Wada Mitsuhiro, Hirose Maki, Kuroki Mana, Ikeda Rie, Sekitani Yui, Takamura Noboru, Kuroda Naotaka, Nakashima Kenichiro
Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki, 852-8521, Japan.
Biomed Chromatogr. 2013 Jun;27(6):708-13. doi: 10.1002/bmc.2848. Epub 2012 Dec 6.
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.
建立了一种用于测定人血浆中含硫氨基酸如高半胱氨酸（Hcy）、蛋氨酸（Met）和半胱氨酸（Cys）的高效液相色谱 - 荧光（FL）法。含硫氨基酸用4 - （N，N - 二甲基氨基磺酰基） - 7 - 氟 - 2，1，3 - 苯并恶二唑（DBD - F）进行标记。1 - 100 μmol/L（Hcy和Met）以及5 - 500 μmol/L（Cys）范围内的校准曲线显示出良好的线性关系（r≥0.998）。信噪比为3时的检测限分别为0.13（Hcy）、0.02（Met）和0.11 μmol/L（Cys）。日内和日间测量的准确度和精密度结果均可接受。所提方法测得的Hcy和Cys结果与传统方法显示出良好的相关性（r>0.911，n = 20）。此外，该方法还应用于分娩后母体血浆（n = 200）中含硫氨基酸的测定。Hcy、Met和Cys的浓度中位数（四分位间距，Q1和Q3）分别为5.37（3.32 - 7.79）μmol/L、25.20（20.10 - 31.06）μmol/L和147.25（102.81 - 189.31）μmol/L。
