紫草总色素β-环糊精包合物的定性鉴别及左旋紫草素含量测定

Qualitative Identification of Total Pigment β-Cyclodextrin Inclusion Complex of Radix Arnebiae and Content Determination of L-Shikonin

目的建立定性鉴别紫草总色素β-环糊精包合物有效成分的薄层色谱(TLC)法和测定左旋紫草素含量的高效液相色谱(HPLC)法。方法采用TLC法进行定性鉴别,展开系统为环己烷-甲苯-乙酸乙酯-甲酸(5∶5∶0. 5∶0. 1,V/V/V/V);采用HPLC法测定包合物中有效成分左旋紫草素的含量,流动相为甲醇-0. 025 mol/L磷酸(82∶18,V/V)。结果 TLC鉴别中,供试品溶液与对照品溶液在相应位置上显相同颜色斑点,且斑点清晰,阴性对照无干扰;HPLC图中,供试品溶液与对照品溶液色谱出峰时间一致,分离度好,阴性对照无干扰,含量测定中有效成分的线性关系良好,平均加样回收率为102. 09%,RSD为1. 87%(n=6)。结论该方法操作简便、结果准确、专属性强、重复性好,可用于该产品的质量控制。

Objective To establish a thin-layer chromatography( TLC) method for the qualitative identification of total pigment β-cyclodextrin inclusion complex of Radix Arnebiae and a high-performance liquid chromatography( HPLC) method for the content determination of L-shikonin. Methods The inclusion compound of total pigment β-cyclodextrin was identified by TLC. The system was developed into cyclohexane-toluene-ethyl acetate-formic acid( 5 ∶ 5 ∶ 0. 5 ∶ 0. 1,V/V/V/V). The effective component L-shikonin of the inclusion compound was determined by HPLC,with the mobile phase methanol-0. 025 mol/L phosphoric acid( 82: 18,v/v).Results In the TLC identification,chromato-graphs of the tested samples showed the same color spots as those of the reference samples;the spots were clear,and the negative samples had no interference. In the HPLC test,the chromatographic peak times of the sample solution and the reference solution were consistent,the chromato-graph showed good separation degree,no interference with the negative sample,and a good linear relationship between the active ingredients in the content determination;the average recovery was 102. 09%,and RSD = 1. 87%( n =6). Conclusion This method is simple,accurate,specific and reproducible,which can be used for the quality control of the product.