不同产地艾叶中总黄酮及其3种主要苷元的含量测定
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摘要
目的:建立HPLC法同时测定9个不同产地艾叶中3种主要黄酮苷元含量的方法,规范艾叶黄酮类化合物的质量标准。方法:采用紫外分光光度法测定不同产地艾叶总黄酮含量,检测波长为510 nm;采用HPLC法同时测定艾叶黄酮中3种主要苷元的含量,色谱柱为戴安Acclaim 120 C_(18)柱(150 mm×4. 6 mm,5μm),流动相为乙腈-0. 2%磷酸溶液(50∶50),柱温为30℃,检测波长为360 nm,进样量为5μl,对比研究含量变化差异。结果:总黄酮含量测定的线性范围为0. 009 7～0. 048 6 mg·ml~(-1),加样回收率为99. 58%(RSD=0. 38%,n=6);异泽兰黄素、棕矢车菊素、山柰酚的检测进样量线性范围分别为0. 190～0. 950μg、0. 047～0. 237μg、0. 049～0. 244μg,加样回收率分别为99. 84%(RSD=0. 99%),100. 04%(RSD=0. 68%),100. 16%(RSD=0. 75%)(n=6)。湖北蕲春产艾叶总黄酮及3种苷元成分含量较高,体现出道地药材特性,但不同产地总黄酮含量变化趋势与其苷元的变化趋势差异较大。结论:该方法可作为不同产地艾叶中山柰酚、棕矢车菊素、异泽兰黄素的含量测定方法,仅通过总黄酮含量并不能全面体现不同产地艾叶黄酮类化合物的特征,结合山柰酚、棕矢车菊素、异泽兰黄素对不同产地艾叶黄酮类化合物的描述更为准确。
Objective: To establish an HPLC method for the simultaneous determination of kaempferol,jaceosidin and eupatilin in Folium Artemisiae Argyi produced in different origins,so as to standardize the quality of flavonoids in Folium Artemisiae Argyi. Methods: The contents of total flavonoids in Folium Artemisiae Argyi from different habitats were determined by ultraviolet spectrophotometry with the detection wavelength at 510 nm. The contents of 3 major aglycones in the flavonoids of Folium Artemisiae Argyi were determined by HPLC,and the column was Diane Acclaim 120 C_(18)( 150 mm × 4. 6 mm,5 μm),the mobile phase was acetonitrile-0. 2%phosphoric acid solution( 50 ∶ 50),the column temperature was 30℃,the detection wavelength was 360 nm,the sample size was 5μl,and the differences in the contents of flavonoids were compared. Results: The linear range of total flavonoids was 0. 009 0-0. 048 6 mg·ml~(-1),and the recovery was 99. 58%( the RSD was 0. 38%,n = 6). The linear range was 0. 190-0. 950 μg for kaempferol,0. 047-0. 237 μg for jaceosidin,and 0. 049-0. 244 μg for eupatilin. The recovery was 99. 84%,100. 04% and 100. 16%( the RSD was0. 99%,0. 68% and 0. 75%,n = 6),respectively. The contents of total flavonoids and three aglycones in Folium Artemisiae Argyi from Hubei Qichun were higher,which reflected the characteristics of genuine medicinal materials,while the change trends of the contents of total flavonoids and aglycones in different habitats were quite different. Conclusion: The method is simple,accurate and repeatable,which can be used for the content determination of kaempferol,jaceosidin and eupatilin in Folium Artemisiae Argyi. In order to reflect the flavonoids in Folium Artemisiae Argyi more accurately,the content of total flavonoids and the contents of kaempferol,jaceosidin and eupatilin should be described more accurately.
Objective: To establish an HPLC method for the simultaneous determination of kaempferol,jaceosidin and eupatilin in Folium Artemisiae Argyi produced in different origins,so as to standardize the quality of flavonoids in Folium Artemisiae Argyi. Methods: The contents of total flavonoids in Folium Artemisiae Argyi from different habitats were determined by ultraviolet spectrophotometry with the detection wavelength at 510 nm. The contents of 3 major aglycones in the flavonoids of Folium Artemisiae Argyi were determined by HPLC,and the column was Diane Acclaim 120 C_(18)( 150 mm × 4. 6 mm,5 μm),the mobile phase was acetonitrile-0. 2%phosphoric acid solution( 50 ∶ 50),the column temperature was 30℃,the detection wavelength was 360 nm,the sample size was 5μl,and the differences in the contents of flavonoids were compared. Results: The linear range of total flavonoids was 0. 009 0-0. 048 6 mg·ml~(-1),and the recovery was 99. 58%( the RSD was 0. 38%,n = 6). The linear range was 0. 190-0. 950 μg for kaempferol,0. 047-0. 237 μg for jaceosidin,and 0. 049-0. 244 μg for eupatilin. The recovery was 99. 84%,100. 04% and 100. 16%( the RSD was0. 99%,0. 68% and 0. 75%,n = 6),respectively. The contents of total flavonoids and three aglycones in Folium Artemisiae Argyi from Hubei Qichun were higher,which reflected the characteristics of genuine medicinal materials,while the change trends of the contents of total flavonoids and aglycones in different habitats were quite different. Conclusion: The method is simple,accurate and repeatable,which can be used for the content determination of kaempferol,jaceosidin and eupatilin in Folium Artemisiae Argyi. In order to reflect the flavonoids in Folium Artemisiae Argyi more accurately,the content of total flavonoids and the contents of kaempferol,jaceosidin and eupatilin should be described more accurately.
引文
1董鹏鹏,梅全喜,戴卫波.不同产地艾叶总黄酮、重金属和硒元素的含量比较研究[J].时珍国医国药,2016,27(1):74-76
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4李琼瑜.山奈酚联合5-氟尿嘧啶抑制大肠癌生长的作用及其机制研究[D].福州:福建中医药大学硕士学位论文,2016
5张琦.棕矢车菊素改善小鼠肥胖模型中胰岛素抵抗的分子机制[D].南京:南京大学硕士学位论文,2012
6王晓娜.异泽兰黄素抑制食管癌细胞TE1增殖的分子机制研究[D].郑州:郑州大学硕士学位论文,2014
7李林,程慧.不同产地艾叶中异泽兰黄素含量比较[J].中国实验方剂学杂志,2013,19(3):121-123
2李华生,骆航,孙兴力,等.加压同步萃取艾叶挥发油、总黄酮和多糖的工艺研究[J].中国食品添加剂,2016,(10):83-89
3中国药典[S].2015年版.一部.89,354
4李琼瑜.山奈酚联合5-氟尿嘧啶抑制大肠癌生长的作用及其机制研究[D].福州:福建中医药大学硕士学位论文,2016
5张琦.棕矢车菊素改善小鼠肥胖模型中胰岛素抵抗的分子机制[D].南京:南京大学硕士学位论文,2012
6王晓娜.异泽兰黄素抑制食管癌细胞TE1增殖的分子机制研究[D].郑州:郑州大学硕士学位论文,2014
7李林,程慧.不同产地艾叶中异泽兰黄素含量比较[J].中国实验方剂学杂志,2013,19(3):121-123