常山胡柚不同生长期果实中3种成分含量的动态变化

Study on the Dynamic Changes of Three Contents in Citrus Changshan Huyou Fruit in Different Growth Periods

目的 建立HPLC同时测定常山胡柚不同生长期果实中柚皮苷、橙皮苷、新橙皮苷含量的方法,并研究常山胡柚不同生长期果实中3种成分含量的动态变化,从而确定常山胡柚作为药用资源的最佳采收时间。方法 采用HPLC测定常山胡柚不同生长期果实中柚皮苷、橙皮苷、新橙皮苷3种成分的含量,色谱柱为安捷伦ZORBAX SB-C18（4.6 mm×250 mm,5μm）;流动相为乙腈-0.2%磷酸（15∶85）;检测波长：284 nm;流速：1.0 m L·min？1;柱温：40℃;峰面积外标法定量。结果 柚皮苷、橙皮苷、新橙皮苷的线性范围分别为28.45~284.50 ng（r=0.999 8）、18.09~180.93 ng（r=0.999 8）、85.86~858.55 ng（r=0.999 6）,平均回收率分别为98.83%（RSD=2.12%）,99.05%（RSD=2.30%）,98.83%（RSD=1.85%）。常山胡柚不同生长期果实中柚皮苷、橙皮苷、新橙皮苷的含量分别为12.65~109.66,0.45~17.50,8.06~218.90 mg·g？1,柚皮苷在6月时最高,橙皮苷和新橙皮苷在5月时最高,其中5~7月间采收的果实柚皮苷和新橙皮苷的含量均符合中国药典枳壳项下含量的要求。结论 本方法操作简便、快速,结果准确、可靠,重复性好,适合作为含量测定的方法,并为常山胡柚作为药用资源的采收提供了参考依据。

OBJECTIVE To establish a method for simultaneous determination of naringin, hesperidin and neohesperid in Citrus changshan-huyou fruit in different growth periods by HPLC, to study the dynamic changes of three contents, and then to determine the best harvest period of Citrus changshan-huyou as the medicinal resource. METHODS The analysis was performed on an Agilent ZORBAX SB-C18 column（4.6 mm×250 mm, 5 μm） with acetonitrile-0.2% aqueous phosphoric acid（15∶85） as the mobile phase at the flow rate of 1.0 m L·min^-1. The detection wavelength was set at 284 nm. And column temperature was 40 ℃. The analysts were quantified by external standard method using peak areas. RESULTS The linear ranges were 28.45？284.50 ng（r=0.999 8） for naringin, 18.09？180.93 ng（r=0.999 8） for hesperidin, and 85.86？858.55 ng（r= 0.999 6） for neohesperidin. The average recoveries were 98.83%（RSD=2.12%） for naringin, 99.05%（RSD=2.30%） for hesperidin and 98.83%（RSD=1.85%） for neohesperidin. The contents of naringin, hesperidin and neohesperidin in Citrus changshan-huyou fruit in different growth periods were determined simultaneously. As a result, the content ranges were 12.65？109.66 mg·g？1 for naringin, 0.45？17.50 mg·g^-1 for hesperidin, and 8.06？218.90 mg·g^-1 for neohesperidin, respectively. Naringin had the highest content in June, the highest in May for hesperidin and neohesperidin. The contents of naringin and neohesperidin in Citrus changshan-huyou harvested during May to July met the requirements of Chinese pharmacopoeia. CONCLUSION The described HPLC method is simple, accurate and reliable, and is suitable for the determination of naringin, hesperidin and neohesperidin. The results provide reference for determining the best harvest period of Citrus changshan-huyou as the medicinal resource.