基于万古霉素手性柱的马来酸扑尔敏对映体分离及其识别机理

Chiral separation of chlorphenamine maleate on vancomycin-based stationary phase and chiral recognition mechanism

药物是环境介质中一类新型污染物,多数药物具有手性对映体特性,药物对映体的分离分析是研究对映体水平药物环境行为和效应的前提.采用商品手性柱Chirobiotic®V,在反相条件下,对马来酸扑尔敏进行了手性分离,考察了不同流速、柱温、pH值和流动相组成对分离的影响,并优化了色谱条件.结果表明:以含0.1%(体积分数)冰醋酸和0.1%(体积分数)三乙胺的水溶液(TEAA)与四氢呋喃(THF)混合为流动相,在所研究范围内,随着流动相中THF含量的增加、pH值增大和流速增大,马来酸扑尔敏对映体的分离效果降低;柱温影响存在峰值现象,即分离效果随温度的升高先增大后减小.优化得到的拆分条件为:流动相TEAA∶THF为95∶5(V/V),pH值为3,流速为0.3 mL·min^(-1),柱温为15℃.利用分子对接技术模拟万古霉素不同区域与扑尔敏对映体间的相互作用,计算得到了最稳定结合能,据此预测对映体的出峰顺序与实验结果完全一致.初步手性识别机理研究结果表明,引起手性识别的主要作用力为氢键作用.

Pharmaceuticals are a large class of emerging pollutants in environmental matrix.It has been reported that more than 50%of these pharmaceuticals are chiral compounds.Thus,the chiral separation and analysis of pharmaceuticals are prerequisites for investigating environmental behavior and impact of drug enantiomers.The commercial vancomycin-bonded chiral stationary phase(CSP)combined with high performance liquid chromatography(HPLC)in reverse mode was used for the enantioseparation of chlorphenamine maleate enantiomers.The flow rate,column temperature,pH value and composition of mobile phase were optimized for the effective separation.The results showed that when TEAA containing 0.1%of glacial acetic acid and 0.1%of triethylamine(in volume ratio)was mixed with tetrahydrofuran(THF)as mobile phase,the reduced enantioresolution and selectivity were observed with an increased THF percentage,pH value and flow rate.The effect of column temperature on separation indicated that maximum values in enantioresolution and selectivity occurred at 15℃.As a result,the optimal chromatographic condition was:TEAA∶THF=95∶5(V/V),pH value of 3,flow rate of 0.3 mL·min-1,temperature of 15℃.Using molecular docking technique and binding energy calculations to simulate the interaction processes between chlorphenamine maleate and vancomycin,the magnitudes of steady combination energy(ΔG)was obtained,which coincided with experimental elution order of the enantiomers.The molecular docking results showed that the main retention mechanisms of chiral chlorphenamine maleate onto vancomycin were hydrogen bonding.