OBJECTIVETo investigated the chemical structures and bioactivity of polysaccharides from Isatidis Radix.

METHODPolysaccharides were extracted and purified by column chromatograph and their chemical structures were identified by UV, IR, NMR, periodic acid oxadation and Smith degradation method and their stimulation effects to macrophage were evaluated by using MTT method.

RESULTFive polysaccharides, polysaccharide A , B, C, D and E were gotten and their molecular weights were 2 000, 1 757.1, 1 34 2.7, 955.6, 11.7 kDa, respectively. Polysaccharide A was composed of arabinose, polysaccharide E was composed of arabinose and galactose, polysaccharides B, C, D were composed of glucose and 1 --> 2, 1 --> 3, 1 --> 4, 1 --> 6 linkages existed in polysaccharides A-E, of A, B, C, D, E were alpha-configurations. Polysaccharides B, C and D showed better bioactivity than polysaccharides A and E with stimulation index (SI) of 5.31, 4.76, 5.17.

CONCLUSIONFive polysaccharides are seperated firstly from Isatidis Radix.

Animals ; Isatis ; chemistry ; Magnetic Resonance Spectroscopy ; Mice ; Polysaccharides ; chemistry ; pharmacology

OBJECTIVETo develop an HPLC method for determination of clemastanin B which has anti-viral activity in Radix Isatidis and compare the contents of clemastanin B in the drugs from different origins.

METHODThe samples were separated on an ZORBAX SB-C18 (4.6 mm x 250 mm, 5 microm) column with the mobile phase of acetonitrile-water (11:89). Flow rate was 1.0 mL x min(-1). The detection wavelength was set at 225 nm. Column temperature was 30 degrees C.

RESULTThe linear range of clemastanin B was 0.0615-1.8441 microg (r = 0.9995), the average recovery was 97.74%, RSD was 1.4% (n=9). The contents of clemastanin B were in the range of 0.269-0.900 mg x g(-1) in Radix Isatidis from different origins.

CONCLUSIONThe method for quantitation of clemastanin B in Radix Isatidis was accurate and reliable, which can be used to evaluate the quality of Radix Isatidis.

China ; Furans ; analysis ; Isatis ; chemistry ; Plant Extracts ; analysis

OBJECTIVETo establish the double HPLC fingerprints of water-soluble composition and amino acids precolumn derivative reagent of 13 batches of Isatidis Radix micropower.

METHODThe gradient elution was adopted with Hypersil BDS C18 column (4.6 mm x 250 mm, 5 microm). Water-soluble ingredients were detected with acetonitrile-0.1% phosphoric acid solution as mobile phase, flow rate 0.5 mL x min(-1), column temperature 20 degrees C, and the injection volume 10 microL. Amino acid ingredient were derived by PITC, and then were detected with mobile phase of 0.1 mol x L(-1) sodium acetate buffer solution (pH 6.5) - acetonitrile, flow rate 1.0 mL x min(-1), column temperature 30 degrees C, and the injection volume 5 microL. Both of the absorption wavelengths were 254 nm. Pharmacopoeia Commission "Chinese chromatographic fingerprint evaluation system (version 2.0)" was adopted to analyse, fingerprints of Isatidis Radix micropower were established, at the same time 4 main ingredients were recognized by the SPSS cluster analysis.

RESULTCommon mode of Fingerprint to water-soluble and amino acids ingredient of Isatidis Radix micropower was established, then adenosine, epigoitrin and 15 amino acids were identified as characteristic peaks. Cluster analysis showed that different kinds of the herbal Isatidis Radix micropower from different areas were different levels in the main ingredients.

CONCLUSIONDouble fingerprints of Isatidis Radix micropower is established. Each peak is optimally separated in chromatogram, which provides a scientific basis for quality control of Isatidis Radix micropower.

Amino Acids ; analysis ; Isatis ; chemistry ; Powders ; Quality Control

AIMTo study the chemical constituents from water extract of Radix isatidis. (Isatis indigotica Fort. ).

METHODSThe water extract was underwent absorption by D101 macroporous resin, the portion eluted by ethanol of different concentrations was isolated and purified on silica gel column repeatedly. The obtained compounds were identified and structurally elucidated by their physico-chemical properties and spectral analysis.

RESULTSFive compounds were isolated from water extract of Radix isatidis, and were partly identified separately: 3-[2'-(5'-hydroxymethyl) furyl] -1 (2H) -isoquinolinone-7-O-beta-D-glucoside (I), lariciresinol-4,4'-di-O-beta-D-glucopyranoside (II), lariciresinol-4-O-beta-D-glucopyranoside (III), 2-hydroxy-1, 4-benzenedicarboxylic acid (IV), mannitol (V).

CONCLUSIONCompound I is a new compound and compounds IV and V were isolated from the plant for the first time.

Isatis ; chemistry ; Magnetic Resonance Spectroscopy ; Plant Extracts ; analysis

AIMTo investigate the chemical constituents of tetraploidy Banlangen (Isatis indigotica Fort.).

METHODSCompounds were separated by chromatography on silica gel. Their structures were determined by spectral analysis and chemical evidence.

RESULTSThree compounds were isolated. Their structures were identified as (E)-2-[(3'-indole) cyanomethylene]-3-indolinone (I), 2-(4-hydroxy-3-methoxyphenyl)-4-[(4-hydroxy-3-methoxy-phenyl)-methyl]-3- hydroxymethyl-tetrahydro-furan (II) and 2-methoxy-4-[tetrahydro-4-[(4-hydroxy-3-methoxy-phenyl)-methyl]-3- hydroxymethyl-2-furanyl] phenyl-1-O-beta-D-glucopyranoside (III).

CONCLUSIONCompound I is a new compound.

Indoles ; chemistry ; Isatis ; chemistry ; genetics ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; genetics ; Polyploidy

Thirty-three compounds were isolated from the root decoction of Isatis indigotica by using a combination of various chromatographic techniques including silica gel, macroporous adsorbent resin, Sephadex LH-20, and reversed-phase HPLC. Their structures were elucidated by spectroscopic data as (+)-dehydrovomifoliol (1), (S)-(+)-abscisic acid (2), vomifoliol (3), cyclo (L-Phe-L-Leu) (4), cyclo(L-Phe-L-Tyr) (5), cyclo(L-Tyr-L-Leu) (6), cyclo(L-Pro-L-Tyr) (7), evofolin B (8), (+)-syringaresinol (9), (-)-(7R,7'R,8S,8'S)-4,4'-dihydroxy-3-methoxy-7,9';7',9-diepoxy-lignan (10), (-)-medioresinol (11), (+) -(7R,7'R,8S,8'S) -neo-olivil (12), (-) -5-methoxyisolariciresinol (13), 1,3-dihydro-2H-indol-2-one (14), isalexin (15), dihydroneoascorbigen (16), indican (17), (-) -(S) -cyanomethyl-3-hydroxyoxindole (18), isoformononetein (19), calycosin (20), stigamast-5-ene-3beta-ol-7-one (21), acetovanillone (22), 3, 5-dimethoxy-4-hydroxyacetophenone (23), dihydroconiferyl alcohol (24), dihyroferulic acid (25), 3-hydroxy-1-(4-hydroxyphenyl) propan-1-one (26), beta-hydroxypropiovanillone (27), 4-aminobenzoic acid (28), 3-(4-hydroxyphenyl) propan-1-ol (29), 4-(2-hydroxyethyl) phenol (30), 2-methoxy-4-vinylphenol (31), pyrocatechol (32), and 4-pentenamide (33). These compounds were isolated from the root of I. indigotica for the first time. In preliminary in vitro assays, compound 19 showed activity against the influenza virus A/Hanfang/359/95 (H3N2), the herpes simplex virus 1 (HSV-1), and Coxsackie virus B3 (Cox-B3), with IC50 values of 2.06, 6.84, and 8.70 micromol x L(-1), respectively, but other compounds were in-active at a concentration of 1.0 x 10 x (-5) mol x L(-1).
Animals ; Cell Line ; Humans ; Isatis ; chemistry ; Plant Extracts ; chemistry ; pharmacology ; toxicity ; Plant Roots ; chemistry

OBJECTIVEFor optimizing the cutting process of Radix Isatidis, the influence of cutting process on the whole chemical constituents was studied by comparing the difference of HPLC fingerprints of Radix Isatidis from various sources before and after cutting.

METHODL9(3(4)) orthogonal table were designed with three factors: The amount of solvent, soaking time and thickness of pieces. According to the amount of water-soluble extraction, orthogonal design was applied to processing; Fingerprints of acetoacetate extraction from 9 batches of Radix Isatidis from various sources before and after cutting were established with a High Performance Liquid Chromatography method based on a Agilent Zorbax SB-C18 column (4.6 mm x 250 mm, 5 microm) with gradient elution solvent system composed of acetonitrile-water solution; the detection wave length was set at 238 nm; sample injection was 10 microL. The fingerprints were compared with similarity evaluation software published by the Committee of China Codex.

RESULTThe best process of cutting was following: the amount of solvent was 0.6 times of amount of Radix Isatidis, soaked for 24 h and the thickness of piece was 2 mm.

CONCLUSIONThe obvious difference is displayed by the comparing of HPLC fingerprints. It is indicated that the constituents in polarity medium lost a lot after cutting process.

Chromatography, High Pressure Liquid ; Isatis ; chemistry ; Plant Extracts ; chemistry ; Solvents ; chemistry

OBJECTIVETo establish an HPLC method for the content determination of epigoitrin in Radix Isatidis and its preparation, and to provide valuable data for quality control of Radix Isatidis and its preparation.

METHODThe samples were separated on a ZORBAX SB-C18 (4. 6 mm x 150 mm, 5 microm) column with the mobile phase of acetonitrile-water-phosphoric acid-triethylamine (8.50 : 90.72 : 0.73 : 0.05) in the flow rate of 0.7 mL x min(-1). The detection wavelength was set at 245 nm. Column temperature was 30 degrees C.

RESULTThe linear range of epigoitrin was 0.0204-0.3060 microg (r = 0.9998), and the average recovery was 98.99% with the RSD was 1.31% (n = 9).

CONCLUSIONThe method for quantitation of epigoitrin in Radix Isatidis and its preparation was accurate and reliable, which can be used to evaluate the quality of Radix Isatidis and its preparation.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Isatis ; chemistry ; Reproducibility of Results

OBJECTIVETo analyze characterization and determination of nitrogen in the preparation of Qingkailing injection and its intermediate products.

METHODHitich amino acid auto analyzer was used, with the packed analysis column (2.6 mm x 50 mm) and the type of was Hitich 2622 SC resin. The speed of buffer solution and ninhydrin colorimetric solution were 0.4 mL x min(-1) and 0. 3 mL x min(-1) respectively. Program heating was used for controlling column temperature, from 57 degrees C (0.0 min) to 65 degrees C (36 min) to 57 (50 min). The reaction temperature was set at 130 degrees C.

RESULTFree and binding amino acid the existenceare the main form of nitrogen is amino acid in Qingkailing injection and its intermediate products. The total contents of amino acid in the preparation of Qingkailing injection and its intermediate products, including hydrolyzed solution which is made from neutralization of Concha Margaritifera solution extracted by diluted sulfuric acid and Cornu Bubali solution extracted by diluted sodium hydroxide, aqueous solution of Radix Isatidis extract, 4-blended solution, 6-blended solution and 8-blenede solution, were 59.56%, 24.88%, 41.84%, 13.49, 14.63% respectively. The type of bonded amino acid was founded in the preparation of Qingkailing injection and its intermediate products, including hydrolyzed aqueous solution of Radix Isatidis extract, 4-blended solution, 6-blended solution and 8-blenede solution, and the contents were 9.33%, 15.07%, 16.85%, 19.94% and 19.55%, respectively.

CONCLUSIONThe main resource of the total nitrogen was Bubalus bubalis L. and Isatis indigotica Fort.

Amino Acids ; analysis ; chemistry ; Animals ; Buffaloes ; metabolism ; Chromatography ; methods ; Drugs, Chinese Herbal ; chemistry ; Isatis ; chemistry ; Nitrogen ; analysis ; chemistry

OBJECTIVETo select the optimum formula fertilization of Isatis indigotica through analyzing the yield and contents of polysaccharide of Radix Isatis for different treatments.

METHODAn orthogonal experiment design on the basis of three factors and four levels was applied for studying the effect of formula fertilization on yield. The contents of polysaccharides were determined with phenol-witriolic colorimetry.

RESULTThe optimum formula fertilization of Radix Isatis was carbamide 869.0 kg x hm(-2), superphosphate 1 428.6 kg x hm(-2) and potassium sulfate 0 kg x hm(-2).

CONCLUSIONSuperphosphate can observably influence the yields of Radix Inditis. while carbamide influence the contents of polysaccharide of Radix Inditis.

Biomass ; Diphosphates ; Fertilizers ; Isatis ; chemistry ; growth & development ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; growth & development ; Polysaccharides ; analysis ; Sulfates ; Urea