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Determination of Pyrazinamide in Human Plasma Samples Containing Fixed Dose Combination Molecules by using Liquid Chromatography Tandem Mass Spectrometry

Chaitanya Krishna A, Saravanan RS, Jeevanantham S, Vignesh R and Karthik P

A rapid, simple, sensitive and compatible liquid chromatography tandem mass spectrometric method has been developed and validated for the estimation of Pyrazinamide. Glipizide was used as an internal standard. Detection was performed using TSQ Quantum Discovery max mass spectrometer with ESI source in positive polarity. The detection transition for Pyrazinamide is 124.100 → 79.160 and for Glipizide is 446.200 → 321.200. Chromatographic separation of analyte and internal standard were carried out using a reverse phase column, Hypersil, Gold, 4.6 X 50 mm, 5 μ at a flow rate of 0.400 mL/min. Mobile Phase is composed of Methanol: 0.1 % FA in 10 mM Ammonium Formate (90:10) v/v. Extracted by Solid Phase Extraction with a sample volume of 200 μL plasma. The assay of Pyrazinamide is linear over the range of 0.935 μg /mL to 60.408 μg/mL with a precision of < 9.86%. Mean extraction recovery for Pyrazinamide and Glipizide were more than 61%. Samples are stable at room temperature for 6 hrs, processed samples were stable at least for 28 hrs and also stable at three freeze–thaw cycles.