Simultaneous determination of rosiglitazone and its metabolites in rat liver microsomal fraction using hollow-fiber liquid-phase microextraction for sample preparation.
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.
Calixto
Leandro Augusto
LA
Department of Physics and Chemistry, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
Bonato
Pierina Sueli
PS
D000818:Animals; D001711:Biotransformation; D005591:Chemical Fractionation / Q000295:instrumentation / Q000379:methods*; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D007004:Hypoglycemic Agents / Q000737:chemistry / Q000378:metabolism; D008110:Liver Extracts / Q000032:analysis*; D008297:Male; D008862:Microsomes, Liver / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D000077154:Rosiglitazone; D012680:Sensitivity and Specificity; D012997:Solvents / Q000737:chemistry; D045162:Thiazolidinediones / Q000737:chemistry / Q000378:metabolism
D007004:Hypoglycemic Agents; D008110:Liver Extracts; D012997:Solvents; D045162:Thiazolidinediones; D000077154:Rosiglitazone