广东紫珠中抗凝血活性成分的薄层色谱鉴别及含量测定

Identification and determination of the anticoagulant active component from Callicarpa kwangtungensis by TLC and HPLC-ELSD

目的:建立不同产地、不同部位的野生与栽培广东紫珠中抗凝血活性成分2α,3α,19α,23-四羟基熊果^(-1)2-烯-28-O-β-D-吡喃葡萄糖苷和2α,3α,19α-三羟基齐墩果^(-1)2-烯-28-O-β-D-吡喃葡萄糖苷的薄层鉴别和水苏碱含量测定的方法。方法:TLC法,展开剂为三氯甲烷-甲醇-甲酸(5∶1∶0.1),显色剂为10%硫酸乙醇溶液;HPLC-ELSD法,采用Hypersil-NH_2色谱柱(250 mm×4.6 mm,5μm),以乙腈-水(17∶83)为流动相,流速1.0 m L·min^(-1),柱温30℃,漂移管温度75℃,载气流速2.0 L·min^(-1)。结果:广东紫珠药材及对照药材中2个三萜的TLC分离效果均较好;HPLC-ELSD法测得盐酸水苏碱进样量在0.091 2～0.912μg范围内与峰面积呈现良好的线性关系(r=0.999 9),平均回收率为98.3%(RSD=1.3%),精密度、稳定性良好。不同产地广东紫珠水苏碱的含量有一定的差异,且叶含量高于茎枝含量,栽培含量高于野生含量。结论:此方法简便易行,重复性好,可作为广东紫珠药材质量控制指标。

Objective：To establish a TLC method for identification of the anticoagulant active components（2α,3α,19α,23-tetrahydroxy-urs-（-1）2-en-28-O-β-D-glucopyranoside and 2α,3α,19α-trihydroxy-olean-（-1）2-en-28-O-β-D-glucopyranoside）,and to establish an HPLC-ELSD determination method for stachydrine in wild and cultivate Callicarpa kwangtungensis Chun from different habitats and different parts.Methods：TLC was performed using chloroform-methanol-formic acid（5∶1∶0.1）as the mobile phase and 10% sulfuric acid ethanol solution as colorimetric agent.HPLC-ELSD was also adopted,and the separation was carried out on a HypersilNH_2（250 mm×4.6 mm,5 μm）with the mobile phase of acetonitrile-water（17∶83）.The flow rate was 1.0 m L·min-（-1）,the column temperature was at 30 ℃,the evaporation temperature was 75 ℃,and carrier gas flow rate was 2.0 L·min-（-1）.Results：The two triterpenoid glucosides in C.kwangtungensis and reference crude drug of Callicarpae Caulis et Folium could be identified by TLC.The linear range of stachydrine hydrochloride was 0.091 2-0.912 μg（r=0.999 9）,the average recovery was 98.3%,RSD was 1.3%,and the precision and stability were good.There were certain differences in contents of stachydrine in C.kwangtungensis among different habitats,and the content in the leaf was higher than that of stem branch,while the content of wild C.kwangtungensis was higher than that of cultivate C.kwangtungensis.Conclusion：The established method,which is specific,accurate and reproducible,can be applied to the quality control of C.kwangtungensis.