American Ginseng Research Articles

Biotransformation of American Ginseng Stems and Leaves by an Endophytic Fungus Umbelopsis sp. and Its Effect on Alzheimer’s Disease Control

Background: Common ginsenosides can be transformed into rare ginsenosides through microbial fermentation, and some rare ginsenosides can prevent Alzheimer’s disease (AD). This study aimed to transform common ginsenosides into rare ginsenosides through solid-state fermentation of American ginseng stems and leaves (AGSL) by an endophytic fungus and to explore whether fermented saponin extracts prevent AD. Methods: The powders of AGSL were fermented in a solid state by endophytic fungus. Total saponins were extracted from fermentation products using the methanol extraction method. The types of saponins were analyzed by liquid chromatography mass spectrometry (LC/MS). The Aβ42 concentration and β-secretase activity were measured by ELISA for the prevention of AD. Results: After AGSL was fermented by an endophytic fungus NSJG, the total saponin concentration of the fermented extract G-SL was higher than the unfermented CK-SL. Rare ginsenoside Rh1 was newly produced and the yield of compound K (561.79%), Rh2 (77.48%), and F2 (40.89%) was increased in G-SL. G-SL had a higher inhibition rate on Aβ42 concentration (42.75%) and β-secretase activity (42.22%) than CK-SL, possibly because the rare ginsenoside Rh1, Rh2, F2, and compound K included in it have a strong inhibitory effect on AD. Conclusion: The fermented saponin extracts of AGSL show more inhibition effects on AD and may be promising therapeutic drugs or nutrients for AD.

Boronate affinity-based surface molecularly imprinted polymer microspheres using polyethyleneimine/dopamine coating for efficient selective recognition and separation of Ginsenoside Rb1

A novel approach combining boronate affinity with surface molecular imprinting was developed for the specific recognition and separation of ginsenoside Rb1. Firstly, the boronic acid-functionalized polymer microspheres were prepared as inner cores via suspension polymerization using methyl methacrylate, ethyleneglycol dimethacrylate, and 4-vinylphenyl boronic pinacol ester (4-VBPE) as polymerization monomers. Then, the Ginsenoside Rb1 (GRb1) template was covalently pre-anchored onto the surface of microspheres through boronate affinity binding after the hydrolysis of 4-VBPE. Finally, dopamine and polyethyleneimine were used as co-coating monomers to form a thickness-controllable imprinting layer onto the polymer microspheres via in-water self-copolymerization. The key factors affecting specific recognition, including the boronic acid content, adsorption media, and polymerization time were carefully optimized. The resulting molecularly imprinted microspheres (MIMs) exhibited strong surface hydrophilicity. In the aqueous media, the optimal adsorption capacity for the template reached 81.45 μmol g−1, and the imprinting factor was 5.32. The specific evaluation showed that the MIMs had a much higher selectivity for GRb1 than its analogues, and the selectivity coefficients were all higher than 2.4. In addition, the MIMs can be applied as adsorbents for solid-phase extraction to selectively separate ginsenosides from American ginseng extract. The eluting recovery ratios of GRb1, GRg2, and GRd reach 92.39%, 88.90%, and 92.49%, respectively.

Comprehensive analysis of different types of ginsenosides in the different parts of American ginseng by targeted and nontargeted MS/MS scanning.

To comprehensively study the ginsenosides distribution in the various tissues of American ginseng, the qualitative and quantitative-targeted and nontargeted mass spectroscopic methods were established using the high-performance liquid chromatography coupled with Qtrap triple quadrupole mass spectrometry (HPLC-QtrapQQQ-MS). The total ginsenosides of the root, stem, and leaf of American ginseng were determined by a colorimetric method, and the contents showed the order from high to low root, stem, and leaf. Eighty-two kinds of ginsenosides were detected in the different parts of American ginseng by enhanced mass scan-information-dependent data acquisition (IDA)-enhanced product ion (EPI) scan mode, including 69 from the root, 62 from the stem, and 48 from the leaf. An HPLC-multiple reaction monitoring (MRM) method was established, and 28 representative ginsenosides were further quantified in the three parts. Nearly all ginsenosides had the highest contents in the root and the lowest content in the leaf. Three types of ginsenosides (protopanaxadiol [PPD]-, protopanaxatiol [PPT]-, and oleanolic acid [OA]-types) were analyzed by precursor ion-IDA-EPI and MRM-IDA-EPI scan modes. Root had the most abundant ginsenosides in PPD- and PPT-type ginsenosides. Meanwhile, the OA-type ginsenosides are significantly enriched in the stem and leaf of American ginseng. The results provided a supplement to the quality assessment of American ginseng. PRACTICAL APPLICATION: The distribution profile of ginsenosides in the parts of American ginseng is different. Except for the root, the stem, and leaf of American ginseng have the most abundant ginsenosides in oleanolic acid type. The results reported herein can help the manufacturers choose appropriate materials to extract the ginsenosides.

Complete composition analysis of polysaccharides based on HPAEC-PAD coupled with quantitative analysis of multi-components by single marker.

Monosaccharide compositions analysis (MCA) is indispensable for structural characterisations and structure-activity relationships of plant polysaccharides. To develop a concise and direct MCA method, we established a quantitative analysis of the multi-monosaccharaides by single marker (QAMS) by high-performance anion-exchange chromatography with pulsed-amperometric detection (HPAEC-PAD) method. A stable and reproducible HPAEC-PAD method for simultaneous determination of aldoses, ketoses and uronic acids (i.e., l-arabinose, d-xylose, d-ribose, l-rhamnose, d-fucose, d-mannose, d-glucose, d-galactose, d-fructose, d-glucuronic acid and d-galacturonic acid) was established by systematic optimisation of stationary phases, column temperatures and elution programmes. On this basis, the QAMS method was proposed through comprehensive investigations of relative correction factor (RCF) variations under different influencing factors, for example, sample concentrations, flow rates, and column temperatures. Using rhamnose as an internal reference standard, the contents of the other monosaccharidecomponents in polysaccharides from Panax quinquefolium L. and Achyranthes bidentata Bl. samples were simultaneously determined by QAMS, and there was no significant difference between the results from the QAMS and external standard method (t test, P > 0.520). In addition, a MCA fingerprinting of 30 batches of P. quinquefolium polysaccharide was established by HPAEC-PAD, and six common peaks were assigned and determined. The established HPAEC-PAD-QAMS method was successfully applied to the MCA of polysaccharides from P. quinquefolium and A. bidentata after optimisation of hydrolysis conditions. HPAEC-PAD-QAMS was proposed and established for MCA of plant polysaccharides for the first time.

Panax quinquefolium saponins attenuates microglia activation following acute cerebral ischemia-reperfusion injury via Nrf2/miR-103-3p/TANK pathway.

Ischemic stroke is one of the leading causes of death and disability among adults worldwide. Intravenous thrombolysis is the only approved pharmacological treatment for acute ischemic stroke. However, reperfusion by thrombolysis will lead to the rapid activation of microglia cells which induces interferon-inflammatory response in the ischemic brain tissues. Panax quinquefolium saponins (PQS) has been proven to be effective in acute ischemic stroke, but there is no unified understanding about its specific mechanism. Here, we will report for the first time that PQS can significantly inhibit the activation of microglia cells in cerebral of MCAO rats via activation of Nrf2/miR-103-3p/TANKaxis. Our results showed that PQS can directly bind to Nrf2 protein and inhibit its ubiquitination, which result in the indirectly enhancing the expression of TANK protein via transcriptional regulation on miR-103-3p, and finally to suppress the nuclear factor kappa-B dominated rapid activation of microglial cells induced by oxygen-glucose deprivation/reoxygenationvitro and cerebral ischemia-reperfusion injury in vivo. In conclusion, our study not only revealed the new mechanism of PQS in protecting against the inflammatory activation of microglia cells caused by cerebral ischemia-reperfusion injury, but also suggested that Nrf2 is a potential target for development of new drugs of ischemic stroke. More importantly, our study also reminded that miR-103-3p might be used as a prognostic biomarker for patients with ischemic stroke.

Effects of rotation and Bacillus on the changes of continuous cropping soil fungal communities in American ginseng

The continuous cropping obstacle is the main factor in leading to difficulty in American ginseng replanting. The dormant microbiota in the soil may be the cause of American ginseng disease and eventually caused continuous cropping obstacles, but there are few studies on the dynamic changes of soil microenvironment after American ginseng planting. In this study, we tracked short-term variation in physicochemical properties, enzyme activities, and fungal communities over time-series in soils with continuous cropping obstacle under crop rotation and probiotic Bacillus treatments. Furthermore, we examined the relationships between the important fungal compositions and the soil properties. The results showed that sucrase, cellulase, urease and acid phosphatase activities were significantly increased, while catalase and dehydrogenase were decreased with treatments time. Rotation treatment significantly affected the diversity, dissimilarity degree and species distribution of soil fungal community with continuous cropping obstacle over a short-term. Moreover, beneficial fungal biomarkers such as Cladorrhinum, Oidiodendron, and Mariannaea were accumulated at 48 h under rotation treatments. Almost all fungal biomarkers were negatively correlated with hydrolases and positively correlated with oxidoreductases and acid phosphatase under crop rotation treatments. This study suggested that compared to probiotic Bacillus, crop rotation can significantly affect soil fungal community structure, especially the enrichment of specific potentially beneficial fungal species. Our findings provide a scientific basis for understanding the dynamic changes of fungal communities and soil properties with continuous cropping obstacle of American ginseng in initial stage of soil improvement.

Research on the dentification of Panax ginseng and Panax quinquefolium using catalysed hairpin assembly technology.

Panax ginseng and Panax quinquefolium are traditional Chinese herb medicines and similar in morphology and some chemical components but differ in drug properties, so they cannot be mixed. However, the processed products of them are often sold in the form of slices, powder, and capsules, which are difficult to identify by traditional morphological methods. Furthermore, an accurate evaluation of P. ginseng, P. quinquefolium and the processed products have not been conducted. This study aimed to establish a catalysed hairpin assembly (CHA) identification method for authenticating products made from P. ginseng and P. quinquefolium based on single nucleotide polymorphism (SNP) differences. By analysing the differences of SNP in internal transcribed spacer 2 (ITS2) in P. ginseng and P. quinquefolium to design CHA-specific hairpins. Establish a sensitive and efficient CHA method that can identify P. ginseng and P. quinquefolium, use the sequencing technology to verify the accuracy of this method in identifying Panax products, and compare this method with high-resolution melting (HRM). The reaction conditions of CHA were as follows: the ratio of forward and reverse primers, 20:1; hairpin concentration, 5 ng/μL. Compared with capillary electrophoresis, this method had good specificity and the limit of detection was 0.5ng/μL. The result of Panax product identification with CHA method were coincidence with that of the sequencing method; the positive rate of CHA reaction was 100%. This research presents an effective identification method for authenticating P. ginseng and P. quinquefolium products, which is helpful to improve the quality of Panax products.

Ecological regulation network of quality in American Ginseng: Insights from macroscopic-mesoscopic-microscopic perspectives

The environment profoundly influences the growth, development and constituents of plants. Nonetheless, a lack of comprehensive studies at various scales hinders our understanding of how environmental factors impact the quality of American ginseng. This study employed an integrated approach, considering macroscopic-mesoscopic-microscopic perspective, to reveal the ecological variables impacting the quality of American ginseng. From the macroscopic perspective, the MaxEnt model revealed that American ginseng is suitable for growth in temperate regions. The temperature seasonality and warmest quarter emerged as crucial bioclimatic factors that influence the distribution of American ginseng. Taking the mesoscopic perspective, the ginsenosides were significantly positively correlated with available phosphorus (AP) and soil water content (SWC), and significantly negatively correlated with ammonium nitrogen (NN), available potassium (AK), and microbial biomass carbon (MBC). From the microscopic perspective, a total of 16 microorganisms, including 14 bacteria and 2 fungi, were identified to have a significant impact on the synthesis of ginsenosides through 16 S and internally transcribed spacer (ITS) rRNA amplicon sequencing combine with machine learning. Notably, this is the first time to construct an ecological network for regulating the quality of American ginseng and identify the key ecological factors, which can serve as valuable references for targeted breeding and domestication of American ginseng. These findings not only established a theoretical foundation for the future investigation of biological fertilizers, but simultaneously provided crucial insights for cultivating high-quality medicinal plants.

Regulatory Effects of Ganoderma lucidum, Grifola frondosa, and American ginseng Extract Formulation on Gut Microbiota and Fecal Metabolomics in Mice.

The bioactivities of Ganoderma lucidum, Grifola frondosa, and American ginseng have been extensively studied and documented. However, the effects of their complexes on the structural properties of intestinal microbiota and fecal metabolism remain unclear. Therefore, this paper aims to present a preliminary study to shed light on this aspect. In this study, an immunocompromised mouse model was induced using cyclophosphamide, and Ganoderma lucidum, Grifola frondosa, and American ginseng extract formulation (referred to as JGGA) were administered via gavage to investigate their modulatory effects on gut microbiota and fecal metabolism in mice. The effects of JGGA on immune enhancement were explored using serum test kits, hematoxylin-eosin staining, 16SrDNA high-throughput sequencing, and UHPLC-QE-MS metabolomics. The findings revealed potential mechanisms underlying the immune-enhancing effects of JGGA. Specifically, JGGA administration resulted in an improved body weight, thymic index, splenic index, carbon scavenging ability, hypersensitivity, and cellular inflammatory factor expression levels in mice. Further analysis demonstrated that JGGA reduced the abundance of Firmicutes, Proteobacteria, and Actinobacteria, while increasing the abundance of Bacteroidetes. Additionally, JGGA modulated the levels of 30 fecal metabolites. These results suggest that the immune enhancement observed with JGGA may be attributed to the targeted modulation of gut microbiota and fecal metabolism, thus promoting increased immunity in the body.

Dietary supplementation with American ginseng dietary fiber ameliorates intestinal mucosal barrier injury in immunosuppressed mice

Dietary fiber is an important component of healthy eating patterns. Although American ginseng (Panax quiquefolium L.) is often used to improve body immunity, less attention has been paid to American ginseng dietary fiber (AGDF) in this regard. The aim of this study was to investigate the protective effect of AGDF on intestinal mucosal barrier injury induced by cyclophosphamide (CTX) in immunosuppressed mice and the underlying mechanism. The results showed that AGDF could increase immune organ index and enhance immunity of immunosuppressed mice, such as DTH response, lymphocyte transformation and proliferation and NK effector cell activity. At the same time, AGDF could restore the proportion of CD4+/CD8+ T lymphocytes induced by CTX to different extents, and promote the secretion of immunoglobulin IgG. More importantly, AGDF could up-regulate the contents of TNF-α, IFN-γ, IL-6 and IL-1β in serum and intestine of immunosuppressed mice, promote the secretion of cytokines and regulate the homeostasis balance between Th1/Th2 cytokines. It also upregulated the expression of tight junction proteins occludin, claudin-1 and ZO-1, and alleviated the degree of CTX-induced damage to the intestinal mucosal barrier. Meanwhile, AGDF could reduce intestinal epithelial cell apoptosis by activating TLR4/NF-κB signaling pathway, improving the phosphorylation of JNK and p38 signaling molecules, which may be the potential mechanism by which AGDF ameliorates intestinal mucosal injury in immunosuppressed mice. Therefore, AGDF may be used as a dietary supplement to prevent and treat intestinal mucosal barrier injury caused by CTX.

Spatial metabolomics method to reveal differential metabolomes in microregions of Panax quinquefolius roots by using ultra-performance liquid chromatography quadrupole/time of flight-mass spectrometry and desorption electrospray ionization mass spectrometry imaging

Panax quinquefolius is a natural homology medicine and food that is rich in bioactive ingredients, such as ginsenosides and polysaccharides. The combination of ultra-performance liquid chromatography quadrupole/time of flight-mass spectrometry (UPLC-Q-TOF/MS) and desorption electrospray ionization mass spectrometry imaging (DESI-MSI) was used for the first time in a spatial metabolomics analysis to comprehensively evaluate the differential components in different microregions of P. quinquefolius. UPLC-Q-TOF/MS and DESI-MSI combined with principal component analysis and orthogonal partial least squares-discriminant analysis were used to screen differential metabolites. UPLC-Q-TOF/MS and DESI-MSI screened 27 and 23 differential metabolites, respectively, among which 15 differential metabolites were identified by both methods. It was found that some components, such as ginsenoside Rg1 and malonyl-ginsenoside Rc, were mainly distributed in P of the transverse slice of P. quinquefolius roots, while ginsenoside Ro and malonyl-ginsenoside Rd were mainly distributed in C. The methods and results of this study could be used to understand the precise localization, biosynthesis, and biological functions of special metabolites in P. quinquefolius.

Application of UPLC-Triple TOF-MS/MS metabolomics strategy to reveal the dynamic changes of triterpenoid saponins during the decocting process of Asian ginseng and American ginseng.

Triterpenoid saponins are the main bioactive components contributed to the nutritional value of ginseng, and different process conditions will affect their content and quality. To study the holistic characterization and dynamic changes of triterpenoid saponins in Asian ginseng (ASG) and American ginseng (AMG) during soaking and decoction, a UPLC-Triple TOF-MS/MS-based metabolomics strategy was used to characterize and discover differential saponin markers. In total, 739 triterpenoid saponins (including 225 potential new saponins) were identified from ASG and AMG in untargeted metabolomics. Based on PCA and OPLS-DA, 51 and 48 saponin markers were screened from soaked and decocted ASG and AMG, respectively. Additionally, targeted metabolomics analysis and HCA of 22 ginsenoside markers suggested that decoction of ASG and AMG for 2h to 4h could significantly increase the contents of rare ginsenosides (G), such as G-Rg3, G-Rg5, G-F4. This study provides a scientific insight that high boiling combined with simmering enriches ASG and AMG extracts with rich rare ginsenosides that are more beneficial to human health.

Panax Quinquefolium Saponins enhances angiogenesis in rats with diabetes and myocardial infarction

Ethnopharmacological relevanceXi Yang Shen (Panax quinquefolium L.) was originally recorded in “Ben Cao Cong Xin” edited by Wu Yiluo during the Qing Dynasty. Panax Quinquefolium Saponins (PQS) is the main component derived from Panax quinquefolium L, and has been wildly used in the treatment of coronary heart disease. Aim of the studyThis study aims to explore the potential role and underlying mechanisms of PQS in promoting angiogenesis in rats with diabetes and myocardial infarction. Materials and methodsEchocardiograms were used to assess cardiac function, while the heart weight to tibia length ratio was calculated to determine cardiac hypertrophy. Hematoxylin and eosin, periodic acid-Schiff and Masson's trichrome staining were used to examine cardiac morphology, myocyte diameter, and myocardial fibrosis. Immunofluorescence staining was employed to evaluate arteriolar density. The transcriptomes were analyzed and bioinformatic analyses were conducted to predict the potential angiogenesis-promoting mechanism of PQS. In addition, RT-PCR and western blotting was utilized to examine the expression of genes and proteins influenced by PQS. ResultsPQS improved blood glucose, ameliorated cardiac function, reduced cardiac hypertrophy, and enhanced myocardial morphology in diabetic rats with myocardial infarction. PQS was also found to decrease myocyte diameter, curtail myocardial fibrosis, and increase arteriolar density. However, the effects of PQS were abolished following the deletion of protein kinase C δ (PKCδ). Molecular docking predicted strong interactions between the major blood components of PQS and PKCδ. Transcriptomic and bioinformatic analyses indicated that PQS may bolster angiogenesis by activating the VEGF/PI3K-Akt/eNOS pathway in rats with diabetes and myocardial infarction. Finally, the study demonstrated that PQS could inhibit the expression of PKCδ and stimulate the activation of the VEGF/PI3K-Akt/eNOS pathway. ConclusionsPQS improves blood glucose, enhances cardiac function, mitigates cardiac damage, and boosts arteriolar density. The angiogenic impact of PQS appears to be, at least partially, due to its modulation of the PKCδ-mediated VEGF/PI3K-Akt/eNOS signaling pathway in rats with diabetes and myocardial infarction.

Rapid Discrimination of Panax quinquefolium and Panax ginseng Using the Proofman-Duplex-LMTIA Technique.

This study aims to establish a rapid identification method based on the Proofman-LMTIA technique for distinguishing between Panax quinquefolium and Panax ginseng. By targeting specific 18S rDNA sequences, suitable primers and Proofman probes labeled FAM or JOE were designed for LMTIA. Initially, single-species-primer Proofman-LMTIA assays were performed separately for each ginseng type to optimize reaction temperature, assess sensitivity and specificity, and determine the detection limit. Subsequently, both sets of primers and their corresponding probes were combined in the same reaction system to further optimize reaction conditions, evaluate sensitivity, and assess stability. Finally, the developed Proofman-duplex-LMTIA technique was employed to detect P. quinquefolium and P. ginseng slices available in the market. Single-plex Proofman-LMTIA assays revealed that the optimal reaction temperature for both P. quinquefolium and P. ginseng was 62 °C. The sensitivity was as low as 1 pg/μL, with a detection limit of 0.1%, and both showed excellent specificity. The optimal temperature for Proofman-duplex-LMTIA assays was 58 °C. This method could simultaneously identify P. quinquefolium and P. ginseng. Testing 6 samples of P. ginseng and 11 samples of P. quinquefolium from the market resulted in a 100% positive rate for all samples. This study successfully established a rapid, simple, sensitive, and specific Proofman-duplex-LMTIA identification method for P. quinquefolium and P. ginseng. It provides an effective means for quality control of P. quinquefolium, P. ginseng, and related products.

Rapid screening and risk assessment of various elements in ultrafine granular powder of Panax quinquefolius L.

IntroductionPanax quinquefolius L. is obtained from a man-shaped root of several species of the genus Panax of the family Araliceae indigenous, which is thought to increase memory, decrease aging, and possess both digestion regulating and liver-protective activity. The widely used in daily life, making its risk assessment a matter of great importance. This research investigates the application of inductively coupled plasma mass spectrometry (ICP-MS) for rapid, non-targeted screening and risk assessment of various heavy metals and toxic agents in Panax quinquefolius L.'s ultrafine granular powder. MethodsThe Panax quinquefolius L. was ground to ultrafine granular powder and subjected to microwave digestion for pre-treatment. ICP-MS was then employed to scan all elements within the full spectrum range, allowing identification of heavy metals and toxic elements using the non-targeted detection method. Subsequently, risk assessment was conducted for Pb, Cu, Ni, Mn, Al, Hg, As, and Cd present in the powder, following the basic steps of risk assessment. ResultsThe concentrations of Pb, Cd, As, Hg, and Cu in ultrafine granular Panax quinquefolius L. powder were found to be below the limits established by the current pharmacopeia. Notably, essential trace elements for the human body, namely Ca, Fe, Mn, Zn, and Mg, were relatively abundant in the powder, with concentrations ranging from 1441.30 to 4016.20, 51.16 to 577.04, 34.78 to 121.99, 17.85 to 83.49, and 1017.69 to 3194.16 mg/kg, respectively. The risk assessment indicated that, in terms of exposure to toxic elements, the order for a person consuming ultrafine granular Panax quinquefolius L. powder would be: Al > Mn > Cu > Ni > Pb > Cd > As > Hg. Importantly, the non-cancer risks associated with Ni, Pb, Al, Mn, Cd, As, Hg, and Cu in the ultrafine granular Panax quinquefolius L. powder were all within acceptable levels. ConclusionThis study successfully establishes a multi-element non-targeted rapid screening method combined with risk assessment techniques, enabling swift determination of harmful target element concentrations and risk profiles in ultrafine granular Panax quinquefolius L. powder. This approach serves as a technical reserve for effectively responding to incidents of toxicity potentially associated with traditional Chinese medicine.

Multiplex genotyping of SNPs in genomic DNA via hydrogel-based assay mediated with MutS and polyethylene glycol

The simultaneous genotyping of multiple single nucleotide polymorphisms (SNPs) in genomic DNA derived from organisms holds significant potential for applications such as precision medicine and food product authentication. However, conventional assay technologies including qPCR-based techniques, microarrays, and hydrogel-based assays face limitations in efficient multiplexing of SNPs, particularly for large-size DNA beyond kilobase scales, due to constraints in multiplex capability, specificity, or sensitivity. In this study, a hydrogel-based multiplex SNP genotyping platform specifically designed for genomic DNA is presented. This platform integrates the ligation detection reaction (LDR) and rolling circle amplification (RCA) techniques within a hydrogel-based multiplex sensing system, enabling adaptable and sensitive SNP genotyping for genomic DNA. To enhance the specificity of the assay, MutS protein and polyethylene glycol are introduced into the protocol, reducing the non-specific ligation and RCA reactions synergistically. With significant specificity improvement of over 10-fold, three types of SNPs within an artificially constructed ∼1000 bp double-stranded DNA (dsDNA) are successfully genotyped with double-digit picomolar sensitivity. Furthermore, the practical applicability of the developed process for the origin identification of raw materials is demonstrated by genotyping three types of SNPs within genomic DNA obtained from two closely related plant species, Korean ginseng (Panax ginseng) and American ginseng (Panax quinquefolius), containing ca. 3.5 gigabase genome size. Of notable significance, this study marks the premiere achievement in PCR-free multiplex genotyping of SNPs in genomic DNA using a single fluorophore.

Safety evaluation of rare ginsenosides of stems and leaves from American ginseng: 90-day exposure toxicity study combined with intestinal flora analysis and metabonomics in rats

Rare ginsenosides have already been widely applied in many fields, including health food and bio-medicine. The human being can expose to rare ginsenosides directly or indirectly increasingly. However, there are few studies on the safety assessment of rare ginsenoside mixtures. In the present study, the sub-chronic toxicity of rare ginsenosides for 90 days on SD rats was performed by combining the intestinal flora analysis and urine metabonomics aiming to illustrate the safety of long-term consumption of rare ginsenosides and the potential damage for liver and intestinal. 48 adult rats were divided into four groups: control (0 mg/kg), low-dose (60 mg/kg), medium-dose (200 mg/kg), and high-dose (600 mg/kg). Rats in the high-dose group showed inflammatory changes in their livers and intestines. The strong bactericidal effect of rare ginsenosides caused intestinal flora disorder and changed the structure of intestinal flora in rats, thus inducing intestinal damage in rats. In the high-dose group, levels of alanine aminotransferase (ALT), lactate dehydrogenase (LDH), and alkaline phosphatase (AKP) increased significantly. As a result of the high-dose treatment, certain metabolic pathways were altered, such as vitamin B6 metabolism, methionine metabolism, glutathione metabolism, and others. These results indicated that high doses of rare ginsenosides induced liver injury by affecting the above metabolic pathways. Rare ginsenosides with no observed adverse effect level (NOAEL) were below 200 mg/kg/day in vivo. Thus, this present study provides insight into the rational use of rare ginsenosides.

Characterization of elements and carbon and nitrogen stable isotopes in American ginseng (Panax quinquefolius L): Determining the geographical origin combining with chemometrics

American ginseng is a valued medicinal and food homologous herbal, which geographical source governs its medicinal and economical values. This work aimed to identify the American ginseng from four regions (including Northeast provinces and Shandong province in China, America, and Canada) using ICP-MS and EA-IRMS. American ginseng harvested in China had significantly higher levels of the essential elements of K (18.7 g/kg), Ca (2.3 g/kg), Na (2.0 g/kg), Mg (1.9 g/kg), P (2.2 g/kg), Mn (59.6 mg/kg), and Cu (12.0 mg/kg), and lower levels of Fe (40.2 mg/kg), total Zn (18.1 mg/kg), B (16.5 mg/kg), and Mo (182.6 μg/kg) than those in America. Similarly, American ginseng harvested in Canada had significantly lower levels of the essential elements of Ca (1.2 g/kg), Fe (7.8 mg/kg), and B (16.5 mg/kg) than those in America. American ginseng from four regions had the significant difference in δ15N value. The highest and lowest δ15N in American ginseng were found in Shandong (3.5‰) and the Canada (-4.4‰), respectively. Multi-elements and δ15N combined with SVM, LDA, RF, and FNN models gave the good classification for American ginseng from four regions. The best FNN model obtained the 100% accuracy. The results showed that elements and δ15N are the useful indictors to identify the geographical origin of American ginseng.

A new ocotillol-type ginsenoside from American ginseng berry

A new ocotillol-type ginsenoside, named pseudoginsenoside F12 (1), was isolated from American ginseng berry, whose structure was elucidated as 6-O-[α-L-2,3-epoxy-rhamnopyranosyl-(1-2)-β-D-glucopyranosyl]-dammar-20S,24R-epoxy-3β, 6α,12β,25-tetraol. In addition, the known alkaloids β-carboline-1-carboxylic acid (2) and anoectochine (3) were isolated for the first time from the Araliaceae family. The new compound 1 was evaluated for cytotoxicity against MDA-MB-231 breast cancer cell line.

Development and application of a visualization method for identification of Panax species with LAMP and a DNAzyme

Panax ginseng and Panax quinquefolium are two valuable Chinese herbal medicines that should not be mixed because they differ in drug properties and efficacy. The traditional identification method is easily affected by subjective factors and cannot effectively distinguish between ginseng products. This study aimed to develop a new chemical analysis method to visually identify P. ginseng and P. quinquefolium. In this method, a large number of sequences containing G-quadruplex were generated by loop-mediated isothermal amplification, and the combination of G-quadruplex and hemin was used to form deoxyribozyme, which catalyzed the color change of H2O2. Artificial simulation of adulteration experiments revealed that this method could detect more than 20% adulterated P. quinquefolium. Compared with the traditional identification methods, this technology was simpler and more efficient, providing a reference for developing rapid visual identification methods and reagents for P. ginseng and P. quinquefolium.