Iwase H, Ozawa S, Ikuta M, Ono I
Ajinomoto Co., Central Research Laboratories, Kawasaki, Japan.
J Chromatogr B Biomed Appl. 1995 Jan 6;663(1):15-24. doi: 10.1016/0378-4347(94)00439-c.
Amino acids in human plasma were determined by liquid chromatography with postcolumn ninhydrin derivatization using a hydroxyapatite cartridge for precolumn deproteination. S-Carboxymethyl-L-cysteine, D-phenylglycine and S-aminoethyl-L-cysteine were found to be suitable internal standards. The proposed method is simple, rapid (deproteination time less than 1 min) and reproducible [relative standard deviation below 3% except for low-level aspartic acid (n = 3)]. The average recovery of 25 amino acids was above 90%. The elution time of amino acids in human plasma was approximately 2 h. Protein binding of tryptophan was also determined by the proposed method. The analytical data for amino acids in human plasma deproteinated using the proposed and published methods (5-sulphosalicylic acid and ethanol) were compared.
采用羟基磷灰石柱进行柱前脱蛋白，通过柱后茚三酮衍生化的液相色谱法测定人血浆中的氨基酸。发现S-羧甲基-L-半胱氨酸、D-苯甘氨酸和S-氨乙基-L-半胱氨酸是合适的内标物。所提出的方法简单、快速（脱蛋白时间少于1分钟）且具有可重复性[除低水平天冬氨酸外（n = 3），相对标准偏差低于3%]。25种氨基酸的平均回收率高于90%。人血浆中氨基酸的洗脱时间约为2小时。还采用所提出的方法测定了色氨酸的蛋白结合率。比较了使用所提出的方法和已发表的方法（5-磺基水杨酸和乙醇）对人血浆进行脱蛋白后氨基酸的分析数据。
