HPLC手性流动相添加剂法拆分药典收载三种含N手性药物及其机制探讨

 目的 采用磺丁基醚-β-环糊精（SBE-β-CD）作为手性流动相添加剂,以反相高效液相色谱技术(RP-HPLC),建立了磷酸氯喹、马来酸氯苯那敏、盐酸维拉帕米3个含N碱性药典品种对映体在RP-HPLC系统中的拆分方法并探讨其机制。方法 色谱柱为Alltima C₁₈（4.6 mm×250 mm,5μm）;流动相：乙腈-水（含磺丁基醚-β-环糊精、磷酸二氢钠,用磷酸调pH值至2.5）;检测波长：332、264、278 nm;流速：0.8 mL·min^-1;柱温：25 ℃。结果 磷酸氯喹、马来酸氯苯那敏、盐酸维拉帕米可在上述色谱条件下分别与其对映体得到分离,同时考察手性流动相添加剂的种类和浓度、pH、缓冲盐的种类和浓度、有机相的种类和比例、柱温对药物对映体拆分效果的影响,选择了最佳的手性分离条件并对其分离机制作了探讨。结论 SBE-β-CD作为一种手性添加剂,用于手性流动相RP-HPLC,可拆分含N碱性的手性药物,其手性分离效果明显优于天然环糊精（β-CD）及衍生化的中性环糊精（HP-β-CD）。

Study on Chiral Separation of Three Drugs Collected Pharmacopoeia N-alkyl Group by HPLC Using Chiral Mobile Phase Additive and Its Mechanism

OBJECTIVE To performe chiral separation of three pharmaceutical racemates collected in Ch.P 2010 containing N-alkyl groups,including verapamil hydrochloride,chlorphenamine maleate,and chloroquine diphosphate by using SBE-β-CD as a mobile-phase additive in an HPLC system.METHODS The analysis was carried out on an Alltima C18 column.The mobile phase consisted of acetonitrile and a phosphate solution added with SBE-β-CD and NaH2PO4,whose pH value was adjusted to 2.5 by phosphoric acid.The flow rate was set at 0.8 mL· min-1,and the column temperature was 25 ℃.The UV detection was carried out at 332,264 and 278 nm,respectively.The effects of different factors on the separation effect were investigated,including CD types(β-CD,HP-β-CD),SBE-β-CD concentration,pH value,oven temperature,and buffer concentration.RESULTS The established method was able to separate the three racemates and the mechanism had been discussed.CONCLUSION Three racemates can be separated on C18 column using SBE-β-CD as a chiral mobile phase additive.This method might be applied to separate various types of biologically important enantiomers which have N-alkyl groups in the chemical structure.