基于特征图谱的金银花与山银花(灰毡毛忍冬)质量表征比较研究

目的阐明金银花与山银花药学质量表征的共性与特性。方法采用HPLC-DAD法,Waters XBridge C18分析柱(250 mm×4.6 mm,5μm),以乙腈-0.4%磷酸水溶液为流动相梯度洗脱,流速1.0 mL/min,检测波长为211 nm,柱温30℃,建立金银花和山银花特征图谱质量表征方法;基于特征图谱中的特征峰的所属化学类型及代表性成分的峰面积和含量对15批金银花和10批山银花饮片进行药学质量表征,并对二者的药学质量特点进行比较分析。结果(1)建立的金银花和山银花的特征图谱及质量表征方法表征了二者质量表征特征值:特征峰、相对保留时间、相对峰面积、代表性成分含量及其相对含量。(2)金银花和山银花均具有特征峰30个,其药学架构涵盖4种化学类型,其中金银花/山银花各类型特征峰数目分别为:环烯醚萜类16/6,酚酸类8/11,黄酮类4/5,三萜皂苷类2/8个。(3)二者具有共有特征峰15个,其中环烯醚萜类6个、酚酸类6个、黄酮类2个、三萜皂苷类1个。(4)在金银花/山银花特征峰中指认了11个代表性成分:环烯醚萜类3/2、酚酸类6/6、黄酮类1/1、三萜皂苷类1/2;其代表性成分含量(mg/g)分别为:断氧化马钱子苷(21.756~29.826)/(6.936~14.643)、獐芽菜苦苷(12.204~30.139)/(26.109~31.608)、当药苷(13.806~23.27/(-),绿原酸(24.686~36.595)/(36.796~42.979)、异绿原酸A(14.010~20.452)/(50.544~58.013)、异绿原酸C(2.896~4.681)/(6.986~7.960)、异绿原酸B(0.935~1.260)/(1.389~1.738)、新绿原酸(0.888~1.560)/(2.025~2.635)、隐绿原酸(0.565~0.852)/(0.947~1.122),木犀草苷(0.579~0.690)/(0.527~0.642),灰毡毛忍冬皂苷乙(14.471~20.243)/(35.487~54.576)、川续断皂苷乙(-)/(31.191~35.731)。结论建立的特征图谱质量表征方法可准确地表征金银花与山银花的药学质量,二者质量表征共性为:具有相同的药学架构及部分共有特征成分;又具有明显的特性:金银花环烯醚萜类特征峰较多,当药苷为其特有环烯醚萜类成分,断氧化马钱子苷和木犀草苷的含量高于山银花;而山银花酚酸类和三萜皂苷类特征峰较多,川续断皂苷乙为其特有三萜皂苷类成分,异绿原酸A、灰毡毛忍冬皂苷乙、绿原酸、獐芽菜苦苷、异绿原酸C、新绿原酸、异绿原酸B和隐绿原酸的含量均高于金银花。研究结果可为金银花与山银花的质量控制及合理应用提供科学依据。

Comparative study on quality characterization of Lonicerae Japonicae Flos and Lonicerae Flos(Lonicera macranthoides Hand.-Mazz.)based on specific chromatogram

Objective To clarify the similarities and characteristics of the quality characterization of Lonicerae Japonicae Flos and Lonicerae Flos.Methods HPLC analysis was performed on a XBridge C18 column(250 mm×4.6 mm,5μm),The mobile phase consisted acetonitrile(A)and 0.4%phosphoric acid aqueous solution(B)with gradient elution.The flow rate was 1 mL/min,the detection wavelength was 211 nm and the column temperature was 30℃.HPLC-PDA method was used to establish a method for quality characterization of Lonicerae Japonicae Flos and Lonicerae Flos specific chromatogram,15 batches of Lonicerae Japonicae Flos and 10 batches of Lonicerae Flos were used to characterize the pharmaceutical quality based on the chemical type of the characteristic peak and the peak area and contents of the representative components in the specific chromatogram,and the pharmaceutical quality characteristics of Lonicerae Japonicae Flos and Lonicerae Flos were analyzed and compared.Results(1)The characteristic values of quality characterization,including characteristic peak,relative retention time,relative peak area,representative component content and its relative content,were characterized by the specific chromatogram and quality characterization method of Lonicerae Japonicae Flos and Lonicerae Flos.(2)Lonicerae Japonicae Flos and Lonicerae Flos both have 30 characteristic peaks,and their pharmaceutical structure covers 4 chemical types,and the number of each type characteristic peaks of Lonicerae Japonicae Flos/Lonicerae Flos is,16/6 of iridoid,8/11 of phenolic acids,4/5 of flavonoid,and 2/8 of triterpenoid saponins,respectively.(3)They both have 15 characteristic peaks in common,including 6 iridoids,6 phenolic acids,2 flavonoids and 1 triterpenoid saponins.(4)11 representative components were identified in the characteristic peaks of Lonicerae Japonicae Flos/Lonicerae Flos,3/2 of iridoids,6/6 of phenolic acids,1/1 of flavonoid,and 1/2 of triterpenoid saponins.The representative component contents(mg/g)are:(21.756~29.826)/(6.936~14.643)of secoxyloganin,(12.204~30.139)/(26.109~31.608)of swereosinosine,(13.806~23.27/(-)of sweroside,(24.686~36.595)/(36.796~42.979)of chlorogenic acid,(14.010~20.452)/(50.544~58.013)of isochlorogenic acid A,(2.896~4.681)/(6.986~7.960)of isochlorogenic acid C,(0.935~1.260)/(1.389~1.738)of isochlorogenic acid B,(0.888~1.560)/(2.025~2.635)of new neochlorogenic acid,(0.565~0.852)/(0.947~1.122)of cryptochlorogenic acid(0.579~0.690)/(0.527~0.642)of galuteolin,(14.471~20.243)/(35.487~54.576)of macranthoidin B,(-)/(31.191~35.731)of dipsacoside.Conclusion The established method of quality characterization of the specific chromatogram can accurately characterize the pharmaceutical quality of Lonicerae Japonicae Flos and Lonicerae Flos.They have the same pharmaceutical structure and some common representative components as common features of their quality characterization.Meanwhile,the differences of their quality characterization are as below:the characteristic peaks of iridoids in Lonicerae Japonicae Flos are the majority,Sweroside is unique iridoids,and the contents of secoxyloganin and cynaroside are higher than that of Lonicerae Flos.The characteristic peaks of phenolic acids and triterpenoid saponins in Lonicerae Flos account for the majority,dipsacoside B is an unique triterpenoid saponin component,and the contents of isochlorogenic acid A、macranthoidin B、chlorogenic acid、swertiamarine、isochlorogenic acid C、neochlorogenic acid、isochlorogenic acid B and cryptochlorogenic acid were higher than those of Lonicerae Japonicae Flos.The research results can provide scientific basis for quality control and reasonable application of Lonicerae Japonicae Flos and Lonicerae Flos.