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Discrimination of Radix Polygoni Multiflori from Different Geographical

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Discrimination of Radix Polygoni Multiflori from Different Geographical Tang et al. Chin Med (2017) 12:34 https://doi.org/10.1186/s13020-017-0155-8 Chinese Medicine RESEARCH Open Access Discrimination of Radix Polygoni Multifori from diferent geographical areas by UPLC‑QTOF/MS combined with chemometrics Jin‑Fa Tang1†, Wei‑Xia Li1†, Fan Zhang1, Yu‑Hui Li1, Ying‑Jie Cao1, Ya Zhao1, Xue‑Lin Li1* and Zhi‑Jie Ma2* Abstract Background: Nowadays, Radix Polygoni Multifori (RPM, Heshouwu in Chinese) from diferent geographical origins were used in clinic. In order to characterize the chemical profles of diferent geographical origins of RPM samples, ultra-high performance liquid chromatography quadrupole time of fight mass spectrometry (UPLC-QTOF/MS) combined with chemometrics (partial least squared discriminant analysis, PLS‑DA) method was applied in the present study. Methods: The chromatography, chemical composition and MS information of RPM samples from 18 geographical origins were acquired and profled by UPLC-QTOF/MS. The chemical markers contributing the diferentiation of RPM samples were observed and characterized by supervised PLS‑DA method of chemometrics. Results: The chemical composition diferences of RPM samples derived from 18 diferent geographical origins were observed. Nine chemical markers were tentatively identifed which could be used as specifc chemical markers for the diferentiation of geographical RPM samples. Conclusions: UPLC-QTOF/MS method coupled with chemometrics analysis has potential to be used for discrimi‑ nating diferent geographical TCMs. Results will help to develop strategies for conservation and utilization of RPM samples. Keywords: UPLC-QTOF/MS, Radix Polygoni Multifori, Diferent geographical origins, Chemical markers, Partial least squared discriminant analysis Background calming the nerves, nourishing blood, activating chan- Radix Polygoni Multifori (RPM, Heshouwu in Chinese) nels and collaterals, tonifying liver and kidneys, and pre- is the dried root tuber of Polygonum multiforum Tunb. venting the premature graying of hair. Many 1000 years (Fam. Polygonaceae). As one of the most popular and of clinical practice of TCM has demonstrated the efect precious traditional Chinese medicines (TCMs), it is of RPM in terms of preventing dementia and improving ofcially documented in the Chinese Pharmacopoeia for memory [1]. As a traditional medicine and dietary sup- plement for health, it has also been considered efective in antiaging and increasing longevity [2, 3]. According to *Correspondence: [email protected]; [email protected]; modern researches, RPM has the pharmacological efects [email protected] of enhancing immunity, anti-atherosclerosis, anti-infam- †Jin-Fa Tang and Wei-Xia Li contributed equally to this work 1 The First Afliated Hospital of Henan University of Chinese Medicine, No. matory, antibacterial, anti-cancer, anti-mutagenic, anti- 19, Renmin Road, Jinshui District, Zhengzhou 450000, People’s Republic oxidation, increasing DNA repair, and improving adipose of China metabolism [4–6]. 2 Beijing Friendship Hospital Afliated to Capital Medical University, No. 95, Yongan Road, Xuanwu District, Beijing 100050, China © The Author(s) 2017. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Tang et al. Chin Med (2017) 12:34 Page 2 of 12 With the extensive application of RPM, its safety has science. Chemometrics combined with liquid chromatog- drawn widespread attention. More and more literatures raphy and other spectrometric methods are widely used showed that RPM and RPM-containing herbal products in many felds concerning TCMs, such as the comparison had the adverse efects of hepatotoxicity [7]. Te RPM of diferent species [18], quality control and moderniza- dose in the Chinese Pharmacopoeia (2005 edition) is tion of TCM [19]. Herein, eighteen RPM samples from 6–12 g [8]. According to the safety considerations, the 10 counties of 4 provinces were analyzed using ultra-high recommended dose of RPM was adjusted to 3–6 g in the performance liquid chromatography quadrupole time of 2010 edition of the Chinese Pharmacopoeia [9]. In addi- fight mass spectrometry (UPLC-QTOF/MS). And partial tion, the safety, quality and efcacy of RPM samples may least squared discriminant analysis (PLS-DA) of chemo- vary greatly because of the diferent geographical origins. metrics approach was applied to classify diferent RPM As well known, medicinal herbs in authentic produc- samples and fnd chemical variables that contribute to ing areas had the best quality, which can produce the the diferentiation of RPMs. Furthermore, the Progenesis best pharmacological efect. Te place where authentic QI software (v2.0, Waters Corporation, Milford, USA) medicinal herbs produced is called the “trueborn area”. with fast, objective, and reliable characteristics was used RPM is widely distributed in China’s southwest, central, for the chemometrics statistical analysis, which had been south, east and other regions, including Sichuan, Yun- already used to found the chemical diferences among the nan, Guizhou, Chongqing, Guangdong, Guangxi, Jiangsu, diferent extracts of RPM and RPM Praeparata [20], but Anhui, Hubei, Hunan, Henan, Jiangxi, Shanxi, Gansu and it only analyzed the diferences between the water and other provinces and cities [10]. Owing to its many ori- ethanol extracts of RPM and RPM Praeparata. Terefore, gins, the “trueborn area” of RPM is still being studied. it would also be feasible to use this method to fnd difer- It is consensus that the efect of Chinese medicine relies ent chemical markers among RPMs from diferent geo- on the role of its multi-component. Tere are large dif- graphical origins. Te results can provide more efective ference among the chemical composition and content of strategy guidance for the utilization and domestication of RPMs because of its diferent species and origins, which RPM. will cause a greater impact on its efcacy. Terefore, the distinction among RPM samples from diferent origins is Methods essential for determining the trueborn area of RPM and Te Minimum Standards of Reporting Checklist (Addi- for selecting good quality RPM to treat diseases. tional fle 1) contains details of the experimental design, Te introduction of new analytical techniques and the and statistics, and resources used in this study. application of novel data analysis methods have greatly promoted the quality assurance of TCM. From the lit- Chemicals and reagents eratures summary, we found that fngerprinting qual- Acetonitrile (HPLC grade) and formic acid were pur- ity control of RPM from diferent geographical origins chased from Merck KGaA (Darmstadt, Germany); ultra- was determined by thin-layer chromatography (TLC) pure water was purifed by a Milli-Q system (Milford, scanning and high-performance liquid chromatography MA, USA). (HPLC) [11, 12]; the quality of various commercial speci- fcations of RPM and its dregs was evaluated by HPLC Plant materials [13]; the quality control of RPM from diferent origins Seventeen species of planted or wild RPM samples were was determined by infrared spectrum (IR), inductively collected from 10 counties, 4 provinces of China; and one coupled plasma-atomic emission spectrometer (ICP- kind of RPM sample (S13) was purchased from pharma- AES) and LC–mass spectrometry (MS), etc. [14–16]. cies (Table 1). All the herbal samples were authenticated However, the authentic correlation between geographical by the authors. Te corresponding voucher specimens distribution regions and chemical variation in RPMs has were stored in the laboratory for drug metabolism and been rarely reported. pharmacokinetics (DMPK) Research of Herbal Medi- It was reported that RPM mainly contains anthraqui- cines, the First Afliated Hospital of Henan University of nones, stilbene glycosides, phospholipids, phenols, fa- Chinese Medicine. vonoids, etc. [17]. Te present study is aimed to classify and characterize RPM samples from diferent geographi- Sample preparation cal origins based on the chemical compounds by chemo- Te 18 RPM samples were sliced, dried, and powdered. metrics. Chemometrics is an interdisciplinary science Te powdered samples were screened trough no. 4 sieve, involving mathematics and statistics, chemistry and com- respectively; and 0.25 g was extracted with 25 mL 70% puter science. In recent years, chemometrics have gained ethanol for 30 min by refux extraction method, cooled more attention along with the development of computer at room temperature and weighted. Te reduced weight Tang et al. Chin Med (2017) 12:34 Page 3 of 12 Table 1 Geographical information of 18 RPM samples No. Source Longitude Latitude S1 Cultivated in Xinzhou town, Huangping county, Guizhou province 107.92213 26.98450 S2 Wild, Xinzhou town, Huangping county, Guizhou province 107.92213 26.99918 S3 Wangsi town, Duyun county, Guizhou province 107.51576 26.30596 S4 Huaxi district, Guiyang city, Guizhou province 107.27755 26.73151 S5 Cultivated in Meitan county, Zunyi city, Guizhou province for 2 years 107.45728 28.02350 S6 Cultivated in Meitan county, Zunyi city, Guizhou province for 3
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