Ultra-Performance Liquid Chromatography Method for Bioanalysis of Teriflunomide using Dried Blood Spot Methodology

Abstract

Plasma slowly loses teriflunomide, an oral immunomodulator used once in a day and majorly it is approved for treating of relapsing-remitting multiple sclerosis. It is not possible to measure the plasma concentrations of teriflunomide at locations without the tools necessary to prepare the processed blood samples. Plasma monitoring can be replaced with DBS technique, dried blood spot sampling, which allows for easier sample storage and travel. A few drops of blood are extracted from the rat tail using a lancet and applied to specially made absorbent filter paper. A UPLC test technique for the measurement of teriflunomide in DBS is created and validated for specificity, accuracy, selectivity, stability and repeatability using blood samples from pharmacokinetic studies. Process efficiency was necessary, method was selective and specific regarding endogenous chemicals, and there was no matrix effect. All concentrations were evaluated for accuracy as well as precision for intra-day and also inter-day analysis. The amount of blood deposited and the punch position within the spot had no bearing on the detection of teriflunomide in the DBS assay, however the hematocrit level had a negligible but tolerable influence on measurement precision. Teriflunomide has a minimum stability of three months at room temperature. With an average ratio of blood to plasma, an association between DBS concentration and plasma concentrations is seen. A straightforward and useful technique for keeping track of teriflunomide concentrations is DBS sampling. The technique has been expanded to the in-vivo determination of teriflunomide in male albino rats and is fully verified in accordance with ICH criteria.