类特异性分子印迹固相萃取/高效液相色谱法分析马尿泡果实中4种托烷类生物碱

Analysis of Four Tropane Alkaloids in Przewalskia Tangutica Maxim.by Analogous-specific Molecularly Imprinted Solid-phase Extraction Combined with High Performance Liquid Chromatography

以樟柳碱（ASD）为模板分子,甲基丙烯酸（MAA）为功能单体,三羟甲基丙烷三甲基丙烯酸酯（TRIM）为交联剂,乙腈为致孔剂,采用沉淀聚合法,合成了对4种托烷类生物碱（樟柳碱、东莨菪碱、山莨菪碱、阿托品）具有类特异性识别能力的聚合物。通过紫外光谱预测法筛选了最佳功能单体与配比。采用振荡平衡吸附实验对印迹和非印迹聚合物进行了吸附性能表征,结果表明,印迹聚合物对4种生物碱的饱和吸附量分别为7.53,10.90,24.27,11.04μg/mg,相对选择性系数分别为3.58,1.49,1.62,2.25。以该分子印迹聚合物为固相萃取柱填料,采用分子印迹固相萃取/高效液相色谱法实现了藏药马尿泡中4种托烷类生物碱的高效富集和快速分离。该方法在2-250μg/m L范围内具有较好的线性关系,相关系数（r^2）为0.999 9,检出限为0.26-0.39μg/m L,在10,50,100μg/g加标浓度下的平均回收率为70.0%-96.3%,相对标准偏差（RSD）≤5.7%。该固相萃取柱重复使用率高,分离效果良好,有效去除了样品中杂质的干扰,大大提高了马尿泡果实中4种托烷类生物碱的萃取效率。

New molecularly imprinted polymers（MIPs） for recognizing four tropane alkaloids（anisodine,scopolamine,anisodamine,atropine） were synthesized by precipitation polymerization using anisodine（ASD） as template,methacrylic（MAA） as functional monomer,trimethylolpropane trimethacrylate（TRIM） as crosslinker and acetonitrile as porogen.The optimal functional monomer and the ratio were selected by UV spectrum method.Comparative determination of the prepared MIP and NIPwere carried out by equilibrium adsorption experiment.The results showed that the MIPs possessed the analogous-specific adsorptivity toward four TAs.Scatchard anilysis displayed that the apparent maximum binding capacities（Qmax） are 7.53,10.90,24.27,11.04 μg /mg,respectively,and relative selectivity coefficients（IF） are 3.58,1.49,1.62,2.25,respectively.By using the MIPs as sorbent in solid-phase extraction column,a molecularly imprinted solid-phase extraction combined with high performance liquid chromatographic（MISPE-HPLC） method was developed for the high performance enrichment and fast separations of four TAs in Przewalskia tangutica Maxim..Good linear calibration curves were obtained in the concentration range of 2-250 μg / m L（r^2≥0.999 9） with LODs of 0.26-0.39 μg / m L.The method gave excellent recoveries（70.0%-96.3%） and precision（RSDs≤5.7%,n = 3） at 10,50,100 μg /g spiked levels.The results indicated that the molecularly imprinted solid-phase extraction（MISPE） with slight matrix interference,highly repeated usage and expectant separation effect,greatly improved extraction efficiencies for four TAs in Przewalskia tangutica Maxim..