高效液相色谱法分离测定DL-天冬氨酸对映体

Separation and Determination of Aspartic Acid Enantiomers by High Performance Liquid Chromatography

用邻苯二甲醛（OPA）、N-乙酰-L-半胱氨酸为衍生试剂对天冬氨酸对映体进行柱前衍生．采用ZORBAX Eclipse AAA4．6×150mm，5μm色谱柱，以0．05mol／LpH5．08的醋酸钠溶液与甲醇按57：43体积比作为流动相，流速0．5mL／min，柱温40℃下，考察高效液相色谱柱前衍生化法分离测定天冬氨酸对映体的效果．结果显示：在3．5min内分离出天冬氨酸对映体．在20～100nmol／mL线性范围内，线性关系良好，相关系数在0．99915以上，相对标准偏差在0．65％～1．70％之间．该法快速、简便、分离度高，能够完全分离DL-天冬氨酸对映体，并对其他氨基酸对映体的分离有良好的参考价值．

Aspartic acids enantiomers were precolumn derivatized with O-phthalaldehyde（OPA） and N- acetyl-L-cysteine through a ZORBAX Eclipse AAA column （4.6 × 150 mm,5μm）.The mobile phase was methanol-sodium acetate solution（adjusting to pH=5.08）= 43 ： 57 （V/V）.The flow rate was 0.5 mL/min.The column temperature was 40 ℃.The influence of precolumn derivatization on the enantiomeric separations of Aspartie acids were investigated.The results were determined as follows. The separation was finished in 3.5minute. The calibration curve was linear in the range of 20 - 100 nmol/mL. The RSD were 0.65% - 1.70%. The analytical method was rapid, simple with high resolution and could be used as a reference of chiral resolution to other amino acid enantiomers .