%	1.5	Pass

Annexure-2: Chromatograms or Spectra Summary

Includes HPLC chromatograms or UV spectra for standard and test samples. All system suitability parameters met.

Annexure-3: Validation Summary Report

The developed method for granules for suspension was validated across ICH Q2(R1) criteria and showed acceptable linearity, accuracy, precision, and robustness. Method suitable for batch release and stability testing.

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
21/05/2025	2.0	Expanded to include UV and HPLC options with validation requirements	Annual Review	Sunita Reddy
20/09/2022	1.0	Initial SOP Release	New SOP	QA Head