OBJECTIVETo study the effects of various planting densities on dynamic growth and root yield of Isatis indigotica.

METHODThe planting samples were collected to measure the growth period of each organ.

RESULT AND CONCLUSIONUnder different planting densities, both main root length and breadth exhibited a trend of "fast-slow" by stages. However, the number of individual plant leaves were showed a trend of "slow-fast" on growth period. Meanwhile, the leaf length and breadth were exhibited a trend of "increase-decrease". The increase of dry leaf, dry root and whole plant dry matter was faster during the period of 65-76 days after seeding. The differences of root and leaf yields under various densities were significant. Planting densities has a great effect on yield of root. It must be shown that there was a positive development between the individual plant and colony. According to the dry matter of root and leaf, treatment B (i.e., 7 cm x 25 cm) was good choice.

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.

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.

OBJECTIVEThe aim of the present study is to investigate the effect of low temperature vernalization on metabolism change of carbon and nitrogen of Isatis indigotica.

METHODThe Yunnan and Beijing I. indigotica seedlings with six leaves were vernalized at 4 degrees C for 25 days, and the metabolism indicators of carbon and nitrogen were measured.

RESULTThere appeared a dramatic increase in the soluble sugar content, reducing sugar content and soluble protein content in response to the low temperature, after termination of vernalization it reached the maximum, however, starch and total nitrogen concentration decreased significantly, after termination of vernalization it reached the minimum.

CONCLUSIONThe high C/N value can promote the low temperature vernalization of I. indigotica.

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.

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.

OBJECTIVETo study the dynamic accumulations of bioactive components in different germplasm Isatis indigotica and compare its quality of medical material, in order to provide a basis for breeding and high yield cultivation of I. indigotica.

METHODThe planting samples were collected during growth period, bioactive components in different germplasm Isatis indigotica were measured.

RESULTThe dynamic accumulations of bioactive components in different germplasm I. indigotica were consistently changed in a field experiment. The differences of bioactive components contents in medical material of I. indigotica were significant.

CONCLUSIONThe germplasm from Gansu Longxi showed a high yield and good quality characters in Fuyang area, and may be applied to production.

In this paper, through the collection and collation of ancient herbs, medical books and prescriptions, combined with modern literature, the historical changes of the name, origin, position, medicinal parts, collection, processing and processing of bluegrass were systematically combed and verified.It can be seen from the research that bluegrass was first used as medicine by the fruit, namely blueberry, which was originally Polygonum tinctorium. Since the Ming and Qing Dynasties, blueberry was rarely used, and it has been no longer used medicinally. In the Wei and Jin Dynasties, the medicinal parts extended to the stems and leaves, and most of them used juice as medicine.Since the Tang Dynasty, origin has been extended to Isatis indigotica, Baphicacanthus cusia, Indigofera tinctoria, Compositae plant Wulan, etc. In the Song Dynasty, the medicinal parts extended to the roots, and the "Banlangen" began to appear, and gradually became the main medicinal parts of blue medicinal materials, the main base of which was B. cusia. Since the Qing Dynasty, I. indigotica, a Cruciferae, has gradually become a genuine indigo root, while B. cusia has become a southern indigo root. It was the first mineral dye imported from abroad for thrush, and then used as medicine, also known as clam powder. Because it was found that it had the same effect with the extract of bluegrass, it was also named indigo naturalis in China, which has lasted till now. The main stream of Isatidis Folium in the past dynasties is the dry stem and leaf of Clerodendrum cyrtophylum. Since the Qing Dynasty, the stem and leaf of Isatis indigotica, P. tinctorium and other blue grasses have been gradually mixed as substitutes and gradually become the mainstream.
China ; Clerodendrum ; Isatis ; Medicine, Chinese Traditional ; Plants, Medicinal

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.

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.