Differences in the Volatile Compositions of Ginseng Species ( Panax sp.)

The volatile compositions in dried white ginseng according to species ( Panax ginseng , Panax notoginseng , and Panax quinquefolius ) were analyzed and compared by applying multivariate statistical techniques to gas chromatography-mass spectrometry data sets. Main volatile compounds of ginseng species in the present study were sesquiterpenes, such as bicyclogermacrene, (E)-β-farnesene, β-panasinsene, calarene, α-humulene, β-elemene, etc. In particular, α-selinene, α-terpinolene, β-bisabolene, β-phellandrene, β-sesquiphellandrene, zingiberene, germacrene D, limonene, α-gurjunene, (E)-caryophyllene, δ-cadinene, (E)-β-farnesene, α-humulene, bicyclogermacrene, longiborn-8-ene, β-neoclovene, and (+)-spathulenol were mainly associated with the difference between P. ginseng and P. notoginseng versus P. quinquefolius species. On the other hand, the discrimination between P. ginseng and P. notoginseng could be constructed by hexanal, 2-pyrrolidinone, (E)-2-heptenal, (E)-2-octenal, heptanal, isospathulenol, (E,E)-2,4-decadienal, 3-octen-2-one, benzaldehyde, 2-pentylfuran, and (E)-2-nonenal.     

Phytochemistry of wild populations of Panax quinquefolius L. (North American ginseng)

A survey of the phytochemistry of Panax quinquefolius L. (North American ginseng) collected from wild populations in Ontario, Quebec, Maine, Vermont, and Wisconsin was undertaken. Reverse-phase HPLC was used to determine the natural variation of levels of ginsenosides Rg1, Re, Rf, Rb1, Rc, Rb2, and Rd and their total in leaf, stem, and root of authentic wild-grown material. The totals in roots varied from 1 to 16%, with the greatest number of individual samples having 4-5% total ginsenosides. The lack of ginsenoside Rf in roots of authentic wild populations confirmed its status as a phytochemical marker differentiating American and Asian ginseng. Ten geographically isolated wild populations were collected, and several showed significant variation in the levels of major ginsenosides. There was no statistical difference in mean ginsenoside content between wild and cultivated P. quinquefolius roots at 4 years of age, suggesting there is no phytochemical justification for wild crafting. Baseline data on total ginsenoside levels for authentic wild P. quinquefolius reported here provide reference levels for quality assurance programs.     

DNA identification of commercial ginseng samples

An investigation was performed with the objective of developing a DNA-based protocol for the identification of commercial samples of the herbal compound ginseng. There are currently two major herbal products referred to as ginseng. They are Korean or Chinese ginseng (Panax ginseng) and American ginseng (Panax quinquefolius). The market for ginseng in the United States is estimated to be approximately $300 million annually. Current tests for ginseng species identification rely on expert botanical identification of fresh plant/root specimens or on biochemical characterization of active and marker compounds (e.g., ginsenosides). For the determination of the feasibility of ginseng identification by DNA analysis, a strategy based on the direct DNA sequence analysis of the nuclear ribosomal internal transcribed spacer region was developed. Other genetic tests included sequence analysis of the chloroplast ribulose 1,5-bisphosphate carboxylase large subunit gene and DNA fingerprinting by the rapid amplification of polymorphic DNA technique. To confirm the results, each ginseng sample was identified using high-performance liquid chromatography. All methods were successful in distinguishing American from Korean ginseng. In addition, the protocol was improved for the isolation of genomic and plastid DNA from commercial ginseng preparations by incorporating an impact homogenization step into the standard column chromatography purification procedure.     

人参EST-SSR标记的建立

从7055条人参EST序列中共搜索出791个SSR。根据引物设计标准,设计了68对EST-SSR引物。在合适的PCR反应体系下,分别以人参品种集安长脖、抚松二马牙的DNA为模板,对设计的EST-SSR引物进行筛选,发现有43对EST-SSR引物能扩增出产物。在9个人参品种、2个西洋参品种和2个刺五加品种中进一步对这些可扩增的引物对进行多态性检测,发现有26对引物显示多态性,占可扩增引物的60.47%,占设计引物总数的38.23%。本研究结果表明,根据人参EST建立EST-SSR标记是一种行而有效的的方法。     

Metabolomic approach for age discrimination of Panax ginseng using UPLC-Q-Tof MS

An ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-Tof MS)-based metabolomic technique was applied for metabolite profiling of 60 Panax ginseng samples aged from 1 to 6 years. Multivariate statistical methods such as principal component analysis and hierarchical clustering analysis were used to compare the derived patterns among the samples. The data set was subsequently applied to various metabolite selection methods for sophisticated classification with the optimal number of metabolites. The results showed variations in accuracy among the classification methods for the samples of different ages, especially for those aged 4, 5, and 6 years. This proposed analytical method coupled with multivariate analysis is fast, accurate, and reliable for discriminating the cultivation ages of P. ginseng samples and is a potential tool to standardize quality control in the P. ginseng industry.     

Simplified extraction of ginsenosides from American ginseng (Panax quinquefolius L.) for high-performance liquid chromatography-ultraviolet analysis

Four methods were tested for extraction and recovery of six major ginsenosides (Rb1, Rb2, Rc, Rd, Re, and Rg1) found in roots of American ginseng (Panax quinquefolius): method A, sonication in 100% methanol (MeOH) at room temperature (rt); method B, sonication in 70% aqueous MeOH at rt; method C, water extraction (90 degrees C) with gentle agitation; and method D, refluxing (60 degrees C) in 100% MeOH. After 0.5-1 h, the samples were filtered and analyzed by high-performance liquid chromatography (HPLC)-UV. A second extraction by methods C and D was done, but 85-90% of ginsenosides were obtained during the first extraction. Lyophilization of extracts did not influence ginsenoside recovery. Method D resulted in the highest significant recoveries of all ginsenosides, except Rg1. Method C was the next most effective method, while method A resulted in the lowest ginsenoside recoveries. Method B led to similar recoveries as method C. All methods used one filtration step, omitted time-consuming cleanup, but maintained clear peak resolution by HPLC, and can be used for quantitative screening of ginsenosides from roots and commercial ginseng preparations.     

Sanchi ginseng (<Emphasis Type="Italic">Panax notoginseng</Emphasis> (Burkill) F. H. Chen) in China: distribution,cultivation and variations

Sanchi ginseng (Panax notoginseng (Burkill) F. H. Chen) has been cultivated in China for more than 400 years, whose root is an important traditional Chinese medicine mainly used to stanch blood, to disperse gore and to reduce pain caused by injury due to falls. The cultivated populations of this crop are distributed in Wenshan mountain area of Yunnan with a narrow habitat, whose location is around N 23.5°, E 104° and altitude ranges from 1,200 to 2,000 m. Although its wild species has not been found, current cultivated populations show rich morphological variations in stem, leaf, root, flower and fruit. Recent studies exhibit that Sanchi root has more active compounds than the roots of P. ginseng and P. quinquefolius. Therefore, this crop needs further utilization.     

Protopanaxatriol-type ginsenosides from the root of Panax ginseng

Ginseng, the root of Panax ginseng C. A. Meyer (Araliaceae), is one of the most important traditional medicines and functional foods. A detailed phytochemical investigation on the roots of P. ginseng led to the isolation of 6 new natural protopanaxatriol (PPT)-type ginsenosides, ginsenosides Re(1)-Re(6) (compounds 1-6), along with 10 known PPT-type ginsenosides. Their structures were elucidated on the basis of chemical and spectroscopic analyses, including high-resolution mass spectrometry (HRMS) and 1D and 2D nuclear magnetic resonance (NMR). The unusual α-D-glucopyranosyl-(1→3)-β-D-glucopyranosyl sugar chain, as found in compounds 1 and 2, is reported in the genus Panax for the first time.     

Analysis of organophosphorus pesticides in dried ground ginseng root by capillary gas chromatography-mass spectrometry and -flame photometric detection

A method was developed to determine organophosphorus pesticides (OPs) in dried ground ginseng root. Pesticides were extracted from the sample using acetonitrile/water saturated with salts, followed by solid-phase dispersive cleanup, and analyzed by capillary gas chromatography with electron ionization mass spectrometry in selective ion monitoring mode (GC-MS/SIM) and flame photometric detection (GC-FPD) in phosphorus mode. The detection limits for most of the pesticides were 0.025-0.05 microg/g using GC-FPD but were analyte-dependent for GC-MS/SIM, ranging from 0.005 to 0.50 microg/g. Quantitation was determined from 0.050 to 5.0 microg/g with r 2 > 0.99 for a majority of the pesticides using both detectors. Recovery studies were performed by fortifying the dried ground ginseng root samples to concentrations of 0.025, 0.1, and 1.0 microg/g, resulting in recoveries of >90% for most pesticides by GC-FPD. Lower (<70%) and higher (>120%) recoveries were most likely from complications of pesticide lability or volatility, matrix interference, or inefficient desorption from the solid-phase sorbents. There was difficulty in analyzing the ginseng samples for the OPs using GC-MS at the lower fortification levels for some of the OPs due to lack of confirmation. GC-FPD and GC-MS/SIM complement each other in detecting the OPs in dried ground ginseng root samples. This procedure was shown to be effective and was applied to the analysis of OPs in ginseng root samples. One particular sample, a ground and dried American ginseng (Panax quinquefolius) root sample, was found to contain diazinon quantified at approximately 25 microg/kg by external calibration using matrix-matched standards or standard addition using both detectors. The advantage of using both detectors is that confirmation can be achieved using GC-MS, whereas the use of a megabore column in GC-FPD can be used to quantitate some of the nonpolar OPs without the use of matrix-matched standards or standard addition.     

三种人参属植物的EST-SSR信息分析及其在三七中的应用

为了解人参属植物EST中SSR分布特点及其在三七SSR标记中的应用,利用生物信息学方法,用primer3软件对dbEST数据库中人参属植物人参、西洋参和三七的EST序列进行搜索,发现人参属植物EST-SSR出现频率为11.54%,平均每4.39kb出现1个SSR。人参属植物单核苷酸重复基元占主导地位,其次为三核苷酸重复基元和二核苷酸重复基元,分别占总SSR的54.48%、17.31%和16.36%。人参属EST-SSR的优势类型为A/T和AT/TA,分别占54.73%和8.21%。根据人参属植物EST中的SSR设计48对引物,在合适的PCR扩增体系下,用5个三七样品的DNA为模板进行PCR扩增,引物有效扩增率85.42%,其中多态性引物占可扩增引物的70.73%。结果表明,利用人参属EST序列开发三七EST-SSR标记是可行的。     

Simultaneous quantification of ginsenosides in American ginseng (Panax quinquefolium) root powder by visible/near-infrared reflectance spectroscopy.

Near-infrared reflectance spectroscopy (NIRS) was examined as a possible alternative to high-performance liquid chromatography (HPLC) for the analysis of ginsenosides from American ginseng (Panax quinquefolium) root powder (n = 26). NIR spectra were collected over 400-2500 nm. For each sample and individual ginsenoside quantified by HPLC, spectral data were regressed against the chemical data to develop prediction equations. The spectral prediction equations produced high correlation coefficient (1-VR) values and low standard errors of cross validation (SECV) values for the determination of individual and total ginsenosides. The contents of individual ginsenosides, Rb(1), Rb(2), Rc, Rd, Re, Rg(1), Ro, m-Rb(1), m-Rb(2), m-Rc, m-Rd, and total ginsenosides (X +/- SECV) were (1.29+/-0.18)%, (0.273+/-0.096)%, (0.298+/-0.052)%, (0.091+/-0.027)%, (1. 015+/-0.114)%, (0.116+/-0.018)%, (0.25+/-0.040)%, (0.776+/-0.116)%, (0.197+/-0.074)%, (0.239+/-0.083)%, (0.143+/-0.042)%, and (4.393+/-0.283)%, respectively. The (1-VR) values of cross validation were 0.877, 0.872, 0.955, 0.834, 0.899, 0.919, 0.325, 0.849, 0.902, 0.877, 0.871, and 0.963, respectively. Results indicated that the NIRS method could be used for the analysis of the major ginsenosides, Rb(1), Re, and m-Rb(1), as well as the total ginsenosides in American ginseng.     

Quantification of two polyacetylenes in Radix Ginseng and roots of related Panax species using a gas chromatography-mass spectrometric method

A sensitive method for quantitating the pharmacologically active polyacetylenes panaxynol and panaxydol in Radix Ginseng was developed using a capillary gas chromatography-mass spectrometric (GC-MS) method. The detection mode of selected ion monitoring (SIM) allowed sensitive and selective quantitation of the two compounds in ginseng. Method validation showed that the GC-MS method has much lower detection and quantitation limits than the high-performance liquid chromatography (HPLC)-UV method. This indicates that GC-MS is particularly useful for the analysis of polyacetylene compounds, which have relatively low abundances compared with ginsenosides in ginseng. Based on the quantitative results of different types of ginseng herbs, it was found that the panaxydol and panaxynol contents were higher in forest ginseng than in cultivated ginseng. This method was further applied to the quantitative analyses of panaxynol and panaxydol in Radix Notoginseng and American ginseng. The ratio of panaxydol to panaxynol can be utilized as a marker for differentiating ginseng, notoginseng, and American ginseng. This study introduces the first GC-MS method for the quantitative analysis of polyacetylenes in herbs of the Panax genus.     

不同生育期同化的光合产物在人参体内的转移和分配

在人参不同生育期,采用(14)~CO_2标记整体植株的方法,测定人参同化(14)~C产物的分配比率和残留率。结果表明,展叶末期同化的光合产物60％用于建造人参营养体,为地上部各器官的生长发育提供碳源和能源;绿果期以后的同化产物是参根中干物质和有效成份构成的主要来源;施用人参复合肥料,4年生参根干重及总皂甙含量较对照提高16.1％和4.3％。     

应用~(14)C示踪法测定人参的光合速率

人参光合速率是研究人参光合生理特性的重要依据。本研究测得人参光饱和强度为10—20klx,适宜光照强度为6—20klx,参棚适宜透光率为25％,蓝膜的光合效率高。     

Genetic diversity and variation of saponin contents in Panax notoginseng roots from a single farm

Radix notoginseng, the root of Panax notoginseng (Burk.) F. H. Chen, has been widely used in traditional Chinese medicine. Its main components, saponins, have been reported to have many pharmacological activities. To test the general assumption that herbs of a single species planted and harvested from a single location are uniform in chemical and genetic makeup, chemical analysis and DNA fingerprinting were carried out. High-performance TLC together with HPLC analysis were used to analyze 17 randomly sampled 3-year-old roots from a single farm for the presence of six saponins. Five roots showed distinct chemical profiles with changed ratios of ginsenosides Rd/Rg1, Re/Rg1, or Rb1/Rg1. The same samples, together with some 1- and 2-year-old samples, were also subjected to fluorescent amplified fragment length polymorphism (AFLP) analysis, and their internal transcribed spacer 2 (ITS 2) regions were sequenced. Fluorescent AFLP analysis was found to be much more polymorphic than the ITS 2 sequence and showed clear evidence of genetic diversity within the tested population. In conclusion, genetic diversity and variation of saponin contents between individual P. notoginseng roots have been detected. We suggest that genetic diversity affects the contents of the six saponins. The saponin contents variation and genetic diversity were also found among P. notoginseng root samples collected from China and Singapore markets. Since variable saponin contents may affect therapeutic efficacy, combining the use of genetic profiling with chemical profiling will help ensure greater uniformity in the quality of P. notoginseng roots. The genetic and chemical diversity within a population also provides the opportunity for breeding new cultivars with more desirable chemical constituents.     

Degradation of ginsenosides in American ginseng (Panax quinquefolium) extracts during microwave and conventional heating

The degradation of ginsenosides in American ginseng (Panax quinquefolium) extracts during microwave and water (oil) bath heating (conventional heating) was investigated. Both the 50% ethanol-water extracts and the aqueous extracts were boiled in a modified laboratory microwave oven and in a water (or oil) bath, respectively. The neutral ginsenosides (Rb(1), Rc, Rd, and Re) and malonyl ginsenosides (m-Rb(1), m-Rc, and m-Rd) were determined by reverse-phase high-performance liquid chromatography. The results showed that the degradation of ginsenosides in 50% ethanol-water extracts was a first-order reaction. The malonyl ginsenosides were much less stable than the corresponding neutral ginsenosides, with the rate constant value of the malonyl ginsenosides being 3-60 times that of the neutral ginsenosides. At the same temperature, the effect of microwave heating on the degradation of ginsenosides was the same as that of conventional heating.     

Weight gain reduction in mice fed Panax ginseng saponin, a pancreatic lipase inhibitor

Roots of the herb Panax ginseng are known to contain high levels of bioactive saponins. Here, we isolated saponins from ginseng root powder and studied their inhibitory effect on the absorption of dietary fat in male Balb/c mice. Consumption of ginseng saponins suppressed the expected increase in body weight and plasma triacylglycerols, following a high-fat diet and observed higher intake. Consumption of ginseng saponins had no effect on the concentration of the total plasma cholesterol in both chow and high-fat diets in mice. The mode by which saponins from ginseng inhibit lipid metabolism was assessed as the in vitro inhibition of pancreatic lipase. Ginseng saponin inhibited pancreatic lipase with an apparent IC50 value of 500 mug/mL. Our results suggest that the anti-obesity and hypolipidemic effects of Ginseng in high-fat diet-treated mice were attributed to the isolated saponin fraction. These metabolic effects of the ginseng saponins may be mediated by inhibition of pancreatic lipase activity.     

Simultaneous determination of ginsenosides and polyacetylenes in American ginseng root (Panax quinquefolium L.) by high-performance liquid chromatography

A method for simultaneous determination of ginsenosides and polyacetylenes in Panax quinquefolium L. (American ginseng) roots was developed. The ginsenosides Rb1, Rb2, Rc, Rd, Re, Rg1, Ro, malonyl-Rb1, malonyl-Rc, and malonyl-Rd and the polyacetylenes falcarinol and panaxydol were extracted from fresh ginseng roots in a sequential extraction process with 100% methanol followed by 80% aqueous methanol and quantified simultaneously in extracts by high-performance liquid chromatography using diode array detection. Separations were achieved with a phosphate buffer-acetonitrile gradient system using an RP-C18 column. Except for Rd, the present extraction method resulted in similar or significantly higher concentrations of both ginsenosides and polyacetylenes in comparison to commonly used extraction methods for these compounds. The contents of polyacetylenes and ginsenosides were determined in the root hairs, lateral roots, and main roots of 6 year old ginseng plants. The total mean concentrations of ginsenosides and polyacetylenes in root hairs were 31.0 g/kg fresh weight (FW) and 2.6 g/kg FW, respectively, whereas the concentrations of these bioactive compounds in the main roots were significantly lower with total mean concentrations of 17.8 g/kg FW for ginsenosides and 0.6 g/kg FW for polyacetylenes. The concentration of individual and total ginsenosides and polyacetylenes did not differ significantly between main roots of different sizes. Consequently, it is possible to do quantitative screening for ginsenosides and polyacetylenes to breed ginseng roots with higher levels of bioactive compounds.     

Panax ginseng extract rich in ginsenoside protopanaxatriol attenuates blood pressure elevation in spontaneously hypertensive rats by affecting the Akt-dependent phosphorylation of endothelial nitric oxide synthase

Nitric oxide (NO) produced by endothelial nitric oxide synthase (eNOS) is a fundamental regulator of systemic blood pressure. Ginsenosides from Panax ginseng have been investigated in vitro for the molecular and biochemical mechanisms by which they stimulate NO release in vascular endothelial cells; however, little research has been done to confirm the physiological relevance of these in vitro studies. To address this research gap, the effects of a P. ginseng extract rich in ginsenosides from protopanaxatriol on spontaneously hypertensive rats (SHRs) was examined. Ginseng extract administration stimulated nongenomic Akt-mediated eNOS activation, enhanced NO production, improved vessel wall thickening, and alleviated hypertension in SHRs, confirming the physiological relevance of previous in vitro studies with ginsenosides.     

Effects of population, age, and cultivation methods on ginsenoside content of wild American ginseng (Panax quinquefolium)

Genotype and environmental effects on ginsenoside content among eight wild populations of American ginseng (Panax quinquefolium) were investigated. Root concentrations of six ginsenosides were determined at the time of collection of plants from the wild (T0) and 2 years (T2) after transplanting roots from each of the eight populations to each of two different forest garden locations. Both location and population had significant effects on root and shoot growth. Overall, ginsenoside Rb1 was most abundant, followed by Rg1 and Re. Concentrations of Rg1 and Re were inversely related among and within populations. The relative ranking of populations differed depending upon the particular ginsenoside and sampling time. The relative importance of genotype and environment was not the same for all ginsenosides. Ginsenoside Re was influenced by population but not location, whereas Rb1, Rc, and Rb2 were influenced only by location (environment), while Rg1 and Rd were influenced by both. Ginsenoside levels were consistently lower, but growth was consistently higher at the more intensively managed garden location.