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Published on:Oct 2016
Indian Journal of Pharmaceutical Education and Research, 2016; 50(4):612-619
Original Article | doi:10.5530/ijper.50.4.14

Picogram Level Quantification of Grazoprevir and Elbasvir with Deuterated Internal Standards in Human Plasma Samples by LC–ESI-MS/MS


Authors and affiliation (s):

Haritha Potluri1*, Sreenivasa Rao Battula2, Sunandamma Yeturu3

1Department of Chemistry, JNTUK, Kakinada-533003, INDIA

2Department of Chemistry, GITAM University, Visakhapatnam-530045,INDIA

3Department of Chemistry, Acharya Nagarjuna University, Nagarjuna nagar-522510, INDIA

Abstract:

A fixed oral dose combination of grazoprevir and elbasvir is used for the treatment of patients with genotypes 1&4 HCV infections. In this manuscript, authors developed a simple, sensitive and specific liquid chromatography–tandem mass spectrometry (LC–MS/MS) method used for quantification of grazoprevir and elbasvir in human plasma. Agilent TC-C18, 4.6 x 75 mm, 3.5 μm, 80 Å column, and 5 mM ammonium acetate: acetonitrile (20:80 v/v) mobilephase was used for Chromatographic separation. MRM positive mode was used to detect the grazoprevir and elbasvir at m/z 767.3/553.2, 883.4/656.3 respectively. Liquid-liquid extraction was employed in the extraction of analytes and internal standard from human plasma. This method is validated over a linear concentration range of 50.0 – 10000.0 pg/ml for grazoprevir and elbasvir. Calibration functions, lower limit of quantitation (LLOQ), stability, intra and inter-day reproducibility, accuracy, and recovery are estimated. Both drugs were stable in plasma samples. This method was free from matrix effects and any abnormal ionization.

Key words: LC-ESI-MS/MS, Grazoprevir, Elbasvir, Human plasma, Bioanalytical.

 

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The Official Journal of Association of Pharmaceutical Teachers of India (APTI)
(Registered under Registration of Societies Act XXI of 1860 No. 122 of 1966-1967, Lucknow)

Indian Journal of Pharmaceutical Education and Research (IJPER) [ISSN-0019-5464] is the official journal of Association of Pharmaceutical Teachers of India (APTI) and is being published since 1967.

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