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目的以HPLC法同时测定青黛中有效成分靛蓝和靛玉红的含量。对中国药典2010年版中青黛含量测定方法的不足之处进行改进。方法用适量DMF超声30 min提取青黛药材中的靛蓝和靛玉红。液相条件如下:色谱柱:Kromasil C18(250 mm×4.6 mm,5μm);流速1 mL/min;流动相:乙腈∶水(70∶30);柱温:30℃;检测波长:290 nm。此法在不影响分析结果的条件下可大大缩短分析时间。结果靛蓝和靛玉红进样量在29.90 ng~298.90 ng和6.71 ng~67.06 ng间与峰面积的线性关系良好(r>0.999,n=6)。靛蓝平均加样回收率:101.05%(RSD=1.81%);靛玉红平均加样回收率:99.78%(RSD=2.52%)。结论本法合理、便捷;能同时测定青黛中靛蓝和靛玉红含量,符合现行药典简便、快速、准确的要求。
Objective To determine the contents of indigo and indirubin, the active ingredients of indigo, by HPLC. On the Chinese Pharmacopoeia 2010 edition of indigo content determination methods to improve the deficiencies. Methods Indigo and indirubin were extracted from indigo naturalis with appropriate amount of DMF for 30 min. The liquid phase conditions were as follows: Column: Kromasil C18 (250 mm × 4.6 mm, 5 μm); flow rate 1 mL / min; mobile phase: acetonitrile: water 70:30; This method can greatly reduce the analysis time without affecting the result of the analysis. Results The linear relationship between peak area and volume of indigo and indirubin was 29.90 ng ~ 298.90 ng and 6.71 ng ~ 67.06 ng (r> 0.999, n = 6). The average recovery of indigo was 101.05% (RSD = 1.81%). The average recovery of indirubin was 99.78% (RSD = 2.52%). Conclusion The method is reasonable and convenient. The simultaneous determination of indigo and indirubin contents in indigo is in line with the simple, rapid and accurate requirements of the current pharmacopoeia.