Development of a HPLC method for the quantification of Docetaxel in the BPMO@DTX injectable formulation

Biodegradable BPMO@DTX (B@D) nanoparticles were successfully synthesized, exhibiting potent anticancer activity while mitigating the systemic toxicity of docetaxel (DTX). To advance the development of an injectable B@D nanoformulation, an accurate and robust method for DTX quantification was required. High-performance liquid chromatography (HPLC) was employed, with sample preparation based on a biodegradation technique, Glutathione (GSH-induced degradation), which was selected as the optimized procedure. Ultrasonication was evaluated only during the method development stage as a comparative approach. Key parameters—including solvent type, sample-to-solvent ratio, and biodegradation time—were optimized, and the method was validated according to International Conference on Harmonization Q2(R2) guidelines. The optimized HPLC conditions comprised a mobile phase of acetonitrile:water (60:40, v/v), PDA detection at 230 nm, a Gemini 5 μm C18 (250 × 4.6 mm) column, 30°C temperature, 1.2 ml/minute flow rate, and 20 μl injection volume, all meeting validation requirements. Ultrasonication resulted in only ~30% DTX recovery, rendering it unsuitable for quantification. In contrast, the biodegradation method—incubating B@D with 20 mM GSH for 48 hours, followed by dichloromethane extraction (1:5 ratio, 5 minutes, 6,000 rpm, repeated five times)—achieved >99% recovery. This validated method provides a reliable analytical tool for DTX quantification in B@D formulations, supporting further pharmaceutical development.