title: Exploring dried blood spot sampling technique for simultaneous quantification of antiretrovirals: lamivudine, stavudine and nevirapine in a rodent pharmacokinetic study.
abstract: A high-performance liquid chromatography/positive ion electrospray tandem mass spectrometry method for the simultaneous quantification of lamivudine, stavudine and nevirapine was developed and validated in dried blood spot (DBS) cards. The analytes were separated using an isocratic mobile phase on a reverse phase column and analyzed by MS/MS in the MRM mode using the respective [M + H]⁺ ions, m/z 230-112 for lamivudine, m/z 225-127 for stavudine, m/z 267-226 for nevirapine, m/z 383-337 for zidovudine (IS). The lower limit of quantification was 1 ng/mL for both lamivudine and stavudine and 10 ng/mL for nevirapine. Acceptable precision and accuracy were obtained for concentrations over the standard curve range. The method was successfully applied to quantify them in a rat pharmacokinetic study in whole blood, plasma and DBS cards after a single oral co-administration at the dose of 10, 2 and 13 mg/kg for lamivudine, stavudine and nevirapine, respectively, to male Wistar rats. Following oral administration the pharmacokinetic results in all the matrices are in close agreement. Thus accomplishment of this method would facilitate the ease of collection of clinical samples on DBS cards for lamivudine, stavudine and nevirapine during human clinical trials and therapeutic drug monitoring.
authors.affiliation: Pharmacokinetics and Drug Metabolism, Discovery Research, Suven Life Sciences Ltd, Serene Chambers, Road 5, Avenue 7, Banjara Hills, Hyderabad 500034, India. ramakrishna_nirogi@yahoo.co.in
authors.forename: Ramakrishna
authors.initials: R
authors.lastname: Nirogi
authors.forename: Vishwottam
authors.initials: V
authors.lastname: Kandikere
authors.forename: Prashanth
authors.initials: P
authors.lastname: Komarneni
authors.forename: Raghupathi
authors.initials: R
authors.lastname: Aleti
authors.forename: Nagasuryaprakash
authors.initials: N
authors.lastname: Padala
authors.forename: Ilayaraja
authors.initials: I
authors.lastname: Kalaikadhiban
authors.forename: Gopinadh
authors.initials: G
authors.lastname: Bhyrapuneni
authors.forename: Nageswararao
authors.initials: N
authors.lastname: Muddana
mesh_terms: D000818:Animals; D044966:Anti-Retroviral Agents / Q000097:blood* / Q000737:chemistry / Q000493:pharmacokinetics; D002851:Chromatography, High Pressure Liquid; D059788:Dried Blood Spot Testing / Q000379:methods*; D004355:Drug Stability; D019259:Lamivudine / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D008297:Male; D019829:Nevirapine / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018119:Stavudine / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D053719:Tandem Mass Spectrometry
chemical_list: D044966:Anti-Retroviral Agents; D019259:Lamivudine; D019829:Nevirapine; D018119:Stavudine