title: Rapid and sensitive ultra-high-pressure liquid chromatography method for quantification of antichagasic benznidazole in plasma: application in a preclinical pharmacokinetic study.
abstract: Benznidazole (BNZ) and nifurtimox are the only drugs available for treating Chagas disease. In this work, we validated a bioanalytical method for the quantification of BNZ in plasma aimed at improving sensitivity and time of analysis compared with the assays already published. Furthermore, we demonstrated the application of the method in a preclinical pharmacokinetic study after administration of a single oral dose of BNZ in Wistar rats. A Waters® Acquity UHPLC system equipped with a UV-vis detector was employed. The method was established using an Acquity® UHPLC HSS SB C18 protected by an Acquity® UHPLC HSS SB C18 VanGuard guard column and detection at 324 nm. The mobile phase consisted of ultrapure water-acetonitrile (65:35), and elution was isocratic. The mobile phase flow rate was 0.55 mL/min, the volume of injection was 1 μL, and the run time was just 2 min. The samples were kept at 25°C until injection and the column at 45°C for the chromatographic separation. The sample preparation was performed by a rapid protein precipitation with acetonitrile. The linear concentration range was 0.15-20 µg/mL. The pharmacokinetic parameters of BNZ in rats were determined and the method was considered sensitive, fast and suitable for application in pharmacokinetic studies.
authors.affiliation: Department of Natural Active Principles and Toxicology, School of Pharmaceutical Sciences, São Paulo State University, Araraquara, São Paulo, Brazil.
authors.forename: Marcelo Gomes
authors.initials: MG
authors.lastname: Davanço
authors.affiliation: Department of Natural Active Principles and Toxicology, School of Pharmaceutical Sciences, São Paulo State University, Araraquara, São Paulo, Brazil.
authors.forename: Michel Leandro
authors.initials: ML
authors.lastname: de Campos
authors.affiliation: Department of Natural Active Principles and Toxicology, School of Pharmaceutical Sciences, São Paulo State University, Araraquara, São Paulo, Brazil.
authors.forename: Rosângela Gonçalves
authors.initials: RG
authors.lastname: Peccinini
mesh_terms: D000284:Administration, Oral; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016014:Linear Models; D008297:Male; D009593:Nitroimidazoles / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014344:Trypanocidal Agents / Q000008:administration & dosage / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics
chemical_list: D009593:Nitroimidazoles; D014344:Trypanocidal Agents; C009999:benzonidazole
keywords: Chagas disease; benznidazole; bioanalytical method; pharmacokinetic parameters; ultra-high pressure liquid chromatography; validation method