title: LC-Q-TOF-MS/MS determination of darunavir and its metabolites in rat serum and urine: application to pharmacokinetics.
abstract: A simple, rapid and reliable liquid chromatography coupled with quadrupole time of flight mass spectrometry (LC-Q-TOF-MS/MS) method was developed and validated for simultaneous determination of darunavir and its metabolites in rat serum and urine. The separation was accomplished on an Agilent RP-18 (250×4.6mm, 5μm) column using 20mM ammonium acetate and methanol (40:60, v/v) as a mobile phase at a flow rate of 1.0mL/min in an isocratic mode. The [M+H](+) ions of darunavir (m/z 548) and metabolites-I (m/z 392) were monitored in positive mode of ionization, while [M-H](-) ion of metabolite-II (m/z 172) in negative mode selectively. The matrix effects of rat serum and urine were found to be negligible and the recoveries were 87-93% for all the analytes. The short and long term stability of darunavir and its metabolites was within acceptable limits and the lower limits of quantification were in the range of 3.63-5.24ng/mL with a linear range of 5-5000ng/mL in rat serum as well as urine. The method exhibited good intra- and inter-day performance in terms of 2.54-8.92% precision and 0-5% accuracy. The method was successfully applied to a single-dose pharmacokinetic study of darunavir boosted with ritonavir in Wistar rats.
authors.affiliation: Analytical Chemistry Division, Discovery Laboratory, Indian Institute of Chemical Technology, Tarnaka, Hyderabad 500607, India. Electronic address: rnrao@iict.res.in.
authors.forename: R
authors.initials: R
authors.lastname: Nageswara Rao
authors.affiliation: Analytical Chemistry Division, Discovery Laboratory, Indian Institute of Chemical Technology, Tarnaka, Hyderabad 500607, India.
authors.forename: K
authors.initials: K
authors.lastname: Guru Prasad
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods; D000069454:Darunavir; D004355:Drug Stability; D051381:Rats; D017208:Rats, Wistar; D012680:Sensitivity and Specificity; D013449:Sulfonamides / Q000097:blood* / Q000493:pharmacokinetics / Q000652:urine*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D013449:Sulfonamides; D000069454:Darunavir
keywords: Darunavir; Pharmacokinetics; Rat serum; Selective ion monitoring; Urine