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Differentiation of Actaea species by NMR metabolomics analysis
Abstract
The ¹H NMR spectra of crude extracts from a total of 33 Actaea samples were acquired and analyzed for their species- and plant part-specific metabolomic characteristics by identifying fingerprint resonances via visual observation as well as a chemometric approach using principal component analysis (PCA). The main study subjects were the roots/rhizomes and aerial parts of three American species, Actaea racemosa (AR), Actaea podocarpa (AP) and Actaea cordifolia (AC). AP exhibited an already visually distinct chemical profile from those of the other two species. The species-characteristic resonances were identified as analytical chemotaxonomic markers. AR and AC exhibited visually similar ¹H NMR spectral profiles that required statistical analysis for differentiation. Several characteristic peaks and peak patterns were identified for each group of samples. Together with the three American Actaea species, the characteristics of the ¹H NMR spectra of Asian species are also discussed. A statistical analysis method using PCA was employed to provide the metabolomic profile for visually minor but analytically significant chemotaxonomic differences. PCA scores allowed differentiation between the three American Actaea species, as well as the ability to differentiate between the various plant parts (aboveground vs. roots/rhizomes).
... As examples, NMR combined with chemometrics was applied to investigate the roots/rhizomes and aerial parts of three Actaea species, A. racemosa (AR), A. podocarpa (AP), and A. cordifolia (AC) [195]. Roots and rhizomes of AR, known as black cohosh, are traditionally used for indications of premenstrual syndrome (PMS), menstrual pain and cramping. ...
... racemosa, A. podocarpa and A. cordifolia) 1 H NMR fingerprinting in combination with chemometric analysis 1 H NMR in combination with PCA was applied for differentiation of three species. [195] Four Crataegus species (C. laevigata, C. douglasii, C. okanaganensis, and C. monogyna) 1 H NMR fingerprinting in combination with chemometric analysis Differentiation of Crataegus spp. was best achieved by focusing on the NMR spectral region containing signals unique to phenolic compounds. ...
Botanicals have played an important role in maintaining human health and well-being throughout history. During the past few decades in particular, the use of botanical health products has gained more popularity. Whereas, quality, safety and efficacy concerns have continuously been critical issues due to the intrinsic chemical complexity of botanicals. Chemical analytical technologies play an imperative role in addressing these issues. Nuclear magnetic resonance (NMR) spectroscopy has proven to be a powerful and useful tool for the investigation of botanical health products. In this review, NMR techniques and methodologies that have been successfully applied to the research and development of botanical health products in all the stages, from plants to products, are discussed and summarized. Furthermore, applications of NMR together with other analytical techniques in a variety of domains of botanical health products investigation, such as plant species differentiation, adulteration detection, and bio-activity evaluation, are discussed and illustrated with typical examples. This article provides an overview of the potential uses of NMR techniques and methodologies in an attempt to further promote their recognition and utilization in the field of botanical health products analysis and quality control.
... For the phytochemical characterization of the complex matrices, tests were carried out to determine total phenolics (using the spectrophotometric quantification method in microplates with the Folin-Ciocalteau reagent), total flavonoids (using the spectrophotometric method in microplates with aluminum chloride -AlCl 3 ), and spectroscopic analysis through Hydrogen Nuclear Magnetic Resonance ( 1 H NMR). Identification of characteristic phenolic peaks was confirmed by comparison with literature data (Dos Santos et al., 2017;Imai et al., 2020). ...
... liquid-liquid extraction, solid phase extraction, or fractionation). Thus, plant NMR metabolomics for detection of secondary metabolite are more common (Sumner, Mendes et al. 2003, Schauer and Fernie 2006, Zulak, Weljie et al. 2008, Kim, Choi et al. 2010, Schripsema 2010, Kroymann 2011, Leiss, Choi et al. 2011, Simmler, Napolitano et al. 2014, Deborde, Moing et al. 2017, Imai, Lankin et al. 2020, Nephali, Piater et al. 2020). An interesting approach to grow large scale amounts of the nematode Caenorhabditis elegans to collect analytical and phenotypic data was taken by using cake pans (Au -Shaver, Au - Gouveia et al. 2021). ...
Confidently, nuclear magnetic resonance (NMR) is the most informative technique in analytical chemistry and its use as an analytical platform in metabolomics is well proven. This chapter aims to present NMR as a viable tool for microbial metabolomics discussing its fundamental aspects and applications in metabolomics using some chosen examples.
... VS methods are rapid and the generated fingerprint spectra can be treated using chemometrics to yield more interpretable results for answering different biological questions (Wang and Yu, 2015;Rohaeti et al., 2021). NMR spectroscopy has also emerged as a powerful analytical technique for the QC of HMs because it can be used for simultaneous and rapid analysis of primary or secondary metabolites with good sensitivity Imai et al., 2020). Chemical fingerprints can also be obtained from MS spectra which readily provide information on the presence of certain metabolites or elements within the HM samples based on their masses (Yang and Deng, 2016). ...
... Thus, two-dimensional (2D) fingerprinting has been introduced to better address the chemical complexity of the herbal preparations [5,6]. Moreover, hyphenated chromatographic and spectrometric approaches, such as HPLC-DAD [7][8][9][10][11][12][13][14], GC-MS [15,16], and HPLC-MS [17,18], can greatly enhance the potential of since in quality control of botanical products, different plant parts can be considered as adulterants of the main material [24,25]. ...
Plant samples are potential sources of physiologically active secondary metabolites and their classification is an extremely important task in traditional medicine and other fields of research. In the production of herbal drugs, different plant parts of the same or related species can serve as adulterants for primary plant material. The use of highly informative and relatively easily accessible tools, such as liquid chromatography and low-resolution mass spectrometry, helps to solve these tasks by means of fingerprint analysis. In this study, to reveal specific plant part features for 20 species from one family (Apiaceae), and to preserve the maximum information content, two approaches are suggested. In both cases, minimal raw data pretreatment, including rescaling of time and m/z axes and cutting off some uninformative regions, was applied. For the support vector machine (SVM) method, tensor unfolding was required, while neural networks (NNs) were able to work directly with squared heatmaps as input data. Moreover, five data augmentation variants are proposed, to overcome the typical problem of a lack of data. As a result, a comparable F1-score close to 0.75 was achieved by SVM and two employed NN architectures. Eight marker compounds belonging to chlorophylls, lipids, and coumarin apio-glucosides were tentatively identified as characteristic of their corresponding sample groups: roots, stems, leaves, and fruits. The proposed approaches are simple, information-saving and can be applied to a broad type of tasks in metabolomics.
... VS methods are rapid and the generated fingerprint spectra can be treated using chemometrics to yield more interpretable results for answering different biological questions (Wang and Yu, 2015;Rohaeti et al., 2021). NMR spectroscopy has also emerged as a powerful analytical technique for the QC of HMs because it can be used for simultaneous and rapid analysis of primary or secondary metabolites with good sensitivity (Kim et al., 2010;Imai et al., 2020). Chemical fingerprints can also be obtained from MS spectra which readily provide information on the presence of certain metabolites or elements within the HM samples based on their masses (Yang and Deng, 2016). ...
Herbal drugs or herbal medicines (HMs) have a long-standing history as natural remedies for preventing and curing diseases. HMs have garnered greater interest during the past decades due to their broad, synergistic actions on the physiological systems and relatively lower incidence of adverse events, compared to synthetic drugs. However, assuring reproducible quality, efficacy, and safety from herbal drugs remains a challenging task. HMs typically consist of many constituents whose presence and quantity may vary among different sources of materials. Fingerprint analysis has emerged as a very useful technique to assess the quality of herbal drug materials and formulations for establishing standardized herbal products. Rather than using a single or two marker(s), fingerprinting techniques take great consideration of the complexity of herbal drugs by evaluating the whole chemical profile and extracting a common pattern to be set as a criterion for assessing the individual material or formulation. In this review, we described and assessed various fingerprinting techniques reported to date, which are applicable to the standardization and quality control of HMs. We also evaluated the application of multivariate data analysis or chemometrics in assisting the analysis of the complex datasets from the determination of HMs. To ensure that these methods yield reliable results, we reviewed the validation status of the methods and provided perspectives on those. Finally, we concluded by highlighting major accomplishments and presenting a gap analysis between the existing techniques and what is needed to continue moving forward.
Highlights
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The handheld NIRS can distinguish spectral features of agarwood oil and its adulterants.
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In 2nd-order derivatized NIR spectra, authentic A. crassna oil shows dominant typical peaks.
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Authentic sample forms a cluster in the PCA of 2nd-order derivatized NIR spectral data.
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The DD-SIMCA training sets of authentic samples are 100 % sensitive with no type-1 error.
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Agarwood oil authenticity can be verified on the spot with handheld NIRS and DD-SIMCA.
Botanical natural medicinal products and dietary supplements are utilized globally for their positive impacts on health and wellness. However, the effectiveness and safety of botanical products can be compromised by unintentional or intentional adulteration. The presence of adulterated botanical ingredients in the global market has been documented in the published literature but a key question, namely what the extent of adulteration is, remains to be answered. This review aims to estimate the prevalence of adulteration in preparations made from black cohosh rhizome, echinacea root or herb, elder berry, ginkgo leaf, and turmeric root/rhizome. According to the information provided in the 78 publications retrieved for this paper, 818 of 2995 samples were reported to be adulterated and/or mislabeled. Ginkgo leaf samples (n = 533) had the highest adulteration rate with 56.7%, followed by black cohosh rhizome (n = 322) samples with 42.2%, echinacea root/herb (n = 200) with 28.5%, elder berry (n = 695) with 17.1%, and turmeric root/rhizome (n = 1247) with 16.5%. Products sold as licensed or registered herbal medicines were found to have a lower risk of adulteration compared to products sold as dietary/food supplements. The data show that the adulteration rate substantially differs from one ingredient to the other. Due to the significant limitations of the available data upon which the estimated extent of adulteration is based, and the rapidly changing botanical dietary supplement market, conclusions from the five herbs examined in this publication cannot be applied to other botanicals traded in the global market. However, the data clearly show that a substantial portion of the botanical dietary supplements do not contain what is claimed on their labels.
Compton, J. A., Culham, A. & Jury, S. L.: Reclassification of Actaea to include Cimicifuga and Souliea (Ranunculaceae): phytogeny inferred from morphology, nrDNA ITS, and cpDNA trn L‐F sequence variation. – Taxon 47: 593‐634. 1998. – ISSN 0040‐0262
Phylogenetic analyses using parsimony were performed on three independent data sets to test generic relationships between Actaea, Cimicifuga, and Souliea. Analyses of morphology and nuclear ribosomal DNA ITS were performed on 23 species of Cimicifuga, 4 species of Actaea, and the single species of Souliea. Analysis of chloroplast DNA trn L‐F was applied to the same species, less two of Cimicifuga. The outgroup taxa Eranthis and Anemonopsis both resolved outside the ingroup in all parsimony analyses, whereas Souliea resolved within it. Jukes‐Cantor pairwise sequence distances confirm Eranthis and Anemonopsis to be most distant. Souliea distances are comparable with those of taxa within the Actaea‐Cimicifuga assemblage. A strongly supported monophyletic clade including all studied species of Actaea, Cimicifuga, and Souliea was found in all analyses. Evidence presented here allows a broader concept of Actaea to be adopted, reverting to the circumscription of Linnaeus in 1753. Seven sections, based on clades found in the total analysis, could be defined by morphological characters: A. sect. Actaea, sect. Podocarpae, sect. Cimicifuga, sect. Dichanthera, sect. Oligocarpae, sect. Pityrosperma, and sect. Souliea. One species, A. taiwanensis, is newly described and 23 new combinations are made in the ranks of section, species, and variety. Keys are provided to identify taxa at all ranks within the revised circumscription of Actaea. Maps showing the distributions of all seven sections and their constituent species are presented. Phytogeographic patterns suggest a Tertiary origin for the newly redefined genus, with species surviving in refugia during the glacial periods of the Pleistocene.
Metabolic fingerprinting techniques have received a lot of attention in recent years and the annual amount of publications in this field has increased significantly over the past decade. This increase in publications is due to improvements in the analytical performance, most notably in the field of NMR and MS analysis, and the increased awareness of the different applications of this growing field. Metabolomic fingerprinting or profiling is continuously being applied to new areas of research such as drug discovery from natural resources, quality control of herbal material, and discovering lead compounds. In this review the current state of the art of metabolic fingerprinting, focussing on NMR and MS technologies will be discussed. The application of these two analytical tools in the quality control of herbal material and phytopharmaceuticals forms the major part of this review. Finally we will look at the future developments and perspectives of these two technologies in the quality control of herbal material.
A comprehensive metabolomic profiling of Catharanthus roseus L. G. Don infected by 10 types of phytoplasmas was carried out using one-dimensional and two-dimensional NMR spectroscopy followed by principal component analysis (PCA), an unsupervised clustering method requiring no knowledge of the data set and used to reduce the dimensionality of multivariate data while preserving most of the variance within it. With a combination of these techniques, we were able to identify those metabolites that were present in different levels in phytoplasma-infected C. roseus leaves than in healthy ones. The infection by phytoplasma in C. roseus leaves causes an increase of metabolites related to the biosynthetic pathways of phenylpropanoids or terpenoid indole alkaloids: chlorogenic acid, loganic acid, secologanin, and vindoline. Furthermore, higher abundance of Glc, Glu, polyphenols, succinic acid, and Suc were detected in the phytoplasma-infected leaves. The PCA of the (1)H-NMR signals of healthy and phytoplasma-infected C. roseus leaves shows that these metabolites are major discriminating factors to characterize the phytoplasma-infected C. roseus leaves from healthy ones. Based on the NMR and PCA analysis, it might be suggested that the biosynthetic pathway of terpenoid indole alkaloids, together with that of phenylpropanoids, is stimulated by the infection of phytoplasma.
The metabolomic analysis of 11 Ilex species, I. argentina, I. brasiliensis, I. brevicuspis, I. dumosavar. dumosa, I. dumosa var. guaranina, I. integerrima, I. microdonta, I. paraguariensis var. paraguariensis, I. pseudobuxus, I. taubertiana, and I. theezans, was carried out by NMR spectroscopy and multivariate data analysis. The analysis using principal component analysis and classification of the (1)H NMR spectra showed a clear discrimination of those samples based on the metabolites present in the organic and aqueous fractions. The major metabolites that contribute to the discrimination are arbutin, caffeine, phenylpropanoids, and theobromine. Among those metabolites, arbutin, which has not been reported yet as a constituent of Ilex species, was found to be a biomarker for I. argentina,I. brasiliensis, I. brevicuspis, I. integerrima, I. microdonta, I. pseudobuxus, I. taubertiana, and I. theezans. This reliable method based on the determination of a large number of metabolites makes the chemotaxonomical analysis of Ilex species possible.
The cycloartane triterpene content in the roots/rhizomes (RR) and aerial parts (PX) of Actaea racemosa (AR), A. podocarpa (AP), and A. cordifolia (AC) have been investigated by quantitative 1H NMR (qHNMR). Thereby, it was demonstrated that qHNMR represents a powerful methodology for the analysis of crude plant extracts as it does not rely on the rarely available identical reference triterpenes. Specifically, the presence of the characteristic C-19 cyclopropane (exo/endo) hydrogen signals made it possible to quantify the less common/not ubiquitously present group of cycloartane triterpenes, directly in extracts. As an example, ARPX and ARRR were shown to contain, 3.8-20.8% ± 8.2% and 7.2-19.3% ± 4.0% of cycloartane triterpenes, respectively. The cycloartane concentration in ACPX and ACRR was 7.5-8.7% ± 0.8% and 13.9-28.5% ± 7.3%, respectively, based on the weight of the extract. AP was shown to contain notably lower amounts of the cycloartane triterpenes as compared to AR and AC in the roots/rhizomes. The content for APPX and APRR was only 2.1-3.3% ± 0.7% and 1.1-4.0% ± 1.5%, respectively. In addition, an example is presented for the identification of specific cycloartanes as marker compounds for AR within crude extracts based on the same qHNMR spectra and 2D NMR methods.
Investigating the phytochemical equivalence of the aerial parts of Actaea racemosa (syn. Cimicifuga racemosa) relative to the widely used roots/rhizomes, this study provides a perspective for the potential use of renewable ("green") plant parts as a source of black cohosh botanical preparations. In addition to the characterization of Nω-methylserotonin as one representative marker of the Actaea alkaloids, nine cycloartane triterpenes were isolated and characterized, including the two new triterpene glycosides (1S,15R)-1,15,25-trihydroxy-3-O-β-d-xylopyranosyl-acta-(16S,23R,24R)-16,23;16,24-binoxoside (1) and 3-O-α-l-arabinopyranosyl-(1S,24R)-1,24,25-trihydroxy-15-oxo-acta-(16R,23R)-16,23-monoxoside (2). Their structures were elucidated by spectroscopic data interpretation. The relative configuration of 1 was deduced by (1)H iterative full-spin analysis (HiFSA), making it the first example of the complete analysis of the complex (1)H NMR spectrum of a triterpene glycoside. In addition to the new compounds 1 and 2, the aerial plant parts were shown to contain the previously known binoxosides 3, 4, 6, and 7, the monoxoside 8, and the binoxols 5 and 9. Overall, the metabolome of the aerial plant parts consists of a variety of Actaea triterpenes, similar to those found in roots/rhizomes, a tendency toward C-1 and C-7 hydroxylation of the cycloartanol skeleton, a greater abundance of aglycones, and the presence of comparable amounts of Nω-methylserotonin.
Actaea racemosa L. is used as a component of drugs or dietary supplements to alleviate the menopause symptoms. Its biological activity is associated with the presence of phenolic compounds. In our work, the analysis of isoflavones and phenolic acids - caffeic acid (CA), ferulic acid and isoferulic acid (iFA) - both free and bonded in two species of Actaea, was conducted using HPLC-PAD technique. Moreover, the antioxidant effect of extracts from different parts of the investigated plants was determined on the basis of DPPH assay. Significant variation of CA and iFA content was observed. The highest content of CA was found in A. racemosa, while Actaea cordifolia contained the highest amount of iFA. Isoflavones were not found in the investigated plants. The antioxidant activity assay showed the high free radical-scavenging ability of the extracts obtained from different parts of the plant.
Floral fragrances in four species of Cimicifuga (Ranunculaceae) were collected by sorption and analyzed by gas chromatography-mass spectrometry. One pollination morph in C. simplex was found to contain three benzenoid compounds absent in the two other morphs. Two of these substances — methyl anthranilate and isoeugenol — induce behavioural modification in pollinating butterflies. The function of a range of terpenes present in small amounts in all analyzed species is unknown, but they may be involved in herbivore deterrence.
Introduction:
Nuclear magnetic resonance (NMR) spectroscopy is increasingly employed in the quantitative analysis and quality control (QC) of natural products (NP) including botanical dietary supplements (BDS). The establishment of QC protocols based on quantitative (1) H NMR (qHNMR) requires method validation.
Objective:
Develop and validate a generic qHNMR method. Optimize acquisition and processing parameters, with specific attention to the requirements for the analysis of complex NP samples, including botanicals and purity assessment of NP isolates.
Methods:
In order to establish the validated qHNMR method, samples containing two highly pure reference materials were used. The influence of acquisition and processing parameters on the method validation was examined, and general aspects of method validation of qHNMR methods discussed. Subsequently, the method established was applied to the analysis of two NP samples: a purified reference compound and a crude mixture.
Results:
The accuracy and precision of qHNMR using internal or external calibration were compared, using a validated method suitable for complex samples. The impact of post-acquisition processing on method validation was examined using three software packages: TopSpin, Mnova and NUTS. The dynamic range of the qHNMR method developed was 5000:1 with a limit of detection (LOD) of better than 10 µm. The limit of quantification (LOQ) depends on the desired level of accuracy and experiment time spent.
Conclusion:
This study revealed that acquisition parameters, processing parameters and processing software all contribute to qHNMR method validation. A validated method with a high dynamic range and general workflow for qHNMR analysis of NP is proposed.
Covering the literature from mid-2004 until the end of 2011, this review continues a previous literature overview on quantitative (1)H NMR (qHNMR) methodology and its applications in the analysis of natural products. Among the foremost advantages of qHNMR is its accurate function with external calibration, the lack of any requirement for identical reference materials, a high precision and accuracy when properly validated, and an ability to quantitate multiple analytes simultaneously. As a result of the inclusion of over 170 new references, this updated review summarizes a wealth of detailed experiential evidence and newly developed methodology that supports qHNMR as a valuable and unbiased analytical tool for natural product and other areas of research.
The genus Actaea (including Cimicifuga) has been the source of ∼200 cycloartane triterpenes. While they are major bioactive constituents of complementary and alternative medicines, their structural similarity is a major dereplication problem. Moreover, their trivial names seldom indicate the actual structure. This project develops two new tools for Actaea triterpenes that enable rapid dereplication of more than 170 known triterpenes and facilitates elucidation of new compounds. A predictive computational model based on classification binary trees (CBTs) allows in silico determination of the aglycone type. This tool utilizes the Me (1)H NMR chemical shifts and has potential to be applicable to other natural products. Actaea triterpene dereplication is supported by a new systematic naming scheme. A combination of CBTs, (1)H NMR deconvolution, characteristic (1)H NMR signals, and quantitative (1)H NMR (qHNMR) led to the unambiguous identification of minor constituents in residually complex triterpene samples. Utilizing a 1.7 mm cryo-microprobe at 700 MHz, qHNMR enabled characterization of residual complexity at the 10-20 μg level in a 1-5 mg sample. The identification of five co-occurring minor constituents, belonging to four different triterpene skeleton types, in a repeatedly purified natural product emphasizes the critical need for the evaluation of residual complexity of reference materials, especially when used for biological assessment.
The effects of climatic conditions on green tea metabolites in three different growing areas of Jeju Island, South Korea, were investigated through global metabolite profiling by (1)H nuclear magnetic resonance (NMR) spectroscopy. Pattern recognition methods, such as principal component analysis (PCA) and orthogonal projection on latent structure-discriminant analysis (OPLS-DA), revealed clear discriminations of green teas from the three different growing areas. Variations of theanine, isoleucine, leucine, valine, alanine, threonine, glutamine, quinic acid, glucose, epicatechin (EC), epigallocatechin (EGC), epigallocatechin-3-gallate (EGCG), and caffeine levels were responsible for the discriminations. Green teas grown in an area with high temperature, long sun exposure time, and high rainfall had higher levels of theanine but lower levels of isoleucine, leucine, valine, alanine, EC, EGC, EGCG, and caffeine than those grown in areas with relatively low temperature, short sun exposure time, and low rainfall. These results indicate that high temperature, long sun exposure, and high preciptation stimulate theanine synthesis in green tea during the spring season. This study highlights how metabolomics coupled with multivariate statistical analysis can illuminate the metabolic characteristics of green tea associated with climatic variables, thereby allowing for the assessment of quality strategy in green tea production.
Three new cyclolanostanol xylosides were isolated from a batch of commercial Cimicifuga Rhizome, cimicifugoside H-1 (1), C35H52O9, mp 260-262 degrees C, [alpha]D -43.5 degrees, cimicifugoside H-2 (2), C35H54O10, mp 227-229 degrees C, [alpha]D -38.8 degrees, and cimicifugoside H-5 (3), C35H52O10, mp 262-264 degrees C, [alpha]D -22.9 degrees, together with known glycosides, actein and 27-deoxyactein. Their structures were determined on the basis of chemical and spectrometric evidence including an X-ray crystallographic analysis. The structure of cimicifugoside H-1 (1) was established as (20R,24R)-24,25-epoxy-11 beta-hydroxy-3-beta-(beta-D- xylopyranosyloxy)-9,19-cyclolanost-7-ene-16,23-dione. Cimicifugoside H-5 (3) is the 15-hydroxylated derivative of 1. Since 1 changed into cimicifugoside H-2 (2) on treatment with p-toluenesulfonic acid, 2 has a 24R,25-diol structure derived from 1 by opening its epoxy ring.
Due to the long-term health risks now associated with hormone replacement therapy, many menopausal women are actively seeking alternative treatments. One such alternative is black cohosh (Actaea racemosa, syn. Cimicifuga racemosa), which has been used in the United States for the treatment of gynecologic complaints for more than 100 years. Review of the published clinical data suggests that black cohosh may be useful for the treatment of menopausal symptoms, such as hot flashes, profuse sweating, insomnia, and anxiety. Results from the most recently published trial, however, indicate that black cohosh is not effective for the treatment of menopausal symptoms in breast cancer survivors being treated with tamoxifen. Because the overall quality of the published clinical trials is low, two new randomized, double-blind, placebo-controlled clinical trials are currently underway in the United States. To date, only one standardized black cohosh extract has been tested clinically; the current recommended dose is 40-80 mg per day. At least 4-12 weeks of treatment may be required before any therapeutic benefits may be apparent. Adverse reactions such as nausea, vomiting, headaches, dizziness, mastalgia, and weight gain have been observed in clinical trials. No drug interactions are reported in the medical literature. The estrogenic effects of black cohosh are controversial, and the more recent data indicate that black cohosh extracts may have an anti-estrogenic activity. Owing to potential effects on sex hormones, however, black cohosh should not be administered to children or during pregnancy and lactation.
The metabolomic analysis of Ephedra species was performed using 1H-NMR spectroscopy and multivariate data analysis. A broad range of metabolites could be detected by 1H-NMR spectroscopy without any chromatographic separation. The principal component analysis used to reduce the huge data set obtained from the 1H-NMR spectra of the plant extracts clearly discriminated three different Ephedra species. The major differences in Ephedra sinica, Ephedra intermedia and Ephedra distachya var. distachya were found to be due to benzoic acid analogues in the aqueous fraction and ephedrine-type alkaloids in the organic fraction. Based on this metabolomic recognition, one of nine commercial Ephedra materials evaluated was shown to be a mixture of Ephedra species. This method will be a useful tool for chemotaxonomic analysis and authentification of Ephedra species including quality control of plant materials.
Black cohosh has become one of the most important herbal products in the US dietary supplements market. It is manufactured from roots and rhizomes of Cimicifuga racemosa (Ranunculaceae). Botanical identification of the raw starting material is a key step in the quality control of black cohosh preparations. The present report summarizes a fingerprinting approach based on high performance liquid chromatography-photodiode array/mass spectrometric/evaporative light scattering detection (HPLC-PDA/MS/ELSD) that has been developed and validated using a total of 10 Cimicifuga species. These include three North American species, Cimicifuga racemosa, Cimicifuga americana, Cimicifuga rubifolia, and seven Asian species, Cimicifuga acerina, Cimicifuga biternat, Cimicifuga dahurica, Cimicifuga heracleifolia, Cimicifuga japonica, Cimicifuga foetida, and Cimicifuga simplex. The chemotaxonomic distinctiveness of the HPLC fingerprints allows identification of all 10 Cimicifuga species. The triterpene glycoside cimigenol-3-O-arabinoside (3), cimifugin (12), and cimifugin-3-O-glucoside (18) were determined to be suitable species-specific markers for the distinction of C. racemosa from the other Cimicifuga species. In addition to identification, the fingerprint method provided insight into chemical interconversion processes occurring between the diverse triterpene glycosides contained in black cohosh. The reported method has proven its usefulness in the botanical standardization and quality control of black cohosh products.