Isolation of a degradation product of ranitidine hydrochloride using the Waters analytical scale purification system

Significance of the Topic

The reliable isolation and characterization of degradation products is essential in pharmaceutical development to ensure drug safety and compliance with regulatory standards. Forced‐degradation studies reveal potential impurities that can affect product quality and patient health.

Objectives and Study Overview

This study aimed to isolate a base‐hydrolysis degradation product of ranitidine hydrochloride using a preparative scale purification system. The workflow included forced degradation under alkaline conditions, scale‐up of an analytical method, and collection of the target impurity for purity assessment and structural analysis.

Methodology and Instrumentation

A forced degradation experiment was conducted on ranitidine drug substance and tablet formulations by treating 1 mg/mL solutions with 0.5 M NaOH at room temperature for up to 24 hours. Samples were neutralized, diluted to 0.1 mg/mL, and analyzed on an analytical HPLC system to detect degradation peaks. The analytical method employed a BEH C18 column (4.6 × 50 mm, 3.5 µm) with a water–methanol gradient (0.1% formic acid), monitored by PDA and QDa mass detectors. The method was scaled to preparative conditions using a 10 mm × 50 mm, 3.5 µm column, increasing flow to 8 mL/min at 30 °C and 7000 psi. Fractions were collected by UV and MS triggers.

Used Instrumentation

• Waters Analytical Scale Purification System with dual quaternary pumps
• Arc Premier HPLC with QSM-R, FTN-R, 2998 PDA Detector
• ACQUITY QDa II Detector for mass‐directed fraction collection
• XBridge Premier BEH C18 analytical column, 4.6 × 50 mm, 3.5 µm
• XBridge Premier Peptide BEH C18 preparative column, 10 × 50 mm, 3.5 µm
• Empower CDS 3.8.0, MassLynx 4.2, FractionLynx software

Main Results and Discussion

Forced degradation under basic conditions generated a prominent impurity with m/z 302. Preparative isolation yielded high‐purity material after collecting UV‐ and MS‐triggered fractions. Analytical reinjection of fractions demonstrated chromatographic purity between 96.3% and 97.5% and spectral homogeneity, confirming successful separation from coeluting components. A loading study showed that injection volumes up to 600 µL could be accommodated with minimal loss of purity.

Benefits and Practical Applications

• Efficient scale‐up from analytical to preparative HPLC without changing column chemistry
• Rapid isolation of degradation products for structural characterization and impurity profiling
• MS‐directed fractionation improves selectivity and reduces solvent usage
• Robust workflow supports QA/QC and regulatory submissions

Future Trends and Applications

Integration of high‐resolution mass spectrometry and automated fraction collection will further enhance impurity identification workflows. Continued development of greener solvents and smaller particle columns may improve throughput and sustainability. Predictive software tools will streamline scale‐up and method development.

Conclusion

The Waters Analytical Scale Purification System enabled the successful isolation of a base‐hydrolysis degradation product of ranitidine HCl with high purity. MS‐directed triggers and matched column chemistry ensured efficient scale‐up and reproducible fractionation, supporting detailed impurity characterization.

Reference

1. United States Pharmacopeia. General Chapter <1086> Impurities in Drug Substances and Drug Products. USP 40 and PF 41, 2015.
2. Dhangar KR, Jagtap RB, Surana SJ, Shirkhedkar AA. Impurity Profiling of Drugs Towards Safety and Efficacy: Theory and Practice. Journal of the Chilean Chemical Society. 62(2):2017.
3. Maziarz M, Cleary RP, Shave D, Harden SN, Rainville PD, Xia J. Isolation of a Degradation Product of Ranitidine Hydrochloride Using the Waters Analytical Scale Purification System. Waters Application Note 720008785, 2025.
4. Waters Technologies Corporation. Waters Preparatory ODB Columns Calculator. 2025.