基于形态和红外光谱分析的云南重楼及近似种的快速鉴别
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摘要
目的基于形态和红外光谱法建立云南重楼及近似种的鉴别方法,并进一步分析其亲缘关系,为资源开发利用提供依据。方法对云南重楼及其近似种进行了原植物形态比较,并对光谱数据进行自动基线校正、自动平滑、纵坐标归一化、二阶求导等预处理,采用主成分分析(principalcomponentanalysis,PCA)、偏最小二乘判别分析(partialleastsquares discriminantanalysis,PLS-DA)及系统聚类分析(hierarchicalclusteranalysis,HCA)分析光谱数据。结果云南重楼与近似种在植株大小、叶片、叶脉、萼片、花瓣、花亭和雄蕊等形态特征上存在典型的鉴别特征,可为其提供形态鉴别依据。6种重楼的二阶导数在3 000～2 000 cm-1和指纹区1 800～500 cm-1波段内差异明显,HCA和PLS-DA均能较好地区分云南重楼及近似种,可为其提供红外光谱鉴别证据。HCA结果表明云南重楼、七叶一枝花、狭叶重楼、矮重楼和多叶重楼亲缘关系较近。结论傅里叶变换红外光谱(FTIR)结合原植物形态鉴定能够快速鉴别云南重楼及其近似种,为指导重楼栽培、临床应用和资源开发提供科学依据。
Objective To explore morphology and infrared spectrum identification evidence of Paris polyphylla var. yunnanensis and its closely relative species, and further analyze their genetic relationship to provide basis for the development and utilization of medicinal plant resources of genus Paris. Methods The morphology and infrared spectrum of P. polyphylla var. yunnanensis and its closely relative species were studied systematically and compared with each other. The original infrared spectra data were pretreated by automatic baseline correction, automatic smoothing, ordinate normalization, and second derivative, and analyzed by principal component analysis(PCA), partial least squares discrimination analysis(PLS-DA), and hierarchical cluster analysis(HCA). Results There were typical characteristics of P. polyphylla var. yunnanensis and its closely relative species, such as plant size, leaves, veins, sepals, petals, flower pavilions and stamens, which could provide morphological identification evidence. There were obviously differences of the second derivative in 3 000—2 000 cm-1 and the fingerprint in 1 800—500 cm-1 in the six types of genus Paris. Both HCA and PLS-DA could better distinguish Paris polyphylla var. yunnanensis and its closely relative species, which could provide an infrared spectral identified evidence. The results of HCA showed that P. polyphylla var. yunnanensis, P. polyphylla var. chinensis, P. polyphylla, P. polyphylla var. stenophylla and P. polyphylla var. nana were relatively close. Conclusion Fourier transform infrared spectroscopy(FTIR) combined with original plant morphological identification can quickly identify P. polyphylla var. yunnanensis and its closely relative species, which will provide a scientific basis for the cultivation, clinical application, and resource development of genus Paris.
Objective To explore morphology and infrared spectrum identification evidence of Paris polyphylla var. yunnanensis and its closely relative species, and further analyze their genetic relationship to provide basis for the development and utilization of medicinal plant resources of genus Paris. Methods The morphology and infrared spectrum of P. polyphylla var. yunnanensis and its closely relative species were studied systematically and compared with each other. The original infrared spectra data were pretreated by automatic baseline correction, automatic smoothing, ordinate normalization, and second derivative, and analyzed by principal component analysis(PCA), partial least squares discrimination analysis(PLS-DA), and hierarchical cluster analysis(HCA). Results There were typical characteristics of P. polyphylla var. yunnanensis and its closely relative species, such as plant size, leaves, veins, sepals, petals, flower pavilions and stamens, which could provide morphological identification evidence. There were obviously differences of the second derivative in 3 000—2 000 cm-1 and the fingerprint in 1 800—500 cm-1 in the six types of genus Paris. Both HCA and PLS-DA could better distinguish Paris polyphylla var. yunnanensis and its closely relative species, which could provide an infrared spectral identified evidence. The results of HCA showed that P. polyphylla var. yunnanensis, P. polyphylla var. chinensis, P. polyphylla, P. polyphylla var. stenophylla and P. polyphylla var. nana were relatively close. Conclusion Fourier transform infrared spectroscopy(FTIR) combined with original plant morphological identification can quickly identify P. polyphylla var. yunnanensis and its closely relative species, which will provide a scientific basis for the cultivation, clinical application, and resource development of genus Paris.
引文
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[18]李恒.重楼属植物[M].北京:科学出版社,1998.
[19]Chen J B,Sun S Q,Ma F,et al.Vibrational microspectroscopic identification of powdered traditional medicines:chemical micromorphology of Poria observed by infrared and Raman microspectroscopy[J].Spectrochim Acta A,2014,128(7):629-637.
[20]Hashim D M,Man Y B C,Norakasha R,et al.Potential use of Fourier transform infrared spectroscopy for differentiation of bovine and porcine gelatins[J].Food Chem,2010,118(3):856-860.
[21]Tan C S,Sing C Y,Loh Y C,et al.Vasorelaxation effect of Glycyrrhizae uralensis through the endotheliumdependent pathway[J].J Ethnopharmacol,2017,199:149-160.
[22]Loh Y C,Tan C S,Chng Y S,et al.Overview of signaling mechanism pathways employed by BPAid in vasodilatory activity[J].J Med Food,2017,20(12):1201-1213.
[23]赖先荣,周邦华,杜明胜,等.6种黄连饮片中6种生物碱的RP-HPLC含量测定及与“治消渴”药效学的谱-效关系分析[J].中国中药杂志,2016,41(24):4579-4586.
[24]Abdi H W L J.Principal component analysis[J].Computation Stat,2010,4(2):433-459.
[25]Agostino D,Ralph B.Principal Components Analysis[M].New York:Springer,2010.
[26]Bassbasi M,Luca M D,Ioele G,et al.Prediction of the geographical origin of butters by partial least square discriminant analysis(PLS-DA)applied to infrared spectroscopy(FTIR)data[J].J Food Compos Anal,2014,33(2):210-215.
[2]杨远贵,张霁,张金渝,等.重楼属植物化学成分及药理活性研究进展[J].中草药,2016,47(18):3301-3323.
[3]Liu Z,Li N,Gao W,et al.Comparative study on hemostatic,cytotoxic and hemolytic activities of different species of Paris L.[J].J Ethonpharmacol,2012,142(3):789-794.
[4]Zhen L,Qi Z,Chen W,et al.Chemosensitizing effect of Paris saponin I on camptothecin and10-hydroxycamptothecin in lung cancer cells via p38MAPK,ERK,and Akt signaling pathways[J].Eur J Med Chem,2017,125:760-769.
[5]Wu X,Wang L,Wang G C,et al.Triterpenoid saponins from rhizomes of Paris polyphylla var.yunnanensis[J].Carbohydr Res,2013,368(5):1-7.
[6]Sun J,Liu B R,Wei J,et al.The extract of Paris polyphylla exerts apoptotic induction and synergic antiproliferative effect with anticancer drugs in SMMC-7721 human liver cancer cells[J].Biomed Prevent Nutr,2011,1(3):186-194.
[7]赵飞亚,陶爱恩,夏从龙.基于国内专利结合资源、应用与开发现状的重楼发展策略探讨[J].中国中药杂志,2018,43(2):404-409.
[8]杨斌,杨丽英,严世武,等.滇重楼授粉特性的初步研究[J].西南农业学报,2008,21(5):1388-1390.
[9]Chen Y,Huang J,Yeap Z Q,et al.Rapid authentication and identification of different types of A.roxburghii by Tri-step FT-IR spectroscopy[J].Spectrochim Acta A,2018,199:271-282.
[10]Song S Y,Lee Y K,Kim I J.Sugar and acid content of Citrus prediction modeling using FT-IR fingerprinting in combination with multivariate statistical analysis[J].Food Chem,2016,190:1027-1032.
[11]Liu Y,Zhang Y,Zhang J,et al.Rapid discrimination of sea buckthorn berries from different H.rhamnoides subspecies by multi-step IR spectroscopy coupled withmultivariate data analysis[J].Infrared Phys Technol,2018,89:154-160.
[12]陈佳乐,金叶,陈红英,等.川芎药材的近红外多指标快速质量评价[J].中草药,2016,47(6):1004-1009.
[13]吴喆,张霁,金航,等.红外光谱结合化学计量学对不同采收期滇重楼的定性定量分析[J].光谱学与光谱分析,2017,37(6):1754-1758.
[14]吴喆,王元忠,张霁,等.基于红外光谱法的云南重楼及其近缘种的亲缘关系研究[J].中草药,2017,48(11):2279-2284.
[15]Liu Y,Finley J,Betz J M,et al.FT-NIR characterization with chemometric analyses to differentiate goldenseal from common adulterants[J].Fitoterapia,2018,127:81-88.
[16]Szymanskachargot M,Chylinska M,Kruk B,et al.Combining FT-IR spectroscopy and multivariate analysis for qualitative and quantitative analysis of the cell wall composition changes during apples development[J].Carbohyd Polym,2015,115:93-103.
[17]吴喆,张霁,左智天,等.红外光谱结合化学计量学快速鉴别云南重楼不同炮制品[J].光谱学与光谱分析,2018,38(4):1101-1106.
[18]李恒.重楼属植物[M].北京:科学出版社,1998.
[19]Chen J B,Sun S Q,Ma F,et al.Vibrational microspectroscopic identification of powdered traditional medicines:chemical micromorphology of Poria observed by infrared and Raman microspectroscopy[J].Spectrochim Acta A,2014,128(7):629-637.
[20]Hashim D M,Man Y B C,Norakasha R,et al.Potential use of Fourier transform infrared spectroscopy for differentiation of bovine and porcine gelatins[J].Food Chem,2010,118(3):856-860.
[21]Tan C S,Sing C Y,Loh Y C,et al.Vasorelaxation effect of Glycyrrhizae uralensis through the endotheliumdependent pathway[J].J Ethnopharmacol,2017,199:149-160.
[22]Loh Y C,Tan C S,Chng Y S,et al.Overview of signaling mechanism pathways employed by BPAid in vasodilatory activity[J].J Med Food,2017,20(12):1201-1213.
[23]赖先荣,周邦华,杜明胜,等.6种黄连饮片中6种生物碱的RP-HPLC含量测定及与“治消渴”药效学的谱-效关系分析[J].中国中药杂志,2016,41(24):4579-4586.
[24]Abdi H W L J.Principal component analysis[J].Computation Stat,2010,4(2):433-459.
[25]Agostino D,Ralph B.Principal Components Analysis[M].New York:Springer,2010.
[26]Bassbasi M,Luca M D,Ioele G,et al.Prediction of the geographical origin of butters by partial least square discriminant analysis(PLS-DA)applied to infrared spectroscopy(FTIR)data[J].J Food Compos Anal,2014,33(2):210-215.