A HIGHLY SPECIFIC, SENSITIVE AND HIGH THROUGHPUT METHOD FOR THE DETERMINATION OF OFLOXACIN IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY COUPLED WITH TANDEM MASS SPECTROMETRY

摘要

A suitable, highly specific and sensitive liquid chromatography tandem mass spectrometric method has been developed and validated for the determination of Ofloxacin from 100μL of human plasma by Protein Precipitation. Ciprofloxacin was used as an internal standard. Quantified by the transition, 362.200→261.420, 332.205→282.500 for Ofloxacin and Ciprofloxacin respectively and detected by TSQ Quantum Ultra triple quadrupole mass spectrometer. Detection was carried out by using ESI source in positive polarity. Chromatographic separation of analyte and internal standard were carried out by reverse phase C18 column at the flow rate of 0.500mL/min with mobile phase of Acetonitrile: 0.02% Formic acid (60:40) v/v. The assay of Ofloxacin was linear over the range of 20.133 ng/mL to 2001.800 ng/mL with a precision of ≤0.45 % and ≤6.88 % respectively, Mean extraction recovery obtained was 89.84%. Samples were stable at room temperature for 6 hrs and also stable at four freeze-thaw cycle. The aim is to develop a suitable, highly specific, and sensitive analytical method for the quantitation of Ofloxacin in the low nanogram range in human plasma.