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Simultaneous determination of simvastatin and simvastatin acid in human plasma by LC‐MS/MS without polarity switch: Application to a bioequivalence study
Author(s) -
Patel Bhavin N.,
Sharma Naveen,
Sanyal Mallika,
Shrivastav Pranav S.
Publication year - 2008
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700367
Subject(s) - chromatography , chemistry , simvastatin , analyte , bioequivalence , extraction (chemistry) , selected reaction monitoring , metabolite , liquid–liquid extraction , solid phase extraction , analytical chemistry (journal) , mass spectrometry , pharmacokinetics , tandem mass spectrometry , pharmacology , biochemistry , medicine
A simple, specific and sensitive LC‐MS/MS assay for simultaneous determination of simvastatin (SV) and its active β‐hydroxy acid metabolite, simvastatin acid (SVA) in human plasma was developed using a statin analog as internal standard (IS). The method was validated over a dynamic linear range of 0.20–100.00 ng/mL for SV and 0.10–50.00 ng/mL for SVA with correlation coefficient r ⪈ 0.9987 and 0.9989, respectively. The analytes and IS were extracted from 500 μL aliquots of human plasma via liquid‐liquid extraction using methyl tert ‐butyl ether and separated through an Aquasil C18 column (100 mm×2.1 mm, 5 μm). Detection of analytes and IS was done by MS/MS with a turbo ion spray interface operating in positive ion and selective reaction monitoring acquisition mode. The total chromatographic run time was 3.0 min. Flash freezing of the aqueous phase was an added advantage during liquid‐liquid extraction, which considerably reduced time and labour. The method was extensively validated for its accuracy, precision, recovery, stability studies and matrix effect. The method was successfully used for bioequivalence study of 40 mg SV tablet formulation in 12 human subjects under fasting condition.