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Use of high-performance liquid chromatography-tandem mass spectrometry to distinguish Panax ginseng C. A. Meyer (Asian ginseng) and Panax quinquefolius L. (North American ginseng).

A liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was developed to distinguish Asian ginseng (Panax ginseng C. A. Meyer) and North American ginseng (Panax quinquefolius L.). The method is based on the baseline chromatographic separation of ginsenoside Rf and 24(R)-pseudoginsenoside F11, two potential chemical markers present in ginseng root methanolic extracts, and their unambiguous on-line identification using tandem mass spectrometry. Consistent with the literature, 24(R)-pseudoginsenoside F11 was detected in abundance in North American ginseng roots in excess of 0.1% (w/w) of the dried root. In contrast to some reports, 24(R)-pseudoginsenoside F11 was also identified in Asian ginseng roots at trace levels using LC-MS-MS but at less than 0.0001% (w/w). Besides showing identical tandem mass spectra to authentic 24(R)-pseudoginsenoside F11, the corresponding compound in Asian ginseng root coeluted with standard under different HPLC conditions, thus confirming this compound as 24(R)-pseudoginsenoside F11. Another ginsenoside often used to distinguish Asian and North American ginseng, ginsenoside Rf, was found in abundance in Asian ginseng roots at more than 0.021% (w/w). In Asian ginseng roots, the ratio of ginsenoside Rf to 24(R)-pseudoginsenoside F11 exceeded 700:1. The limit of detection of ginsenoside Rf or 24(R)-pseudoginsenoside F11 was 120 pg injected on-column, and the limit of quantification was 240 pg on-column. In summary, LC-MS-MS analysis of ginseng products for the presence and ratio of ginsenoside Rf and 24(R)-pseudoginsenoside F11 may be used for the unambiguous identification of Asian and North American ginsengs.

Asia↗

Ultrasound-assisted extraction of ginseng saponins from ginseng roots and cultured ginseng cells.

Ultrasound-assisted extraction was evaluated as a simpler and more effective alternative to conventional extraction methods for the isolation of ginsenosides (saponins) from various types of ginseng. The ginseng samples were extracted with different solvents, under either direct sonication by an ultrasound probe horn or indirect sonication in an ultrasound cleaning bath. The ultrasonic extraction was compared with the conventional method of refluxing boiling solvents in a soxhlet extractor, on the yields of both the total saponin isolated by thin-layer chromatography and the individual ginsenosides by high performance liquid chromatography. It was found that the sonication-assisted extraction of ginseng saponins was about three times faster than the traditional extraction method. The ultrasonic extraction was not only more efficient but also convenient for the recovery and purification of the active ingredients of plant materials. In addition, the sonication-assisted extraction can be carried out at lower temperatures which are favorable for the thermally unstable compounds.

Chromatography, Thin Layer↗

Epidemiological study on cancer prevention by ginseng: are all kinds of cancers preventable by ginseng?

In the light of experimental results, two case-control studies and one cohort study in a population of ginseng cultivation area were conducted to confirm whether ginseng has any anticarcinogenic effect on human cancers. All participants were interviewed using a standardised questionnaire to obtain the information on demographics, cigarette smoking, alcohol consumption and ginseng intake. In 905 pairs case-control study, 62% had a history of ginseng intake compared to 75% of the controls, a statistically significant difference (p<0.01). The odds ratio (OR) for cancer in relation to ginseng intake was 0.56. In extended case-control study with 1987 pairs, the ORs for cancer were 0.37 in fresh ginseng extract users, 0.57 in white ginseng extract users, 0.30 in white ginseng extract users, 0.30 in white ginseng powder users, and 0.20 in red ginseng users. Those who took fresh ginseng slices, fresh ginseng juice, and white ginseng tea, however, did not show decrease in the risk. Overall, the risk decreased as the frequency and duration of ginseng intake increased. With respect to the site of cancer, the ORs for cancers of the lip, oral cavity, pharynx, esophagus, stomach, colorectum, liver, pancreas, larynx, lung and ovary were significantly reduced by ginseng intake. Smokers with ginseng intake showed lower ORs for cancers of lung, lip, oral cavity and pharynx and liver than those without ginseng intake. In 5 yr follow- up cohort study conducted in the ginseng cultivation area, Kangwha-eup, ginseng intakers had significantly lower risk than non-intakers. As for the type of ginseng, cancer risk significantly decreased among intakers of fresh ginseng extract, alone or together with other ginseng preparations. Among 24 red ginseng intakers, no cancer death occurred during the follow-up period. The risk for stomach and lung cancers was significantly reduced by ginseng intake, showing a statistically significant dose-response relationship in each follow-up year. In conclusion, Panax ginseng C.A. Meyer has been established as non-organ specific cancer preventive, having dose response relationship. These results warrant that ginseng extracts and its synthetic derivatives should be examined for their preventive effect on various types of human cancers.

Antineoplastic Agents, Phytogenic↗

Proteome of Oriental ginseng Panax ginseng C. A. Meyer and the potential to use it as an identification tool.

Oriental ginseng (Panax ginseng C. A. Meyer) and American ginseng (Panax quinquefolius) are two widely used valuable traditional Chinese medicines (TCM). Previously, the identification of ginseng was mainly performed by analyzing the ginsengnosides using high performance liquid chromatography and amplification of polymorphic DNA using polymerase chain reaction. However, these methods cannot be used to distinguish TCM samples which are from different parts (main root, lateral roots, rhizome head and skin) of ginseng and ginseng culture cells from wild-grown ginseng. The present study aimed to identify different species of ginseng, different parts of the same ginseng and cultured cells of ginseng using a proteomic approach. Two-dimensional electrophoresis (2-DE) maps were established from the American ginseng main root, different parts (main root, lateral roots, rhizome head and skins) of Oriental ginseng and Oriental ginseng culture cells. Our results show that the 2-DE maps of different ginseng samples contain sufficient differences to permit easy discrimination. We have also identified common and specific protein spots in the 2-DE maps of different ginseng samples. The use of these "marker proteins" may help to speed up the identification process.

Chromatography, High Pressure Liquid↗

Null and opposing effects of Asian ginseng (Panax ginseng C.A. Meyer) on acute glycemia: results of two acute dose escalation studies.

OBJECTIVE: We have repeatedly reported that a batch of American ginseng with a specific ginsenoside (glycosidal saponin) profile decreases acute postprandial glycemia. We investigated whether Asian ginseng is able to replicate this glycemia-lowering efficacy in two separate acute dose escalation studies. METHODS: Each study was conducted in a separate sample of 11 healthy subjects (gender: 8M:3F and 6M:5F, age: 29 +/- 2y and 27 +/- 3y, BMI: 28.5 +/- 2.1 kg/m(2) and 26.9 +/- 1.4 kg/m(2)) using a randomized, single-blind, placebo-controlled, multiple-crossover design. Treatments consisted of 0 (placebo), 1, 2, and 3 g of Asian ginseng for the first study and 0 (placebo), 3, 6, and 9 g Asian ginseng for the second study administered 40 minutes before a 75g-OGTT protocol with blood drawn at -40, 0, 15, 30, 45, 60, 90, and 120 minutes. Ginsenosides were analyzed by HPLC-UV. RESULTS: Neither the main effect of pooled-treatment, nor dose, nor either factors interaction with time was significant for incremental plasma glucose and insulin. But the diagnostically and therapeutically relevant two-hour plasma glucose (2h-PG) value was significantly higher for pooled Asian ginseng treatment than placebo (5.46 +/- 0.31 versus 4.99 +/- 0.30 mmol/L, p = 0.050). Ginsenoside analyses showed that the Asian ginseng contained up to 96% lower and sevenfold higher quantities of various ginsenosides and their ratios than our previous efficacious batch of American ginseng. CONCLUSIONS: Asian ginseng showed both null and opposing effects on indices of acute postprandial plasma glucose and insulin. This is in contrast to our findings with American ginseng. One explanation may be the marked ginsenoside differences. Practitioners and consumers should be aware of ginseng's variable effects.

Acute Disease↗

Ginseng and ginsenoside Rg3, a newly identified active ingredient of ginseng, modulate Ca2+ channel currents in rat sensory neurons.

There is increasing evidence that ginseng influences pain modulation. In spite of extensive behavior studies, the detailed mechanism of ginseng actions at the cellular level and the identity of the active substance have not been elucidated yet. Whole-cell patch-clamp recordings were used to examine the modulation of high-voltage-activated Ca2+ channel currents by ginseng total saponins and its various individual ginsenosides in rat dorsal root ganglion neurons. Application of ginseng total saponins suppressed Ca2+ channel currents in a dose-dependent manner. Occlusion experiments using selective blockers revealed that ginseng total saponins could modulate L-, N-, and P-type currents. The co-application of ginseng total saponins and the gamma-opioid receptor agonist, D-Ala(2), N-MePhe(4), Gly(5)-ol-enkephalin (DAMGO), produced non-additive effects in most cells tested and each effect was significantly relieved by a depolarizing prepulse. Overnight treatment of cells with pertussis toxin profoundly reduced the inhibition. Furthermore, we now report that ginsenoside Rg3, among the major fractions of ginseng saponins, is a newly identified active component for the inhibition. These results suggest that the modulation of Ca2+ channels by ginseng total saponins, in particular by ginsenoside Rg3, could be part of the pharmacological basis of ginseng-mediated antinociception.

Analgesics, Opioid↗

Predicting color kinetics during red Asian ginseng (Panax ginseng) preparation.

A red Asian ginseng preparation was prepared as follows: fresh Asian ginseng (Panax ginseng) roots were first steamed for half an hour to several hours, and then the steamed roots were dried into died roots (red Asian ginseng, 10% moisture content, on a dry basis). During steaming, the color of ginseng (white) turned yellow and brown during subsequent steaming. Color is an important index for grading red Asian ginseng. In this study, the effects of drying time and temperature on the surface color formation (L, a/b) of the Asian ginseng, and the color formation (L, a/b) of the red Asian ginseng powder were investigated. The value of L decreased, while the value of a/b increased as drying proceeded; the value of L was slightly influenced by the drying temperature, but the value of a/b was markedly influenced. With respect to the color of the final product (red ginseng), the value of L was slightly influenced by steaming time and temperature, while the value of a/b was increased as steaming time and drying temperature increased. Based on the nth-order rate equation and Arrhenius equation, a kinetic model for describing these effects was established, and the results were satisfactorily fitting.

Algorithms↗

Influence of simulated microgravity environmental factor on ginseng cell growth and ginseng saponin content.

When ginseng (Panax ginseng C.A. Meyer) cells were subjected horizontal rotation on a clinostat, their growth and ginseng saponin content differed from those cultured in normal gravitational environments (control). Both fresh and dry weights of ginseng cells rotating on clinostat were higher than those of the control, and the difference in dry weight was particularly obvious. After 3 weeks of cultivation, saponin content in ginseng was 10% higher under the horizontal rotation treatment on the clinostat than that of the control. When ginseng cells were cultured on Ca2(+)-deprived medium and clinostatted for 3 weeks, their ginseng saponin content was almost twice of that of the control. Besides, in our experiments, the higher the Ca2+ concentration in medium, the lower the ginseng saponin content in the ginseng cells cultured.

Biomass↗

Effect of Asian and Siberian ginseng on serum digoxin measurement by five digoxin immunoassays. Significant variation in digoxin-like immunoreactivity among commercial ginsengs.

Asian and Siberian ginsengs contain glycosides with structural similarities to digoxin. We studied potential interference of ginseng in 5 digoxin immunoassays in 3 Asian (2 liquid extracts, 1 capsule) and 3 Siberian ginseng preparations (1 liquid extract, 2 capsules). With the fluorescence polarization immunoassay (FPIA), we observed apparent digoxin activity in 1 Asian liquid preparation and in the liquid extract and 1 capsule form of Siberian ginseng. In mice fed ginseng, we observed digoxin activities in the serum (Asian, 0.48-0.68 ng/mL [0.6-0.9 nmol/L]; Siberian, 0.20-0.47 ng/mL [0.3-0.6 nmol/L]), indicating that such interferences also occur in vivo. Serum pools prepared from samples from patients receiving digoxin and then supplemented with Asian or Siberian ginseng showed falsely increased digoxin values using the FPIA (e.g., for Asian ginseng, 1.54 ng/mL [2.0 nmol/L] vs control value, 1.10 ng/mL [1.4 nmol/L]) and falsely decreased values using the microparticle enzyme immunoassay (MEIA; 0.73 ng/mL [0.9 nmol/L] vs control value, 1.04 ng/mL [1.3 nmol/L]). Digoxin-like immunoreactive substances (DLISs) showed synergistic effects with ginsengs in interfering with the FPIA and MEIA for digoxin. No interference was observed with 3 other digoxin assays, even in the presence of elevated DLISs.

Animals↗

[Comparison of ITS sequences between wild ginseng DNA and garden ginseng DNA].

OBJECTIVE: To compare the genetic differences between wild ginseng and garden ginseng (Panax ginseng). METHOD: The sequences of ITS1 and ITS2 of wild ginsengs were determined on LKB DNA sequencing station through Si-liver Sequence DNA Sequencing System. The sequencies were aligned with DNA SIS software. RESULTS AND CONCLUSION: The ITS1 and ITS2 of Panax were 220-221 and 222-224 bases in length respectively. In Panax ginsehg, the seqences of ITS1 were very stable, but ITS2 were changeable. The ITS2 sequences of No. 87 and No. 110 of the wild ginseng collected from Fusong Heilongjiang (China) were exactly the same as those of No. U41680(Jun Wen) and No. U41682(Jun Wen) of garden ginseng collected from Heilongjiang Province (China) and Korea respectively, but different from those of No. U41681(Jun Wen) from Hubei Province (China) in three bases (447, 449, 450) The result implies that the cultivated ginsengs may have been introduced from two different populations of the wild ginseng.

Base Sequence↗

Differentiation and authentication of Panax ginseng, Panax quinquefolius, and ginseng products by using HPLC/MS.

An LC/MS-based method is established for the differentiation and authentication of specimens and commercial samples of Panax ginseng (Oriental ginseng) and Panax quinquefolius (American ginseng). This method is based on the separation of ginsenosides present in the ginseng methanolic extracts using high-performance liquid chromatography (HPLC), followed by detection with electrospray mass spectrometry. Differentiation of ginsenosides is achieved through simultaneous detection of intact ginsenoside molecular ions and the ions of their characteristic thermal degradation products. An important parameter used for differentiating P. ginseng and P. quinquefolius is the presence of ginsenoside Rf and 24-(R)-pseudoginsenoside F11 in the RICs of Oriental and American ginsengs, respectively. It is important to stress that ginsenoside Rf and 24(R)-pseudoginsenoside F11, which possess the same molecular weight and were found to have similar retention times under most LC conditions, can be unambiguously distinguished in the present HPLC/MS method. The method developed is robust, reliable, reproducible, and highly sensitive down to the nanogram level.

Chromatography, High Pressure Liquid↗

[Systemic breeding of the new variety of Biantiao ginseng (Panax ginseng)].

OBJECTIVE: To breed new varieties of Biantiao ginseng for high yield and fine quality. METHODS: Systemic breeding methods were applied. About 3,000 outstanding Biantiao ginseng roots were selected and planted in breeding field, and self-crossed for four generations. During the course, inferior lines or plants were rejected. Then strain comparison, identification of resistance to black-speck disease, and analysis of active compositions were carried out. RESULTS: "Biantiao 1" (BT1), the first new variety of Biantiao ginseng, with green stems and thick, long, elegant roots and median resistance to black-speck disease, has been harvested since 20 years. The percentage of Biantiao ginseng roots and yield were 15% and 30% higher than the control's respectively. The content of total ginsenosides and the main monomers was 1.8%-2.5% higher than the control's. The characteristics of overground part and root of BT1 were uniform and stable. CONCLUSIONS: BT1, a new excellent ginseng variety, has a good potential value to be generalized in ginseng production.

Breeding↗

The physical map of the chloroplast DNA from Korean ginseng (Panax ginseng C.A. Meyer).

To compare the gene order of the chloroplast genome among dicotyledonous plants, we constructed a physical map of chloroplast DNA (cpDNA) of Korean ginseng (Panax ginseng C.A. Meyer) with four restriction enzymes, BamHI, HindIII, EcoRI, and PstI. The restriction enzyme recognition sites of the physical map were also confirmed by Southern hybridization of total ginseng cpDNA with homologous and heterologous probes. The cpDNA of Korean ginseng was determined as a circular molecule with a total size of about 154 kb, which contain two inverted repeats of 23 kb each that disrupt the rest of the molecule into a large (90 kb) and a small single copy region (18 kb). The genome structure of Korean ginseng cpDNA was similar in size and gene order to that of tobacco cpDNA. The cpDNA of Korean and American ginseng (P. quinquefolius) showed very similar restriction patterns.

Chromosome Mapping↗

Dissecting the anti-obesity components of ginseng: How ginseng polysaccharides and ginsenosides target gut microbiota to suppress high-fat diet-induced obesity.

INTRODUCTION: Ginseng demonstrates therapeutic potential in treating obesity, with both experimental and clinical studies suggesting its anti-obesity effects are mediated by gut microbiota. Nonetheless, the specific chemical components responsible for this effect remain largely unidentified. OBJECTIVES: This study aims to investigate the anti-obesity effects and mechanisms of ginseng polysaccharides (GP) and ginsenosides (GS), the primary chemical components of ginseng, with a focus on their impact on gut microbiota. METHODS: The impact of GP and GS on high-fat diet (HFD)-induced obesity was assessed using a mouse model. Molecular mechanisms were explored through a combination of chemical analysis, metagenomics, RT-qPCR, ELISA, and biochemical assays. RESULTS: GP or GS administration effectively prevented adiposity in HFD-fed mice, and both effects were mediated by gut microbiota. Chemical analysis revealed diverse glycosyl groups in GP and GS. Metagenomics data suggested that GP-enriched species, e.g., Bacteroides stercorirosoris and Clostridiales bacterium encoded carbohydrate-active enzymes GH35, GH43 and PL9_1, while GS-enriched Sulfurospirillum halorespirans encoded GH16_5. These enzymes facilitated the utilization of glycosyl groups in GP and GS, selectively stimulating bacterial growth and reshaping the gut microbiota. Furthermore, bacterial species enriched by GP or GS encoded specific functional genes involved in short-chain fatty acid (SCFA) synthesis (K00625 and K00925 for GP; K18118, K00100, and K18122 for GS) and intestinal gluconeogenesis (IGN) (K01678, K00024, and K01596 for GP; K18118 and K00278 for GS). Consequently, the SCFA-GLP-1/PYY signaling and IGN were activated by both GP and GS to ameliorate obesity phenotypes. CONCLUSION: GP and GS, containing diverse glycosyl groups, selectively stimulate specific gut bacteria, triggering mechanisms involved in SCFA-GLP-1/PYY signaling and IGN activation to reduce adiposity in HFD-fed mice. The study enhances understanding of the chemical components crucial for the gut microbiota-mediated anti-obesity effect of ginseng. The mechanistic understanding provides valuable insights for developing ginseng-based drugs or health products to combat obesity.

Gastrointestinal Microbiome↗