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Determination of tributyrin and its metabolite butyrate in Wistar rat plasma samples by gas chromatography/mass spectrometry
Author(s) -
Su Jie,
Zhang Ningning,
Ho Paul C.
Publication year - 2004
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1607
Subject(s) - tributyrin , chemistry , chromatography , metabolite , protein precipitation , detection limit , mass spectrometry , gas chromatography , butyrate , analyte , gas chromatography–mass spectrometry , enzyme , biochemistry , lipase , fermentation
A gas chromatographic (GC) method with mass spectrometric (MS) detection was developed for the determination of tributyrin and its metabolite butyrate in rat plasma. Following precipitation of plasma protein with acetonitrile, the analytes in the samples were separated on a DB‐5ms capillary column with helium as carrier gas. Phenylmethylsulfonyl fluoride (PMSF), an inhibitor for serine proteases, papain and acetylcholinesterase, was found to be essential to inhibit the activity of enzyme(s) responsible for the hydrolysis of tributyrin in both rat and human blood samples. The enzyme inhibitor in 5 mM (final concentration) was added immediately into the blood samples after collection to prevent the hydrolysis. The linear concentration ranges for tributyrin and butyrate were 0.1–2.0 and 1–20 μM, respectively. The coefficients of variation for intra‐day and inter‐day assays for tributyrin were all <10%, and those for butyrate were also <10%, except for the lowest concentration (1 μM), which was less than 20%. The accuracy of all concentration determinations ranged from 96.0–110.0%. The limit of quantification (LOQ) was 0.1 μM for tributyrin and 1.0 μM for butyrate. This method could detect tributyrin and butyrate simultaneously, and represents an improvement in sensitivity for the detection of tributyrin compared with the previous gas chromatography‐flame ionization detection (GC‐FID) method. Copyright © 2004 John Wiley & Sons, Ltd.