酸橙和甜橙2种枳实药材HPLC指纹图谱研究及其柚皮苷和辛弗林含量分析

目的:研究酸橙和甜橙2种枳实药材的高效液相色谱 (HPLC) 指纹图谱并测定药材中柚皮苷和辛弗林的含量, 以建立枳实药材有效的质量评价方法。方法:采用高效液相色谱 (HPLC) 法建立指纹图谱和测定有效成分含量, 对来自20个产地的2个种的枳实药材进行分析。色谱柱为AlltimaC18 (250mm×4.6mm, 5μm) , 流动相为乙腈-水 (含0.1%磷酸或加上0.1%十二烷基硫酸钠) , 流速为1.0mL·min-1, 指纹图谱检测波长为224nm, 柚皮苷和辛弗林含量测定的检测波长分别为283、224nm。结果:得到分离度、重复性均较好的枳实药材HPLC指纹图谱。对20批不同产地枳实药材 (包含2个种) HPLC指纹图谱的相似度评价结果显示, 酸橙和甜橙2种枳实的指纹图谱差异显著, 但同种枳实中不同产地的差异不大;含量测定结果显示, 不同产地和不同种枳实药材中柚皮苷和辛弗林含量差异较大。结论:该方法简便、准确、灵敏度高、重复性好, 可作为枳实药材的质量控制方法, 此方法已被《香港中药材标准》采用。不同种和产地的枳实药材有效成分 (柚皮苷和辛弗林) 的含量具有明显差异, 临床处方应当区别应用。OBJECTIVE:To study the HPLC fingerprints of Citrus aurantium and Citrus sinensis and to determine the contents of naringin and synephrine in order to establish the quality evaluation method of Aurantii Fructus Immaturus.METHODS:HPLC method was use to establish fingerprints and determine content of active component.2 kinds of Aurantii Fructus Immaturus from 20 habitats were evaluated.The samples were separated on Alltima C18 column (250 mm×4.6 mm, 5μm) using acetonitrile-water (0.1% phosphoric acid or 0.1% SDS) as mobile phase at the flow rate of 1.0 mL·min-1.The detection wavelength for fingerprint was set at 224 nm, and the wavelength for content determination of naringin and synephrine were set at 283 nm and 224 nm, respectively.RESULTS:HPLC fingerprint of Aurantii Fructus Immaturus was established with good separation and repeatability.The results showed that the HPLC fingerprints were significantly different between C.aurantium and C.sinensis, but not in different habitats.The contents of naringin and synephrine from different species and sources of Aurantii Fructus Immaturus were obviously different from each other.CONCLUSION:The method is simple, accurate, sensitive and repeatable, and it can be used as quality control for Aurantii Fructus Immaturus and has been included in Hong Kong Chinese Materia Medica Standards.The contents of naringin and synephrine vary significantly in Aurantii Fructus Immaturus from different habitats and species, which suggest that more attention should be paid to clinical use of different Aurantii Fructus Immaturus in practice.