Preparation and Identification of Angelica sinensis Polysaccharide-iron Complex

Angelica sinensis polysaccharide(ASP) was extracted from Angelica sinensis by boiling water. An Angelica sinensis polysaccharide-iron complex(APC) was prepared under the alkaline condition by adding a ferric chloride solution to the ASP solution. Then some identifiable properties of the complex were studied. The content of iron (Ⅲ) in the complex was determined with iodometry. The thermal property, the microscopic structure, the spectral characteristics, and N, C, H contents of the complex were examined by a variety of techniques including DSC, TEM, IR,NMR, and elemental analysis. The content of iron (Ⅲ) in the complex ranges from 10% to 40%. The DSC result shows that the melting point of the complex is about 450 ℃. The TEM result shows that the complex has an iron(Ⅲ)core(β-FeOOH core) linked by hydroxy and oxy bridges, with the polysaccharide chains attached to the surface of the core. The IR and NMR results also show that there is aβ-FeOOH core in the complex. The elemental analysis shows that the contents of N, C , H in the complex are, respectively, lower than those of N, C, H in ASP. All our studies indicate that the APC consists of aβ-FeOOH core surrounded by ASP.     

Molecular Weight and Proposed Structure of the Angelica sinensis Polysaccharide‐iron Complex

The molecular weight and the proposed structure of the Angelica sinensis polysaccharide‐iron complex (APIC) were studied. Fourier transform infrared spectroscopy, X‐ray powder diffraction, differential scanning calorimetry, transmission electron microscopy, electron paramagnetic resonance, thermogravimetric analysis, atomic force microscopy, and gel filtration chromatography were used to characterize APIC, which is a macromolecule complex composed of Angelica sinensis polysaccharide (ASP) and iron. The structure of APIC was proposed to be a polynuclear ferrihydrite core chelated firmly by an encircling framework of ASP chains, forming a core molecule, which is surrounded by a removable outer protective sheath of colloidal ASP. And the molecular formula of APIC was proposed to be {[(Fe₂O₃·2.2H₂O)₁₀₄₃(ASP)₃₂](ASP)₁₂}, with MW=270000 Da.     

当归多糖ASP3的甲基化分析

通过部分酸水解和甲基化分析, 结合GC-MS测试手段, 对当归多糖ASP3的糖链结构进行了研究. 采用0.2 mol/L三氟乙酸(Trifluoroacetic acid, TFA)对ASP3进行了部分酸水解, 对水解前后的多糖组分进行甲基化分析. 结果显示: ASP3的糖残基主要由D-GalpA, D-Galp, L-Araf和L-Rhap组成, 主链由1,4-D-GalpA连接, 形成“光滑区”半乳糖醛酸聚糖, 由1,4-D-GalpA通过O-4位与1,2-; 1,2,4-L-Rhap的O-2位交替连接形成含有较多分支的“毛发区”鼠李半乳糖醛酸聚糖. 58.8%的Rhap残基发生O-4位取代(1,2,4-L-Rhap). 由T-, 1,5-, 1,3,5-Araf和T-, 1,3-, 1,3,6-, 1,4-, 1,4,6-D-Galp聚合形成的阿拉伯半乳聚糖、半乳聚糖以及阿拉伯聚糖是ASP3侧链的主要组成, 通过Rhap残基的O-4位与主链相连.     

A new procedure for the preparative separation and isolation of Z-ligustilide from the roots of Angelica sinensis

A new procedure for the separation and isolation of Z-ligustilide from the roots of Angelica sinensis (AS) was developed, and the storage conditions for Z-ligustilide were optimized. Using the present procedure, Z-ligustilide was enriched by decomposing the Z-ligustilide dimers yielding Z-ligustilide and dissolving the polar impurities in hot water. Then, the crude Z-ligustilide was further purified by a semipreparative HPLC system. The spiked and nonspiked samples were used for the evaluation of the proposed procedure. Recoveries obtained varied from 86.2 to 90.7% and RSDs from 4.0 to 6.6%. The yield and purity of the isolated Z-ligustilide were found to be 4.57 mg/g and 99.6%, respectively. The results of stability tests have shown that the presence of oxidant contributes to Z-ligustilide degradation, therefore argon was chosen as a shielding gas for storage. The overall procedure is efficient and convenient which is considered suitable for the preparative separation of Z-ligustilide from AS.     

A New Triterpene from Cephalomappa sinensis

Cephalomappa sinensis (Chun et How) Kosterm belongs to the family Euphorbiaceae. It is distributed in southwest and west of Guangxi Zhung Autonomous Region1. Its fresh leaves are used as feeding stuff for cattle, horse, and sheep and can make them strong1. Chemical constituents of this species have not been reported previously. In our research, a new triterpene was isolated from the ethanolic extract of its roots. Compound 1 was obtained as colorless needles, mp 235-237(C. The EI-mass sp…     

Urinary metabolomics analysis reveals the effect of volatile oil from Angelica sinensis on LPS‐induced inflammation rats

Lipopolysaccharide (LPS)‐induced inflammation occurs commonly and volatile oil from Angelica sinensis (VOAS) can be used as an anti‐inflammatory agent. The molecular mechanisms that allow the anti‐inflammatory factors to be expressed are still unknown. In this paper, we applied gas chromatography–mass spectrometry (GC–MS) and high‐performance liquid chromatography–time‐of‐flight mass spectrometry (LC‐Q/TOF–MS) based on a metabolomics platform coupled with a network approach to analyze urine samples in three groups of rats: one with LPS‐induced inflammation (MI); one with intervention with VOAS; and normal controls (NC). Our study found definite metabolic footprints of inflammation and showed that all three groups of rats, MI, intervention with VOAS and NC have distinct metabolic profiles in urine. The concentrations of 48 metabolites differed significantly among the three groups. The metabolites in urine were screened by the GC–MS and LC‐Q/TOF–MS methods. The significantly changed metabolites (p < 0.05, variable importance in projection > 1.5) between MI, NC and VOAS were included in the metabolic networks. Finally, hub metabolites were screened, including glycine, arachidonic acid, l ‐glutamate, pyruvate and succinate, which have high values of degree (k). the Results suggest that disorders of glycine, arachidonic acid, l ‐glutamate, pyruvate and succinate metabolism might play an important part in the predisposition and development of LPS‐induced inflammation. By applying metabolomics with network methods, the mechanisms of diseases are clearly elucidated.     

The Dynamic Accumulation Rules of Chemical Components in Different Medicinal Parts of Angelica sinensis by GC-MS

The chemical components and medicinal properties of different medicinal parts of Angelica sinensis are often used as medicine after being divided into the head, body and tail of Angelica sinensis. In this study, the chemical components of different medicinal parts in different periods were analyzed by GC-MS for the first time, and the differences of the accumulation rules of chemical components in different medicinal parts of Angelica sinensis were obtained. This study demonstrated that the differences of composition accumulation in different medicinal parts of Angelica sinensis were mainly reflected in the types and relative contents of compounds. The study found that the number of compounds in different medicinal parts of Angelica sinensis in each period were different and the change rules of the same compound in different medicinal parts were also different. The number of compounds in the tail of Angelica sinensis was the least in April, and the largest in October. The content of ligustilide in the body of Angelica sinensis was higher in April and was the highest in the tail in October. The relative content of butylidenephthalide in the head was the highest in October. The relative contents of senkyunolide A and butylphthalide in the head were decreased in October, while the contents in the body and tail increased, indicating that the compounds that accumulate in the head may transfer to the body and tail in later stages of growth. This study clarified the differences in the accumulation of chemical components in different medicinal parts of Angelica sinensis, which could provide a theoretical basis for the reasons for the differences of chemical components in the different medicinal parts.     

A Novel Dimeric Eremophilane from Ligularia virgaurea spp. oligocephala

A novel dimeric eremophilane, ligulolide B, was isolated from the alcoholic extract of the whole plant of Ligularia virgaurea spp. oligocephala. The structure was elucidated by various spectroscopic methods including intensive 2D NMR techniques （^1H-^1H COSY, gHMQC, gHMBC and ^1H-^1H NOESY） and HR-ESI-MS.     

当归多糖ASP3及其水解产物的NMR光谱分析

对当归多糖ASP3的糖链结构进行分析. 分别采用0.2 mol/L三氟乙酸(Trifluoroacetic acid, TFA)和内切-α-(1→4)-聚半乳糖醛酸酶(EndoPG)对ASP3进行部分酸水解和酶水解, 并对水解前后多糖组分的1D和2D NMR光谱特征进行分析. 实验结果表明, ASP3是一种果胶多糖. GalpA和Rhap位于多糖分子的主链, 由1→4-D-GalpA相连形成的“光滑区”(半乳糖醛酸聚糖)是其主要组成部分; 由α-(1→4)-GalpA通过O4位与α-(1→2)-和α-(1→2,4)-Rhap的O2位交替连接所形成的重复单元[→4)-α-GalpA-(1→2)-α-Rhap-(1→]构成具有较高分支的“毛发区”(富含中性糖侧链的鼠李半乳糖醛酸聚糖). Galp和Araf是中性糖侧链的主要组成, 通过Rhap残基的O4位与主链相连. 非还原性末端T-β-Galp, β-(1→3)-, β-(1→3,6)-, β-(1→4)-, β-(1→4,6)-Galp聚合形成以β-(1→3,6)-Galp为分支点的β-(1→6)-半乳聚糖和以β-(1→4,6)-Galp为分支点的β-(1→4)-半乳聚糖. T-α-Araf, α-(1→5)-Araf和α-(1→3,5)-Araf聚合形成以α-(1→3,5)-Araf为分支点的α-(1→5)-阿拉伯聚糖. 此外, 由α-(1→5)-阿拉伯聚糖通过α-(1→3)连接与β-(1→6)-半乳聚糖末端聚合形成阿拉伯半乳聚糖.     

Quick and selective extraction of Z‐ligustilide from Angelica sinensis using magnetic multiwalled carbon nanotubes

A facile and highly efficient magnetic solid‐phase extraction method has been developed for Z‐ligustilide, the major therapeutic agent in Angelica sinensis. The solid‐phase adsorbent material used was prepared by conjugating carbon nanotubes with magnetic Fe₃O₄ nanoparticles via a hydrothermal reaction. The magnetic material showed a high affinity toward Z‐ligustilide due to the π–π stacking interaction between the carbon nanotubes and Z‐ligustilide, allowing a quick and selective exaction of Z‐ligustilide from complex sample matrices. Factors influencing the magnetic solid‐phase extraction such as the amount of the added adsorbent, adsorption and desorption time, and desorption solvent, were investigated. Due to its high extraction efficiency, this method was proved highly useful for sample cleanup/enrichment in quantitative high‐performance liquid chromatography analysis. The proposed method had a linear calibration curve (R² = 0.9983) over the concentration between 4 ng/mL and 200 μg/mL Z‐ligustilide. The accuracy of the method was determined by the recovery, which was from 92.07 to 104.02%, with the relative standard deviations >4.51%.     

Artselenoide, a New Dimeric Guaianolide from Artemisia selengensis

Inaprelindnary.ommunication',wehavereportedonanewguaianolideartseleninisolatedfromArtemisiaselengensisTurcz(Compositae).Inthepresentpaperwedescribethestructureelucidationofartselenoide1obtainedasanewdimericguaianolidefromthesamepart.1wasobtainedascolorlesspowders,mpl85-l86D,[a]." l25(cO.l,MeOH).TheIRspectrumexhibitedabsorptionbandsduetohydroxyl(3435cm-'),Ylactone(l764cm-')anddoublebond(l648,l635,l456cm-')groups.Themolecularformu1aCstheooof1wasdeducedfromthedataofESIMS(Inlz5l9[M Na] ,53…     

A new Compound from Chaenomeles sinensis (Thouin) Koehne

A new dihydrochromone derivative,named chaenomone,was isolated from the twigs of Chaenomeles sinensis (Thouin)Koehne .The structure was determined by spectroscopic methods.     

A New Triterpene from Cynanchum forrestii

Cynanchum forrestii Schlechter (Asclepiadaceae) is widely distributed in Yunnan, Xizang, Gansu, Sichuan, Guizhou Provinces, etc. in China and its roots have been used as an antifebrile, diuretic and analgetic in China1. In the previous investigation, we i…     

超高效液相色谱-串联质谱法对比11种净化方式对当归中50种农药残留检测的影响

采用超高效液相色谱-串联质谱仪(UPLC-MS/MS)同时测定当归药材中50种农药残留,对比了11种净化方式对检测结果的影响,筛选出最有效的基质净化方法。 以回收率为考察指标,评估了不同基质净化方法对当归中多农残的提取净化效果,最终确定样品经乙腈提取,NaCl净化,在电喷雾正负离子扫描、依赖保留时间的动态多反应监测模式(dMRM)下,以基质匹配内标法定量。 结果表明,50种农药在NaCl净化法下回收率最高,在各自的浓度范围内线性关系良好(R²>0.99);3个添加水平(10、50、100 μg/kg)下,大多数农药的回收率为70.1%~117.7%,相对标准偏差(RSD,n=6)不大于 20.0%,50种农药的定量限为1.0~20.0 μg/kg。 该方法为当归中准确、高效、经济的检测目标物提供了可靠依据。     

Cool Temperature Enhances Growth,Ferulic Acid and Flavonoid Biosynthesis While Inhibiting Polysaccharide Biosynthesis in Angelica sinensis

Angelica sinensis, a perennial herb that produces ferulic acid and phthalides for the treatment of cardio-cerebrovascular diseases, prefers growing at an altitude of 1800–3000 m. Geographical models have predicted that high altitude, cool temperature and sunshade play determining roles in geo-authentic formation. Although the roles of altitude and light in yield and quality have been investigated, the role of temperature in regulating growth, metabolites biosynthesis and gene expression is still unclear. In this study, growth characteristics, metabolites contents and related genes expression were investigated by exposing A. sinensis to cooler (15 °C) and normal temperatures (22 °C). The results showed that plant biomass, the contents of ferulic acid and flavonoids and the expression levels of genes related to the biosynthesis of ferulic acid (PAL1, 4CLL4, 4CLL9, C3H, HCT, CCOAMT and CCR) and flavonoids (CHS and CHI) were enhanced at 15 °C compared to 22 °C. The contents of ligustilide and volatile oils exhibited slight increases, while polysaccharide contents decreased in response to cooler temperature. Based on gene expression levels, ferulic acid biosynthesis probably depends on the CCOAMT pathway and not the COMT pathway. It can be concluded that cool temperature enhances plant growth, ferulic acid and flavonoid accumulation but inhibits polysaccharide biosynthesis in A. sinensis. These findings authenticate that cool temperature plays a determining role in the formation of geo-authentic and also provide a strong foundation for regulating metabolites production of A. sinensis.     

Metabolomics study on promoting blood circulation and ameliorating blood stasis: Investigating the mechanism of Angelica sinensis and its processed products

Angelica sinensis (Danggui, DG) parched with alcohol (Jiu Danggui, JDG) and charred DG are the main processed products of DG, which are used to treat blood stasis syndrome (BSS). However, their therapeutic effect and mechanisms are still unclear. Based on an acute rat BSS model, the intervention effects of DG and its processed products (DGPPs) were evaluated by the hemorheology and coagulation function parameters. Meanwhile, plasma and urine metabolites were detected and analyzed by liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry and multivariate statistical analysis method. The results of hemorheology, coagulation function parameters and metabolomics all showed that the BSS model was successfully established, DGPPs intervention could significantly relieve rats BSS and the therapeutic effect of JDG was best. Moreover, 23 differential metabolites (14 in plasma and nine in urine) were identified that were closely related to the BSS, involving seven potential target metabolic pathways. DGPP intervention showed different degrees of reverse effect on these metabolites. JDG was the most effective owing to extensive regulation effect on differential metabolites. This study provides a reference for understanding the pathological mechanism of BSS and the mechanism of DGPPs, which lays a theoretical foundation for the rational use of DGPPs in clinical practice.     

Simultaneous determination of eight components in Angelica sinensis based on UHPLC‐ESI‐MS/MS method for quality evaluation

A method employing ultra‐high performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) for determination of eight components including ferulic acid, senkyunolide A, butylphthalide, ligustilide, butylidenephalide, senkyunolide I, senkyunolide H and levistolide A in Angelica sinensis was established. The separation was carried out using a Waters ACQUITY UHPLC BEH C₁₈ column with gradient elution with 0.1% formic acid aqueous and acetonitrile at a flow rate of 0.4 mL/min. Good linearity was attained with R² of 0.9983–0.9998 in wide concentration ranges. The method had limit of detection (LOD) and quantitation (LOQ) in the range of 0.42–6.98 ng/mL and 1.39–23.28 ng/mL, respectively. Intra‐ and inter‐day precisions varied with relative standard deviations (RSDs) from 0.33% to 0.88% and 0.37% to 1.04%, respectively. Moreover, the average recoveries were in a satisfactory range of 92.7%–102.1% with RSDs of less than 3.60%. Finally, the method was successfully applied to analyze 19 batches of A. sinensis samples grown in Min County, Gansu province, China, as well as that collected in other regions. The findings indicated that the established method is reliable and may thus be applied as a powerful tool for qualitative and quantitative analysis of components in A. sinensis, which has its implications in quality control of A. sinensis.     

Metabonomic analysis of the anti‐inflammatory effects of volatile oils of Angelica sinensis on rat model of acute inflammation

Metabonomics based on GC‐MS was used to study the possible anti‐inflammatory mechanisms of volatile oils of Angelica sinensis (VOAS) in rats with acute inflammation. Acute inflammation was induced by subcutaneous injection of carrageenan in rats. The levels of prostaglandin E₂ (PGE₂), histamine (HIS) and 5‐hydroxytryptamine (5‐HT) in the inflammatory fluid were detected. Principal component analysis and orthogonal partial least squares‐discriminant analysis models were performed for pattern recognition analysis. After the administration of VOAS, the levels of PGE₂, HIS, and 5‐HT returned to levels observed in normal group. According to GC‐MS analysis, the intervention of VOAS in rats with acute inflammation induced substantial and characteristic changes in their metabolic profiles. Fourteen metabolite biomarkers, namely, lactic acid, malic acid, citric acid, trans‐dehydroandrosterone, aldosterone, linoleic acid, hexadecanoic acid, pregnenolone, octadecenoic acid, myristic acid, l ‐histidine, octadecanoic acid, arachidonic acid (AA) and l ‐tryptophan, were detected in the inflammatory fluid. The levels of all biomarkers either increased or decreased significantly in model groups. VOAS possibly intervened in the metabolic process of inflammation by altering histidine metabolism, tryptophan metabolism, AA metabolism, steroid hormone biosynthesis, fatty acid metabolism and energy metabolism. Metabonomics was used to reflect an organism's physiological and metabolic state comprehensively, and it is a potentially powerful tool that reveals the anti‐acute‐inflammatory mechanism of VOAS. Copyright © 2014 John Wiley & Sons, Ltd.