Identification of Chemical Markers for the Discrimination of Radix Angelica sinensis Grown in Geoherb and Non-Geoherb Regions Using UHPLC-QTOF-MS/MS Based Metabolomics.

This research aimed to discover chemical markers for discriminating radix Angelica sinensis (RAS) from different regions and to explore the differences of RAS in the content of four active compounds and anti-inflammatory activities on lipopolysacchride (LPS)-induced RAW264.7 cells and calcium antagonists on the HEK 293T cells of RAS. Nine compounds were selected as characteristic chemical markers by ultra-high-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UHPLC-QTOF-MS/MS), based on metabolomics, in order to rapidly discriminate RAS from geoherb and non-geoherb regions. The contents of senkyunolide I and butylidenephthalide in geoherb samples were higher than those in non-geoherb samples, but the contents of ferulic acid and levistolide A were lower in the geoherb samples. Furthermore, the geoherbs showed better nitric oxide (NO) inhibitory and calcium antagonistic activities than the non-geoherbs. These results demonstrate the diversity in quality of RAS between geoherbs and non-geoherbs.

Angelica sinensis in China-A review of botanical profile, ethnopharmacology, phytochemistry and chemical analysis.

ETHNOPHARMACOLOGICAL RELEVANCE: Angelica sinensis (Oliv.) Diels, known as Dang Gui (in Chinese), is a traditional medicinal and edible plant that has long been used for tonifying, replenishing, and invigorating blood as well as relieving pain, lubricating the intestines, and treating female irregular menstruation and amenorrhea. A. sinensis has also been used as a health product and become increasingly popular in China, Japan, and Korea. AIM OF THE REVIEW: This paper aims to provide a systemic review of traditional uses of A. sinensis and its recent advances in the fields of phytochemistry, analytical methods and toxicology. In addition, possible trends, therapeutic potentials, and perspectives for future research of this plant are also briefly discussed. MATERIALS AND METHODS: An extensive review of the literature was conducted, and electronic databases including China National Knowledge Infrastructure, PubMed, Google Scholar, Science Direct, and Reaxys were used to assemble the data. Ethnopharmacological literature and digitalised sources of academic libraries were also systematically searched. In addition, information was obtained from local books and The Plant List (TPL, www.theplantlist.org). RESULT: This study reviews the progress in chemical analysis of A. sinensis and its preparations. Previously and newly established methods, including spectroscopy, thin-layer chromatography (TLC), gas chromatography (GC), high-performance liquid chromatography (HPLC), ultra-performance liquid chromatography(UPLC), and nuclear magnetic resonance analysis (NMR), are summarized. Moreover, identified bioactive components such as polysaccharides, ligustilide and ferulic acid were reviewed, along with analytical methods for quantitative and qualitative determination of target analytes, and fingerprinting authentication, quality evaluation of A. sinensis, and toxicology and pharmacodynamic studies. Scientific reports on crude extracts and pure compounds and formulations revealed a wide range of pharmacological activities, including anti-inflammatory activity, antifibrotic action, antispasmodic activity, antioxidant activities, and neuroprotective action, as well as cardio- and cerebrovascular effects. CONCLUSIONS: Within the published scientific literature are numerous reports regarding analytical methods that use various chromatographic and spectrophotometric technologies to monitor various types of components with different physicochemical properties simultaneously. This review discusses the reasonable selection of marker compounds based on high concentrations, analytical methods, and commercial availabilities with the goal of developing quick, accurate, and applicable analytical approaches for quality evaluation and establishing harmonised criteria for the analysis of A. sinensis and its finished products. Compounds isolated from A. sinensis are abundant sources of chemical diversity, from which we can discover active molecules. Thus, more studies on the pharmacological mechanisms of the predominant active compounds of A. sinensis are needed. In addition, given that A. sinensis is one of the most popular traditional herbal medicines, its main therapeutic aspects, toxicity, and adverse effects warrant further investigation in the future.

Distinguishing Radix Angelica sinensis from different regions by HS-SFME/GC-MS.

An automated headspace solvent free microextraction (HS-SFME) based gas chromatography/mass spectrometry (GC/MS) was developed for discrimination of Radix Angelica sinensis (RAS) from different cultivation regions. The MS data were subjected to principal component analysis (PCA) and hierarchical clustering analysis (HCA) to rapidly find the potential characteristic components of RAS from top-geoherb region and non top-geoherb region. Totally, fifty-one volatile organic compounds (VOCs) were identified, in which ß-ocimene, α-pinene, 3-methylbutanal, heptanes, butanal were identified as potential markers for distinguishing RAS from top-geoherb region and non top-geoherb region. Sulphur dioxide was detected in some commercial RAS samples, which implied that sulphur-fumigation might be the main reason for the quality inconsistencies of commercial RAS samples. These results suggested that RAS from top-geoherb region and non-top geoherb region could be discriminated by the method. And characteristic chemical markers found in current study can be used for ensuring consistent quality of top-geoherb of RAS.

Simultaneous determination of ferulic acid and phthalides of Angelica sinensis based on UPLC-Q-TOF/MS.

The radix of Angelica sinensis (AS) is one of the most commonly used as a herbal medicine. To investigate the geoherbalism and quality evaluation of AS, an ultra performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (UPLC-ESI-Q-TOF/MS) method was established to analyze and identify ferulic acid and phthalides in AS. The results showed that among samples collected in four regions, the relative contents of ferulic acid and phthalides were highest in samples collected in Gansu, and the samples from the four different regions were apparently classified into four groups. Meanwhile, the relative content in non-fumigated root was higher than after sulfur-fumigation and the sulfur-fumigated and non-fumigated samples were obviously divided into two groups by PCA. The paper establishes a systematic and objective evaluation system to provide a scientific basis for evaluating the quality of AS.

[AFLP analysis on genetic diversity of Angelica sinensis].

OBJECTIVE: To explore the genetic diversity and relationship of different germplasm of Angelica sinensis. METHODS: Amplified fragment length polymorphism (AFLP) markers were developed to analyze genetic polymorphism in 12 populations 117 samples of Angelica sinensis. The amplified fragments were used for cluster analysis among the different germplasm of Angelica sinensis and to construct the genetic phylogenetic tree with NTSYSpc 2. 11 software. RESULTS: Eight primer combinations selected from 64 primer combinations were used for amplification and a total of 815 fragments were obtained. Analysis identified 812 polymorphic fragments, accounting for 99.63% of the total detected fragments. Different phenotypes and germplasm of Angelica sinensis could be divided by genetic phylogenetic tree analysis. CONCLUSION: AFLP molecular markers can indicate the significant polymorphism and genetic diversity among germplasm resources of Angelica sinensis. The cultivated purple-stemmed and green-stemmed phenotypes of Angelica sinensis may have different genotypes. The results can provide theoretical evidence for reasonable utilization and breeding new cultivar in molecular level.

[Quick identification of sun-dried and sulfur-fumigated angelicae sinensis radix by Fourier transform infrared spectroscopy].

OBJECTIVE: To develop a quick identification method for the sun-dried and sulfur-fumigated Angelicae Sinensis Radix used by Fourier transform infrared spectroscopy (FTIR) combined with second derivative infrared spectroscopy. METHOD: The alcoholic and aqueous extracts of sun-dried and sulfur-fumigated Angelicae Sinensis Radix were analyzed by using FTIR, the further analysis was used by second derivative infrared spectroscopy. RESULT: There existed differences between their infrared spectra either extracted by ethanol or water, while the distinctions were more obvious after analyzing their alcoholic and aqueous extracts through high resolution of second derivative infrared spectroscopy. Infrared spectra showed that the absorption peaks of Angelicae Sinensis Radix were significantly reduced and a new absorption peak appeared after sulfur-fumigated process in alcoholic extracts, while both of them changed markedly in the "fingerprint region" ranging from 1 000 to 400 cm(-1) in aqueous extracts. Second derivative spectra showed that the absorption peaks of sulfur-fumigated Angelicae Sinensis Radix extracted by ethanol weakened and disappeared at about 3 578 cm(-1) and 3 541 cm(-1), while both of them differed significantly from each other ranging from 1 400 to 1 200 cm(-1) as well as 800 cm(-1) to 600 cm(-1), difference also existed between them extracted by water ranging from about 3 900 to 3 850 cm(-1) and 3 800 to 3 750 cm(-1). CONCLUSION: The FTIS method combined with second derivative can be utilized to distinguish sun-dried and sulfur-fumigated Angelicae Sinensis Radix efficiently, conveniently and accurately, and provide a basis for identification and quality control of Angelicae Sinensis Radix.

[Studies on comparison and classification of Angelica sinensis from different areas in Gansu province].

OBJECTIVE: To compare and classify the samples of Angelica sinensis from 36 different areas in Gansu province. METHOD: The HPLC was used to detect samples, and the computer aided similarity evaluation was used to analyze the fingerprints. Similarity combined with principal component analysis (PCA) and multidimensional pattern recognition of Euclidean distance were used to compare and classify the samples of A. sinensis in different areas. RESULT: It was found that the quality of A. sinensis is closely related to the growth environment. CONCLUSION: This method could be used to classify the samples of A. sinensis from a variety of sources.

[Molecular characteristic marker method of 27 breeds of common Umbelliferae Chinese herb medicinal plants by sequencing rDNA].

OBJECTIVE: To probe a molecular marker method of accrediting fingerprinting of 27 kinds of common Umbelliferae Chinese herb medicinal plants by sequencing rDNA. METHOD: The rDNA sequences of the 27 breeds of common Umbelliferae Chinese herb medicinal plants were amplified, and were digested by restriction endonuclease, and were seperated via polypropylene electrophoresis, at last 6 breeds of them were sequenced. RESULTS: The rDNA sequence fragment we gained concluded ITS1, ITS2, 5.8S complete sequence and 18S, 26S part sequence. On the electrophoresis map of PCR products digested by restriction endonuclease MSP I, 27 breeds appeared 16 kinds of characteristic map, 11 of them differ from others; and PCR products digested by restriction endonuclease HaeIII, there appeared 5 kinds of characteristic map among 27 breeds, 3 of them differ from others. The sequenced result of 6 breeds showed genes whose length extented from 652bp to 656bp were acquired. These sequences of 3 breeds which showed the same electrophoresis map after digested by restriction endonuclease HaeIII exhibited great similarity according to similar phylogenetic tree constructed on rDNA sequence. CONCLUSION: The rDNA sequence character is effective molecular marker for classifying the different Umbellerae Chinese herb medicinal plants. And the method of sequencing rDNA surpassed that of restriction fragment long polymorphism (RFLP).

Molecular genetic and chemical assessment of radix Angelica (Danggui) in China.

The roots of Angelica sinensis (Danggui), a traditional Chinese medicine, have been used for invigorating blood circulation for over 2000 years in China. Three common species of Angelica roots are found in Asia: A. sinensis from China, A. acutiloba from Japan, and A. gigas from Korea. By using a molecular genetic approach, the 5S-rRNA spacer domains of the three species of Angelica were amplified, and their nucleotide sequences were determined. Diversity in DNA sequences among various species was found in their 5S-rRNA spacer domains, which could serve as markers for authentic identification of Angelica roots. In chemical analyses, the main constituents of Angelica roots including ferulic acid and Z-ligustilide were determined by HPLC; roots of A. sinensis were clearly distinct in that they contained approximately 10-fold higher levels of ferulic acid and Z-ligustilide as compared to roots of A. acutiloba and A. gigas. In addition, the amounts of main constituents in roots of A. sinensis varied according to different regions of cultivation and different methods of preservation. The chemical profile determined by HPLC therefore could serve as a fingerprint for quality control of Angelica roots.