Premium
Quantification of 1‐ 13 C‐ l ‐methionine in rat serum with hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry and its application in pharmacokinetic study
Author(s) -
Xu Ying,
Huang Xiao,
Nie Xiuli,
Yang Li,
Yan Weili,
Wang Zhengtao,
Wang Changhong,
Hu Zhibi
Publication year - 2011
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.1563
Subject(s) - chemistry , chromatography , hydrophilic interaction chromatography , formic acid , tandem mass spectrometry , ammonium acetate , selected reaction monitoring , mass spectrometry , liquid chromatography–mass spectrometry , detection limit , acetonitrile , high performance liquid chromatography
Abstract A rapid, selective and sensitive hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass spectrometry (MS/MS) method was developed to determine 1‐ 13 C‐ l ‐methionine in rat serum. Proteins in serum were precipitated using acetonitrile and the supernatant was separated after centrifugation. 1‐ 13 C‐ l ‐phenylalnine was used as the internal standard. HILIC–tandem mass spectrometry analysis was performed on a hydrophilic interaction silica column (TSK‐GEL AMIDE‐80) using a linear gradient elution system, acetonitrile−5 m m ammonium acetate containing 0.1% formic acid and multiple reaction monitoring mode for 1‐ 13 C‐ l ‐methionine and 1‐ 13 C‐ l ‐phenylalnine. The assay was validated with a linear range between 10 and 150 ng mL −1 ( r ≥ 0.99) and a lower limit of quantification of 10 ng mL −1 , calculated with weighted (1/ x 2 ) least squares linear regression. The RSD of intra‐day precision was smaller than 3.6% and the inter‐day RSD less than 6.5%, while the average recovery was 100.48% with an RSD of accuracy within 2.9%, determined from quality control samples. The HILIC‐MS/MS method was fully validated and successfully applied to the in vivo pharmacokinetic study of stable‐isotope 1‐ 13 C‐ l ‐methionine in rats. Copyright © 2011 John Wiley & Sons, Ltd.