A NOVEL METHOD FOR THE DETERMINATION OF SECNIDAZOLE IN HUMAN PLASMA BY USING LIQUID CHROMATOGRAPHY-ELECTRO SPRAYS IONIZATION TANDEM MASS SPECTROMETRY
Ramesh Adepu*, Dr. Leela Mohan Kumar Pallapothu and Dr. A. Ashok Kumar
ABSTRACT
Objective: A novel rapid, specific and sensitive liquid chromatography tandem mass spectrometry (LCMS/MS) method was developed for the determination of Secnidazole in K2EDTA human plasma. Methods: The method involves simple, liquid-liquid extraction procedure and separation with an Inertsil® ODS-3V (5μm, 4.6 × 150 mm) with Acetonitrile / 10mM Ammonium acetate [70/30, V/V] isocratic elution at a flow-rate of 1.0 mL/min with a total run time of 2.50 minutes. Labeled isotope Secnidazole D6 was used as the internal standard. The protonate of analyte’s were quantitated in positive ionization by multiple reaction monitoring with a mass spectrometer. The mass transitions m/z 185.95 →127.73 and m/z 192.04 →127.76 were used to measure Secnidazole and Secnidazole D6 respectively. Results: The method was developed and validated using 100 μL of plasma, over a concentration range of 0.200 – 40.032 μg/mL. The intra and inter day Precision and Accuracy values were found <3.77 and 103.48%. The overall extraction efficiency was observed 66.79%. All the stability parameters were within the assay variability limits as per regulatory guidelines. Conclusion: This is a novel and sensitive and cost effective method. Total run time was 2.50 min only for each sample analysis. By using this method 250 samples analyzed per day. The method can be successfully applied to a human pharmacokinetic study and has the potential to be useful for bioequivalence studies and routine therapeutic drug monitoring.
Keywords: Secnidazole, tandem mass spectrometry, HPLC, LC-MS/MS, Human plasma.
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