高效液相色谱法同时测定青黛中色胺酮、靛蓝及靛玉红的含量

Simultaneous Determination of Tryptanthrin,Indigo and Indirubin in Indigo Naturalis by High Pressure Liquid Chromatography

为了建立高效液相色谱法同时测定青黛中色胺酮、靛蓝及靛玉红的含量,以色胺酮、靛蓝和靛玉红对照品为对照,对三批青黛药材中色胺酮、靛蓝及靛玉红的含量进行测定,采用色谱条件为:Shim-pack VP-ODS C18(4.6mm×150mm,5μm)色谱柱,流动相为乙腈-水(53∶47,V∶V),流速:1mL/min,检测波长290nm。实验结果表明色胺酮在0.02～0.16μg/mL时,线性关系良好(R2=0.999 5);靛蓝在8～64μg/mL时,线性关系良好(R^2=0.999 5);靛玉红在0.5～4μg/mL时,线性关系良好(R^2=0.999 1),方法的平均回收率为97.63%,相对标准偏差(RSD)=1.61%、回收率为100.4%,相对标准偏差(RSD)=2.21%、回收率为97.50%,相对标准偏差(RSD)=2.13%。三批青黛中色胺酮,靛蓝和靛玉红三种成分的质量浓度分别为0.065～0.10、17～31、2.4～3.9mg/g。实验操作简单,灵敏度高,重现性良好,结果准确可靠,可以同时测定青黛有效成分的含量。

In order to establish high pressure liquid chromatography(HPLC)method for simultaneous determination of tryptanamine,indigo and indirubin in indigo naturalis,the contents of the above three index components in three batches of indigo naturalis were determined by comparing with tryptophan,indigo and indirubin.The following chromatographic conditions were used:Shim-pack VP-ODS C18(4.6 mm×150 mm,5μm)column,mobile phase:acetonitrile-water(53∶47,V∶V),flow rate:1 mL/min,detection wavelength:290 nm.The experimental results showed that a linear relationship was very good between0.02~0.16μg/mL of tryptanthrin(R2=0.999 5),indigo was good at a linear range of 8~64μg/mL(R^2=0.999 5)and indirubin has a good linear relationship at 0.5~4μg/mL(R^2=0.999 1),The average recoveries were 97.63%(relative standard deviation=1.61%),100.4%(relative standard deviation=2.21%)and 97.50%(relative standard deviation=2.13%),respectively.The contents of tryptanthrin,indigo and indirubin were used in the three batches of indigo naturalis are 0.065~0.10,17~31 and 2.4~3.9 mg/g,respectively.The experiment is simple in operation,with high sensitivity,good reproducibility and accurate and reliable results,and it can determine the content of active ingredients in indigo naturalis.