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Simultaneous determination of colchicine and febuxostat in rat plasma: Application in a rat pharmacokinetic study
Author(s) -
Gabani Bhavesh Babulal,
Saini Neeraj Kumar,
Jairam Ravi Kumar,
Shrinivas Pavan,
Trivedi Ravi Kumar,
Srinivas Nuggehally R.,
Mullangi Ramesh
Publication year - 2020
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.4939
Subject(s) - febuxostat , chemistry , chromatography , formic acid , colchicine , ammonium acetate , ammonium formate , pharmacokinetics , extraction (chemistry) , analyte , high performance liquid chromatography , pharmacology , biochemistry , uric acid , hyperuricemia , medicine
A selective, sensitive and rapid LC–MS/MS method has been developed and validated as per US Food and Drug Administration regulatory guidelines for the simultaneous quantitation of colchicine and febuxostat in rat plasma. Colchicine and febuxostat were extracted from the rat plasma using 10% tert ‐butyl methyl ether in ethyl acetate using colchicine‐ d 6 as an internal standard (IS). The chromatographic separation of colchicine, febuxostat and the IS was achieved using a mobile phase comprising 5 m m ammonium formate and 0.025% formic acid in acetonitrile (20:80, v/v) in isocratic mode on an Eclipse XDB‐C 18 column. The injection volume and flow rate were 5.0 μl and 0.9 ml/min, respectively. Colchicine and febuxostat were detected by positive electrospray ionization in multiple reaction monitoring mode using transition pairs (Q1 → Q3) of m/z 400.10 → 358.10 and 317.05 → 261.00, respectively. The assay was linear in the ranges of 0.25–254 and 2.60–622 ng/ml for colchicine and febuxostat, respectively. The inter‐ and intra‐day precision values were 0.58–13.0 and 1.03–4.88% for colchicine and febuxostat, respectively. No matrix or carryover effects were observed during the validation. Both analytes were stable on the bench‐top, in the autosampler and in storage (freeze–thaw cycles and long‐term storage at −80 ° C). A pharmacokinetic study in rats was performed to show the applicability of the validated method.