一种基于全波长UV建立HPLC指纹图谱的质量控制模式:以不同等级的玄参为例Establishment of quality control model for high performance liquid chromatography fingerprint based on ultraviolet full-wavelength scanning spectrum: case study on different grades of Scrophulariae Radix
刘眴蒂;邓静;龚梦佳;盛瀚萱;林丽美;夏伯候;
LIU Shun-di;DENG Jing;GONG Meng-jia;SHENG Han-xuan;LIN Li-mei;XIA Bo-hou;Key Laboratory for Quality Evaluation of Bulk Herbs of Hunan Province, Hunan University of Chinese Medicine;
摘要(Abstract):
HPLC-UV指纹图谱是目前《中国药典》用于中药材质量控制的重要手段之一。然而,关于紫外(UV)具体的波段选择存在一定主观性,不能较好地表征中药内在的质量差异。因此,该研究以不同等级的玄参为研究对象,提出一种新的基于紫外全波长扫描图谱建立高效液相指纹图谱的质量控制模式。首先,采集不同等级的玄参样品,利用紫外光谱(200~400 nm)建立所有样品的紫外全波长吸收光谱。通过光谱数据的预处理和主成分分析(principal component analysis, PCA)以及偏最小二乘法判别分析(partial least squares discrimination analysis, PLS-DA)等多元统计分析方法建立差异分析模式,得到251 nm的紫外波段可能是区分不同等级玄参质量差异的主要贡献的波段。进一步,利用该波段建立其HPLC指纹图谱,通过多元统计方法分析可得,基于差异紫外波段筛选得到的HPLC指纹图谱较原来的UV指纹图谱有更显著的分类趋势,可用于判别不同等级的玄参样品,能够较好的体现各等级之间的差异性。该研究建立的方法对于中药特异性HPLC指纹图谱的建立及其质量控制具有较大的指导和参考意义。
High performance liquid chromatography-ultraviolet(HPLC-UV) fingerprint is one of the most important methods for the quality control of Chinese medicines in Chinese Pharmacopoeia. However, certain subjectivity is present in selection of specific band of UV, and the inherent quality differences of Chinese medicine can′t be well characterized by this method. Therefore, with different grades of Scrophulariae Radix were taken as the research object in this study, a new quality control model of HPLC-UV was established in this study based on the ultraviolet full-wavelength scanning spectrum. Firstly, different grades of Scrophulariae Radix samples were collected, and the full-wavelength ultraviolet absorption spectra of all the samples were established at the bands of 200-400 nm. In order to analyze the differences among samples, the analysis model was built following multivariate statistical analysis methods such as principal component analysis(PCA) and partial least squares discrimination analysis(PLS-DA) after the pretreatment of spectral data. The result showed that the ultraviolet band at 251 nm may contribute most to distinguish the quality differences among different grades of samples. Then, the HPLC fingerprints of samples were established with the band at 251 nm. The multivariate statistical analysis showed that there was a more significant classification trend in HPLC fingerprints than that in the original UV fingerprints, which could be used to distinguish different grades of samples, and could better reflect the differences among different grades. The method reported in this study can be of a great guidance and reference for the establishment of specific fingerprints of Chinese medicines as well as for the quality control of Chinese medicine.
关键词(KeyWords):
玄参;质量控制;紫外全波长图谱;高效液相色谱
Scrophulariae Radix;quality control;ultraviolet full-wavelength spectra;high performance liquid chromatography
基金项目(Foundation): 湖南省自然科学基金项目(2017JJ4045);; 湖南省教育厅科研项目(19B416);; 湖南省中药学一流学科项目;; 湖南省大学生研究性学习和创新性实验计划项目(1021-0001017174)
作者(Author):
刘眴蒂;邓静;龚梦佳;盛瀚萱;林丽美;夏伯候;
LIU Shun-di;DENG Jing;GONG Meng-jia;SHENG Han-xuan;LIN Li-mei;XIA Bo-hou;Key Laboratory for Quality Evaluation of Bulk Herbs of Hunan Province, Hunan University of Chinese Medicine;
Email:
DOI: 10.19540/j.cnki.cjcmm.20200706.301
参考文献(References):
- [1]LIANG Y Z,XIE P,CHAN K.Quality control of herbal medicines[J].J Chromatogr B,2004,812(1):53.
- [2]谢培山.中药色谱指纹图谱[M].北京:人民卫生出版社,2005:8.
- [3]石上梅.逐步完善中药质量标准体系和质量控制模式---解读2015年版《中国药典》(一部)[J].中国药学杂志,2015,50(20):1752.
- [4]孙波,孙涛.高效液相色谱法在中药指纹图谱中的应用现状及分析[J].时珍国医国药,2006,17(1):109.
- [5]谢培山.中药制剂色谱指纹图谱(图像)鉴别[J].中成药,2000,22(6):3.
- [6]国家药典委员会.中华人民共和国药典.一部[S].北京:中国医药科技出版社,2015:117.
- [7]缪希雍.神农本草经疏[M].北京:中国中医药出版社,1997:1171.
- [8]FILZOMSER P.Introduction to multivariate statistical analysis in chemometrics[J].Appl Spectrosc,2010,64(4):112.
- [9]DUNN W B,ELLIS D I.Metabolomics:current analytical platforms and methodologies[J].Trac-Trend Anal Chem,2005,24(4):285.
- [10]TRYGG J,HOLMES E,LUNDSTEDT T.Chemometrics in metabonomics[J].J Proteome Res,2007,6(2):469.
- [11]WORLEY B,HALOUSKA S,POWERS R.Utilities for quantifying separation in PCA/PLS-DA scores plots[J].Anal Biochem,2013,433(2):102.
- [12]ZHANG D,DUAN X,DENG S,et al.Fingerprint analysis,multi-component quantitation,and antioxidant activity for the quality evaluation of Salvia miltiorrhiza var.alba by highperformance liquid chromatography and chemometrics[J].J Sep Sci,2015,38(19):3337.
- [13]PAULI E D,BRUNS R E,SCARMINIO I S.UV-Vis spectral fingerprinting and chemometric method applied to the evaluation of Camellia sinensis leaves from different harvests[J].Anal Methods-UK,2016,8(41):7537.
- [14]张春泥,王英姿,孙欣光,等.HPLC-CAD结合化学计量学的川楝子饮片指纹图谱研究[J].药学学报,2017,52(3):456.
- [15]WOLD S,ESBENSEN K,GELADI P J C,et al.Principal component analysis[J].Chemometr Intell Lab,1987,2(1):37.
- [16]余欣尉,吴谦,吕望,等.基于液相色谱-质谱联用技术的肺癌细胞代谢组学分析[J].色谱,2013,31(7):691.
- [17]张召强,李明.玄参的化学成分及药理作用的研究进展[J].中国医药指南,2013,11(26):49.
- [18]喻欢欢,钟猛,丁锐,等.一测多评法测定玄参中7种有效成分的含量[J].中国中药杂志,2017,42(14):2719.
- [19]BALLABIO D,CONSONNI V.Classification tools in chemistry.Part 1:linear models.PLS-DA[J].Anal Method,2013,5(16):3790.
- [20]BOULESTEIX A L,STRIMMER K.Partial least squares:a versatile tool for the analysis of high-dimensional genomic data[J].Brief Bioinform,2007,8(1):32.
- [21]SZYMAN'SKA E,SACCENTI E,SMILDE A K,et al.Doublecheck:validation of diagnostic statistics for PLS-DA models in metabolomics studies[J].Metabolomics,2012,8(1):3.
- [22]WESTERHUIS J A,HOEFSLOOT H C J,SMIT S,et al.Assessment of PLSDA cross validation[J].Metabolomics,2008,4(1):81.
- [23]石佳,沈立,邓哲,等.质量常数评价方法应用于玄参饮片等级评价研究[J].中国中药杂志,2019,44(9):1744.
- [24]许洪波,高春晖,蔡兴航,等.玄参饮片标准汤剂制备及质量标准研究[J].中国中药杂志,2019,44(12):2493.
- [25]许一鸣,吴啟南,乐巍,等.玄参芦头片对玄参药材整体质量的影响研究[J].天然产物研究与开发,2016,28(9):1414.
- [26]陈鹏,李硕,邱佳,等.玄参药材质量控制研究进展[J].中国实验方剂学杂志,2015,21(18):220.
- 玄参
- 质量控制
- 紫外全波长图谱
- 高效液相色谱
Scrophulariae Radix - quality control
- ultraviolet full-wavelength spectra
- high performance liquid chromatography
- 刘眴蒂
- 邓静
- 龚梦佳
- 盛瀚萱
- 林丽美
- 夏伯候
LIU Shun-di- DENG Jing
- GONG Meng-jia
- SHENG Han-xuan
- LIN Li-mei
- XIA Bo-hou
- Key Laboratory for Quality Evaluation of Bulk Herbs of Hunan Province
- Hunan University of Chinese Medicine
- 刘眴蒂
- 邓静
- 龚梦佳
- 盛瀚萱
- 林丽美
- 夏伯候
LIU Shun-di- DENG Jing
- GONG Meng-jia
- SHENG Han-xuan
- LIN Li-mei
- XIA Bo-hou
- Key Laboratory for Quality Evaluation of Bulk Herbs of Hunan Province
- Hunan University of Chinese Medicine