• S. SATHESHKUMAR Department of Pharmaceutics, College of Pharmacy, Vinayaka Mission’s College of Pharmacy, Salem 636008, Tamilnadu, India
  • V. MURUGANANTHAM Department of Pharmaceutics, College of Pharmacy, Vinayaka Mission’s College of Pharmacy, Salem 636008, Tamilnadu, India


Objective: The current research work focus to simple and rugged bioanalytical method development and validation of brivudine in human plasma using high-performance liquid chromatography.

Methods: The analyte (Brivudine) and internal standard (Sofosbuvir) were extracted using the Solid Phase Extraction (SPE) technique. The chromatographic separation was accomplished by using Zorbax eclipse XDB-C18 Column (150×4.6 mm, 5 μm) with a mobile phase consisted of Methanol: 0.5% Ortho-phosphoric acid (65:35%, v/v) respectively, at a flow rate of 0.7 mL/min. The developed method was validated by performing system suitability, carryover effect, linearity, selectivity, sensitivity, precision, accuracy, recovery, ruggedness, and stability studies. The method was validated as per USFDA guidelines.

Results: The selected chromatographic condition was found to efficiently separated brivudine (RT-3.55 min) and ISTD (RT-7.87 min). The assay demonstrated a linear dynamic range of 85.205 to 4500.246 ng/ml for brivudine in human plasma with r2>0.99. Demonstrated the lowest limit of detection at 85.205 ng/ml. This method established an intra-run and inter-run precision within the range of 2.99-6.31%CV and 3.67-5.80%CV, respectively. Additional intra-run and inter-run accuracy were within the range of 97.55-105.37% and 99.27-102.15%, respectively. The mean percentage recovery of brivudine and ISTD studies proved good extraction efficiency and the robustness was also evaluated.

Conclusion: A simple, accurate, precise, linear and rugged RP-HPLC method was developed and validated for the estimation of brivudine in human plasma with K2EDTA anticoagulant and suitable for conducting BA/BE and TDM.

Keywords: Brivudine, Human Plasma, Liquid Chromatography, Solid Phase Extraction


Download data is not yet available.


1. Rabasseda X. Brivudine a herpes virostatic with rapid antiviral activity and once-daily dosing. Drug Today 2003;39:359-71.
2. Salvaggio MR, Gnann. JW JR. Drugs for herpesvirus infections. Anti-Infective Therapy 2004;153:1309-17.
3. De Clercq E, Lia G. Approved antiviral drugs over the past 50 Y. Clin Microbiol Rev 2016;29:696-47.
4. Wassilew SW. Brivudin compared with famciclovir in the treatment of herpes zoster effects in acute disease and chronic pain in immune-competent patients. A randomized, double-blind, multinational study. Eur Acad Dermatol Venereol JEADV 2005;19:47–55.
5. European Medicines Agency Report for brivudine. Committee for Orphan Medicinal Products EMA/COMP/768295/2009; 2010.
6. https://www.drugbank.ca/drugs/DB03312 [Last accessed on 10 Jun 2019]
7. Public Assessment Report for brivudine. ZosterGalen 125 mg Tabletten, DE/H/5591/001/DC; 2019.
8. Binu V, Sangeetha S. Method development and validation of brivudine by UV spectrophotometric method. Int J Curr Adv Res 2018;5:12276-80.
9. Olgemoller J, Hempel G, Boos J, Blaschke G. Determination of (E)-5-(2-bromovinyl)-29-deoxyuridine in plasma and urine by capillary electrophoresis. J Chromatogr B 1999;726:261-8.
10. Reeuwijk HJEM, Tjaden UR, Greef JVD. Development and validation of a bioanalytical assay for (E)-5-(2-promo vinyl)-29-deoxyuridine in plasma by capillary zone electrophoresis. J Chromatogr B 1999;726:269-76.
11. Ayisi NK. High-performance liquid chromatographic analysis of (E)-5-(2-bromovinyl)-Z’deoxyuridine and its metabolites in serum, urine and herpes simplex virus type-l infected cells. J Chromatogr 1986;375:423-30.
12. Reeuwijk HJEM, Lingeman H, Tjaden UR, Bruijn EAD, Keizer HJ, Greef JVD. Bioanalysis of (e)-5-(2-bromovinyl)-2’–deoxyuridine. J Chromatogr 1988;428:93-102.
13. Peng XD, Tan ZR, Wu HZ, Zhou G, Guo CX, Pei Q, et al. LC–MS–MS quantitative determination of brivudine in human plasma and its application to pharmacokinetic studies. Chromatographia 2011;73:1089–95.
14. Food and drug Administration. Guidance for Industry: bioanalytical method validation. Rockville MD: US Department of Health and Human Services, Food and Drug Administration. Centre for Drug Evaluation and Research; 2018.
15. Tijare LK, Rangari NT, Mahajan UN. A review on bioanalytical method development and validation. Asian J Pharm Clin Res 2016;9:6-10.
16. Dyade GK, Sawant RL. Simple bioanalytical quantification method for simultaneous estimation of simvastatin and ezetimibe in human plasma by reverse-phase high-performance liquid chromatography technique. Asian J Pharm Clin Res 2020;13:160-5.
90 Views | 118 Downloads
How to Cite
SATHESHKUMAR, S., and V. MURUGANANTHAM. “A SIMPLE AND RUGGED BIOANALYTICAL METHOD DEVELOPMENT AND VALIDATION OF BRIVUDINE IN HUMAN PLASMA BY USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 12, no. 7, May 2020, pp. 45-50, doi:10.22159/ijpps.2020v12i7.37703.
Original Article(s)