Simultaneous determination of homocysteine,methionine and cysteine in maternal plasma after delivery by HPLC‐fluorescence detection with DBD‐F as a label |
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Authors: | Mitsuhiro Wada Maki Hirose Mana Kuroki Rie Ikeda Yui Sekitani Noboru Takamura Naotaka Kuroda Kenichiro Nakashima |
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Institution: | 1. Graduate School of Biomedical Sciences, Nagasaki University, , Nagasaki, 852‐8521 Japan;2. Faculty of Pharmaceutical Sciences, Nagasaki International University, , Sasebo, Nagasaki, 859‐3298 Japan |
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Abstract: | An HPLC‐fluorescence (FL) method for determination of sulfur‐containing amino acids such as homocysteine (Hcy), methionine (Met) and cysteine (Cys) in human plasma was developed. The sulfur‐containing amino acids were labeled with 4‐(N,N‐dimethylaminosulfonyl)‐7‐fluoro‐2,1,3‐benzoxadiazole (DBD‐F). Calibration curves in the range of 1–100 µm (Hcy and Met) and 5–500 µm (Cys) indicated good linearities (r ≥ 0.998). The limits of detection at a signal‐to‐noise ratio of 3 were 0.13 (Hcy), 0.02 (Met) and 0.11 µm (Cys), respectively. Acceptable results for accuracy and precision of intra‐ and inter‐day measurements were obtained. The results of Hcy and Cys obtained by the proposed method indicated good correlations with the conventional method (r > 0.911, n = 20). Furthermore, the method was applied to determination of the sulfur‐containing amino acids in maternal plasma (n = 200) after delivery. The concentrations of Hcy, Met and Cys as a median (inter quartile range, Q1 and Q3) were 5.37 (3.32–7.79) μm , 25.20 (20.10–31.06) μm and 147.25 (102.81–189.31) μm , respectively. Copyright © 2012 John Wiley & Sons, Ltd. |
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Keywords: | homocysteine methionine cysteine HPLC‐fluorescence plasma 4‐(N N‐dimethylaminosulfonyl)‐7‐fluoro‐2 1 3‐benzoxadiazole |
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