2,2-二甲基环丙甲酰胺的HPLC测定及其在手性拆分反应监测中的应用

目的 建立2,2-二甲基环丙甲酰胺的HPLC检测方法,并以该非手性的HPLC方法监测2,2-二甲基环丙甲酰胺的手性拆分反应.方法 采用C_(18)反相色谱柱,柱温为40℃,以50mmol/L,pH3.0的磷酸二氢钠:甲醇=1:1(V/V)为流动相,流速为0.5mL/min,检测波长为210nm.结果 2,2-二甲基环丙甲酰胺在10-1000μg/mL浓度范围内线性关系良好,2,2-二甲基环丙甲酰胺、2,2-二甲基环丙甲酸的分离度为14.28;以本法监控酰胺酶的手性拆分反应液,当HPLC测定结果显示反应液中底物不再降低时,手性气相检测表明(S)-2,2-二甲基环丙甲酰胺的ee值达到99.926%.结论 HPLC方法灵敏度高、线性关系良好;HPLC测定结果可以很好地用于2,2-二甲基环丙甲酰胺手性拆分反应的监测.

HPLC method for 2,2-dimethylcyclopropanecarboxamide determination and its use in monitoring chiral resolution of 2,2-dimethylcyclocarboxamide

Objective To establish a HPLC method for the determination of 2, 2-dimethylcyclopropanecarbox amide, and use this method to monitor the chiral resolution reaction of 2, 2-dimethylcyclopropanecarboxamide for the production of (S)-2, 2-dimethylcyclopropanecarboxamide. Methods C_(18) reversed-phase column Hypersil ODS2 was used.The column temperature was 40℃, the mobile phase was 50% 50mmol/L pH3.0 sodium phosphate buffer and 50% methanol, the flow rate was 0.5mL/min, and the detection wavelenth was 210nm. Results The calibration curve was linear in the range of 10-1000μg/mL, and the resolution between 2, 2-dimethylcyclopropanecarboxamide and 2, 2-dimethylcyclopropanecarboxylic acid was 14.28. This method was also used for the analysis of chiral resolution reaction of 2, 2-dimethylcyclocarboxamide with amidase. When the concentration of substrate determined by HPLC method did not decrease, the (S)-2, 2-dimethylcyclocarboxamide ee value of the unreacted substrate was 99.926%. Conclusion This achiral HPLC method was very sensitive with high linear relationship, and could be used for the chiral resolution reaction for the monitor in the production of (S)-2, 2-dimethylcyclopropanecarboxamide.