药材唐古特瑞香中4种主要成分的含量测定与聚类分析

Content determination and cluster analysis of 4 components in Daphne tangutica Maxim

目的建立唐古特瑞香药材中4种主要成分的HPLC测定方法,并进行聚类分析。方法:采用高效液相色谱法。色谱柱为Teckmeck C_(18)(5μm,4.6 mm×250 mm);流动相A:乙腈,流动相B:0.4%磷酸,梯度洗脱(0～40 min,A:15%→30%;40～45 min,A:30%→15%;45～50 min,A:15%);检测波长为266 nm,流速1.0 mL·min^(-1),柱温为35℃,进样量为20μL。结果祖师麻甲素、芫花素、紫丁香苷、7-羟基香豆素检测质量浓度线性范围分别为0.4112～205.6μg·m L^(-1)(r=0.9999)、0.4348～217.40μg·m L^(-1)(r=0.9999)、0.4104～205.2μg·m L^(-1)(r=0.9999)、0.4276～213.80μg·m L^(-1)(r=0.9999);精密度、稳定性、重复性试验的RSD均<2.0%;加样回收率为97.4%～101.3%(RSD=0.80%～1.3%,n=6)。10批次样品可聚为2类,产地为青海海北州门源县与青海湟中县样品所含成分含量相近。结论建立的含量测定方法操作简单,精密度、稳定性、重复性好,可用于同时测定唐古特瑞香药材中祖师麻甲素、芫花素、紫丁香苷、7-羟基香豆素的含量;不同产地的唐古特瑞香药材成分含量差异较大。

Objective To establish a method for the simultaneous determination of 4 components in Daphne tangutica Maxim and to conduct cluster analysis. Methods HPLC method was adopted. The determination was performed on Teckmeck C_（18）（5 μm, 4.6 mm×250 mm） column with mobile phase consisting of acetonitrile-0.4% phosphate acid solution（gradient elution） at the flow rate of 1.0 mL·min^-1. The detection wavelength was 266 nm and the column temperature was 35℃. The injection volumn was 20 μL. Results The linear ranges of daphnetin, genkwanin, syringin, and 7-hydroxycumarin were 0.4112-205.6 μg·mL^-1（r =0.9999）, 0.4348-217.40 μg·mL-（-1）（r = 0.9999）, 0.4104-205.2 μg·mL-（-1）（r = 0.9999）, and 0.4276-213.80 μg·mL-（-1）（r = 0.9999）, respectively. The RSDs of precision, stability and reproducibility tests were all lower than 2.0%. The recoveries were 97.4%-101.3%（RSD = 0.80%-1.3%, n = 6）. The 10 batches of medicinal materials could be divided into 2 categories. Conclusion The method is simple, precise, stable and reproducible, and it can be used for the simultaneous determination of 4 components in Daphne tangutica Maxim. The quality of Daphne tangutica Maxim from different regions is quite different.