HPLC fingerprinting-based multivariate analysis of chemical components in Tetrastigma Hemsleyanum Diels et Gilg: Correlation to their antioxidant and neuraminidase inhibition activities
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).
References (31)
- et al.
Rapid characterization of chemical constituents in Radix Tetrastigma, a functional herbal mixture, before and after metabolism and their antioxidant/antiproliferative activities
J. Funct. Foods
(2015) - et al.
The research progresses and future prospects of Tetrastigma hemsleyanum Diels et Gilg: a valuable Chinese herbal medicine
J. Ethnopharmacol.
(2021) - et al.
A comprehensive review on traditional uses, chemical compositions, pharmacology properties and toxicology of Tetrastigma hemsleyanum
J. Ethnopharmacol.
(2021) - et al.
Chemical profiling and quantitation of bioactive compounds in Platycladi Cacumen by UPLC-Q-TOF-MS/MS and UPLC-DAD
J. Pharm. Biomed. Anal.
(2018) - et al.
The phytochemical composition, metabolites, bioavailability and in vivo antioxidant activity of Tetrastigma hemsleyanum leaves in rats
J. Funct. Foods
(2017) - et al.
Phenolic profiles and antioxidant activities of free, esterified and bound phenolic compounds in walnut kernel
Food Chem.
(2021) - et al.
Design, synthesis and biological activity of thiazolidine-4-carboxylic acid derivatives as novel influenza neuraminidase inhibitors
Bioorg. Med. Chem. Lett.
(2011) - et al.
Monitoring batch-to-batch reproducibility using direct analysis in real time mass spectrometry and multivariate analysis: a case study on precipitation
J Pharmaceut Biomed
(2013) - et al.
Structural characterization and discrimination of Ophiopogon japonicas (Liliaceae) from different geographical origins based on metabolite profiling analysis
J. Pharm. Biomed.
(2020) - et al.
Molecular docking analysis of rutin reveals possible inhibition of SARS-CoV-2 vital proteins
Afr. J. Tradit. Complement. Altern. Med.
(2021)
Inhibition of influenza virus replication by plant-derived isoquercetin
Antiviral Res.
Pharmacology, and quality control of Tetrastigma hemsleyanum Diels & Gilg in China: a review
Front. Pharmacol.
Analysis on chemical constituents in Tetrastigma hemsleyanum by UPLC-Triple - TOF/MS
Chin. Trad. Herb. Drug.
New C-glycosylflavones from Tetrastigma hemsleyanum (Vitaceae)
Acta Bot. Sin.
A novel method for geographical origin identification of Tetrastigma hemsleyanum (Sanyeqing) by near-infrared spectroscopy
Anal Methods-Uk
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These authors contributed equally to this work.