Analytical discrimination between sources of ginseng using Raman spectroscopy

Ginseng is a widely used medicinal product that grows mainly in Korea, China and America. American ginseng is classified as an endangered species, and so the import and export of this type of ginseng is illegal in certain countries. Due to this restriction it is becoming increasingly important to be able to distinguish between different types of ginseng. FT-Raman spectroscopy has the ability to discriminate between ginseng specimens according to the country of origin and the effects of processing on the ginseng material. The ginsenoside content of ginseng differs in both conformation and concentration depending on the source of the ginseng, which means that ginseng grown in different countries should express unique spectral features. The presence or absence of these features, therefore, could indicate the geographical origin of the sample. Several spectral features were identified for a range of ginsengs, such as a peak at 980 cm(-1) that was only found in Chinese ginseng, and the different wavenumber positions of characteristic ginseng bands near 1600 cm(-1). This indicates that Raman spectroscopy can be used to pinpoint the origin of an unknown ginseng sample and that it would provide a rapid nondestructive analytical technique for formally discriminating between restricted and permitted imports.     

Determination of polyacetylenes and ginsenosides in Panax species using high performance liquid chromatography

A new HPLC method was developed to separate and identify three polyacetylenes (panaxynol, panaxydol and 1,8-heptadecadiene-4,6-diyne-3,10-diol) found in Panax species. The mobile phase was a linear gradient of 2 : 1 : 3 to 2 : 1 : 1 (v/v/v) methanol/acetonitrile/water in 40 min. HPLC analysis was performed at a flow rate of 1.5 ml/min with UV detection at 254 nm. The contents of the polyacetylenes and ginsenosides in Panax ginseng (white ginseng and red ginseng), P. quinquefolium, P. japonicus, and P. noteginseng were determined using these methods. The species containing the highest polyacetylene content (0.080%) was P. quinquefolium cultivated in Nagano, Japan. Meanwhile, the species with the highest ginsenoside content (9.176%) was P. noteginseng cultivated in Yunnan, China.     

Rapid authentication of ginseng species using microchip electrophoresis with laser-induced fluorescence detection

Ginseng is one of the most expensive Chinese herbal medicines and the effectiveness of ginseng depends strongly on its botanical sources and the use of different parts of the plants. In this study, a microchip electrophoresis method coupled with the polymerase chain reaction (PCR)–short tandem repeats (STR) technique was developed for rapid authentication of ginseng species. A low viscosity hydroxypropyl methylcellulose (HPMC) solution was used as the sieving matrix for separation of the amplified STR fragments. The allele sizing of the amplified PCR products could be detected within 240 s or less. Good reproducibility and accuracy of the fragment size were obtained with the relative standard deviation for the allele sizes less than 1.0% (n=11). At two microsatellite loci (CT 12, CA 33), American ginseng had a different allele pattern on the electropherograms compared with that of the Oriental ginseng. Moreover, cultivated and wild American ginseng can be distinguished on the basis of allele sizing. This work establishes the feasibility of fast genetic authentication of ginseng species by use of microchip electrophoresis.     

Liquid chromatography/mass spectrometry of malonyl-ginsenosides in the authentication of ginseng

Different negative ion electrospray (ES) source conditions are required to concentrate the ion current in [M-H](-) for malonylated and non-malonylated ginsenosides. However, both can be ionised optimally in a single liquid chromatography/mass spectrometry (LC/MS) analysis by employing switchable voltages in the post-source ion optics of a quadrupole ion trap mass spectrometer. Coupled with automatic MS/MS scanning and post-acquisition neutral loss data analysis, this method provides a means of profiling the malonylated and acetylated ginsenosides in ginseng extracts. Analyses revealed numerous malonylated ginsenosides that could be partially characterised by serial MS/MS experiments. The ratio of mRb(1) to other isomeric forms present and to mRb(2) and mRc appears to show consistent differences among Panax ginseng (Asian ginseng), P. quinquefolius (American ginseng) and P. notoginseng (Sanchi ginseng). The ratio of malonylated to non-malonylated ginsenosides is reduced in the red form of Asian ginseng compared with the white form and there is a concomitant increase in the levels of the corresponding acetylated ginsenosides. The ability to analyse malonylated ginsenosides is an important contribution to the range of chemical characteristics that can be used to authenticate the different species of ginseng and will assist in quality control and standardisation.     

Neuroprotective Effect and Antioxidant Potency of Fermented Cultured Wild Ginseng Root Extracts of Panax ginseng C.A. Meyer in Mice

Wild ginseng has better pharmacological effects than cultivated ginseng. However, its industrialization is limited by the inability to grow wild ginseng on a large scale. Herein, we demonstrate how to optimize ginseng production through cultivation, and how to enhance the concentrations of specific ginsenosides through fermentation. In the study, we also evaluated the ability of fermented cultured wild ginseng root extract (HLJG0701-β) to inhibit acetylcholinesterase (AChE), as well as its neuroprotective effects and antioxidant activity. In in vitro tests, HLJG0701-β inhibited AChE activity and exerted neuroprotective and antioxidant effects (showing increased catalyst activity but decreased reactive oxygen species concentration). In in vivo tests, after HLJG0701-β was orally administered at doses of 0, 125, 250, and 500 mg/kg in an animal model of memory impairment, behavioral evaluation (Morris water maze test and Y-maze task test) was performed. The levels of AChE, acetylcholine (ACh), blood catalase (CAT), and malondialdehyde (MDA) in brain tissues were measured. The results showed that HLJG0701-β produced the best results at a dose of 250 mg/kg or more. The neuroprotective mechanism of HLJG0701-β was determined to involve the inhibition of AChE activity and a decrease in oxidative stress. In summary, both in vitro and in vivo tests confirmed that HJG0701-β administration can lead to memory improvement.     

人参栽培研究进展

人参作为经济作物在中国有着数千年的种植历史。人参具有多种药理活性,在食品、医药和化妆品等领域有着广泛的应用。然而,目前人参的实际产量并不能满足市场需求。提升人参生产效率仍是当前阶段人参种植研究工作的首要目标。近些年,在人参选地、种子处理、播种、搭棚、培育和采收等生产环节中,涌现了许多新模式、新技术、新方法和新仪器。本文从栽培模式和技术角度综述人参栽培的研究现状,希望为进一步提升人参生产效率提供参考。     

气相色谱-三重四极杆质谱分析人参炮制品中的挥发性成分

人参炮制的化学成分变化研究主要集中在皂苷和糖类,本文首次从挥发性成分角度阐释了人参不同炮制品的物质基础。利用气相色谱-质谱联用(GC-MS/MS)方法,对鲜参、生晒参和红参中挥发性成分及其衍生规律进行研究。采用TG-5SILMS非极性气相色谱柱,以He为载气,通过NIST MS Spectral Database对挥发性成分进行检测并鉴定。鲜参、生晒参、红参中分别检出30、33和34种挥发性成分,其中生晒参中(-)-斯巴醇含量为鲜参含量的31.98倍,辛醛等8种挥发性成分为鲜参中含量的3倍以上,红参中有环癸等10种挥发性成分为鲜参中含量的3倍以上。生晒参和红参中各有4种挥发性成分在鲜参中未检出。     

Differentiation of <Emphasis Type="BoldItalic">Panax quinquefolius</Emphasis> grown in the USA and China using LC/MS-based chromatographic fingerprinting and chemometric approaches

American ginseng (Panax quinquefolius) is one of the most commonly used herbal medicines in the world. Discriminating between P. quinquefolius grown in different countries is difficult using traditional quantitation methods. In this study, a liquid chromatographic mass spectrometry fingerprint combined with chemometric analysis was established to discriminate between American ginseng grown in the USA and China. Fifteen American ginseng samples grown in Wisconsin and 25 samples grown in China were used. The chromatographic fingerprints, representing the chemical compositions of the samples, made it possible to distinguish samples from the two locations. In addition, it was found that some ginsenosides varied widely from P. quinquefolius cultivated in these two countries. P. quinquefolius grown in the USA is higher in ginsenoside R_c, ginsenoside R_d, quinquenoside III/pseudo-ginsenoside RC₁, malonyl ginsenoside R_b1, and ginsenoside R_b2, but lower in ginsenoside R_b1 compared with P. quinquefolius grown in China. These ginsenosides may be responsible for the class separation seen using fingerprinting and chemometric approaches.     

矿物元素指纹图谱的建立及在西洋参产地鉴别中的应用研究

采用电感耦合等离子体质谱法测定了山东、吉林、美国和加拿大4个产地西洋参中50种矿物元素的含量,研究了不同产地西洋参矿物元素的差别和转换系数,构建了西洋参的矿物元素指纹图谱。以各产地矿物元素含量的平均值构建了山东、吉林、美国和加拿大产西洋参的矿物元素标准指纹图谱。采用SPSS 20.0计算了各西洋参矿物元素指纹图谱与其矿物元素标准指纹图谱的相似度,确定了山东、吉林、美国和加拿大产西洋参矿物元素指纹图谱的相似度阈值分别为0.93、0.91、0.98和0.93。通过比较未知产地西洋参矿物元素指纹图谱与矿物元素标准指纹图谱的相似度,进行西洋参的产地判别。采用20批未知产地西洋参样品验证模型的准确性,正确率为85%。此外,研究表明,不同生长年限和不同部位西洋参样品对所建立的西洋参产地鉴别方法无影响。     

Low-density lipoprotein protects Vibrio vulnificus-induced lethality through blocking lipopolysaccharide action

Lipoprotein plays a role in the host defense against bacterial infection, and its serum level has been demonstrated to be an important prognosis factor of survival. We have previously demonstrated that LDL directly inactivates the hemolytic activity of Vibrio vulnificus cytolysin (VVC) in vitro. The object of this study was therefore to examine whether the LDL-mediated inactivation of VVC leads to protection against lethal infection of V. vulnificus in vivo, using wild and VVC-deficient V. vulnificus strains. Unexpectedly, we found that LDL protects mouse lethality induced by VVC-deficient as well as wild V. vulnificus strain. We also demonstrated that LDL blocks V. vulnificus LPS-induced lethality in mice. These results suggest that LDL preferentially act on endotoxin rather than exotoxin in the protection against V. vulnificus-induced mice lethality.     

Discrimination among Panax species using spectral fingerprinting

Spectral fingerprints of samples of three Panax species (P. quinquefolius L., P. ginseng, and P. notoginseng) were acquired using UV, near-infrared (NIR), and MS. With principal component analysis, all three methods allowed visual discrimination among the three species. All three methods were able to discriminate between white and red ginseng, and showed distinctive subgroupings of red ginseng related to root quality (age/size). Analysis of variance was used to evaluate the relative variance arising from the species, run, and analytical uncertainty, and was used to identify the most information-rich portions of the spectrum for NIR and UV. Accurate classification of the three species was obtained by using partial least squares-discriminant analysis and a fuzzy rule-building expert system. Relatively poor accuracy was obtained using soft independent modeling of class analogy when a single component was used.     

Distinguishing Ontario ginseng landraces and ginseng species using NMR-based metabolomics

The use of ¹H-NMR-based metabolomics to distinguish and identify unique markers of five Ontario ginseng (Panax quinquefolius L.) landraces and two ginseng species (P. quinquefolius and P. ginseng) was evaluated. Three landraces (2, 3, and 5) were distinguished from one another in the principal component analysis (PCA) scores plot. Further analysis was conducted and specific discriminating metabolites from the PCA loadings were determined. Landraces 3 and 5 were distinguishable on the basis of a decreased NMR intensity in the methyl ginsenoside region, indicating decreased overall ginsenoside levels. In addition, landrace 5 was separated by an increased amount of sucrose relative to the rest of the landraces. Landrace 2 was separated from the rest of the landraces by the increased level of ginsenoside R_b1. The Ontario P. quinquefolius was also compared with Asian P. ginseng by PCA, and clear separation between the two groups was detected in the PCA scores plot. The PCA loadings plot and a t-test NMR difference plot were able to identify an increased level of maltose and a decreased level of sucrose in the Asian ginseng compared with the Ontario ginseng. An overall decrease of ginsenoside content, especially ginsenoside R_b1, was also detected in the Asian ginseng’s metabolic profile. This study demonstrates the potential of NMR-based metabolomics as a powerful high-throughput technique in distinguishing various closely related ginseng landraces and its ability to identify metabolic differences from Ontario and Asian ginseng. The results from this study will allow better understanding for quality assessment, species authentication, and the potential for developing a fully automated method for quality control.

Brain Concentration of Ginsenosides and Pharmacokinetics after Oral Administration of Mountain‐cultivated Ginseng

Mountain‐cultivated ginseng (MCG ) can be considered a mimic of wild ginseng, whose seeds are sowed artificially, cultivated in the natural environment, and returned to the wild state before being used clinically. Cultivated ginseng (CG ) and MCG are mainly used as the commercial material for clinical applications. However, MCG is much more expensive but effective than CG . The aim of this study is to explore the differences in the pharmacokinetics and brain concentration of Rg1, Re, Rb1, and Rd after oral administration of MCG , CG, and pure ginsenosides. An ultra‐performance liquid chromatography‐tandem mass spectrometry method was developed and validated to determine the concentration of the four ginsenosides in rat plasma and brain tissue. Compared with the pure group, the area under the curve (AUC) of all the four ginsenosides for CG and MCG increased. The mean brain concentrations of the four ginsenosides were found to be 10‐ to 15‐fold lower than the corresponding contents in the plasma, and the poor permeability of ginsenosides into blood–brain barrier was indicated by the low concentrations of the four ginsenosides.     

Analysis of the neutral ginsenosides of wild and plantation roots of Panax ginseng growing in maritime territory

A procedure is proposed for determining ginsenosides with the aid of a Milikhrom liquid chromatograph. A comparative and quantitative analysis has been made of the neutral ginsenosides of the total glycoside fractions obtained from the roots of wild and plantation ginseng (Panax ginseng C. A. Meyer) growing in Maritime Territory. It has been shown that the amount of each ginsenoside and their total vary over a wide range in both wild and plantation roots.Pacific Ocean Institute of Bioorganic Chemistry, Far-Eastern Scientific Center, Russian Academy of Sciences, Vladivostok. Translated from Khimiya Prirodnykh Soedinenii, No. 2, pp. 237–242, March–April, 1993.     

HPLC-MS结合多元统计分析区分人参产地及筛选皂苷类标志物

利用高效液相色谱-质谱联用(HPLC-MS)技术结合多元统计分析方法, 区分中国人参主产区5个不同产地的45个人参样本, 筛选出差异性皂苷类标志物. 根据人参总皂苷在反相C₁₈色谱柱中的洗脱顺序, 结合串联质谱分析和标准品比对, 在提取的人参总皂苷中鉴定出15种原人参三醇型、 24种原人参二醇型和2种齐墩果酸型共41种皂苷. 对人参总皂苷的HPLC-MS全扫描数据进行了多元统计分析. 正交偏最小二乘-判别分析(OPLS-DA)结果表明, 所建立的分析模型具有良好的数据描述能力和预测能力. 所有人参样本能够根据产地被区分, 并筛选得到同时区分5个产地的差异性皂苷类组分18种; 能够区分任意2个产地人参样本的差异性组分主要为在人参中含量较高的人参皂苷Rb1, Rg1, Re, Rc, Rd, Ro和m-Rb1等. 分层聚类分析(HCA)结果显示, 黑龙江和吉林两省的样本能够独自聚类, 但是绥化市的样本更接近于吉林省. 初步推断原因为绥化市地理位置较接近吉林省, 两地人参生长环境相似并可能存在种质资源交换.     

加拿大一枝黄花的研究及应用现状

加拿大一枝黄花(Solidago Canadensis L.),是原产于北美的一种菊科多年生草本植物,20世纪30年代开始进入我国,现在这种植物已成为我国大部分境内迅速扩散传播的恶性杂草。目前,国内外的很多研究者已经利用各种方法从加拿大一枝黄花中提取出了倍半萜烯类、氨基酸、醇类等化学物质,对其生物性能进行了研究,并且对于控制这种植物的生长提出了各种解决方案。同时,加拿大一枝黄花已经在提炼精油、医学制药等方面得到广泛的应用。     

Metabolomics approach to growth-age discrimination in mountain-cultivated ginseng (Panax ginseng C. A. Meyer) using ultra-high-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry

Mountain-cultivated ginseng is typically harvested after 10 years, while ginseng aged over 15 years is considered wild ginseng. This study aims to differentiate mountain-cultivated ginseng by age, as the fraudulent practice of selling low-aged cultivated ginseng disguised as high-aged one is damaging the market. In this study, LC-MS analyzed 98 ginseng samples, and multivariate statistical analysis identified patterns between samples to select influential components. Machine learning models were developed to identify ginseng samples of different ages. The untargeted metabolomic analysis clearly divided samples aged 4-20 years into three age groups. Twenty-two potential age-dependent biomarkers were discovered to differentiate the three sample groups. Three machine learning models were used to predict new samples, and the optimal model was selected. Some biomarkers could determine age phases according to the differentiation of mountain-cultivated ginseng samples. These biomarkers were thoroughly analyzed for variation trends. The machine learning models established using the screened biomarkers successfully predicted the age group of new samples.     

A method for the analysis of ginsenosides, malonyl ginsenosides, and hydrolyzed ginsenosides using high-performance liquid chromatography with ultraviolet and positive mode electrospray ionization mass spectrometric detection

A high-performance liquid chromatographic separation coupled to diode array absorbance and positive mode electrospray mass spectrometric detection has been developed for the analysis of ginsenosides, malonyl ginsenosides, and hydrolyzed ginsenosides in extracts of Asian ginseng (Panax ginseng) and American ginseng (P. quinquefolius). The method is capable of separating, identifying, and quantifying the predominant ginsenosides found in heated alcoholic extracts of Asian and American ginseng roots routinely sold as nutraceuticals. It also separates and identifies the malonyl ginsenosides often found in cold alcoholic extracts of ginseng root and has the potential to quantify these compounds if pure standards are available. Furthermore, it can separate and identify ginsenoside hydrolysis products such as those readily produced in situations mimicking gastric situations, including those used for dissolution studies (i.e., 0.1 N HCl, 37 degrees C).     

Classification of cultivation area of ginseng by near infrared spectroscopy and ICP-AES

A rapid and nondestructive near infrared (NIR) method using soft independent modeling of class analogy (SIMCA) for the classification of cultivation area (Korea and China) was evaluated and confirmed. Raw, first, and second derivative NIR spectra were compared to develop a robust classification rule. The chemical properties of ginseng samples were also investigated to find out the differences between Korean samples and Chinese samples. These differences make NIR spectroscopic method viable. The average value of each Korean and Chinese ginseng sample for crude fiber, crude protein, starch, and 10 inorganic constituents were measured and compared with F-test and t-test. The inorganic constituents were also measured by induced coupled plasma-atomic emission spectroscopy (ICP-AES). It could be found that the amount of starch and ten inorganic elements for example Na, Mg, P, K, Ca, Mn, Fe, Ni, Cu and Zn in ginseng samples are considerably different based on cultivation area. SIMCA has been applied to the inorganic data to investigate the possibility of ICP-AES as classification tool. However, it was observed that the result was not equal to than NIR spectra data. The overall results showed the availability of NIR method using SIMCA would be adequate for classification of cultivation of ginseng, since NIR spectra includes useful and various information on chemical properties in spite of broad and overlapped bands.