Rapid Ultra High Performance Liquid Chromatography–Tandem Mass Spectrometry Method for Quantification of Ethacrynic Acidin Human Plasma
Journal Title: IOSR Journal of Biotechnology and Biochemistry (IOSR-JBB - Year 2018, Vol 4, Issue 1
Abstract
A sensitive and high throughput high performance liquid chromatography tandem mass spectrometry (HPLC–MS/MS) method has been developed for the determination of ethacrynic in human plasma. Sample preparation involved liquid–liquid extraction for ethacrynic and ethacrynic D5 as the internal standard (IS) in ethyl acetate from 200 μL human plasma. The chromatographic separation is achieved on a shimadzu HPLC and Gemini-NX 5μ C18 110 A° (50 X 4.6, mm) analytical column using an isocratic mobile phase, consisting of 0.1 % formic acid in water–acetonitrile (30:70, v/v). Detection was carried out on a tandem mass spectrometer inmultiple reaction monitoring mode using a negative electro pressure ionization interface.The method has been validated in linear range of 4.997-1000.283 ng/mL. The method is rugged and rapid with a total run time of 3.0 min and is applied to a bioequivalence study of 25 mg tablet formulation in 48 healthy Indian male subjects under fasting condition.
Authors and Affiliations
Smith Christian, Rashmikant Patel, Shivprakash .
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