Validation of high performance liquid chromatography mass spectrometric method in negative ion mode for the estimation of Monomethyl fumarate in human plasma using Monomethyi fumarate D3 as internal standard

Abstract

The present project work was to develop a robust, rapid, simple and sensitive liquid chromatography mass spectrometry (LC-MS/MS) assay method for the quantification of monomethyl fumarate in human plasma. An analytical method employing LC-MS/MS using human plasma was developed and fully validated for the estimation of Monomethyl fumarate in human plasma by using monomethyl fumarateD3 as internal standarad. The solid phase extraction technique (SPE) was used for the extraction of the drug and Internal standard (IS). The chromatographic separation was achieved on a Zodiac C18 column by using a 70:30 (v/v) mixture of acetonitrile and 0.1% formic acid as the mobile phase at a flow rate of 0.5 mL/min. Totally five precision and accuracy batches were performed during the entire validation and intra-day and inter-day precision and accuracy were proved. KEYWORDS: Monomethyl fumarate, Human plasma, LC–MS/MS, Pharmacokinetics, Quantification.