Development and validation of QbD approach in RP-HPLC analytical method for separation and identification of anti-tuberculosis drug Rifapentine

Abstract

Background: Tuberculosis (TB) poses both a global health and economic burden. Timely treatment is essential but costly and slow. HPLC, enhanced by QbD, ensures precise drug analysis, improving efficiency and reliability in TB treatment.

Objective: The present study aims to develop the QbD approach RP-HPLC method for the separation and identification of the anti-tuberculin drug rifapentine.

Methodology: The current study focused on optimising RT-HPLC conditions, including the mobile phase, flow rate, and wavelength, for the analysis of rifapentine. 20 µg of rifapentine was prepared using HPLC-grade methanol. The chromatographic conditions were optimised with BBD Design Expert Software v13. Analysis was performed using a C18 reversed-phase column (250 mm x 4.6 mm, 5.0 µm) with a diode array detector (DAD). The mobile phase was a mixture of methanol and 1% orthophosphate acid (81:19, v/v), with a flow rate of 1.1 mL and an injection volume of 20 µL. The validation and data analysis were conducted at room temperature with a retention time of 5 minutes at 252 nm.

Results: The developed method demonstrated linearity with r²=0.1705 and statistical significance (p<0.0001). The chromatography peal purity confirmed the absence of coeluting peaks with the rifapentine peak. The system suitability parameters, including the tailing factor and theoretical plate, were 1.15 and 7965, respectively. Additionally. The method’s validation factors were within the limits specified by ICH guidelines.

Conclusion: The QbD approach for developing the rifapentine analytical method proved accurate and specific, meeting ICH regulatory norms. RP-HPLC by QbD is a reliable method that ensures validity and high-quality drug production, supports effective TB treatment, and facilitates quality checks and bulk production of the pharmaceutical formulation