HPLC特征指纹图谱结合化学计量学比较川牛膝及其混淆品、掺混品化学成分差异Comparison of chemical constituents between Cyathula offinalis and adulterant and their admixture by HPLC characteristic chromatogram fingerprint combined with chemometrics
施崇精;王姗姗;程中琴;刘小妹;袁强华;刘莉;呼梅;宋英;
SHI Chong-jing;WANG Shan-shan;CHENG Zhong-qin;LIU Xiao-mei;YUAN Qiang-hua;LIU Li;HU Mei;SONG Ying;Chengdu University of Traditional Chinese Medicine;Teaching Hospital of Chengdu University of Traditional Chinese Medicine;
摘要(Abstract):
川牛膝的主要混淆品为同属植物头花杯苋的根,研究报道其在性味和药理活性上存有明显差异,仅可作为地方习惯用药,不得代替川牛膝入药。但因头花杯苋外观形态上与川牛膝十分相似,且杯苋甾酮含量也能达到现行药典限量,导致市场上多以冒充或掺混形式出售,这严重地影响了川牛膝的临床用药安全和道地药材开发。鉴于此,该文采用HPLC特征指纹图谱技术试图揭示川牛膝、混淆品头花杯苋、掺混川牛膝在多组分下的化学特征成分差异,以期为川牛膝区别用药和真伪鉴别提供有力支撑。通过测得川牛膝及其混淆品、掺混品色谱图,得出65个不同组分,以建立26×65的峰面积数据矩阵,并进行特征峰差异分析、相似度分析、聚类分析和主成分分析。特征峰差异分析得出头花杯苋及其掺混品特征峰较川牛膝多且响应值高,筛选出的9个特征峰用以鉴别伪品头花杯苋,其中2个可用以鉴别出掺混5%川牛膝,且9个特征峰的光谱图大多类似于皂苷类成分的末端吸收光谱图,表明头花杯苋可能含有大量皂苷类成分。以建立的川牛膝特征指纹图谱为参照,相似度分析得出川牛膝相似度均大于0.942,而头花杯苋、掺混川牛膝相似度分别小于0.383,0.399。聚类分析树状图和主成分得分三维图均能将川牛膝、头花杯苋、掺混川牛膝明显的分为3类。以上研究结果均表明川牛膝、头花杯苋、掺混川牛膝在化学组分上表现出明显的差异,当区别用药,不可混淆。
Cyathula capitate is the main adulterant of C. offinalis. According to the literature reported,there are obvious differences in properties,taste and pharmacological activity between C. capitate and C. offinalis. Therefore,C. capitate can only be used as a local conventional medicine and can't be a substitute for C. offinalis. Since the appearance of C. capitata is very similar to the C. offinalis and the content of cyasterone also can reach the limit of the current pharmacopoeia standard,the C. capitata is mostly sold in the form of impersonation oradmixture,which seriously affected the safety of the clinical medication and the development of the genuine crude drugs. In view of this,HPLC characteristic fingerprint was used to reveal the difference of multi-ingredients of C. offinalis,C. capitata and their admixture. According to the HPLC chromatogram of C. offinalis,C. capitata. and their admixture,65 different components were obtained to set up a peak area data matrix of 26 × 65,which was applied to perform the characteristic peak difference analysis,similarity analysis,hierarchical clustering analysis HCA and principal component analysis( PCA). Characteristic peak difference analysis showed that the characteristic peaks of C. capitata and their admixture are more and higher respond than those of C. offinalis.The 9 characteristic peaks were used to distinguish C. capitata,2 of which were used to distinguish C. offinalis mixed with 5% C.capitata. UV spectra of 9 characteristic peaks are mostly similar to the end absorption spectra of saponins,indicating that C. capitata may contain a large amount of saponins. By the reference fingerprint of C. offinalis established,the similarity analysis showed that the similarity degree of C. offinalis are higher than 0. 942,while the similarity degree of C. capitata,C. offinalis mixed with 5% C. capitata are less than 0. 383 and 0. 399. C. offinalis,C. capitata,C. offinalis mixed with 5% C. capitata could be obviously divided into3 classes by HCA and PCA. These results showed that there are obvious difference in the chemical composition of C. offinalis,C. capitata and their admixture,which could provide evidence for their identification.
关键词(KeyWords):
川牛膝;头花杯苋;混淆品;掺混品;HPLC指纹图谱;化学计量学;化学成分差异
Cyathula offinalis;C.capitata;adulterant;admixture;HPLC chromatogram fingerprint;chemometrics;chemical composition difference
基金项目(Foundation): 国家中医药管理局国家中药标准化项目(ZYBZH-Y-SC-41);; 四川省中医药管理局川产道地药材综合开发与区域发展(川牛膝)项目(2017ZY001)
作者(Author):
施崇精;王姗姗;程中琴;刘小妹;袁强华;刘莉;呼梅;宋英;
SHI Chong-jing;WANG Shan-shan;CHENG Zhong-qin;LIU Xiao-mei;YUAN Qiang-hua;LIU Li;HU Mei;SONG Ying;Chengdu University of Traditional Chinese Medicine;Teaching Hospital of Chengdu University of Traditional Chinese Medicine;
Email:
DOI: 10.19540/j.cnki.cjcmm.2018.0071
参考文献(References):
- [1]中国药典.一部[S].2015.
- [2]李成,马新建.道地药材之怀牛膝、川牛膝的本草考证[J].中医学报,2011,26(11):1336.
- [3]施崇精,王姗姗,刘小妹,等.川产道地药材川牛膝现状探讨[J].亚太传统医药,2017,13(22):55.
- [4]何光星,舒光明,宾雪英,等.川牛膝及其混淆品活血化瘀作用比较[J].时珍国医国药,2015,26(10):2336.
- [5]宾雪英.川牛膝及其混淆品药理作用研究[D].泸州:泸州医学院,2013.
- [6]范巧佳,李方安,李杰勤,等.川牛膝与其伪品头花杯苋及其杂交后代同工酶的研究[J].中药研究与信息,2004(4):24.
- [7]陈诗晴,蒲沁琳,陈小军,等.基于ITS序列分析技术鉴定川牛膝与常见伪品麻牛膝[J].中药与临床,2014,5(4):4.
- [8]赵华杰,舒光明,周先健,等.我国川牛膝资源分布及生产状况调查[J].资源开发与市场,2012,28(5):414.
- [9]赵磊.川牛膝及其常见混用品鉴别研究[J].亚太传统医药,2017,13(16):32.
- [10]肖佳佳,黄红,雷有成,等.天麻HPLC指纹图谱建立及判别分析[J].中国中药杂志,2017,42(13):2524.
- [11]许灿新.延胡索及其伪品的鉴别研究[D].广州:广州中医药大学,2016.
- [12]王少平,孙乙铭,姜艳,等.基于指纹图谱及柚皮素含量对铁皮石斛及伪品的鉴定[J].浙江农业学报,2015,27(12):2199.
- [13]童凯,李昭玲,闫燊,等.基于HPLC指纹图谱比较分析川牛膝及其混淆品的化学成分特征[J].时珍国医国药,2015,26(10):2424.
- 川牛膝
- 头花杯苋
- 混淆品
- 掺混品
- HPLC指纹图谱
- 化学计量学
- 化学成分差异
Cyathula offinalis - C.capitata
- adulterant
- admixture
- HPLC chromatogram fingerprint
- chemometrics
- chemical composition difference
- 施崇精
- 王姗姗
- 程中琴
- 刘小妹
- 袁强华
- 刘莉
- 呼梅
- 宋英
SHI Chong-jing- WANG Shan-shan
- CHENG Zhong-qin
- LIU Xiao-mei
- YUAN Qiang-hua
- LIU Li
- HU Mei
- SONG Ying
- Chengdu University of Traditional Chinese Medicine
- Teaching Hospital of Chengdu University of Traditional Chinese Medicine
- 施崇精
- 王姗姗
- 程中琴
- 刘小妹
- 袁强华
- 刘莉
- 呼梅
- 宋英
SHI Chong-jing- WANG Shan-shan
- CHENG Zhong-qin
- LIU Xiao-mei
- YUAN Qiang-hua
- LIU Li
- HU Mei
- SONG Ying
- Chengdu University of Traditional Chinese Medicine
- Teaching Hospital of Chengdu University of Traditional Chinese Medicine