This study is to improve the quality standard and supply the scientific basis for Anemarrhenae Rhizoma and its raw processed products. Steroidal saponin including timosaponin BⅡ, timosaponin AⅢ and flavonoids including neomangiferin and mangiferin were selected as the indicative components. Silica gel G thin layer chromatography(TLC) and polyamide TLC were used to detect the two types of compounds, respectively. The contents of timosaponin BⅡ and timosaponin AⅢ were determined by HPLC-ELSD and the content of neomangiferin, mangiferin and isomangiferin were determined by HPLC-UV. Moisture, total ash and acid insoluble ash were determined according to Chinese Pharmacopoeia(2015 edition). And 80% ethanol was selected as the solvent and the content determination of total extract were determined. The fingerprints of Anemarrhenae Rhizoma and its raw processed products were established by HPLC-UV and HPLC-ELSD. The results showed that the methods of TLC and HPLC have been successfully stablished. There are 2 and 3 peaks which have been identified by HPLC-ELSD and HPLC-UV, respectively. The HPLC fingerprint methods are specific and can be used to identify and quality control for Anemarrhenae Rhizoma and its raw processed products in the mass. Comparing to Chinese Pharmacopoeia(2015 edition), the TLC identification and content determination were revised and the total extract determination and HPLC fingerprints were added in the present study. Our results can be used as the scientific basis of quqlity control for Anemarrhenae Rhizoma and its raw processed products.
Anemarrhena ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Reference Standards ; Rhizome

AIMTo investigate the chemical constituents of the rhizomes of Anemarrhena asphodeloides Bunge.

METHODSThe compounds were separated by means of solvent extraction, chromatography on absorbent resin SP825 and silica gel C18 repeatedly, and their structures were elucidated on the basis of chemical methods and spectral analyses (FAB-MS, 1H NMR, 13C NMR, 1H-1H COSY).

RESULTSSix steroidal saponins were isolated from the rhizomes of Anemarrhena asphodeloides Bunge. They were identified as (25S)-26-O-beta-D-glucopyranosyl-22-hydroxy-5beta-furostane-2beta, 3beta, 26-triol-3-O-beta-D-glucopyranosyl-(1 --> 2)-beta-D-galactopyranoside (timosaponin N, 1), timosaponin E1 (2), (25S)-26-O-beta-D-glucopyranosyl-22-methoxy-5beta-furostane-2beta, 3beta, 26-triol-3-O-beta-D-glucopyranosyl-(1 --> 2)-beta-D-galactopyranoside (timosaponin O, 3) , timosaponin E2 (4), (25R) -26-O-beta-D-glucopyranosyl-22-hydroxy-5alpha-furostane-2alpha, 3beta, 26-triol-3-O-beta-D-glucopyranosyl-(1 --> 2)-[beta-D-xylpyranosyl-(1 --> 3)]-beta-D-glucopyranosyl-(1 --> 4)-beta-D-galactopyranoside (purpureagitosid, 5) and marcogenin-3-O-beta-D-glucopyranosyl-(1 --> 2)-beta-D-galactopyranoside (6).

CONCLUSIONCompound 1 and compound 3 are new compounds, and compound 5 was isolated from the rhizomes of Anemarrhena asphodeloides Bunge for the first time.

Anemarrhena ; chemistry ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Saponins ; chemistry ; isolation & purification

OBJECTIVETo establish an HPLC-ELSD method for determination of Anemarsaponin C and Anemarsaponin A III in Anemarrhenae Rhizoma.

METHODKromasil C18 column(4.6 mm x 250 mm, 5 microm) was used as stationary phase. Mobile phase was methanol-water gradient with the flow rate of 1 mL x min(-1); the temperature of the drift tube and evaporation was 50 degrees C and 70 degrees C respectively. The gas pressure was 1.03 x 10(5) Pa.

RESULTThere are good linearity in the range 0.310-3.10 microg of anemarsaponin C (lgA = 1.254 2lgM + 5.734 7, r = 0.999 5) and in the range 0.323-3.23 microg (lgA = 1.328 41gM + 5. 937, r = 0.999 6) of anemarsaponin A III. The average recovery of anemarsaponin C and anemarsaponin A III was 98.1% with RSD 2.1% and 97.3% with RSD 1.5% (n = 6) respectively.

CONCLUSIONThe method is rapid and accurate. It is suitable for quality control of Anemarrhenae Rhizoma. The result of determination reveals that the quality of Anemarrhenae Rhizoma from different places of north China are of notable difference.

Anemarrhena ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Saponins ; analysis ; Triterpenes

Type 2 diabetes mellitus (T2DM) is a global metabolic disease with potentially life- threatening complications. Liver metabolism plays a vital role in the occurrence and development of diabetes mellitus. It has been reported that the Chinese medicinal Anemarrhenae rhizoma (AR) can relieve insulin resistance and diabetes mellitus. However, the effect on abnormal liver metabolism in diabetes mellitus is still unclear. Therefore, we extracted liver proteins of T2DM rats induced by high-fat diet (HFD) and streptozotocin (STZ), T2DM rats treated with AR extract (ARE), obesity rats (fed with HFD), and normal control rats (fed with normal diet). Then, through tandem mass tag (TMT) labeling combined with mass spectrometry (MS), we obtained the quantitative proteomic data. Bioinformatics software was used for hierarchical cluster analysis and principal component analysis of the data in each group. The volcano map for differentially expressed proteins (P < 0.05, fold change > 1.5) was plotted. It was found that the treatment group was closer to the normal control group, indicating that the quantitative proteomic data of liver tissue can reflect the therapeutic effect of ARE on T2DM rats. Key protein clusters closely related to the treatment of ARE were screened out. The Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the protein clusters were analyzed by David, and the result showed that AR's alleviation of abnormal fatty acid metabolism in livers of T2DM rats may be related to the regulation of the expression of key proteins Ndufa6 and Prkar2b.
Rats ; Animals ; Diabetes Mellitus, Type 2/metabolism* ; Proteomics/methods* ; Drugs, Chinese Herbal/therapeutic use* ; Anemarrhena ; Liver/metabolism*

OBJECTIVETo compare the quality of cultivated and wild Anemarrhena Rhizome from Yi County (Xiling Zhimu) based on contents analysis of active constituents.

METHODSamples of cultivated Anemarrhena Rhizome from most townships of Yi County were analyzed and compared with wild ones. Six indexes belonged to three kinds active constituents of saponin, flavornoid and polysaccharide were adopted. HPLC-ELSD method with cholesterol as internal standard was adopted to determine the content of sarsasapongenin. HPLC-ELSD method was used to simultaneously determine the contents of anemasaponin C and Anemasaponin A III. Contents of neomangiferin and mangiferin were determined by HPLC-UV method. Total polysaccharide was determined by phenol sulfate method.

RESULTThe mean content of sarsasapongenin in cultivated Anemarrhena Rhizome samples is slightly lower than the wild. The mean contents of anemasaponin C and Anemasaponin A III in cultivated Anemarrhena Rhizome samples are higher than the wild. There is no notable difference of these three index between the cultivated and the wild. The cultivated Anemarrhena Rhizome samples have a lower content of neomangiferin and a higher content of mangiferin than the wild. While the total content of these two flavonoids have no notable difference. The cultivated Anemarrhena Rhizome samples have a higher content of total polysaccharide than the wild samples.

CONCLUSIONContents of active constituents in cultivated Anemarrhena Rhizome from Yi County (Xiling Zhimu) are not notably different with the wild Anemarrhena Rhizome. They have similar good quality as the wild ones.

Anemarrhena ; chemistry ; growth & development ; China ; Chromatography, High Pressure Liquid ; Gardening ; methods ; Plant Extracts ; analysis ; Rhizome ; chemistry ; growth & development

This study is to establish an HPLC-DAD-ELSD method for simultaneous determination of 5 flavones and saponins in Rhizoma Anemarrhenae including neo-mangiferin, mangiferin, timosaponin B II, timosaponin B III and timosaponin A III. Samples were analyzed on a Merck Purospher STAR column(4.6 mm x 250 mm, 5 μm). The mobile phase consisted of acetonitrile( A) and 0. 1% formic acid (B) with gradient elution at a flow rate of 1.0 mL · min(-1). The column temperature was set at 40 °C. The DAD detector wavelength was set at 254 nm. The ELSD conditions were as follows: the nebulizing gas flow rate was 2.0 L · min(-1) and temperature of drift tube was 105 °C. The volume was 10 μL. The five compounds were well separated with good linear correlations. The mean recoveries were between 102.0%-104.0%. This method was quick and reliable which provides a foundation for quality control of R. Anemarrhenae.
Anemarrhena ; chemistry ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Rhizome ; chemistry ; Saponins ; analysis

During a screening program to search the anticolitic herbal medicines, 80% ethanol extract of the rhizome of Anemarrhena asphodeloides (AA) was found to potently inhibit the expression of proinflammatory cytokines TNF-alpha and IL-1beta, as well as the activation of NF-kappaB in LPS-stimulated colonic macrophages, followed by that of the rhizome of C. chinensis (CC). AA also potently inhibited TNBS-induced colitic markers, shortening of the colon and increase of macroscopic score, myeloperoxidase activity, TNF-alpha, IL-1beta, and IL-6, in mice. The synergistic effect of CC against the anticolitic effect of AA was investigated. CC synergistically inhibited the anticolitic effect of AA. AC-mix (AA+CC, 1:1) potently inhibited them. AC-mix also inhibited the activation of NF-kappaB, as well as the expression of TNF-alpha, IL-1beta, IL-6, iNOS and COX-2. The effects of AC-mix against oxazolone-induced colitis were investigated in mice. AC-mix also potently inhibited oxazolone-induced inflammatory markers, colon shortening, macroscopic score, myeloperoxidase activity, NF-kappaB activation and proinflammatory cytokines. Overall, the anti-colitic effect of AC-mix was superior to that of mesalazine. Based on these findings, AC-mix may improve colitis by inhibiting NF-kappaB activation.
Anemarrhena* ; Animals ; Colitis ; Colon ; Cytokines ; Ethanol ; Interleukin-6 ; Macrophages ; Mass Screening ; Mesalamine ; Mice* ; NF-kappa B ; Peroxidase ; Rhizome* ; Tumor Necrosis Factor-alpha

Five compounds were isolated from the fibrous roots of Anemarrhena asphodeloides by silica gel, Sephadex LH-20 and semi-HPLC column chromatography. On the basis of physic-chemical properties and spectroscopic data analysis, these compounds were identified as methyl 2-[2,4-dihydroxy-3-(4-hydroxybenzoyl)-6-methoxyphenyl]acetate(1), 4-[formyl-5-(methoxymethyl)-1H-pyrrol-1-yl]butanoate(2), perlolyrine(3),syringaresinol-4'-O-β-D-glucoside(4) and 4',6-dihydroxy-4-methoxybenzophenone-2-O-(2″),3-C-(1″)-1″-desoxy-α-L-fructofuranoside(5). Among them, 1 was a new benzophenone. Compounds 2-5 were isolated from this plant for the first time. Compound 1 was tested for neuroprotective effects against H_2O_2-induced damage in SH-SY5 Y cells.
Anemarrhena ; chemistry ; Benzophenones ; isolation & purification ; pharmacology ; Cell Line ; Chromatography, High Pressure Liquid ; Humans ; Neuroprotective Agents ; isolation & purification ; pharmacology ; Phytochemicals ; isolation & purification ; pharmacology ; Plant Roots ; chemistry

Two new steroidal saponins, named timosaponin P (1) and timosaponin Q (2), were isolated from the rhizome parts of Anemarrhena asphodeloides Bunge using various chromatographic methods. Their structures and absolute configurations were elucidated by a combination of spectroscopic and spectrometric data, including 1D, 2D NMR, HR-ESI-MS and ECD calculations, and this is the first time the absolute configuration of C-23 of steroidal saponin was confirmed by ECD calculations.
Anemarrhena ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Molecular Structure ; Saponins ; chemistry ; isolation & purification ; Steroids ; chemistry ; isolation & purification

Investigations into the development of new therapeutic agents for lung inflammatory disorders have led to the discovery of plant-based alternatives. The rhizomes of Anemarrhena asphodeloides have a long history of use against lung inflammatory disorders in traditional herbal medicine. However, the therapeutic potential of this plant material in animal models of lung inflammation has yet to be evaluated. In the present study, we prepared the alcoholic extract and derived the saponin-enriched fraction from the rhizomes of A. asphodeloides and isolated timosaponin A-III, a major constituent. Lung inflammation was induced by intranasal administration of lipopolysaccharide (LPS) to mice, representing an animal model of acute lung injury (ALI). The alcoholic extract (50–200 mg/kg) inhibited the development of ALI. Especially, the oral administration of the saponin-enriched fraction (10–50 mg/kg) potently inhibited the lung inflammatory index. It reduced the total number of inflammatory cells in the bronchoalveolar lavage fluid (BALF). Histological changes in alveolar wall thickness and the number of infiltrated cells of the lung tissue also indicated that the saponin-enriched fraction strongly inhibited lung inflammation. Most importantly, the oral administration of timosaponin A-III at 25–50 mg/kg significantly inhibited the inflammatory markers observed in LPS-induced ALI mice. All these findings, for the first time, provide evidence supporting the effectiveness of A. asphodeloides and its major constituent, timosaponin A-III, in alleviating lung inflammation.
Acute Lung Injury ; Administration, Intranasal ; Administration, Oral ; Alcoholics ; Anemarrhena* ; Animals* ; Bronchoalveolar Lavage Fluid ; Herbal Medicine ; Humans ; Lung* ; Mice ; Models, Animal* ; Plants ; Pneumonia* ; Rhizome*