Development and application of a validated stability-indicating Ultra-Performance Liquid Chromatography (UPLC) method for the determination of dantrolene and its related impurities

Abstract

A rapid, sensitive, and specific ultra-performance liquid chromatographic (UPLC) method was developed for the simultaneous determination of dantrolene and its potential degradation impurities. Chromatographic separation was achieved on a Waters Acquity UPLC system using a Waters BEH C₁₈ (2.1 × 100 mm, 1.7 µM) analytical column and Waters BEH C₁₈ (2.1 × 5 mm, 1.7 µM) guard column. The compounds were eluted with a linear acetonitrile gradient (25–75%) over 3 min with a buffer composition of sodium acetate for method development, quantitation, and forced degradation studies. The flow rate was maintained at 0.5 mL/min. Column temperature was maintained at 35°C. Injection volume was 4 µL, and analysis was detected by a photodiode array detector at 375 nm. The method was validated according to USP Category I requirements for dantrolene. Forced degradation of dantrolene was conducted under the conditions of hydrolysis, oxidation, photolysis, and stability-indicating UPLC method was developed and validated. Two degradation products (related compound B and C) were formed in 0.1 N NaOH and 0.1 N HCl, respectively. The dantrolene was stable to oxidative decomposition. The degradation behavior under UV light was similar to 0.1 N HCl conditions. The method was used successfully for the quality assessment of dantrolene and its three impurities.

Keywords::

• Dantrolene
• UPLC
• impurity
• degradation
• USP related compound A, B, C

Acknowledgements

This scientific contribution is intended to support regulatory policy development. The views presented in this article have not been adopted as regulatory policies by the Food and Drug Administration at this time.

Declaration of interest

The authors declare that there are no conflicts of interest. The authors alone are responsible for the content and writing of this paper.