Drug Res (Stuttg) 2013; 63(12): 644-649
DOI: 10.1055/s-0033-1358665
Original Article
© Georg Thieme Verlag KG Stuttgart · New York

A Validated Method for Quantifying Entecavir in Biological Matrices and Its Application in a Pharmacokinetic Study in Rats and Dogs

X. Huang
1   Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing, P. R. China
2   Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing, P. R. China
,
Z.-Z. Jiang
1   Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing, P. R. China
,
W.-W. Tang
1   Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing, P. R. China
,
Y. Xiao
1   Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing, P. R. China
,
L.-Y. Zhang
1   Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing, P. R. China
,
Z.-J. Zhang
2   Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing, P. R. China
› Author Affiliations
Further Information

Publication History

received 02 May 2013

accepted 05 October 2013

Publication Date:
07 November 2013 (online)

Abstract

A sensitive and specific liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) method that uses ganciclovir as an internal standard (IS) has been developed and validated for quantifying entecavir in rat and dog plasma. Following solid-phase extraction (SPE), the analytes were separated on an Inertsil® ODS-3 (5 μm, 150 mm×2.1 mm i.d.) column and analyzed in selected ion monitoring (SIM) mode with a positive electrospray ionization (ESI) source using the [M+H]+ ions, 278.1 for entecavir and 256.0 for ganciclovir. The method was validated over the concentration range of 0.01–9 μg/mL for entecavir. All precisions (RSD) within and between batches were less than 10%, and accuracies ranged from 98.1 to 102.5%. The lower limit of quantification was 0.01 µg/mL. The extraction recovery averaged 93.9–96.7%. The validated method was used for a pharmacokinetic study of entecavir in rats and dogs. The following pharmacokinetic parameters were obtained for rats and dogs, respectively: the area under the plasma concentration vs. time curves from time 0 to 24 h (AUC0–24) were 15.4±4.5 and 23.4±7.2 μg∙h/mL; the mean maximum plasma concentration (Cmax) were 2.4±0.8 and 5.0±0.9 μg/mL; the mean time to reach the maximum plasma concentrations (Tmax) were 1.7±0.7 and 1.5±0.4 h; and the mean elimination half-life (t1/2) were 5.3±1.4 and 3.8±1.3 h.

 
  • References

  • 1 Hu P, Jiang J, Wang H et al. Single-dose and multiple-dose pharmacokinetics and safety of telbivudine after oral administration in healthy Chinese subjects. J Clin Pharmacol 2006; 46: 999-1007
  • 2 Mohamed R, Desmond P, Suh DJ et al. Practical difficulties in the management of hepatitis B in the Asia-Pacific region. J Gastroenterol Hepatol 2004; 19: 958-969
  • 3 Merican I, Guan R, Amarapuka D et al. Chronic hepatitis B virus infection in Asian countries. J Gastroenterol Hepatol 2000; 18: 1356-1361
  • 4 Zhou XJ, Fielman BA, Lloyd DM et al. Pharmacokinetics of telbivudine in healthy subjects and absence of drug interaction with lamivudine or adefovir dipivoxil. Antimicrobial Agents And Chemotherapy 2006; 50: 2309-2315
  • 5 Yan JH, Bifano M, Olsen S et al. Entecavir pharmacokinetics, safety and tolerability after multiple ascending doses in healthy subjects. J Clin Pharmacol 2006; 46: 1250-1258
  • 6 Albert D, Jules L. Dienstag. Oral antivirals for chronic hepatitis B. Clin Liver Dis 2007; 1: 851-868
  • 7 Ferreira MS, Borges AS. Advances in the treatment of hepatitis B. Rev Soc Bras Med Trop 2007; 40: 451-462
  • 8 Challa BR, Awen BZ, Chandu BR et al. LC-ESI-MS/MS method for the quantification of entecavir in human plasma and its application to bioequivalence study. Journal of Chromatography B 2011; 879: 769-776
  • 9 Zhang D, Fu Y, Gale JP et al. A sensitive method for the determination of entecavir at picogram per milliliter level in human plasma by solid phase extraction and high-pH LC-MS/MS Journal of Pharmaceutical and Biomedical Analysis 2009; 49: 1027-1033
  • 10 Zhang QH, Yang J, He Y et al. Food effect on the pharmacokinetics of entecavir from dispersible tablets following oral administration in healthy Chinese volunteers. Arzneimittelforschung 2010; 60: 640-644
  • 11 US Department of Health and Human Services. Food and Drug Administration, Center for Drug Evaluation and Research (CDER), Guidance for Industry, Bioanalytical Method Validation. 2001
  • 12 Li Z, Huang X, Jiang Z et al. A sensitive and specific liquid chromatography – mass spectrometry method for determination of metacavir in rat plasma. Journal of Chromatography B 2008; 864: 9-14
  • 13 Novakova L, Gottvald T, Vlckova H et al. Highly sensitive fast determination of entecavir in rat urine by means of hydrophilic interaction chromatography-ultra-high-performance liquid chromatography-tandem mass spectrometry. J Chromatogr A 2012; 1259: 237-243
  • 14 Li FY, Hao HP, Hao K et al. Effect of diammonium glycyrrhizinate on entecavir pharmacokinetics in rats. Chin J Nat Med 2013; 11: 0309-0313
  • 15 Wang DW, Li F, Li P et al. Validated LC-MS/MS assay for the quantitative determination of clematichinenoside AR in rat plasma and its application to a pharmacokinetic study. Biomed Chromatogr 2012; 26: 1282-1285