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 |
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Authors: | Xu Ying Huang Xiao Nie Xiuli Yang Li Yan Weili Wang Zhengtao Wang Changhong Hu Zhibi |
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Institution: | The Ministry of Education Key Laboratory for Standardization of Chinese Medicines and the State Administration of TCM Key Laboratory for New Resources and Quality Evaluation of Chinese Medicines, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201210, China. |
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Abstract: | A rapid, selective and sensitive hydrophilic interaction liquid chromatography (HILIC) coupled with tandem mass spectrometry (MS/MS) method was developed to determine 1‐13C‐l ‐methionine in rat serum. Proteins in serum were precipitated using acetonitrile and the supernatant was separated after centrifugation. 1‐13C‐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 mm ammonium acetate containing 0.1% formic acid and multiple reaction monitoring mode for 1‐13C‐l ‐methionine and 1‐13C‐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/x2) 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 1‐13C‐l ‐methionine in rats. Copyright © 2011 John Wiley & Sons, Ltd. |
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Keywords: | 1‐13C‐l‐methionine hydrophilic interaction liquid chromatography tandem mass spectrometry pharmacokinetics |
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