Chin-Chung Lin

Specific, sensitive and accurate liquid chromatographic–tandem mass spectrometric method for the measurement of ribavirin in rat and monkey plasma

Ribavirin is a purine nucleoside analog with broad spectrum activity against a spectrum of DNA and RNA viruses. To facilitate pharmacokinetics studies, a LC-MS-MS method for the analysis of ribavirin in rat and monkey plasma was developed and validated. The method involved the addition of acyclovir as an internal standard and protein precipitation with acetonitrile followed by separation by an Intertsil Silica column and quantification by a MS-MS equipped with a positive electrospray ionization in the multiple reaction monitoring mode. The MS-MS reaction was selected to monitor the 245-->113 and 226-->152 transitions for ribavirin and internal standard, respectively. The calibration curve was linear over a concentration range of 10-5000 ng/ml. The lower limit of quantitation was 10 ng/ml, the coefficient of variation (CV) was 8-11%, and the bias was 1-3%. Intra-day and inter-day analysis of QC samples at 30, 1500 and 3500 ng/ml indicate that the method was precise (CV<18%) and accurate (bias<13%). Ribavirin in rat and monkey plasma was stable at 5 degrees C for at least 24 h, 0 degrees C for at least 4 h, and after three freeze-thaw cycles. This specific, accurate and precise assay is useful in the study of the pharmacokinetics of this compound.

Pharmacokinetics and Metabolism of [14C]Ribavirin in Rats and Cynomolgus Monkeys

ABSTRACT Absorption, pharmacokinetics, distribution, metabolism, and excretion of [14C]ribavirin were studied in rats (30 mg/kg of body weight) and cynomolgus monkeys (10 mg/kg) after intravenous (i.v.) and oral administration. The oral absorption and bioavailability were 83 and 59%, respectively, in rats and 87 and 55%, respectively, in monkeys. After i.v. administration, the elimination half-life (t[1/2]) was 9.9 h in rats and 130 h in monkeys and the total body clearance was 2,600 ml/h/kg in rats and 224 ml/h/kg in monkeys. The apparent volume of distribution was 11.4 liter/kg in rats and 29.4 liter/kg in monkeys. There was extensive distribution of drug-derived radioactivity into red blood cells and extensive metabolism of ribavirin in rats and a lesser degree of metabolism in monkeys. Excretion of total radioactivity in urine from rats accounted for 84% of the i.v. dose and 83% of the oral dose, whereas that from monkeys accounted for 47% of the i.v. dose and 67% of the oral dose. Several metabolites were observed in plasma and urine from both species. The amount of unchanged ribavirin in urine from both species was quite small after either i.v. or oral administration.

Specific, sensitive and accurate LC–MS/MS method for the measurement of levovirin in rat and monkey plasma

Levovirin is a guanosine nucleoside analogue and the L-enantiomer of ribavirin. Levovirin has a better safety profile than ribavirin, exerts similar immunomodulatory effects in a mouse efficacy model, and may provide a better therapeutic option than ribavirin in patients with chronic hepatitis C virus (HCV) infection. To facilitate pharmacokinetic studies, a LC-MS/MS method for the analysis of levovirin in rat and monkey plasma was developed and validated. The method involved adding ICN 10537 as an internal standard, protein precipitation with acetonitrile followed by separation on an Intersil Silica column, and quantification by a MS/MS system equipped with positive electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode. The MS/MS reaction was selected to monitor the 245-->113 and 259-->128 transitions for levovirin and internal standard, respectively. The calibration curve was linear over a concentration range of 10-5000 ng/ml. The limit of quantitation was 10 ng/ml, the coefficient of variation (CV) was 3-5%, and the bias was 3-6%. Intra- and inter-day analysis of QC samples at 30, 1500 and 3500 ng/ml indicated that the method was precise (CV<6%) and accurate (bias <9%). Levovirin in rat and monkey plasma was stable at 5 degrees C for at least 24 h, 0 degrees C for at least 4 h, and after three freeze-thaw cycles. This specific, accurate and precise assay is useful in the study the pharmacokinetic characteristics of this compound.