Field survey of Glycyrrhiza plants in Central Asia (3). Chemical characterization of G. glabra collected in Uzbekistan

The chemical characteristics of Glycyrrhiza glabra L. were investigated at a habitat in Uzbekistan. HPLC analysis of the underground parts indicated that glycyrrhizin contents varied from 3.3 to 6.1% of dry weight, and that glabridin, a species-specific flavonoid for G. glabra, was detected in all underground samples (0.08-0.35% of dry weight). HPLC analysis of the leaves indicated that G. glabra plants collected in the present study could be divided into two types, RT-type and IQ-type, according to their major flavonol glycosides, rutin or isoquercitrin, respectively.     

Screening and characterization of natural antioxidants in four Glycyrrhiza species by liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry

Licorice, derived from the dried roots and rhizomes of several species of genus Glycyrrhiza L. (Leguminosae family), has been traditionally used in herbal medicine for over 4000 years. In recent years, the interest in antioxidative constituents in licorice has greatly increased. In this work, a new method based on 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) spiking test combined with HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS) analysis was developed to screen and identify the antioxidants in licorice. The results of the method validation indicated that the developed method was reliable and repeatable. Compared with DPPH on-line method, the HPLC-Q-TOF MS/MS method combined with DPPH spiking test offered much higher sensitivity and resolution. Using this method, 35 radical scavengers were screened from four Glycyrrhiza species (G. inflata, G. glabra, G. pallidiflora and G. uralensis), and 21 of them were unambiguously or tentatively identified by HPLC-Q-TOF MS/MS. Among the 21 identified flavonoids, 10 compounds had been reported to possess antioxidative activities in the previous studies, and the radical scavenging activities of the other 11 compounds were reported for the first time. The effects of six purified flavonoids on DPPH radical and lipid peroxidation were evaluated for validation of the developed method. The results indicated that HPLC-Q-TOF MS/MS coupled with DPPH treatment is an efficient and powerful method to discover the potential antioxidative compounds from the complex natural product mixtures. In this study, the identified components with free radical scavenging activity, would help to explain the therapeutic benefit of licorice in the treatment of human disease associated with oxidative stress.     

马钱子与甘草配伍前后生物碱成分的变化规律

采用电喷雾质谱技术和高效液相色谱法分别对马钱子配伍甘草前后主要生物碱的变化规律进行了系统研究。实验结果表明,马钱子配伍甘草后其主要生物碱士的宁和马钱子碱的含量均有不同程度的降低,其中士的宁的含量下降显著。电喷雾质谱的实验结果与高效液相色谱的结果相吻合,为进一步阐明甘草解马钱子类药物毒性和马钱子合理配伍用药提供了科学的实验依据。     

乌拉尔甘草叶中氨基酸的反相高效液相色谱快速测定

用邻苯二甲醛柱前荧光衍生梯度洗脱的方法分析了乌拉尔甘草叶中14种氨基酸的含量，为利用甘草叶作为饲料提供了科学参考；该法操作简单，衍生反应迅速，灵敏度高，重复性好。     

Identification of Glycyrrhiza species by direct analysis in real time mass spectrometry

DART (Direct Analysis in Real Time)-MS is a novel mass spectrometric ion source, and allows the analysis of most compounds at ambient pressure and ground potential by producing [M+H]+ molecular ion species. Using this method, we examined the compounds characteristic of several kinds of licorices. For the analysis of Glycyrrhiza inflata Batalin, the peak at m/z 339 originates mainly from [M+H]+ of licochalcone A (LA), a species-specific compound. This peak was hardly detected in G. glabra Linné and G. uralensis Fischer. These results indicate that G. inflata can be differentiated from the other two species by detection of LA peaks using DART-MS analysis.     

Chemical analysis of the Chinese herbal medicine Gan-Cao (licorice)

Gan-Cao, or licorice, is a popular Chinese herbal medicine derived from the dried roots and rhizomes of Glycyrrhiza uralensis, G. glabra, and G. inflata. The main bioactive constituents of licorice are triterpene saponins and various types of flavonoids. The contents of these compounds may vary in different licorice batches and thus affect the therapeutic effects. In order to ensure its efficacy and safety, sensitive and accurate methods for the qualitative and quantitative analyses of saponins and flavonoids are of significance for the comprehensive quality control of licorice. This review describes the progress in chemical analysis of licorice and its preparations since 2000. Newly established methods are summarized, including spectroscopy, thin-layer chromatography, gas chromatography, high-performance liquid chromatography (HPLC), liquid chromatography/mass spectrometry (LC/MS), capillary electrophoresis, high-speed counter-current chromatography (HSCCC), electrochemistry, and immunoassay. The sensitivity, selectivity and powerful separation capability of HPLC and CE allows the simultaneous detection of multiple compounds in licorice. LC/MS provides characteristic fragmentations for the rapid structural identification of licorice saponins and flavonoids. The combination of HPLC and LC/MS is currently the most powerful technique for the quality control of licorice.     

Studies on Triterpenoids and Flavones in Glycyrrhiza uralensis Fisch. By HPLC-ESI-MSn and FT-ICR-MSn

应用高效液相色谱质谱联用方法(HPLC-ESI-MSn)研究了甘草提取物中的七种化合物,四种三萜类化合物和三种黄酮类化合物。通过多极串联质谱(ESI-MSn)和多极串联傅里叶变换回旋共振质谱(FT-ICR-MSn)法研究了它们的碎裂规律。通过比较保留时间和质谱数据对上述七种化合物进行了归属,并阐述了其可能的质谱裂解途径。以上结果显示ESI-MSn和FT-ICR-MSn是非常有效的分析三萜类化合物和黄酮类化合物结构的工具。     

A novel biflavonoid from roots of Glycyrrhiza uralensis cultivated in China

A novel biflavonoid named licobichalcone was isolated from the roots of Glycyrrhiza uralensis cultivated in China, along with twelve known compounds, including five chalcones, two isoflavones, two flavanones, two flavones and one pterocarpan. Their structures were respectively elucidated on the basis of chemical and spectroscopic evidence.     

The isolation and structure elucidation of minor isoflavonoids from licorice of Glycyrrhiza glabra origin

Two new isoflavanone and one new 3-arylcoumarin derivatives, along with a known compound 3,4-didehydroglabridin, were isolated from commercially available licorice of Glycyrrhiza glabra origin, and their structures were elucidated on the basis of both the chemical and spectroscopic evidence.     

Lipids and Essential Oil from Roots of Glycyrrhiza glabra Growing in Uzbekistan

Chemistry of Natural Compounds - Lipids and essential oil from roots of licorice (Glycyrrhiza glabra L., Fabaceae) growing in Uzbekistan were investigated. Neutral, glyco-, and phospholipids and...     

Occurrence of methylated arsenic species in parts of plants growing in polluted soils

Arsenic compounds were determined in extracts of branches, leaves and roots from plants growing in a mining contaminated area. The selected species were Dryopteris filix-max, Quercus pubescens, Dipsacus fullonum, Alnus glutinosa, Buxus sempervirens and Brachythecium cf. reflexum. Total arsenic content in the subsamples was analysed by ICPMS after acidic digestion. In general, concentrations in the plant parts followed the gradient roots?>?branches?>?leaves indicating that they are arsenic-resistant plants. Arsenic species were determined in water/methanol (9?+?1, v/v) extracts by HPLC-ICPMS. Different levels of organoarsenicals were found depending on plant part and plant species. Higher percentages of organoarsenic compounds were recorded in branches and leaves (up to 35% in the boxtree sample), than in roots (0.7–5.2% in the same plant species). The absence of organic arsenic species in the soil where the plants were collected and the low levels of organoarsenicals found in the roots, indicate that the studied plants have the ability to accumulate or synthesise organoarsenic compounds in relatively high percentages, and this information contributes to enlarge the knowledge of arsenic uptake and speciation in plants.     

New prenylated isoflavanones from the roots of <Emphasis Type="Italic">Glycyrrhiza glabra</Emphasis>

Two new prenylated isoflavanones have been isolated from the roots of Glycyrrhiza glabra L. along with the known compounds cetoleic acid, β-sitosterol, stigmasterol, lanast-5,24-dien-3β-D-glucuronopyranoside, and glucuronic acid. The structures of the prenylated isoflavanones have been established as 8-isoprenyl-7,4′-dihydroxylicoisoflavanone (glabraisoflavanone A) and 7,3′-dihydroxy-8-isoprenyl-4′-cyclogeranioloxyisoflavanone (glabraisoflavanone B) on the basis of spectral data analyses and chemical reactions.     

One step isolation and purification of liquiritigenin and isoliquiritigenin from Glycyrrhiza uralensis Risch. using high-speed counter-current chromatography

High-speed counter-current chromatography (HSCCC) technique in semi-preparative scale has been successfully applied to the separation of bioactive flavonoid compounds, liquiritigenin and isoliquiritigenin in one step from the crude extract of Glycyrrhiza uralensis Risch. The HSCCC was performed using a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-acetonitrile-water (2:2:1:0.6:2, v/v). Yields of liquiritigenin (98.9% purity) and isoliquiritigenin (98.3% purity) obtained were 0.52% and 0.32%. Chemical structures of the purified liquiritigenin and isoliquiritigenin were identified by electrospray ionization-MS (ESI-MS) and NMR analysis.     

A novel neutral polysaccharide having activity on the reticuloendothelial system from the root of Glycyrrhiza uralensis

A neutral polysaccharide, named glycyrrhizan UC, was isolated from the root of Glycyrrhiza uralensis Fischer. It was homogeneous on electrophoresis and gel chromatography, and its molecular mass was estimated to be 69,000. Glycyrrhizan UC is composed of L-arabinose:D-galactose:D-glucose:L-rhamnose in the molar ratio of 10:30 27:1. Methylation analysis, carbon-13 nuclear magnetic resonance and periodate oxidation studies indicated its structural feature as an arabino-3,6-galacto-glucan type polysaccharide.     

Molecular Cloning of a Glycyrrhiza uralensis F. Aquaporin GuPIP1 Up-regulated in Response to Drought, Salt and ABA Stress

IntroductionWater is an ubiquitous and indispensable moleculefor plant growth and development[1]. According to thecomposite water flux model[2], water is transported intoplant tissues by three pathways: apoplastic, symplas-tic, and transcellular. The latt…     

Comparative analysis of radical scavenging and antioxidant activity of phenolic compounds present in everyday use spice plants by means of spectrophotometric and chromatographic methods

Comparative analysis of radical scavenging and antioxidant activities of phenolic compounds present in everyday use spice plants was carried out by means of spectrophotometric and chromatographic methods. Six spice plant samples, namely onion (Allium cepa), parsley (Petroselinum crispum) roots and leaves, celery (Apium graveolens) roots and leaves and leaves of dill (Anethum graveolens) were analyzed. Total amount of phenolic compounds and radical scavenging activity (RSA) was the highest in celery leaves and dill extracts and was the lowest in celery roots. Comparing commonly used spectrophotometric analysis of 2,2-diphenyl-1-picrylhydrazyl (DPPH) RSA of extracts with the results obtained using reversed-phase chromatographic separation with on-line post-column radical scavenging reaction detection, good correlation was obtained (R(2)=0.848). Studies using HPLC system with electrochemical detector showed that bioactive phytochemicals can be separated and antioxidant activities of individual compounds evaluated without the need of a complex HPLC system with reaction detector. The results obtained using electrochemical detection correlate with the RSA assayed using spectrophotometric method (R(2)=0.893).     

Tracking antiangiogenic components from Glycyrrhiza uralensis Fisch. based on zebrafish assays using high-speed countercurrent chromatography

Natural products are some of the most important sources of lead compounds for drug discovery. The advanced isolation technique of lead compounds of natural origin using therapeutically relevant bioassays is capable of enhancing work efficiency from complex multiconstituent extracts. In the present study, a bioassay-guided isolation strategy combined with bioactivity screening was used to identify novel angiogenesis inhibitors from licorice (Glycyrrhiza uralensis Fisch.) based on the zebrafish model and rapid preparative separation by high-speed countercurrent chromatography. Zebrafish embryos at 24 h postfertilization were chosen as the angiogenesis inhibition model for bioactivity screening. A solvent system (n-hexane-ethyl acetate-methanol-water) with different ratios was optimized and applied in the high-speed countercurrent chromatography separation of two fractions, Fr5 and Fr6, from the ethyl acetate extract of licorice. Blood circulation and vascular outgrowth in intersegmental vessels were found to be simultaneously inhibited by isoliquiritigenin and isolicoflavonol in a dose-dependent manner. Thus, these two compounds were identified and considered as active inhibitors against angiogenesis. These experimental results indicate that zebrafish bioassays combined with high-speed countercurrent chromatography may provide an alternative pathway for the rapid isolation of bioactive natural products.     

转换移动窗口因子分析法用于中药组合前后成分变化分析

以正十四烷为内标,采用气相色谱-质谱(GC/MS)联用技术对中药甘草(GUF)、甘遂(EKL)单味药材,以及其组成的药对(GUF-EKL)的挥发成分进行测定,基于一种新的化学计量学方法——转换移动窗口因子分析法(AMWFA)和直观推导式演进特征投影法(HELP)对产生的二维色谱/质谱数据同时进行分辨和解析,获取各个组分的纯色谱曲线和质谱,根据分辨得到的纯质谱在质谱库中进行相似检索以实现对组分的定性。利用AMWFA对组合前后的成分进行归属分析,然后采用面积归一法进行定量。分别在甘草、甘遂及其药对中鉴定出52、51和63个成分,占各自挥发油成分的84%、90%和75%。     

Application of capillary electrophoresis to the simultaneous determination of betaines in plants.

The simultaneous determination of betaines, the key compounds for osmotic regulation in plants, was established by capillary electrophoresis (CE). After four betaines, glycine betaine (GB), beta-alanine betaine (AB), proline betaine (PB), and 2-hydroxyproline betaine (HPB), were esterified with p-bromophenacyl bromide, the esters were electrophoresed in 100 mM sodium phosphate at pH 3.0. A low-pH condition in CE and esterification gave an advantage of resolving each of the ester peaks as well as those of the unreacted reagent and other components. Furthermore, the addition of 4% polyethylene glycol (PEG) gave a better resolution of 4 peaks, resulting in the separation of the overlapped peaks of PB and AB. It was found from the standard addition method being applied to barley leaves that the GB content in plants could be evaluated by using a calibration curve of the GB standard solution. The extraction of GB from plant leaves was also improved by adopting water as the extraction solvent instead of a mixture of organic solvents. The present method was suitably applied to actual plant specimens collected from a saline area of China.     

A Fast Isolation Method for Glycyrrhizic Acid,the Bioactive Marker of Glycyrrhiza glabra,and Its Quantitative Evaluation in Some Single and Multiherbal Formulations Using High-Performance Thin-Layer Chromatography

JPC – Journal of Planar Chromatography – Modern TLC - Glycyrrhizic acid, the major metabolite of Glycyrrhiza glabra, is a ‘Hot Molecule’ in view of its sweet tasting...