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Quantification of metronidazole in healthy subjects' feces using an LC–MS/MS method
Author(s) -
Vanol Pravin G.,
Sanyal Mallika,
Shah Priyanka A.,
Shrivastav Pranav S.
Publication year - 2018
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4265
Subject(s) - chromatography , chemistry , formic acid , extraction (chemistry) , mass spectrometry , electrospray ionization , ethyl acetate , liquid chromatography–mass spectrometry , triple quadrupole mass spectrometer , feces , human feces , solid phase extraction , selected reaction monitoring , tandem mass spectrometry , microbiology and biotechnology , biology
Abstract A highly selective and sensitive liquid chromatography–tandem mass spectrometry (LC MS/MS) method was developed for the quantification of metronidazole (MTZ) in human feces. The analyte was recovered from feces after liquid–liquid extraction with ethyl acetate and separated on Waters Symmetry® C 18 (100 × 4.6 mm, 5μm) column using 0.1% formic acid in water and acetonitrile (40:60, v /v) as the mobile phase. A stable‐deuterated internal standard metronidazole‐d4 (MTZ‐d4) was used in the study. Mass analysis was performed on a triple quadrupole mass spectrometer in the positive electrospray ionization mode. A linear response function of MTZ was established in the concentration range of 0.50–250 ng/g, based on dry mass. The mean extraction recovery of MTZ (97.28%) and MTZ‐d4 (96.76%) from spiked feces samples was consistent at higher as well as lower concentrations. Post‐column infusion analysis showed no ion‐suppression/enhancement effects and the mean IS‐normalized matrix factor ranged from 0.986 to 1.013. Spiked feces samples stored at −20 and − 70°C for long‐term stability were stable for at least 3 months, while extracted samples (dry and wet extracts) were stable up to 24 h. The method was applied to determine MTZ in feces of 12 healthy Indian subjects.