Comparison of mouse plasma and brain tissue homogenate sample pretreatment methods prior to high-performance liquid chromatography for a new 1,2,4-triazole derivative with anticonvulsant activity

The focus of the study was to develop a bio-analytical assay for a 1,2,4-triazole derivative from plasma and brain tissue homogenate samples. The goal was to compare analytical techniques that facilitate high accuracy with simplified sample processing. In this study, commonly used standard protein precipitation and solid-phase extraction methods utilizing C-18 and cartridges of Hybrid technology were compared in terms of their ability for sample pretreatment and removal of biological matrices before high-performance liquid chromatography quantification. Fast classical reversed-phase chromatography on a C18 column paired with selective sample preparation using Hybrid solid-phase extraction technology resulted in the most precise bio-analytical determination of the hydrophobic 1,2,4-triazole derivative in both biological samples studied. The obtained recovery values were above 95% with the coefficient of variation lower than 5%.