Quantification of Sofosbuvir in Human Plasma

RP-HPLC Method Development and Validation

Rahul U. Rathod, K. S. Navyasree, Krishnamurthy Bhat

Research output: Contribution to journalArticle

Abstract

Sofosbuvir, a potential inhibitor of the nucleotide analog of hepatitis C virus (HCV) NS5B polymerase, is used to treat hepatitis C and marketed under brand name Sovaldi. This study was aimed to develop and validate a simple, sensitive and precise RP-HPLC method for the estimation of sofosbuvir in human blood plasma using quinine sulfate as internal standard (ISTD). Separation was achieved using Shimadzu-HPLC system equipped with Gracesmart RP-C18 (250 mm × 4.6 mm, 5 μm) column eluted at a flow rate of 0.5 mL/min and detected at 260 nm. The mobile-phase composition was optimized to be ammonium formate (pH 3.0; 10 mM) and acetonitrile (67: 33, % v/v). The retention times of sofosbuvir and quinine sulfate were found to be 13.60 and 8.39 min, respectively. The proposed method was linear over a range from 20 to 1200 ng/mL. Liquid–liquid extraction technique was applied for the extraction of drug and ISTD from human plasma using TBME – DCM (80: 20, % v/v) mixture. The proposed method was validated as per bioanalytical method validation guidelines for industry.

Original languageEnglish
Pages (from-to)663-673
Number of pages11
JournalPharmaceutical Chemistry Journal
Volume52
Issue number7
DOIs
Publication statusPublished - 01-10-2018

Fingerprint

High Pressure Liquid Chromatography
formic acid
Quinine
Hepatitis C
Hepacivirus
Names
Industry
Nucleotides
Guidelines
Sofosbuvir
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Pharmacology
  • Drug Discovery

Cite this

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abstract = "Sofosbuvir, a potential inhibitor of the nucleotide analog of hepatitis C virus (HCV) NS5B polymerase, is used to treat hepatitis C and marketed under brand name Sovaldi. This study was aimed to develop and validate a simple, sensitive and precise RP-HPLC method for the estimation of sofosbuvir in human blood plasma using quinine sulfate as internal standard (ISTD). Separation was achieved using Shimadzu-HPLC system equipped with Gracesmart RP-C18 (250 mm × 4.6 mm, 5 μm) column eluted at a flow rate of 0.5 mL/min and detected at 260 nm. The mobile-phase composition was optimized to be ammonium formate (pH 3.0; 10 mM) and acetonitrile (67: 33, {\%} v/v). The retention times of sofosbuvir and quinine sulfate were found to be 13.60 and 8.39 min, respectively. The proposed method was linear over a range from 20 to 1200 ng/mL. Liquid–liquid extraction technique was applied for the extraction of drug and ISTD from human plasma using TBME – DCM (80: 20, {\%} v/v) mixture. The proposed method was validated as per bioanalytical method validation guidelines for industry.",
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Quantification of Sofosbuvir in Human Plasma : RP-HPLC Method Development and Validation. / Rathod, Rahul U.; Navyasree, K. S.; Bhat, Krishnamurthy.

In: Pharmaceutical Chemistry Journal, Vol. 52, No. 7, 01.10.2018, p. 663-673.

Research output: Contribution to journalArticle

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