Simultaneous determination of rosiglitazone and its metabolites in rat liver microsomal fraction using hollow‐fiber liquid‐phase microextraction for sample preparation

Abstract

A three-phase hollow-fiber liquid-phase microextraction method for the analysis of rosiglitazone and its metabolites N-desmethyl rosiglitazone and ρ-hydroxy rosiglitazone in microsomal preparations is described for the first time. The drug and metabolites HPLC determination was carried out using an X-Terra RP-18 column, at 22°C. The mobile phase was composed of water, acetonitrile and acetic acid (85:15:0.5, v/v/v) and the detection was performed at 245 nm. The hollow-fiber liquid-phase microextraction procedure was optimized using multifactorial experiments and the following optimal condition was established: sample agitation at 1750 rpm, extraction for 30 min, hydrochloric acid 0.01 mol/L as acceptor phase, 1-octanol as organic phase, and donor phase pH adjustment to 8.0. The recovery rates, obtained by using 1 mL of microsomal preparation, were 47-70%. The method presented LOQs of 50 ng/mL and it was linear over the concentration range of 50-6000 ng/mL, with correlation coefficients (r) higher than 0.9960, for all analytes. The validated method was employed to study the in vitro biotransformation of rosiglitazone using rat liver microsomal fraction.

In vitro characterization of rosiglitazone metabolites and determination of the kinetic parameters employing rat liver microsomal fraction

“…In this study, the incubation times were: 3, 5, 10, 20, 30, 45 and 60 min (n = 3). Of note is that in a previous stability study, it was confirmed that during this time and temperature, there was no degradation of the analyzed analytes (Calixto and Bonato 2010).…”

“…Sample preparation was performed by hollow-fiber liquid-phase microextraction (HF-LPME), according to a method previously developed in our laboratory (Calixto and Bonato 2010). n-octanol, used as extraction solvent, was purchased from Sigma-Aldrich (St. Louis, MO, USA); sodium hydroxide, perchloric acid and hydrochloric acid were obtained from Spectrum (New Brunswick, NJ, USA), Nuclear (São Paulo, SP, Brazil), and Mallinckrodt (Xalostoc, Mexico), respectively.…”

“…The stability under basic conditions was reported in a previous paper describing the development and validation of an LC method for the analysis of rosiglitazone and its metabolites in microsomal fraction. During the validation of this method, we evaluated the stability of the drug and metabolites in basic conditions used for the extraction (Calixto and Bonato, 2010). Our results showed that the drug and metabolites were stable.…”

Application of Three-phase Hollow Fiber LPME using an Ionic Liquid as Supported Phase for Preconcentration of Malachite Green from Water Samples with HPLC Detection

“…15 As an alternative for analysis of MG in water, hollow fiber liquid-phase microextraction (HF-LPME) has the merits of simplicity, efficiency, negligible volume of solvents needed and excellent sample cleaning ability. [16][17][18] HF-LPME could be classified into three-phase or two-phase mode, the former was suitable for pre-concentration of acidic or basic compounds, especially for the sample with complex matrix. In order to make the method more effective and environmental friendly, room temperature ionic liquids (ILs) was introduced to the analyzing system, which possess many significant advantages, such as low vapor pressure, wide range of miscibility with other organic solvents, good thermal stability, moderate dissolvability of organic compounds and dual natural polarity.…”

Avaliação de técnicas miniaturizadas de preparação de amostras na análise enantiosseletiva de fármacos quirais com diferentes características ácido-base em meio microssomal e aplicação em estudos de metabolismo in vitro

“…Idealmente, o solvente orgânico deve ser compatível com a membrana, imiscível na amostra, estável e ter alto poder de solubilização dos analitos a serem extraídos (CALIXTO; BONATO, 2010). A ISR é um fármaco neutro, com logP 3,7 (SCIFINDER, 2012), portanto, a HF-LPME na configuração "duas fases" é a recomendada.…”