一测多评法测定罗布麻叶中6个黄酮类成分的含量

目的：建立罗布麻叶中6个活性成分（芦丁、金丝桃苷、异槲皮苷、紫云英苷、槲皮素和山柰酚）同时测定的一测多评含量测定方法。方法：采用Waters XBridge C₁₈色谱柱（250 mm&#215;4.6 mm，5 μm），以乙腈-0.2%磷酸溶液为流动相，梯度洗脱，流速0.8 mL&#183;min^-1，检测波长256 nm，柱温35℃。以金丝桃苷为参照物建立与其他成分的相对校正因子，并计算其含量，实现一测多评。同时采用外标法测定该6个成分的含量，并比较二者的差异，以验证一测多评法的准确性和可行性。结果：金丝桃苷、芦丁、异槲皮苷、紫云英苷、槲皮素、山柰酚线性范围分别为11.36~284、0.416~10.4、9.2~230、4.08~102、0.812~20.3、0.272~6.8 μg&#183;mL^-1，金丝桃苷与芦丁、异槲皮苷、紫云英苷、槲皮素、山柰酚的相对校正因子值分别为1.378、0.974、1.197、0.867、1.114，且在不同实验条件下重复性良好（相对校正因子的RSD<3.0%），并与常规外标一点法比较，20批罗布麻叶中芦丁、金丝桃苷、异槲皮苷、紫云英苷、槲皮素、山柰酚的含量测定结果分别为0.066 57~0.178 4、2.766~3.576、2.794~3.334、0.271 9~0.668 5、0.078 65~0.256 5、0.018 72~0.106 2 mg&#183;g^-1，6个黄酮类成分的含量之和范围为6.343~7.732 mg&#183;g^-1。一测多评法计算值与实测值间均无显著性差异（RSD<2.0%）。结论：以金丝桃苷建立的5个黄酮类成分相对校正因子可用于罗布麻叶的定量分析及质量评价。

Objective:To develop a method of quantitative analysis of multi-components by single marker(QAMS)for simultaneous determination of six bioactive constituents(rutin,hyperoside,isoquercitrin,astragalin,quercetin and kaempferol)in Apocyni Veneti Folium.Methods:A Waters XBridge C₁₈ column(250 mm&#215;4.6 mm,5 μm) was used for the separation,with gradient elution of acetonitrile and 0.2% phosphoric acid as the mobile phase at a flow rate of 0.8 mL&#183;min^-1.The detection wavelength was set at 256 nm.The column temperature was 35℃.Hyperoside was selected as the internal reference substance,the relative correction factors(RCFs) between hyperoside and the other five flavanoids were established,and the contents of these six constituents in samples were calculated to realize QAMS.At the same time,the external standard method was used to quantify contents of the six constituents,the two methods were compared to validate the accuracy and feasibility of the new method.Results:The linear ranges of hyperoside,rutin,isoquercetin,astragaloside,quercetin and kaempferol were 11.36-284,0.416-10.4,9.2-230,4.08-102,0.812-20.3 and 0.272-6.8 μg&#183;mL^-1,respectively.RCFs of rutin,isoquercitrin,astragalin,quercetin and kaempferol with reference to hyperoside were 1.378,0.974,1.197,0.867 and 1.114,respectively,and repeatability was good in different experimental conditions(RSDs of RCFs were less than 3.0%).The contents of rutin,hyperoside,isoquercitrin,astragaloside,quercetin and kaempferol in 20 batches of Apocyni Veneti Folium were 0.066 57-0.178 4,2.766-3.576,2.794-3.334,0.271 9-0.668 5,0.078 65-0.256 5 and 0.018 72-0.106 2 mg&#183;g^-1,respectively.The range of total content of six flavonoids was 6.343-7.733 mg&#183;g^-1.No significant differences was found in the quantitative results of the six flavonoids in 20 batches of Apocyni Veneti Folium determined by external standard method and QAMS(RSD<2.0%).Conclusion:RCFs of 5 flavones with reference to hyperoside can be used for assay and quality evaluation of Apocyni Veneti Folium.

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