RELIABLE ESTIMATION OF PREDNISOLONE, A GLUCOCORTICO STEROID IN HUMAN PLASMA BY LCMS/MS

Pritesh Contractor; Pranav S. Shrivastav

Volume : V, Issue : VIII, August - 2016

RELIABLE ESTIMATION OF PREDNISOLONE, A GLUCOCORTICO STEROID IN HUMAN PLASMA BY LCMS/MS

Pritesh Contractor, Pranav S. Shrivastav

Abstract :

An accurate and precise method was developed and validated using LC–MS/MS to quantify prednisolone in human plasma. The merits of this method include solid phase extraction with high extraction recovery using deuterated internal standard (IS) and short chromatographic run time. The analyte and prednisolone–d6 (IS) were extracted from 500 �L plasma volume using solid–phase extraction on OASIS WCX (1cc, 30mg) cartridges. Chromatographic analysis was performed on a Gemini C18 (150 × 4.6 mm, 5 �m) column using acetonitrile–0.5 % acetic acid in water (50:50, v/v) as the mobile phase. Tandem mass spectrometry in positive ionization mode was used to quantify prednisolone and prednisolone–d6 by multiple reaction monitoring. Entire data processing was done using Watson LIMSTM software which provided excellent data integrity, high throughput with improved operational efficiency. The caliation graph was linear in the range of 2.0–1000 ng/mL. The accuracy and precision values for intra– and inter–batch values ranged from 94.7 to 103.8 % and 1.1 to 6.3 %, respectively. The mean overall recovery across all quality control levels was ≥ 80 % for both analyte and IS, while matrix factors ranged from 1.00 to 1.06 which were evaluated for the assessment of matrix effect.