Optimization and validation of HPLC methods for in vitro and ex vivo analyses of bosentan monohydrate in FDA-recommended and biorelevant media

Abstract

Background and Aims: Bosentan (BOS) is an endothelin receptor antagonist indicated for the treatment of pulmonary arterial hypertension. It is a BCS Class II drug. This study aimed to apply and method validation the new HPLC methods for FDArecommended and biorelevant media. These methods were used to assess the quantification of BOS in in vitro and ex vivo studies on lipid-based drug delivery systems (BOS-loaded self-nanoemulsifying drug delivery systems (SNEDDS), and solid self-nanoemulsifying drug delivery systems (S-SNEDDS)) compared to commercial products (Tracleer®). In vitro studies include assessments of content uniformity and dissolution in FDA-recommended and biorelevant media. The stability of S-SNEDDS tablets was evaluated in an FDA-recommended medium. The ex vivo study assessed the permeability of BOS-loaded SNEDDS, S-SNEDDS tablets, and reference tablets in biorelevant media. Methods: HPLC was operated using buffer solution: acetonitrile (45:55) with a flow rate of 1.5 mL/min at 220 nm. The injection volume was set at 100 μL. Separation was carried out using a Waters XSelect® HSS C18 column (250x4.6 mm, 5μm) at 25°C. Results: HPLC methods were validated using ICH Q2(R2) and FDA guidelines. Retention times were found to be between 4.7 and 5.5 in different media. The validated methods were proved to be sensitive, simple, reproducible, rapid, and precise for determining BOS in pharmaceutical formulations and dosage forms. Conclusion: These new HPLC methods were successfully applied and validated for FDA-recommended and biorelevant media in in vitro, ex vivo, and quality control tests of BOS-loaded SNEDDS, S-SNEDDS tablets, and reference tablets.

Keywords

• Bosentan monohydrate
• HPLC
• FDA-recommended and biorelevant media
• Pharmaceutical dosage forms
• Pharmaceutical formulations

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