Systematic Method Development and Validation of Paclitaxel and Tamoxifen by RP-HPLC for Simultaneous Estimation in Nanoformulations

Abstract

Background: Paclitaxel (PTX) is well known anticancer drug used for treatment of a wide range of cancers. Tamoxifen (TMX) is a gold-standard drug for the treatment of breast cancer. They show synergistic action against breast cancer. Quantitative analytical methods are still not available for simultaneous quantification of these drugs. So for the first time, we developed and validated a new systematic analytical technique for simultaneous estimation of PTX and TMX in nanoformulations. Objective: Systematic method development and validation of Paclitaxel and Tamoxifen by RP-HPLC for simultaneous estimation in nanoformulations. Methods: PTX and TMX were successfully separated and quantified using RP-HPLC. The detection was done based on the isobastic point of both drugs. Thermo Scientific Company's C18 column, measuring 2.5 cm × 4.5 cm x 5 µm, was utilized for separation. A 50:50 (vol/vol) ratio of ACN: Phosphate buffer solution (pH 3.0) was employed as the mobile phase, with a flow rate of 1 mL/minute. The detection was done at 235nm. The method was developed and validated as per the criteria. The developed method was then evaluated for applicability by detecting both analytes in prepared SLN and LPHNPs. Results: The well-resolved separate peaks of both analytes were found and distinct RT for PTX and TMX. The linear relationship of both the analytes against concentration was found over the concentration range of 0.25-9 µg/mL. The PTX RT was 10.358 minutes and that of TMX was 12.57 minutes. Every criterion for method validation fell within the acceptable range. The method is precise with inter-day and intra-day precision and shows a %RSD of < 2%.