毛冬青HPLC-UV指纹图谱与化学模式识别
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摘要
为了建立毛冬青的高效液相(HPLC-UV)指纹图谱的质量评价方法。该文采用Phenomenex Luna C18色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.05%磷酸水为流动相,梯度洗脱,流速为0.8 m L·min~(-1),柱温30℃,进样量10μL。检测波长210 nm,根据相似度评价,结合化学计量学方法对毛冬青进行质量评价。结果建立了16批毛冬青HPLC-UV指纹图谱,标定共有峰29个,通过对照品指认了12个共有峰,其中14批样品指纹图谱的相似度均>0.92,通过聚类分析可将样品聚为3类,结合主成分分析、正交偏最小二乘法-判别分析发现其中7个成分是造成不同批次样品差异性的主要标记物。根据色谱峰的紫外吸收特征,可将毛冬青药材的指纹图谱分为3个区:A区域色谱峰主要为木脂素及酚酸类成分;B区域色谱峰主要为皂苷类成分;C区域色谱峰主要为其他类成分。该文所建立的液相指纹图谱特征性和专属性强,可作为毛冬青药材内在质量控制的有效方法。
The present study is to establish the fingerprints for the quality evaluation of Ilicis Pubescentis Radix by HPLC-UV. The chromatographic conditions were defined as Phenomenex Luna C18( 4. 6 mm × 250 mm,5 μm). Mobile phase was acetonitrile-0. 05%phosphoric acid in gradient elution,and the flow rate was 0. 8 m L·min~(-1). Column temperature was 30 ℃ and the injection volume was10 μL. The detection wavelength was 210 nm. According to the similarity evaluation,the chemometric method was used to assess the quality of Ilicis Pubescentis Radix. The fingerprints of 16 batches of Ilicis Pubescentis Radix were established. There were 29 common peaks in the fingerprints and 12 common peaks were identified by reference substances. Fingerprints similarity of samples were greater than 0. 92. The samples were classified into three groups by hierarchical cluster analysis combined with principal component analysis( PCA) and orthogonal partial least squares-discriminant analysis( OPLS-DA),and seven components were the main markers that cause differences in the different batches of samples. By comparing the on-line UV spectra of chromatographic peaks,the chromatographic fingerprint was divided into three regions: region A showed seventeen main peaks( mainly lignans and phenolic acids); region B showed eight main peaks,which were proved as saponins; region C showed four main peaks,which were proved as other components. The established HPLC-UV fingerprint is highly specific,and can be used to evaluate the quality consistency of different batches of Ilicis Pubescentis Radix.
The present study is to establish the fingerprints for the quality evaluation of Ilicis Pubescentis Radix by HPLC-UV. The chromatographic conditions were defined as Phenomenex Luna C18( 4. 6 mm × 250 mm,5 μm). Mobile phase was acetonitrile-0. 05%phosphoric acid in gradient elution,and the flow rate was 0. 8 m L·min~(-1). Column temperature was 30 ℃ and the injection volume was10 μL. The detection wavelength was 210 nm. According to the similarity evaluation,the chemometric method was used to assess the quality of Ilicis Pubescentis Radix. The fingerprints of 16 batches of Ilicis Pubescentis Radix were established. There were 29 common peaks in the fingerprints and 12 common peaks were identified by reference substances. Fingerprints similarity of samples were greater than 0. 92. The samples were classified into three groups by hierarchical cluster analysis combined with principal component analysis( PCA) and orthogonal partial least squares-discriminant analysis( OPLS-DA),and seven components were the main markers that cause differences in the different batches of samples. By comparing the on-line UV spectra of chromatographic peaks,the chromatographic fingerprint was divided into three regions: region A showed seventeen main peaks( mainly lignans and phenolic acids); region B showed eight main peaks,which were proved as saponins; region C showed four main peaks,which were proved as other components. The established HPLC-UV fingerprint is highly specific,and can be used to evaluate the quality consistency of different batches of Ilicis Pubescentis Radix.
引文
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[2]昌艳艳,郭阳.中药毛冬青对心血管疾病的影响[J].现代中西医结合杂志,2002,11(21):2198.
[3]邓六勤,钟鸣,马志玲.毛冬青化学成分、药理作用及应用研究进展[J].中国中医药现代远程教育,2006,4(10):24.
[4]冯锋,朱明晓,谢宁.毛冬青化学成分研究[J].中国药学杂志,2008,43(10):732.
[5]农新维,梁冰丽,李颖,等.壮药毛冬青配方颗粒一测多评的定量研究[J].中国现代应用药学,2017,34(8):1118.
[6]童国勇,陈汀波,强皎,等.HPLC法测定毛冬青中毛冬青苷B的含量[J].中药新药与临床药理,2013,24(5):503.
[7]李英,吕晔,林丽萍.HPLC-ELSD法同时测定毛冬青中3种五环三萜的含量[J].中药材,2014,37(3):451.
[8]倪晨,陈汀波,陈军,等.HPLC法测定毛冬青中毛冬青苷元A的含量[J].中药新药与临床药理,2011,22(1):89.
[9]余成霞,柳文媛,冯锋,等.毛冬青药材的指纹图谱[J].中国天然药物,2008,6(6):439.
[10]应鸽,丁平,代蕾,等.毛冬青根和茎的HPLC指纹图谱研究[J].华西药学杂志,2013,28(5):506.
[11]Peng Q,Tian R,Chen F,et al.Discrimination of producing area of Chinese Tongshan kaoliang spirit using electronic nose sensing characteristics combined with the chemometrics methods[J].Food Chem,2015,178:301.
[12]夏伯候,严东,曹艺,等.不同剂型夏桑菊颗粒HPLC指纹图谱及其模式识别分析[J].中国中药杂志,2016,41(3):416.
[13]徐小娜.中药色谱指纹图谱研究及化学计量学方法的应用[D].长沙:中南大学,2010.
[14]陈永婧,王小华,黄真真,等.膀胱癌血清及尿液代谢组学研究[J].分析化学,2012,40(9):1322.
[15]Liang J,Wu W Y,Sun G X,et al.A dynamic multiple reaction monitoring method for the multiple components quantification of complex traditional Chinese medicine preparations:Niuhuang Shangqing pill as an example[J].J Chromatogr A,2013,1294:58.