HPLC fingerprinting-based multivariate analysis of chemical components in Tetrastigma Hemsleyanum Diels et Gilg: Correlation to their antioxidant and neuraminidase inhibition activities

https://doi.org/10.1016/j.jpba.2021.114314Get rights and content

Highlights

  • Four of the forty-two components from Tetrastigma Hemsleyanum Diels et Gilg were identified for the first time.

  • Different origin showed different antioxidant and neuraminidase inhibitory activities.

  • Four components were identified as vital compounds related to bioactivities and quantified.

Abstract

Tetrastigma Hemsleyanum Diels & Gilg (TDG) has attracted growing attention in China; however, there were few studies on its bioactive components. Herein, the characteristic chemical components and dual antioxidant and neuraminidase inhibitory activities of fifteen batches of TDG from different places of origin and their relevance were investigated. The HPLC fingerprint was first established and the marker components were identified by using UPLC-Q-TOF-MS/MS. Catechin-5-O-β-d-glucopyranoside, tartaric acid, (1R, 2R, 4S)-2-hydroxy-1, 8-cineole-β-d-glucopyranoside, and phlorizin were identified for the first time. The result of multivariate statistical analysis indicated that multiple components have a significant contribution to the classification of TDG, such as chlorogenic acid, saccharumoside C/D, robinin, procyanidin B2, rutin, isoquercitrin, etc. Then, the antioxidant and neuraminidase inhibitory activities of fifteen batches of TDG were measured. The result of grey relationship analysis showed that the contents of rutin, isoquercitrin, kaempferol-3-rutinoside, and astragalin were positively correlated with these two activities with correlation coefficients more than 0.8. The quantitative analysis of these four bioactive compounds was performed by using HPLC-DAD. The recovery rate of the method varied from 98.02% to 100.21%, the RSD values of precision, stability and repeatability were between 1.32–3.15 %, and the R value of the linear equation was above 0.9990. To sum up, this study is valuable in the quality control of TDG.

Introduction

Tetrastigma hemsleyanum Diels & Gilg (TDG), a perennial liana plant from the Vitaceae family, shows functions of clearing heat and detoxification, activating blood circulation and relieving pain, dispelling wind and resolving phlegm [1]. Now, TDG is commonly used as functional foods and medicinal herbs in traditional Chinese medicine (TCM). People are increasingly consuming it as a functional soup/juice/nutritional food to keep healthy by boiling/squeezing/extracting the herb [2]. Modern research showed that it could prevent some chronic diseases, enhance the immunity, antioxidant and anti-influenza virus capacity of the body, and protect the liver from chemical damage [3]. TDG contains numerous chemical components, such as flavonoids, phenolic acids and their derivatives, triterpenoids, organic acids, fatty acids, and son on [4,5]. These compounds may contribute to the bioactive effects of TDG [1,6]. With the gradual recognition of its medicinal value, TDG from wild resources have also been exhaustively exploited. Thus, TDG has become a rare and endangered medicinal plant, and large-scale planting has become an effective substitute for wild resources. In 2018, Zhejiang province designated it as the new “Zhebawei” cultivar of traditional Chinese medicine.

The efficacy of TCM has a great correlation with its chemical compositions and the content of active ingredients [7]. However, analysis of the chemical components of TCM is far more complex than that of chemical drugs, because of a variety of factors affecting the chemical components of TCM, such as germplasm resources, places of origin, and processing methods [8]. However, differences lying in the chemical components and bioactivities of TDG from different places of origin were still unclear. In previous studies, different pattern recognition methods combined with HPLC fingerprints were used for the identification of TDG from different origins and twenty-three characteristic peaks were identified through multivariate analysis [9]. These results were helpful to the quality control and grading of TDG to some extent. However, the new challenge was to identify the most closely related marker chemical components from dozens of chemical components, so as to improve the applicability and scientificity of TDG quality control methods.

In this study, the characteristic chemical components in fifteen batches of TDG from different places of origin were qualitatively analyzed by UPLC-Q-TOF-MS/MS. Moreover, various statistical models, including principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), similarity analysis (SA), and heatmap hierarchical clustering analysis (heatmap HCA), were used to evaluate the difference of TDG from different places of origin. Furthermore, the antioxidant, assessed via the scavenging rate of 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH), and the neuraminidase inhibition activities of fifteen batches of TDG were measured. Grey relational analysis (GRA) and partial least square (PLS) analysis were used to evaluate the correlation between the characteristic chemical components and these two activities. Finally, the contents of marker chemical components, including rutin, isoquercitrin, kaempferol-3-rutinoside, and astragalin were determined simultaneously by HPLC.

Section snippets

Reagents and herbal medicine samples

Kaempferol-3-rutinoside was purchased National Institutes for Food and Drug Control (Beijing, China). Rutin, isoquercitrin, and astragalin were purchased from Shanghai Yuanye Bio-Technology Co., Ltd (Shanghai, China). The purity of four reference materials was reported in the certificate of analysis as ≥ 98.0 % (by HPLC). DPPH was purchased from Baomanbio (Shanghai, China). HPLC-grade acetonitrile and acetic acid were purchased from Thermo Fisher Scientific Company (MA, USA). Ultra-pure water

Analysis of HPLC fingerprints and similarities

Based on the fingerprints of TDG with different places of origin, the chromatogram of S7 was taken as a reference spectrum as it contains the most identified chemical components, and fourteen common peaks were further marked in the chromatographic fingerprint (Fig. 1). Considering the higher response, better separation, and appropriate retention time than the others, it was selected as the reference peak (S), which was further identified to be kaempferol-3-rutinoside. The stacking chromatogram

Conclusion

In the present study, starting from two chromatographic techniques (HPLC fingerprint and UPLC-Q-TOF–MS/MS), forty-two characteristic chemical components were found in TDG, and four of them were identified for the first time. HPLC fingerprints showed that the similarity of fifteen batches of TDG from different places of origin were 0.866−0.988. Fifteen batches of TDG showed different antioxidant and neuraminidase inhibitory activities, mainly ascribed to rutin, isoquercitrin,

CRediT authorship contribution statement

Yu-li Jiang: Conceptualization, Methodology, Software, Writing - original draft. Zi-jin Xu: Methodology, Writing - original draft. Yi-feng Cao: Writing - review & editing. Fang Wang: Formal analysis, Software. Chu Chu: Resources, Validation. Cheng Zhang: Project administration. Yi Tao: Supervision, Formal analysis. Ping Wang: Supervision, Funding acquisition.

Declaration of Competing Interest

The authors have declared no conflict of interest.

Acknowledgments

This work was financially supported by the Special Project of International Technology Cooperation of One Belt and One Road (No. 2017C04009) and the key projects of International Scientific and Technological Innovation Cooperation between Governments (No. 2017YFE0130100).

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