BIOANALYTICAL LC-MS METHOD FOR DETECTION AND QUANTIFICATION OF NIFURTIMOX IN HUMAN PLASMA

Abstract

Objective: Nifurtimox is a type of nitrofuran derivative employed for treating sleeping sickness as well as Chagas disease. The aim is to develop and validate a precise and specific liquid chromatography mass spectrometric technique for the estimation of Nifurtimox in human plasma.Material and Method: Chromatographic separation had been accomplished on reverse-phase C18 (4.6 mm × 100 mm, 3.5 µm) column employing benznidazole as an internal standard along mobile phase made up of methanol and 5 mM aqueous ammonium acetate in a 75:25 (v/v) ratio at a flow rate of 0.8 ml/min in isocratic mode. A quadrupole mass spectrometer running in positive mode and with numerous reaction monitoring settings was employed to quantify the procedure.Result and Discussion: The developed analytical method exhibited good recovery and was validated over a calibration range of 0.5–400 ng/ml. In the ESI spectra, the major ion peaks for Nifurtimox and the internal standard, benznidazole, were observed at m/z 288 and m/z 261, respectively, corresponding to their [M+H]+ ions. The method exhibited LOD, LOQ, and LLOQ of 0.2 ng/ml, 1.0 ng/mL and 0.5 ng/ml respectively. Precision at intra and interday and accuracy were evaluated at low (1 ng/ml), medium (50 ng/ml), and high (400 ng/ml) quality control levels, showing a %RSD below 2. The percentage recoveries exceeded 95%, with %RSD values under 2. Overall, the validated method is suitable for routine quantification of Nifurtimox in biological samples.