Analytical Method Development: HPLC Dissolution Profile Method Development – V 2.0

SOP for Development of HPLC Method for Dissolution Profile Evaluation

Department	Analytical Method Development
SOP No.	SOP/AMD/076/2025
Supersedes	SOP/AMD/076/2022
Page No.	Page 1 of 14
Issue Date	19/05/2025
Effective Date	20/05/2025
Review Date	19/05/2026

1. Purpose

This SOP describes the procedure for developing High Performance Liquid Chromatography (HPLC) methods for the quantitative analysis of drug release in dissolution studies. It is intended to ensure reproducible and accurate profiling of drug dissolution over multiple time points, meeting regulatory standards.

2. Scope

This SOP is applicable to the Analytical Method Development (AMD) department and covers HPLC method development for dissolution profiling of tablets, capsules, and other solid oral dosage forms across multiple media types (e.g., acidic, buffer, water).

3. Responsibilities

Analytical Chemist: Performs method trials, prepares calibration standards, collects dissolution aliquots, analyzes samples, and compiles data.
Method Reviewer: Reviews chromatographic data, ensures proper profiling accuracy and precision, and verifies method suitability.
QA Officer: Ensures that the method complies with pharmacopeial and ICH guidelines and that documentation is complete.
Head – AMD: Approves the method for use in formal dissolution studies and regulatory submissions.

4. Accountability

The Head of Analytical Method Development is accountable for the scientific and regulatory soundness of HPLC dissolution profile methods before validation and transfer.

5. Procedure

5.1 Pre-Development Considerations

Review formulation characteristics and expected release mechanism (immediate, delayed, sustained).
Select appropriate dissolution media and apparatus as per pharmacopoeia (USP/BP/IP).
Define dissolution time points (e.g., 5, 10, 15, 30, 45, 60 min or extended to 24 hours for sustained release).
Document initial plan in Annexure-1: Dissolution Method Planning Sheet.

5.2 Chromatographic Condition Selection

Choose a reversed-phase column (C18 preferred) with appropriate dimensions (e.g., 150 mm × 4.6 mm, 5 µm).
Select a UV detection wavelength based on API λmax.
Mobile phase:
- Commonly water with acetonitrile/methanol; use buffer if pH control is required.
- Mobile phase pH should match the dissolution medium or optimize peak shape.
Prepare mobile phase and degas. Record in Annexure-2: Mobile Phase Preparation Log.

5.3 Standard and Sample Preparation

Standard:
- Prepare stock solution of API in diluent.
- Dilute to expected concentrations covering 80–120% of expected release.
Sample:
- Withdraw dissolution samples at defined time points.
- Filter samples immediately to avoid degradation.
- Use autosampler-compatible vials labeled with time point and ID.
Document in Annexure-3: Sample and Standard Preparation Log.

5.4 Chromatographic Evaluation

Inject blank, standard, and dissolution samples sequentially.
Check peak shape, retention time, and area consistency.
Ensure no interference at API RT in blank and placebo samples.
RSD of peak area from replicate injections ≤ 2.0%.
Document results in Annexure-4: Chromatographic Evaluation Sheet.

5.5 Linearity and Calibration

Prepare at least 5 calibration levels (e.g., 50, 75, 100, 125, 150% of expected concentration).
Plot concentration vs peak area and determine R² value (≥ 0.999).
Calculate sample concentration using calibration curve equation.
Record in Annexure-5: Calibration Curve and Linearity Log.

5.6 Dissolution Profile Plotting

Calculate % drug release at each time point using labeled claim.
Prepare time vs % release profile using spreadsheet or software (e.g., Excel, Empower).
Overlay profiles for different batches or media types if applicable.
Record final profile in Annexure-6: Dissolution Profile Summary.

5.7 Specificity and Robustness

Check placebo interference at API RT.
Assess peak purity using PDA if available.
Test small variations in mobile phase composition, flow rate, or column temperature.
Document observations in Annexure-7: Method Specificity and Robustness Log.

6. Abbreviations

HPLC: High-Performance Liquid Chromatography
API: Active Pharmaceutical Ingredient
RSD: Relative Standard Deviation
PDA: Photodiode Array
SOP: Standard Operating Procedure

7. Documents

Dissolution Method Planning Sheet – Annexure-1
Mobile Phase Preparation Log – Annexure-2
Sample and Standard Preparation Log – Annexure-3
Chromatographic Evaluation Sheet – Annexure-4
Calibration Curve and Linearity Log – Annexure-5
Dissolution Profile Summary – Annexure-6
Method Specificity and Robustness Log – Annexure-7

8. References

ICH Q2(R1) – Validation of Analytical Procedures
USP General Chapter <711> – Dissolution
USP <621> – Chromatography
FDA Guidance for Industry – Dissolution Testing of Immediate Release Solid Oral Dosage Forms

9. SOP Version

Version: 2.0

10. Approval Section

	Prepared By	Checked By	Approved By
Signature
Date
Name
Designation
Department

11. Annexures

Annexure-1: Dissolution Method Planning Sheet

Product	Media	Apparatus	RPM	Time Points
XYZ Tablet	900 mL 0.1N HCl	USP II (paddle)	50	5, 10, 15, 30, 45, 60

Annexure-2: Mobile Phase Preparation Log

Aqueous Phase	Organic Phase	pH	Filtered	Prepared By
Water (10 mM phosphate)	Acetonitrile	3.0	Yes	Sunita Reddy

Annexure-3: Sample and Standard Preparation Log

ID	Type	Concentration	Diluent	Filtered	Prepared By
STD-001	Standard	100 µg/mL	Media	Yes	Rajesh Kumar

Annexure-4: Chromatographic Evaluation Sheet

Sample ID	RT	Area	Tailing	Resolution
T5	3.42	145689	1.12	2.8

Annexure-5: Calibration Curve and Linearity Log

Level	Concentration	Area
50%	50 µg/mL	78420
75%	75 µg/mL	115634
100%	100 µg/mL	151204

Annexure-6: Dissolution Profile Summary

Time (min)	% Release
5	18%
15	46%
30	72%
60	98%

Annexure-7: Method Specificity and Robustness Log

Parameter	Variation	Observation	Status
Flow Rate	±0.1 mL/min	No significant change	Pass

Revision History:

Revision Date	Revision No.	Details	Reason	Approved By
04/05/2025	2.0	Expanded annexures, added specificity and robustness checks	Annual Review