Development and validation of a stability indicating RP-HPLC-DAD method for the determination of bromazepam

Area of Science:

• Analytical Chemistry
• Pharmaceutical Analysis

Background:

• Bromazepam (BMZ) is a widely used anxiolytic medication.
• Ensuring the quality and safety of pharmaceutical products requires accurate quantification of active ingredients and potential impurities.
• Stability-indicating methods are crucial for assessing drug degradation under various conditions.

Purpose of the Study:

• To develop and validate a reliable, selective, and sensitive stability-indicating RP-HPLC assay for bromazepam (BMZ).
• To quantify a key degradant and potential impurity, 2-(2-amino-5-bromobenzoyl) pyridine (ABP).
• To assess the stability of BMZ under forced degradation conditions.

Main Methods:

• RP-HPLC assay using a C18 column with a methanol-water mobile phase.
• Detection by photodiode array (DAD) detector at 230 nm.
• Method validation according to ICH guidelines, including specificity, sensitivity, precision, linearity, accuracy, and robustness.

Main Results:

• The method demonstrated high specificity, sensitivity, precision, linearity, and accuracy for both BMZ and ABP.
• Calibration curves were linear within the 1-16 μg mL-1 range with excellent recovery percentages.
• Bromazepam was found to degrade easily under acidic and alkaline conditions, indicating its instability in these environments.

Conclusions:

• The developed RP-HPLC method is suitable for quantifying bromazepam and its impurity, ABP, in pure samples and pharmaceutical dosage forms.
• The method is validated and robust, meeting ICH guidelines for analytical procedures.
• The study provides a reliable tool for quality control and stability assessment of bromazepam-containing products.