HPLC法测定不同pH条件下余甘子中4种成分的含量变化  

HPLC Method to Determine the Content of 4 Components of Phyllanthus Emblica under Different pH Conditions

作　　者：黄宽 向润清 林艾和 张艳娇 范源 HUANG Kuan;XIANG Runqing;LIN Aihe;ZHANG Yanjiao;FAN Yuan(Yunnan University of Chinese Medicine,Kunming 650500,China;The Second Affiliated Hospital of Yunnan University of Chinese Medicine,Kunming 650216,China)

机构地区：[1]云南中医药大学,云南昆明650500 [2]云南中医药大学第二附属医院,云南昆明650216

出　　处：《云南中医学院学报》2019年第6期14-19,共6页Journal of Yunnan University of Traditional Chinese Medicine

基　　金：国家自然科学基金地区基金项目(81360652);云南省科技厅-云南中医学院应用基础研究联合专项(2017FF117(-008));云南省高校中西医结合防治甲状腺病重点实验室项目。

摘　　要：目的建立同时测定余甘子中4种成分含量的HPLC法,并研究在不同pH条件下4种成分含量变化。方法采用高效液相色谱法测定余甘子中焦性没食子酸(Pyrogallic acid,PA)、没食子酸(Gallic acid,GA)、1,3,6-O-三没食子酰基葡萄糖(1,3,6-tri-O-galloyl-β-D-glucose,TGG)和1,2,3,4,6-O-五没食子酰葡萄糖(1,2,3,4,6-penta-O-galloyl-β-D-glucose,β-PGG)的含量。色谱柱为Agilent Zorbax C18键合硅胶柱(150mm×4.6 mm,5μm),流动相为甲醇-0.1%磷酸梯度洗脱,流速为1.0 mL/min,检测波长为280 nm,柱温为25℃,进样量为10μL,测定不同pH条件下4种成分的含量。结果PA、GA、TGG和β-PGG分别在0.0229~0.2290mg/mL(r=0.9996,n=6),0.0148~0.1480 mg/mL(r=0.9991,n=6),0.0018~0.0180 mg/mL(r=0.9998,n=6),0.0031~0.0310 mg/mL(r=0.9993,n=6)范围内呈良好的线性关系,加样回收率均符合含量测定要求。余甘子中的PA在pH为2时含量最高;GA在pH为5时含量最高;TGG在pH为4时含量最多;β-PGG在pH为3时含量最高。结论该方法简便、准确,可以用于余甘子中PA、GA、TGG和β-PGG 4个成分的含量测定。通过研究不同pH条件下余甘子中的PA、GA、TGG和β-PGG的含量变化,可为余甘子中相关成分的质控提供一定理论参考。Objective An HPLC method was established for simultaneous determination of the contents of four components in Phyllanthus Emblica,and the changes of the contents of the four components under different pH conditions were studied.Methods The contents of PA,GA,TGG andβ-PGG in Phyllanthus Emblica were determined by HPLC.The column was an Agilent Zorbax C18 bonded silica column(150 mm×4.6 mm,5μm).The mobile phase was methanol-0.1%phosphoric acid gradient and the flow rate was 1.0 mL/min.The detection wavelength was 280 nm at 25℃,the injection volume was 10μL.Then the contents of four components under different pH conditions were determined.Results PA,GA,TGG,andβ-PGG range from are 0.0229-0.2290 mg/mL(r=0.9996,n=6),0.0148-0.1480 mg/mL(r=0.9991,n=6),0.0018-0.0180 mg/mL(r=0.9998,n=6),0.0031-0.0310 mg/mL(r=0.9993,n=6)respectively,showed a good linear relationship,and the recovery rates of the samples all met the requirements for content determination.The content of PA in Phyllanthus emblica was highest at pH 2,GA was highest at pH 5,TGG was highest at pH 4,β-PGG was highest at pH 3.Conclusion This method is simple and accurate,and can be used to determine the content of PA,GA,TGG andβ-PGG in Phyllanthus Emblica.By studying the content changes of PA,GA,TGG andβ-PGG in Phyllanthus Emblica under different pH conditions,it can provide a certain theoretical reference for the quality control of related ingredients in Phyllanthus Emblica.

关 键 词：HPLC 余甘子 焦性没食子酸 没食子酸 1 3 6-O-三没食子酰基葡萄糖 1 2 3 4 6-O-五没食子酰葡萄糖 

分 类 号：R286.0[医药卫生—中药学]