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

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

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)。

OBJECTIVE To performe chiral separation of three pharmaceutical racemates collected in Ch. P 2010 containing N-al- kyl 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 Cls column. The mobile phase consisted of acetonitrile and a phosphate solution added with SBE-fl-CD and NaH2PQ , 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 （^-CD, HP-fl- CD）, SBE-/3-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 impor- tant enantiomers which have N-alkyl groups in the chemical structure.