PHARMACOKINETIC DETERMINATION OF METHYLPREDNISOLONE IN INDIAN HEALTHY VOLUNTEERS BY LC-MS/MS

Das Dibya, Halder Dhiman, Bose Anirbandeep, Maji Himangshu Sekhar*, Pal Tapan Kumar

JIS University, Kolkata, West Bengal, India

ABSTRACT

Methylprednisolone is a synthetic glucocorticoid. It relieves inflammation and uses to treat thyroid, colitis, allergies, asthma, and many more. This study aims to develop a bio-analytical method for measuring methylprednisolone in human plasma, accurate, sensitive, rapid, precise and simple. In this study, Propranolol used as an Internal Standard (IS). Liquid Chromatography with tandem mass spectrometry (LC-MS/MS) instrument was used to identify and quantify the drug. Gradient conditions were used to establish the method; with a flow rate of 0.5 ml/min, we used 0.1 % formic acid in Milli-Q water and 0.1 % formic acid in methanol as mobile phases. A C18 Phenomenex Kinetex (50x3 mm, 5µ) column was used to isolate the analyte and the IS. The total chromatographic run time 7.0 min. The extraction of analyte and IS from plasma is done by a simple protein precipitation technique. The method was very selective and sensitive. Lower limit of quantification was 12.50 ng/ml. Linearity range between 12.50 ng/ml - 800ng/ml. The precision and accuracy results from 5 validation batches at five different concentration levels were well within acceptable ranges. The analyte was stable under different stability conditions. The developed method was found reproducible and very simple. The Retention Time of the analyte and IS was 3.53 min and 3.24 min, respectively. The methodology was validated in accordance with EMEA and USFDA regulations. The particular study has been carried out expertly on the pharmacokinetic study and BA/BE study in healthy human male volunteers. No Serious Adverse Events were noted in the clinical phase.

Keywords:

Methylprednisolone, LC-MS/MS, Method Development and Validation, Healthy human volunteers, Application in BA/BE Studies.