正交试验法优选“武当二号金银花”藤中4种有效成分的提取条件及其含量测定

Optimization of Extraction Conditions for Four Major Effective Components in Wudang Ⅱ Flos lonicerae Caulis by Orthogonal Design and Content Determination

目的:研究"武当二号金银花"藤中主要有效成分含量测定方法,为评价其品质奠定基础。方法:采用超声法考察提取溶剂、超声时间、超声功率及料液比4个因素对样品中芦丁和木犀草苷成分提取的影响,并使用4因素3水平的正交试验方法优化提取条件。含量测定色谱条件:色谱柱:绿原酸为C18柱(Phenomenex Luna,250 mm×4.6 mm,5μm);马钱苷、芦丁与木犀草苷均为苯基柱(Fortis Xi phenyl,250 mm×4.6 mm,5μm);流动相:绿原酸、马钱苷为乙腈-0.4%磷酸溶液(15∶85);芦丁与木犀草苷为乙腈-0.5%冰醋酸溶液梯度洗脱;柱温:30℃;检测波长分别为327,237,354,348 nm。结果:提取"武当二号金银花"藤中芦丁与木犀草苷成分的最佳条件:提取溶剂均为60%乙醇;料液比均为1∶30,超声功率分别为350,250 w;超声时间分别为50,60 min。样品中有效成分绿原酸、马钱苷、芦丁和木犀草苷的平均含量分别为10.27,6.33,0.401,0.450 mg·g-1。结论:该方法简便、准确、重现性好,为其质量控制及进一步开发利用提供参考。

Objective： To study the content determination method for the effective components in Wudang H Flos lonicerae Caulis to lay foundation for the quality evaluation. Methods： An ultrasonic method was used. The effects of extraction solvent, ultrasonic time, ultrasonic power and ratio of solid to liquid on the contents of rutin and mignonette nucleoside were studied, and the extraction conditions were optimized by a 4-factor and 3-level orthogonal experiment. The chromatographic conditions were as follows ： a Phenomenex Luna-C18（250mm×4.6mm,5μm） column was adopted for chlorogenic acid, and a Fortis Xi Phenyl column （250mm×4.6mm,5μm） was used for rutin, loganin and luteoloside ; the mobile phase was acetonitrile （B） -0.4% phosphoric acid （C） solution （ 15 ： 85 ） for chlorogenic acid and loganin, and acetonitrile （B） -0.5% glacial acetic acid aqueous solutjion （D） with gradient elution for rutin and luteoloside; the column temperature was 30℃, and the detection wavelength was 327,237,354 and 348 nm, respectively. Results ： The optimum extraction conditions for rutin and luteoloside from Wudang Ⅱ Flos lonicerae Caulis were as follows ： the extraction solvent was 60% ethanol, the solid-liquid ratio was 1 ： 30, the ultrasonic power and the ultrasonic time were 350 W and 50 min for rutin, and 250W and 60rain for luteoloside, The content of chlorogenic acid, loganin, rutin and luteoloside was 10.27, 6.33, 0. 401 and 0. 450 mg · g^-1 in the samples, respectively. Conclusion： The method is simple and convenient, accurate and reproducible, which can be used to control the quality of Wudang Ⅱ Flos lonicerae Caulis and provide reference for the further development.