HPLC Analysis for Rebamipide in Accordance with Japanese Pharmacopoeia

Importance of the Topic

Rebamipide is a widely used gastroprotective agent requiring rigorous purity testing in pharmaceutical quality control. Accurate analysis of related substances ensures safety, efficacy, and compliance with pharmacopoeial standards.

Objectives and Study Overview

This study aimed to develop and validate a high performance liquid chromatography method for determining rebamipide purity and related substances in accordance with the Japanese Pharmacopoeia 18th edition. An integrated HPLC system was used and system suitability tests were specified to confirm method performance.

Methodology and Instrumentation

The analysis was performed on an LC-2050C 3D HPLC system using a Shim-pack GIST C18 column (250 mm × 4.6 mm, 5 µm). The mobile phase consisted of sodium 1-decanesulfonate aqueous solution, methanol, and phosphoric acid mixed at 100:100:1 by volume. Column temperature was 40 °C, flow rate 1.0 mL/min, injection volume 10 µL, and detection by UV at 232 nm.

• Sample preparation involved dissolving 40 mg of rebamipide in sample preparation solvent and performing serial dilutions for standard and system suitability solutions.
• System suitability covered detectability, performance (resolution), and repeatability (six replicate injections).

Main Results and Discussion

In the detectability test, the diluted standard solution peak area ratio was 9.9 percent, meeting the 7–13 percent requirement. System performance yielded a resolution of 9.3 between rebamipide and 4-chlorobenzoate, exceeding the minimum of 8. The repeatability test showed a relative standard deviation of 0.12 percent, well below the 2 percent threshold. Chromatograms confirmed rebamipide o-chloro isomer and the debenzoyl impurity with relative retention times of approximately 0.53 and 0.68, respectively, aligning with pharmacopoeial specifications.

Benefits and Practical Applications

• The validated HPLC method enables reliable purity testing of rebamipide according to official standards.
• The integrated HPLC system simplifies routine quality control workflows in pharmaceutical laboratories.
• Shim-pack GIST C18 column provides effective separation of rebamipide from related substances.

Future Trends and Potential Applications

Emerging trends include ultra high performance liquid chromatography to reduce run time and solvent consumption, integration of mass spectrometric detection for enhanced sensitivity and impurity profiling, and automation of sample preparation and data analysis to improve throughput and reproducibility.

Conclusion

The described HPLC procedure using LC-2050C 3D and Shim-pack GIST C18 column meets the Japanese Pharmacopoeia criteria for detectability, resolution, and repeatability in rebamipide purity testing. The method provides a reliable and efficient tool for pharmaceutical quality assurance.

References

• Japanese Pharmacopoeia, 18th Revised Edition, The Japanese Pharmacopoeia Committee, 2024