多波长HPLC法同时测定青黛药材中靛蓝和靛玉红的含量

目的:利用多波长高效液相色谱法,建立同时测定青黛药材中2种有效成分(靛蓝和靛玉红)含量的方法,并对其质量特征进行分析.方法:采用HPLC法,Phenomenex C18色谱柱(250 mm×4.6 mm,5 μm),柱温30 ℃;以甲醇-水溶液(70∶30)为流动相洗脱,流速1.0 ml/min;检测波长286 nm(检测靛蓝)、292 nm(检测靛玉红),同时测定不同产地青黛药材中靛蓝和靛玉红含量.结果:在20 min内青黛中靛蓝和靛玉红2种有效成分被完全分离.靛蓝在5.19～83.04 μg/ml范围内线性关系良好(r=0.999 9),平均回收率为100.46％,RSD为2.18％;靛玉红在0.364～5.825 μg/ml范围内线性关系良好(r=0.999 9),平均回收率为101.57％,RSD为2.22％.结论:本方法合理、便捷、快速简便、准确、重复性好,能同时测定青黛中靛蓝和靛玉红含量,符合现行《中国药典》简便、快速、准确的要求.

Simultaneous determination of indigo and indirubin in indigo naturalis by HPLC with multiple UV wavelength detection

Objective: To establish an HPLC method with multiple UV wavelengths detection to simultaneously determine the contents of indigo and indirubin in indigo naturalis and to characterize the quality of indigo naturalis.Methods: The samples were separated on a Phenomenex column (250 mm×4.6 mm, 5μm).The column temperature was set at 30 ℃.The mobile phase was methanol-water (70∶30) and the flow rate was 1.0 ml/min.A multiple detector was used for determination of indigo at 286 nm and of indirubin at 292 nm.Result: The complete separation was obtained within 20 min for the two compounds (indigo and indirubin).The linear correlation was safisfied in the range from 5.19 to 83.04 μg/ml (r=0.999 9)for indigo and from 0.364 to 5.825 μg/ml (r=0.999 9)for indirubin.The average recoveries of the low, middle and high concentrations were 98.78％,100.44％, 104.35％, respectively for indigo;96.20％, 98.63％, 104.03％ respectively for indirubin.Twenty-seven batches of samples were divided into three categories, which reflected the inherent quality characteristics in different regions of indigo naturalis.Conclusion: Compared with the method specified in VolⅠChinese Pharmacpeia 2015, the method is simple and the result is reliable with good repeatability and can be used for the simultaneous determination of indigo and indirubin.