基于ITS/ITS2和HPLC指纹图谱技术的款冬花质量评价

doi:10.5246/jcps.2021.01.006

中国药学(英文版) ›› 2021, Vol. 30 ›› Issue (1): 58-68.DOI: 10.5246/jcps.2021.01.006

基于ITS/ITS2和HPLC指纹图谱技术的款冬花质量评价

卢紫娟¹, 贾岩¹, 邢婕¹, 肖淑贤², 王玉龙², 秦文杰³, 秦雪梅¹, 李震宇¹^,^*()

1. 山西大学中医药现代研究中心, 山西太原 030006
2. 山西振东道地药材开发有限公司, 山西长治 047100
3. 北京振东光明药物研究院, 北京 100085

收稿日期:2020-07-03 修回日期:2020-08-13 接受日期:2020-08-23 出版日期:2021-01-29 发布日期:2021-01-29
通讯作者: 李震宇
作者简介:
+ Tel.: +86-351-7018379, E-mail: lizhenyu@sxu.edu.cn
基金资助:
National Natural Science Foundation China (Grant No. 31270008 and 81973466), and National Standardization Program of Traditional Chinese Medicine (Grant No. ZYBZH-Y-JIN-34).

Quality evaluation of Farfarae Flos by ITS/ITS2 and HPLC fingerprint analysis

Zijuan Lu¹, Yan Jia¹, Jie Xing¹, Shuxian Xiao², Yulong Wang², Wenjie Qin³, Xuemei Qin¹, Zhenyu Li¹^,^*()

1 Modern Research Center for Traditional Chinese Medicine, Shanxi University, Taiyuan 030006, China
2 Shanxi Zhendong Genuine Medicinal Materials Development Co., Ltd., Changzhi 047100, China
3 Beijing Zhendong Guangming Pharmaceutical Research Institute Co., Ltd., Beijing 100085, China

Received:2020-07-03 Revised:2020-08-13 Accepted:2020-08-23 Online:2021-01-29 Published:2021-01-29
Contact: Zhenyu Li

摘要/Abstract

摘要：

款冬花为菊科植物款冬( Tussilago farfara L.)的干燥花蕾, 临床用于治疗咳嗽、支气管炎和哮喘。本研究收集了来自中国北方五个省的26批款冬花样本, 并采用ITS-PCR与高效液相色谱(HPLC)指纹图谱相结合用于款冬花的质量评价。ITS序列分析结果表明这些样品之间具有很高的相似度。与ITS1相比, ITS2序列的分子多样性较低, 并且ITS的变异与产地或栽培方法之间无明显的相关性。HPLC指纹图谱与相似度分析结果展示了26批款冬花样品中苯丙素类化合物和黄酮类化合物的相对含量变化趋势, 并且这两类成分在不同产地之间无明显差异。苯丙素类化合物和黄酮类化合物显示出相似的变化趋势, 且两者之间存在正相关。本研究表明, 来自不同产地的款冬花呈现较高的相似度, 且ITS-PCR结合HPLC指纹图谱分析的方法可以作为款冬花质量评价的一种有效方法。

关键词: 款冬花, ITS, ITS2, HPLC指纹图谱, 多元分析

Abstract:

The flower bud of Tussilago farfara L., also known as Farfarae Flos (FF), is commonly used in the traditional Chinese medicine (TCM) for the treatment of cough, bronchitis and asthmatic disorders. In this study, 26 samples from five provinces across northern China were collected, and internal transcribed spacer-polymerase chain reaction (ITS-PCR) coupled with high performance liquid chromatography (HPLC) fingerprint profiling was used for the quality evaluation of FF. The results of the ITS sequence analysis showed the high similarity values among these samples. ITS2 sequence exhibited less molecular diversity compared with ITS1, and no obvious correlation was found between the variation of ITS with the production areas or cultivation methods. The results of HPLC fingerprints in combination with the similarity analysis suggested that relative contents of phenylpropanoids and flavonoids varied among the 26 FF samples, and there was also no obvious difference among the different habitats. The phenylpropanoids and the flavonoids showed similar fluctuating patterns, and positive correlation among them was also observed. The results presented in this study suggested that FF from different habits showed high similarities, and ITS-PCR coupled with HPLC fingerprint profiling could be used as a valuable approach for the quality evaluation of FF.

Key words: Farfarae Flos, ITS, ITS2, HPLC fingerprints, Multivariate analysis

Supporting:

卢紫娟, 贾岩, 邢婕, 肖淑贤, 王玉龙, 秦文杰, 秦雪梅, 李震宇. 基于ITS/ITS2和HPLC指纹图谱技术的款冬花质量评价[J]. 中国药学(英文版), 2021, 30(1): 58-68.

Zijuan Lu, Yan Jia, Jie Xing, Shuxian Xiao, Yulong Wang, Wenjie Qin, Xuemei Qin, Zhenyu Li. Quality evaluation of Farfarae Flos by ITS/ITS2 and HPLC fingerprint analysis[J]. Journal of Chinese Pharmaceutical Sciences, 2021, 30(1): 58-68.

图/表 6

Table 1. Details of samples.

Figure 1. Nine variant sites in ITS1 sequence of FF samples. The length of the alignment of ITS1 was approximately 259 bp.

Figure 2. NJ tree constructed using the ITS sequences of 26 varieties samples.

Table 2. Similarity value of 14 common peaks and total peak area collected from different locations.

Figure 3. PCA (A) scores plot, lines of the peak area of phenylpropanoids (B), flavonoids (C), and sesquiterpenes (D) in 26 FF samples.

Figure 4. Correlation network of 26 FF samples from various locations was conducted based on Pearson's correlation coefficients.

参考文献 22

[1]	Gao, H.; Huang, Y.N.; Gao, B.; Xu, P.Y.; Inagaki, C.; Kawabata, J. α-Glucosidase inhibitory effect by the flower buds of Tussilago farfara L. Food Chem. 2008, 106, 1195–1201.
[2]	Wu, D.; Zhang, M.; Zhang, C.F.; Wang, Z.T. Flavonoids and phenolic acid derivatives from flos farfarae. China J. Chin. Mater. Med. 2010, 35, 1142–1144.
[3]	Liu, Y.F.; Yang, X.W.; Wu, B. Studies on chemical constituents in the buds of Tussilago farfara. China J. Chin. Mater. Med. 2007, 32, 2378–2381.
[4]	Yaoita, Y.; Suzuki, N.; Kikuchi, M. Structures of new sesquiterpenoids from Farfarae Flos. Chem. Pharm. Bull. 2001, 49, 645–648.
[5]	Kong, W.J.; Zhao, Y.L.; Xiao, X.H.; Jin, C.; Li, Z.L. Quantitative and chemical fingerprint analysis for quality control of Rhizoma Coptidischinensis based on UPLC-PAD combined with chemometrics methods. Phytomedicine. 2009, 16, 950–959.
[6]	Li, Z.Y.; Zhi, H.J.; Zhang, F.S.; Sun, H.F.; Zhang, L.Z.; Jia, J.P.; Xing, J.; Qin, X.M. Metabolomic profiling of the antitussive and expectorant plant Tussilago farfara L. by nuclear magnetic resonance spectroscopy and multivariate data analysis. J. Pharm. Biomed. Anal. 2013, 75, 158–164.
[7]	Li, D.Z.; Gao, L.M.; Li, H.T.; Wang, H.; Ge, X.J.; Liu, J.Q.; Chen, Z.D.; Zhou, S.L.; Chen, S.L.; Yang, J.B.; Fu, C.X.; Zeng, C.X.; Yan, H.F.; Zhu, Y.J.; Sun, Y.S.; Chen, S.Y.; Zhao, L.; Wang, K.; Yang, T.; Duan, G.W. Comparative analysis of a large dataset indicates that internal transcribed spacer (ITS) should be incorporated into the core barcode for seed plants. Proc. Natl. Acad. Sci. USA. 2011, 108, 19641–19646.
[8]	Luo, K.; Chen, S.L.; Chen, K.L.; Song, J.Y.; Yao, H.; Ma, X.Y.; Zhu, Y.J.; Pang, X.H.; Yu, H.; Li, X.W.; Liu, Z. Assessment of candidate plant DNA barcodes using the Rutaceae family. Sci. China Life Sci. 2010, 53, 701–708.
[9]	Selvaraj, D.; Sarma, R.K.; Shanmughanandhan, D.; Srinivasan, R.; Ramalingam, S. Evaluation of DNA barcode candidates for the discrimination of the large plant family Apocynaceae. Plant Syst. Evol. 2015, 301, 1263–1273.
[10]	Chen, X.C.; Liao, B.S.; Song, J.Y.; Pang, X.H.; Han, J.P.; Chen, S.L. A fast SNP identification and analysis of intraspecific variation in the medicinal Panax species based on DNA barcoding. Gene. 2013, 530, 39–43.
[11]	Abu Almakarem, A.S.; Heilman, K.L.; Conger, H.L.; Shtarkman, Y.M.; Rogers, S.O. Extraction of DNA from plant and fungus tissues in situ. BMC Res. Notes. 2012, 5, 1–11.
[12]	Qiao, S.L. Studies on similarity evaluation of chromatographic fingerprint for TCM. Second Military Med. Univ. 2014.
[13]	Zhang, Z.P.; Zhang, Y.; Zhang, Z.; Yao, H.; Liu, H.T.; Zhang, B.G.; Liao, Y.H. Comparative analysis of DNA barcoding and HPLC fingerprint to trace species of phellodendri cortex, an important traditional Chinese medicine from multiple sources. Biol. Pharm. Bull. 2016, 39, 1325–1330.
[14]	Li, D.Z.; Liu, J.Q.; Chen, Z.D.; Wang, H.; Ge, X.J.; Zhou, S.L.; Gao, L.M.; Fu, C.X.; Chen, S.L. Plant dna barcoding in China. J. Syst. Evol. 2011, 49, 165–168.
[15]	Yang, P.; Li, X.W.; Zhou, H.; Hu, H.; Zhang, H.; Sun, W.; Wang, Y.T.; Yao, H. Molecular identification of Chinese materia medica and its adulterants using ITS2 and psbA-trnH barcodes: a case study on rhizoma menispermi. Chin. Med. 2014, 5, 190–198.
[16]	Yu, H.; Wu, K.Y.; Song, J.Y.; Zhu, Y.J.; Yao, H.; Luo, K.; Dai, Y.; Xu, S.J.; Lin, Y.L. Expedient identification of Magnoliaceae species by DNA barcoding. Plant Omics. 2014, 7, 47–53.
[17]	Ma, X.Y.; Xie, C.X.; Liu, C.; Song, J.Y.; Yao, H.; Luo, K.; Zhu, Y.J.; Gao, T.; Pang, X.H.; Qian, J.; Chen, S.L. Species identification of medicinal pteridophytes by a DNA barcode marker, the chloroplast psbA-trnH intergenic region. Biol. Pharm. Bull. 2010, 33, 1919–1924.
[18]	Deng, W.P.; Xu, T.T.; Yang, M.; Cui, Y.J.; Guo, D.A. Improved chromatographic fingerprinting combined with multi-components quantitative analysis for quality evaluation of Penthorum chinense by UHPLC-DAD. Nat. Prod. Commun. 2015, 10, 71–76.
[19]	Zhang, W.; Yang, T.S.; Wang, X.P.; Dang, W.X. HPLC determination of chlorogenic acid and rutin contents in Flos Farfarae. Chin. J. Pharm. Anal. 2008, 28, 2106–2108.
[20]	Viapiana, A.; Wesolowski, M. HPLC fingerprint combined with quantitation of phenolic compounds and chemometrics as an efficient strategy for quality consistency evaluation of sambucus nigra berries. Nat. Prod. Commun. 2016, 11, 1449-1454.
[21]	Goodarzi, M.; Russell, P.J.; Vander Heyden, Y. Similarity analyses of chromatographic herbal fingerprints: a review. Anal. Chimica Acta. 2013, 804, 16–28.
[22]	Yudthavorasit, S.; Wongravee, K.; Leepipatpiboon, N. Characteristic fingerprint based on gingerol derivative analysis for discrimination of ginger (Zingiber officinale) according to geographical origin using HPLC-DAD combined with chemometrics. Food Chem. 2014, 158, 101–111.

基于ITS/ITS2和HPLC指纹图谱技术的款冬花质量评价

Quality evaluation of Farfarae Flos by ITS/ITS2 and HPLC fingerprint analysis

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 6

参考文献 22

相关文章 9

编辑推荐

Metrics

本文评价

[1]	苏静华, 张超, 孙磊, 马双成, 邢以文. 基于多元统计分析和模式识别技术建立浙贝母和川贝母的品种鉴别体系[J]. 中国药学(英文版), 2023, 32(5): 406-416.
[2]	杨红帅, 刘秋怡, 殷卫东, 解满江, 吉翔, 邵双宇, 梁鸿, 屠鹏飞, 张庆英. 酸枣仁和滇枣仁的HPLC指纹图谱分析及鉴别[J]. 中国药学(英文版), 2020, 29(4): 252-259.
[3]	林珠灿, 张龙, 张睿卓, 黄安玉, 郭素华. 基于炎症细胞模型的景天三七醋酸乙酯部位抗炎活性及其HPLC指纹图谱的研究[J]. 中国药学(英文版), 2015, 24(10): 647-653.
[4]	刘毅, 王伟, 赵玉英, 陈虎彪, 梁鸿, 张庆英. 红花岩黄芪药材的化学指纹分析[J]. 中国药学(英文版), 2015, 24(10): 654-659.
[5]	陈金凤, 屠鹏飞, 姜勇. 加压制备色谱技术结合HPLC指纹图谱导向快速分离制备红花提取物中的主要黄酮类成分[J]. 中国药学(英文版), 2014, 23(7): 490-495.
[6]	马晓庆, 邓伟峰, 杨东辉^*, 刘广学, 洪浩, 白光男, 蔡少青 . 秦艽生药中龙胆苦苷和落干酸的含量及其HPLC指纹图谱测定[J]. , 2010, 19(4): 243-250.
[7]	刘玉峰, 杨秀伟^*, 武滨. 款冬花的化学成分[J]. , 2007, 16(4): 288-293.
[8]	刘玉峰, 杨秀伟^*, 武滨. 款冬花挥发油成分的GC-MS分析[J]. , 2006, 15(1): 10-14.
[9]	石巍, 韩桂秋^*. 款冬花的化学成分[J]. , 1996, 5(2): 63-67.