手性固定相HPLC法拆分苏氨酸和赖氨酸对映体

摘要

Objective To establish chiral stationary phases (CSPs) combining with prc-column derivation method for the chiral separation of thrconinc (Thr) and lysinc (Lys). Method The cnantiomcrs of thrconinc and lysinc were derivatized with reagent 4-chlo-ro-7-nitro-2,1,3-bcnzoxadiazolc( NBD-C1). The diastcrcoisomcrs produced were separated on a Sumichiral OA-2500S CSP column with a mobile phase consisting of mcthanol (including 5 mmol · L-1 citric acid)-acctonitrilc (60:40) and wavelength was set at 470 nm. Results The calibration curves of D -thrconinc, and D -lysinc were linear in the concentration range of 2. 0-16. 9 μg · mL-1. The average recoveries of D -thrconinc and D -lysinc were 101. 0% , with RSD of 1. 7 % and 0. 37% , respectively. Conclusions Under the optimized chromatographic conditions,baseline separations of these amino acid cnantiomcrs were obtained. The method is sensitive and reproducible.%目的 建立了柱前衍生结合手性固定相HPLC法拆分了苏氨酸和赖氨酸2种氨基酸对映体.方法 利用4-氯-7-硝基-2,1,3-苯并噁二唑(NBD-Cl)柱前衍生,采用Sumichiral OA-2500S色谱柱,以甲醇-乙腈(60∶40,含5 mmol L-1柠檬酸)为流动相,检测波长为470 nm.结果 应用该法检查L- 氨基酸原料药中的D- 型异构体,D- 苏氨酸和D- 赖氨酸在2.0～16.0 μg mL-1质量浓度范围内线性关系良好,回收率均为101.0%,RSD分别为1.7%和0.37%.结论 在最佳色谱条件下,2种氨基酸对映体均达到了基线分离.建立的对映体杂质检查方法灵敏度高,重复性好.