European Journal of Biomedical and Pharmaceutical Sciences

NEW DETERMINATION AND QUANTIFICATION OF POSACONAZOLE IN HUMAN PLASMA BY LIQUID CHROMATOGRAPHY WITH TANDEM MASS SPECTROMETRY: IT’S APPLICATION TO A PHARMACOKINETIC STUDY

Venkata Ramu Derangula and Venkateswarlu Ponneri*

ABSTRACT

A simple, rapid and sensitive liquid chromatography with tandem mass spectrometry(LC-MS/MS) assay method has been proposed for the determination of Posaconazole in human plasma samples using Posaconazole D5 as internal standard (IS). Analyte and the IS were extracted from the 100 μL of K2 EDTA human plasma by simple Protein Precipitation extraction. The chromatographic separation was achieved on a Zorbax Eclipse XDB-C18 column by using a mixture of Acetonitrile and 5mM ammonium acetate in 0.1% formic acid buffer (85:15, v/v) as the mobile phase at a flow rate of 1.0 mL/min. The calibration curve obtained was linear (r2  0.99) over the concentration range of 5.01 – 2502.29 ng/mL. The Mass detection of Posaconazole involves m/z - 701.20 (parent) and 683.30 (product) and Posaconazole D5 involves m/z - 706.30 (parent) and 687.30 (product) as internal standard in Positive ion mode. Method validation was performed as per FDA guidelines and the results met the acceptance criteria. The intra-day and inter-day precision (%CV) and accuracy results in three validation batches across six concentration levels were well within the acceptance limits. A run time of 2.40 min for each sample made it possible to analyze more number of samples in short time, thus increasing the productivity. The proposed method successfully applied to a pharmacokinetic study of Posaconazole 40 mg/mL oral suspension formulation in healthy, adult, human male subjects under fed condition.

Keywords: Posaconazole, human plasma, Protein Precipitation extraction, LC-MS/MS.