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Background Dried and red ginseng are well-known types of processed ginseng and are widely used as healthy food. The dried and red ginseng quality may vary with the storage period of raw ginseng. Therefore, herein, the effect of the storage period of fresh ginseng on processed ginseng quality was evaluated through multicomponent quantification with statistical analysis. Methods A method based on ultrahigh performance liquid chromatography coupled to triple quadrupole mass spectrometry in multiple-reaction monitoring mode (UPLC-MRM-MS) was developed for quantitation of ginsenosides and oligosaccharides in dried and red ginseng. Principal component analysis and partial least squares discriminant analysis were conducted to evaluate the dynamic distributions of ginsenosides and oligosaccharides after different storage periods. Results Eighteen PPD, PPT and OLE ginsenosides and nine reducing and nonreducing oligosaccharides were identified and quantified. With storage period extension, the ginsenoside content in the processed ginseng increased slightly in the first 2 weeks and decreased gradually in the following 9 weeks. The content of reducing oligosaccharides decreased continuously as storage time extending, while that of the nonreducing oligosaccharides increased. Chemical conversions occurred during storage, based on which potential chemical markers for the storage period evaluation of fresh ginseng were screened. Conclusion According to ginsenoside and oligosaccharide distributions, it was found that the optimal storage period was 2 weeks and that the storage period of fresh ginseng should not exceed 4 weeks at 0 °C. This study provides deep insights into the quality control of processed ginseng and comprehensive factors for storage of raw ginseng.

Soil sickness of ginseng (Panax ginseng C. A. Mey.) has become a major limiting factor in ginseng cultivation. We found that Total Ginsenoside in Ginseng Root (TSPG) significantly decreased the activities of soil urease, acid phosphatase and laccase. Its high concentration of TSPG (10.00 mg L-1) significantly reduced the activity of soil sucrose. Besides, the TSPG can inhibited the growth of ginseng and increased the incidence of disease. Therefore, allelopathic effects of TSPG may be one of the main causes of Soil Sickness in ginseng.

The basic properties of herbal medicines are cold, hot, warm, and cool. The differentiation of these properties is important for the diagnosis and treatment of diseases. Ginseng and American ginseng possess opposite properties of warm and cool, respectively. At present, the mechanisms and the influence of steaming leading to the differences in their properties are not clear and require further investigation. Therefore, nontargeted metabonomics based on liquid chromatography-mass spectrometry was applied to investigate the effects of ginseng, American ginseng, and their variants on the changes in endogenous metabolites in rat urine. A total of 19 potential biomarkers were screened out and identified, of which 17, 7, and 5, were respectively related to warm, cool, and both warm and cool properties with opposite effects. The metabolic pathways corresponded to fatty acids, lipids, glycolysis, and energy metabolisms. The warm and tonic effects of red ginseng are stronger than those of ginseng and consistent with the theory of traditional Chinese medicine. The red American ginseng has cool property; however, the degree of coolness is less than that of American ginseng. This study provides a reference methodology to understand the effects of processing and mechanisms associated with the differences in the properties of herbal medicines.

Two polysaccharides were obtained from steamed ginseng via ultrafiltration, and their physical–chemical properties, solution properties and antifatigue activities were studied. WSGP-S3 and WSGP-G3 were acid heteropolysaccharides with the molecular weights of 2.03 × 104 and 4.86 × 104, respectively. They were composed of different molar ratios of the monosaccharides Rha, GlcA, GalA, Glc, Gal, and Ara. The results of size-exclusion chromatography–multiangle laser light scattering analysis, Conge red staining and Circular dichroism spectroscopy revealed that WSGP-S3 exhibited a random conformation of branched clusters in solution. By contrast, WSGP-G3 exhibited an ordered conformation, including helix-like conformations in aqueous solution. Antifatigue activity tests proved that WSGP-S3 markedly prolonged the exhaustive swimming time of fatigued mice; increased liver and muscle glycogen levels and superoxide dismutase, catalase, glutathione peroxidase activities and decreased blood lactic acid, nitrogen and malondialdehyde levels compared with the control treatment. Moreover, it enhanced spleen cell proliferation in fatigued mice. By contrast, WSGP-G3 had no significant effect on fatigued mice. The results showed that WSGP-S3 might have a major contribution to the antifatigue effects of steamed ginseng polysaccharides and could be a potential anti-fatigue polysaccharide.

Conventional strategies for screening of protein binders cannot be used for complicated samples such as ligand libraries created by combinatorial chemistry or from natural product extracts. In the current study, we developed a novel method in a competitive binding configuration for screening protein binders from complicated samples by a combination of streptavidin-coated 96-well plate format in conjunction with ultra-high-performance liquid chromatography coupled with Orbitrap mass spectrometry (UHPLC-Orbitrap-MS). The concanavalin A (Con A) modified 96-well plate and lysozyme modified 96-well plate (as control) were incubated with oligosaccharide standards respectively, and the compounds with the decreased peak areas in experimental group compared to those in the control group were detected as binders by UHPLC-ESI-MS. The factors such as incubation time, incubation temperature, and buffer, which might affect the binding affinity and reproducibility were optimized. The potential of the approach is examined using the extracts of Radix ginseng cruda and American ginseng. The relative binding degrees (RBDs) of the detected disaccharides were relatively high in the extracts of Radix ginseng cruda, and those of the trisaccharides were similar in the extracts of the two kinds of ginseng. To our knowledge, it’s the first time to reveal the differences and analogies in lectin peanut agglutinin (PNA)-binding capabilities of oligosaccharides between the extracts of radix ginseng cruda and American ginseng, indicating the efficiency of the method for analysis of complicated samples.

The study aims to clarify the structural domain required for the immune enhancement of ginseng neutral polysaccharide (GPN). GPN was first obtained through water extraction and ion-exchange chromatography from Panax ginseng. GPN was hydrolyzed by α-amylase for 24 h and fractionated through gel permeation chromatography to give two final fragments GPNE-I and GPNE-II, with molecular weight of 8.03 × 104 Da for GPNE-I and 3.15 × 104 Da for respectively. FT-IR, methylation and 1D/2D NMR analysis demonstrated that GPNE-I was a heteropolysaccharide consisting mainly of a glucan domain and type I and II arabinogalactans (AG-I and AG-II). GPNE II was a glucan consisting of (1 → 4)-α- d -Glcp backbone with a substitution at O-6 on every two residues. (1 → 3)-α- d -Glcp and (1 → 6)-α- d -Glcp were located at the branches. In the two fractions, both α- and β-t-Glcp as reductive terminals and →4)-α-Glcp as a non-reducing end were detected. The branching degrees of GPNE-I and GPNE-II were 38.17% and 50.78%, respectively. Immunological experiments revealed that GPNE-I exhibited more effectively stimulated lymphocyte proliferation than GPN and GPNE-II, indicating the former showed potential for immunomodulators applications, indicating that GPNE-I might be the core active domain and necessary for GPN to promote lymphocyte proliferation.

Background: The structural conversions in ginsenosides induced by steaming or heating or acidic condition could improve red ginseng bioactivities significantly. In this paper, the chemical transformations of red American ginseng from fresh Panax quinquefolium L. under steaming were investigated, and the possible mechanisms were discussed. Methods: A method with reversed-phase high-performance liquid chromatography coupled with linear ion trap mass spectrometry (HPLC-MSn)-equipped electrospray ionization ion source was developed for structural analysis and quantitation of ginsenosides in dried and red American ginseng. Results: In total, 59 ginsenosides of protopanaxadiol, protopanaxatriol, oleanane, and ocotillol types were identified in American ginseng before and after steaming process by matching the molecular weight and/or comparing MSn fragmentation with that of standards and/or known published compounds, and some of them were determined to be disappeared or newly generated under different steaming time and temperature. The specific fragments of each aglycone-type ginsenosides were determined as well as aglycone hydrated and dehydrated ones. The mechanisms were deduced as hydrolysis, hydration, dehydration, and isomerization of neutral and acidic ginsenosides. Furthermore, the relative peak areas of detected compounds were calculated based on peak areas ratio. Conclusion: The multicomponent assessment of American ginseng was conducted by HPLC-MSn. The result is expected to provide possibility for holistic evaluation of the processing procedures of red American ginseng and a scientific basis for the usage of American ginseng in prescription. Keywords: ginsenoside conversions, HPLC-MSn, multicomponent assessment, Panax quinquefolium L

Two acid polysaccharides were obtained from steamed ginseng (GPS-1 and GPS-2) through water extraction, ion-exchange chromatography and gel chromatography. The structural features and ability of the polysaccharides to inhibit lipid accumulation in oleic acid-induced HepG2 cells were studied. GPS-1 consisted of type I arabinogalactans (AG-I), arabinogalactans-II (AG-II) and rhamnogalacturonan I (RG-I) domains. GPS-2 was a pectin-like polysaccharide consisting mainly of the homogalacturonan (HG) domain and a small amount of AG domain. Both GPS-1 and GPS-2 had branches attaching on O-3 of (1 → 6)-GalA or O-4 of (1 → 2)-Rha and terminated by either Ara or Gal. An in vitro experiment revealed that GPS-1 treatment at 50–400 μg/ml could dose-dependently decrease intracellular lipid accumulation and cholesterol (TC) and triglycerides (TG) levels. GPS-1 exerted a more serious hypolipidemic effect than GPS-2 did. Moreover, GPS-1 considerably increased the phosphorylation of AMP-activated protein kinase (AMPK) and affected the expression of AMPK downstream targets, including the inhibition of the protein expression of sterol regulatory element-binding protein 1c (SREBP-1c) and activation of Acetyl-CoA carboxylase (ACC). Results suggest that GPS-1 could inhibit lipid accumulation via the AMPK the signalling pathway.

Rust rot is a severe Panax ginseng disease caused by Ilyonectria robusta . The severity of the disease is related to the content of residual ginsenoside content in the soil. Herein, the effects of different concentrations of ginsenoside Rg1 on ginseng rust rot and its pathogens were studied, and the growth and sporulation of fungi cultured in vitro were determined. The results showed that Rg1 significantly promoted the mycelial growth, and sporulation compared to the control, and aggravated the disease symptoms of Panax ginseng . A total of 6,708 transcripts out of 213,131 annotated genes identified from global transcriptomic analysis were differentially expressed in I.robusta grown in the Rg1 treatment. These genes were found to be related to the carbon-nitrogen metabolism, transport, and assimilation. Many of these genes were also associated with pathogenicity based on the Phi-base database. Several transcription factors were related to specific biological processes, such as nitrogen utilization. The current results revelation that Rg1 plays a major role in the development of rust rot by promoting fungal cell growth and affecting the expression of genes required for pathogenesis. Rg1 could aggravate the invasion of I.robusta on ginseng root, which preliminarily revealed the reason for the aggravation of rust rot in ginseng c ontinuous cropping obstacle.

Ethnopharmacological relevance Spleen-Qi deficiency is a syndrome of traditional Chinese medicine. Panax ginseng (ginseng) is well known as the key herb for replenishing Qi and tonifying Spleen. However, the pathogenesis of Spleen-Qi deficiency syndrome and therapeutic mechanism of ginseng on Spleen-Qi deficiency constitution have not yet been entirely elucidated. Aim of the study The aim of this work was to investigate the pathogenesis of Spleen-Qi deficiency syndrome and therapeutic mechanism of ginseng on Spleen-Qi deficiency constitution. Materials and methods The urinary metabonomics was investigated before and after ginseng treatment in Spleen-Qi deficiency subjects by rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-QTOF-MS). The spectra of metabolites were analyzed by principal component analysis (PCA) and the score showed significant difference among the three different groups. The serum biochemical parameters creatine kinases (CK), lactate dehydrogenase (LDH), interferon-γ (INF-γ), Interleukin-4 (IL-4), superoxide dismutase (SOD) and malondialdehyde (MDA) were determined by assay kits. In addition, ultraweak photon emission (UPE) intensity was measured at dazhui point in each subject. Results According to the orthogonal partial least-squares discriminant analysis (OPLS-DA) results and biochemistry databases searching, 15 potential biomarkers were identified to be involved in Spleen-Qi deficiency syndrome and ginseng influenced Spleen-Qi deficiency. The metabonomics and biological experiment data indicated that metabolism pathways were corresponding to energy metabolism, amino acid, carbohydrate, lipid and pyrimidine metabolisms, oxidative stress urea cycle, and intestinal flora metabolism. And the UPE intensity decreased significantly after the treatment. Conclusions Ginseng could promote the related metabolisms in Spleen-Qi deficiency constitution and the metabolites and UPE intensity may be useful as potential biomarkers for diagnosing and monitoring for Spleen-Qi deficiency syndrome.

Ginsenoside Rb3 is one of major ginsenosides in Panax ginseng with effect on cardio-vascular and central nervous system. The aim of this study is to develop a rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (RRLC-Q-TOF-MS) method for pharmacokinetic study of ginsenoside Rb3 and simultaneous determination of metabolites in rats. The results showed that the concentration-time profile of ginsenoside Rb3 conformed to a two-compartment pharmacokinetic model after intravenous administration at the dosage of 2.0 mg/kg for rats. The mean plasma elimination half-lives were 13.77 ± 1.23 min and 2045.70 ± 156.20 min for the distribution and exterminate phases t1/2α and t1/2β. In the metabolic study, prototype ginsenoside Rb3 and deglycosylation metabolites were characterized by comparison with the retention time of the standard compounds, accurate mass measurement and the characteristic fragment ions obtained from MS/MS. Two major metabolites Mb1 and M2' were tentatively identified in rat urine samples after intravenous administration, and four possible metabolites Mb1, F2, M2' and CK were detected in rat feces samples after oral administration. The deglycosylation was found to be the major metabolic pathways of ginsenoside Rb3 in rat. The in vivo metabolic pathway of ginsenoside Rb3 was summarized.

Ginsenosides in Panax quinquefolium L. were determined using developed ultra-performance liquid chromatography coupled to high resolution mass spectrometry (UPLC-HRMS) method with electrospray ionization and orbitrap MS analyzer in negative ionization mode. Optimal UPLC separation was achieved using a mixture of acetonitrile and water with 0.1% formic acid as the mobile phase in linear gradient elution. The MS parameters were optimized for reliable detection with enhanced selectivity and sensitivity, and improved identification and quantification of ginsenosides. The applicability of this method was demonstrated on ginsenosides from Panax quinquefolium L. (American ginseng), Panax ginseng (Chinese ginseng) and Panax notoginseng (Sanchi) roots and products. The differences between Chinese and Northern American Panax quinquefolium L., main roots and hair roots, and products from different pharmacy were investigated. The results were also confirmed by principal component analysis and partial least squares discriminatory analysis. It indicated that the strategy can be extended to rapid and accurate authentication of Panax genus.

Context: Panax ginseng is well known for its various bioactivities, but specific knowledge on ginseng proteins (GPs) is limited. Aims: Protein components were extracted from ginseng and antitumor activity in human breast cancer cell line MCF-7 was investigated. Settings and Design: Five methods were applied to extract GP and the antitumor effects and mechanism of GP on MCF-7 were explored, including proliferation, cell cycle, apoptosis, and migration. Subjects and Methods: Five extraction methods were employed. MCF-7 cell proliferation was measured using a cell counting kit-8 with different GP concentrations (0, 0.25, 0.5, 1, 2, and 4 mg/mL) and half inhibitory concentration values were calculated. Cell cycle, morphology, and apoptosis were investigated using immunofluorescence staining and flow cytometry. Migration was probed by scratch wound healing and transwell assays. Quantitative-polymerase chain reaction and western blotting were performed to analyze apoptosis-associated gene/protein expression. Statistical Analysis: Experimental data were analyzed by Microsoft Excel and SPSS software. Results: Acetone extraction achieved the highest GP purity and yield. GP inhibited proliferation of MCF-7 in a time-dependent manner and induced cell cycle arrest at G1/S and apoptosis. Scratch wound healing and transwell assays showed that cell migration was also inhibited by GP and expression levels of Bcl-2 and Bax were affected. Conclusion: GP elicits antitumor activity by inhibiting cell proliferation and migration and inducing cell cycle arrest and apoptosis in MCF-7 cells and may act via the Bcl-2 and Bax apoptotic pathway.