RP-UPLC-MS Assay Method for Eltrombopag: Application in Pharmaceuticals, Human Plasma and Urine Samples

摘要

A simple, accurate, rapid and precise method was developed using UPLC coupled to mass detector (UPLC-MS) for stability and estimation of eltrombopag (EMP) in its bulk and dosage forms, human plasma and urine samples. Instrumentation used for this method was highly sophisticated and of superior technology in terms of speed, separation, solvent usage and analysis cost. The efficient separation of drug from its degradation products was achieved by using Acquity UPLC BEH column C_(18) (50 x 2.1 mm. 1.7 urn) with linear gradient elution of mobile phase (0.1 % formic acid in water & acetonitrile) 97:03 % v/v at initial gradient and flow rate of 0.6 mL/min. The chromatogram was analyzed at a wavelength of 230 nm (λ_(max)) using PDA detector and further confirmed by mass (m/z) using waters SQD (Single Quadrapole Detector-3100 mass range 100 amu-1000 amu) with a run time of 4.5 min. The linearity range was performed from 20 to 100 μg/mL (R~2 = 0.999) with regression of y = 433.88x + 35.4. The variation of precision study was found to be < 1 %. Eltrombopag was showing more sensitivity towards the acidic degradation. The formed degradative products were identified by mass spectrometry (UPLC-MS) and the mode of degradation was reported. The dosage form and bulk drug samples were subjected to forced degradation and analyzed as per ICH Q1A (R2) and Q2 (R1) guidelines.