HPLC法测定(1R,2R)-(-)-1,2-环己二胺中S,S对映异构体

Determination of S,S-enantiomer in(1R,2R)-(-)-1,2-cyclohexanediamine by HPLC

目的建立高效液相色谱法测定(1R,2R)-(-)-1,2-环己二胺中S,S对映异构体含量的方法。方法柱前衍生反相高效液相色谱法,利用间甲基苯甲酰氯为衍生化试剂,采用Chiral-AGP(4.0 mm×150 mm)色谱柱,以异丙醇-磷酸盐缓冲液(pH值=6.1)(15∶85)为流动相、流速0.7 mL.min-1、检测波长为214 nm。结果 1,2-环己二胺的R,R与S,S对映异构体衍生物达到基线分离;S,S-对映异构体的定量限为30.1 ng、检测限12.4 ng;S,S-对映异构体在0.015～0.09μg.mL-1浓度范围内线性良好,平均回收率为98.3%,RSD(n=9)为9.36%。结论建立的间甲基苯甲酰氯柱前衍生高效液相色谱法适用于(1R,2R)-(-)-1,2-环己二胺中S,S对映异构体含量的测定。

Objective To establish an HPLC method for the determination of S, S - enantiomer in （ 1 R, 2R ） - （ - ） - 1,2 - cyclohexanediamine. Methods Pre - column derivatization of RE - HPLC was used with m - toluoyl chloride as derivatization reagent and Chiral -AGP （4.0 mm × 150 mm） chromatographic column was adopted. The mobile phase was isopropanol -phos- phate buffer solution （ pH = 6.1 ） （ 15：85 ） and the flow rate was 0.7 mL .min^-1. The detection wavelength was 214 nm. Results The baseline resolution was achieved between the derivants of S, S - and R, R - enantiomers of 1,2 - cyclohexanedia - mine. The limit of quantitation and detectability of S, S - enantiomer were 30.1 ng and 12.4 ng. The linear range of S, S - enantiomer was 0.015 - 0. 09 μg.mL^ -1 , and the average recovery was 98.3 % （ RSD = 9. 36% , n = 9 ）. Conclusion Pre - column derivatization of RE - HPLC with m - toluoyl chloride as derivatization reagent was suitable for the determination of S, S - enantiomer in （1R,2R） - （ - ） - 1,2 - cyclohexanediamine.