柱前衍生－反相高效液相色谱分析复方氨基酸注射液

Analysis of Compound Amino-acid Injection by Precolumn Derivatization with o-Phthaldialdehyde and RP-HPLC

采用邻苯二甲醛柱前衍生、反相高效液相色谱和荧光检测分析了复方氨基酸注射液。于国产ＹＷＧ－Ｃ１８柱上，以甲醇－四氢呋喃－醋酸钠缓冲液之三元体系的６段台阶梯度洗脱。于λｅｘ＝３３５ｎｍ和λｅｍ＝４３０ｎｍ处荧光检测，１５种氨基酸衍生物在５０ｍｉｎ内均能得到良好的分离和测定。各氨基酸的浓度在０．１～１．６μｇ／ｍｌ范围内，线性相关系数大于０．９９，检出限为ｐｇ（ｆｍｏｌｅ）水平。本方法应用于测定３种复合氨基酸注射液中氨基酸的含量，已获得满意的结果。

Compound amino-acid injections were analysed as their precolumn derivatives with o-phthaldialdehyde by reversed-phase high-performance liquid chromatographic separation and fluorescence detection. On a Chinese-made YWG-C18 column, using methanol-tetrahydrofuran-acetate buffer (pH6.0) ternary system mobile phase by 6-stage gradient eluation, the derivatives of 15 amino acids with o-phthaldialdehyde can be separated with a stable base line within 50 min and detected fluorescently at λex 335 nm and λem 430nm. The calibration curves were linear over the range 0.1～1.6μg/ml for each of amino acids, the regression coefficients were higher than 0.99 and the limits of detection were found to be fmole level. The method has been applied to determine amino acids in three kinds of compound amino-acid injections with satisfactory results.