Stability indicating Modern HPLC-UV method development and authentication of human made Allopregnanolone used to treat postpartum depression – Brexanolone injection

Abstract

Background and Objective: Allopregnanolone is a naturally occurring neurosteroid which is synthesized in our body from progesterone hormone. Brexanolone, a synthetic form of allopregnanolone inhibitory neurosteroid, a class of Anti-depressants, finds uses in treating severe postpartum depression. The present study aimed for the development and validation of RP-HPLC method, the detection of Brexanolone followed by forced degradation studies.

Materials and Methods: Brexanolone was separated on an Agilent C18, 150x 4.6mm, 5µm, 180Ao column using with isocratic elution of mobile phase of 0.1% ortho phosphoric acid and acetonitrile in the ratio of 60:40 at 30°C. The chromatographic detection was done using UV-VIS spectrophotometer at 245.0 nm. The developed method was validated for accuracy, precision, selectivity, linearity, solution stability, LOD and LOQ as per ICH guidelines.

Results: The method precision was found to be 99.58% with an RSD of 0.68%. The established LOD and LOQ are 1.10 and 3.33 µg/mL respectively. Oxidation of drug with 20% hydrogen peroxide (H2O2), acid degradation with 2N hydrochloric acid, alkali degradation using 2N sodium hydroxide, dry heat degradation, neutral degradation, and photo stability studies were evaluated as part of degradation studies with %recovery more than 98%.

Conclusion: current method is accurate and easy for the estimation in labs and validated for all the parameters as per ICH guidelines with a high percentage recovery under various conditions of degradation