Study On Good Agricultural Practice Of Herba DesmodⅡ Styracifolii

Herba DesmodII Styracifolii is one of the special Chinese herbs in South China. Its original plant is Desmodium styracifolium (Osb.) Merr., and is considered to be good in increasing micturition, Clearing overfull heat and water in the body. It is used in preventing and curing kidney stone, urethra inflammation and so on.Desmodium styracifolium (Osb.). Merr. is mainly distributes in Guangdong, Guangxi, Hainan and Yunan provinces. It is abroad used in the folk and clinic. In the past Desmodium styracifolium (Osb.) Merr. grows mainly in wildness. After years of rooted out, its resource has been profoundly damaged. Now there have some in miniature planting in Guangdong and Guangxi provinces. But there are two problem can't solved.: medicinal materials supplying isn't stabilization, and their quality is instability. To ensure a sustainable development of Desmodll Styracifolii resource, To ensure the quality, we establish d a standardized plant basement for Desmodium styracifolium (Osb.) Merr. in Pingyuan county Guangdong province and study of the medical material quality according to the Quality Control Standard on Production of Medicinal Herb(Draft), a Good Agricultural Practices (Abbreviated as GAP) of DesmodII Styracifolii was studied in detail.The whole study included four aspects: selecting plant basement and analyzing environmental factors;species research;standardized cultivation technique research and quality control research. Based on the research we established the Quality Standard and Standardized Operation Practice(SOP) of Herba Desmodll Styracifolii.I Selecting Cultivation Basement and Analyzing Environmental Factors 1. Investigating of Environment Conditions of Cultivation BasementThe investigation showed that Pingyuan county was rich in plant and animal resources and had a long history of Traditional Chinese Herb Medicine planting. The local government strongly supported its development.An investigation on the environment condition showed that the geography location, weather condition and soil of this area fitted well to the growth requirement of Desmodiumstyracifolium (Osb.) Merr.2. Analysis of Air,Soil and WaterThe samples of air, soil and water of Pingyuan were analyzed, the results showed that their qualities were much better than the national standard setted by the Quality Control Standard on Production of Medicinal Herb (Draft). Therefor, Pingyuan forest farmland was chosen as planting basement of Desmodium styracifolium (Osb.) Merr.II Study on Species and Resource of Desmodium styracifolium (Osb.) Merr.After investigating its original plant species and scientific name, confusions over source of Herba Desmodll Styracifolii were clarified. Its origin was Desmodium styracifolium (Osb.) Merr. the resource distribution of Desmodium styracifolium (Osb.) Merr. and acknowledged that there are resources in Southeast part, Southwest part . it's distributed in Guangdong, Guangxi, Hainan and Yunan provinces ect.HI Study on the Standardized Cultivation of Desmodium styracifolium (Osb.) Merr.1. study on quality of the seedsWe carried out the study on quality inspection including 1000-seed weight, water content, vigor detection, germination capacity etc. the result showed that vigor of the maturation seeds is the highest. With the temperature higher, germination periods is shorter. And the best seeding density is 0.5~lg/m2.2. Seeding Technique3. Plant Technique4. Fertilization StudyTo find out a suitable fertilizer for Desmodium styracifolium (Osb.) Merr. ,a comparative study of different fertilizers(biological-organic, compound chemical, farmers' home-produced) was carried out by monofactorial design.^Results showed that all the 3 fertilizations had no distinct difference if enough fertilizer was give before plant.5. Investigation and Prevention of harmful Insects and Diseases of Desmodium styracifolium (Osb.) Merr.Investigation of disease and harmful insects showed that epidemic disease is the commonest disease, while armyworm, caterpillar, grasshopper were the commonest harmful insects. Methods of prevention include agricultural, biological and chemical preventions.To find out the effects of pesticide on plants of Desmodium styracifolium (Osb.) Merr. acomparative study of different pesticides(biological-pesticide A , biological-pesticide B, chemical pesticide) was carried out by monofactorial design. The results showed that all 3 pesticide are effective and safe. 6. Collection and elaboration6.1 Time of colletingThe study showed: Desmodium styracifolium (Osb.) Merr. is the eugonicest from August to September ,the output is the highest, total flavones content is the highest at the begin of anthesis.6.2 Part of collectingTotal flavones content and amylose content in the leaf and the stem was determine. Result showed: total flavones content in leaf is higher than stem. And amylose content in stem is higher than leaf.We suggest the best collecting time is at the begin of anthesis.IV Study on quality of medical material1. Study on chemical compositionWe have found 4 chemical compositions from the medical material by gel silica column chromatography repeatedly, two compounds have identified by four optical spectra, there are O-hydroxybenzoic acid and bis(2-ethylhexyl) phthalate. The two are the first time discovered in Desmodium styracifolium (Osb.) Merr.2. Analysis of GC-MSGC-MS was used to analyze the volatile components extracted by distillation. The results showed the essential oil components consist of n-Hexadecanoic acid, 9,12-octadecadienoic acid(Z,Z)-,acid, Lup-20(29)-en-3-one, Lupeol an so on . 3.Study on TLC, HPLC and FTIR qualitative analysisTLC, HPLC and FTIR methods were applied for distinguishing Desmodium styracifolium (Osb.) Merr. The preparation of sample, mobile phase, the HPLC column, elution mode and gradient program were optimized in order to obtain high quality TLC and HPLC fingerprint profile. The TLC system consisted of two solvents system: benzene-acetic ether-formic acid (1:8:1) and - petroleum ether-acetic ether-formic acid (3:1:0.2);detection wavelength: 366nm;standard substance: O-hydroxybenzoic acid and isovitexin;The HPLC system consisted of a Dionex-P680 and a 996 photodiode-array detector(DAD). HPLC analysis was performed on a Diamonsil? LC-18 column(250X 4. 6nm, 5 u m) with the mobile phase in isocratic elution : methanol(A):H2O(0.2% phosphoric acid) (28 '■ J2) A the column temperature was setup at 40°C and the flow-rate was I.Oml/min, tA substance: isovitexin. FTIR method had the advantage of high speed, conveniencenot to destroy sample. The power was determined direct.Result: Establishing the fingerprint of TLC and HPLC chromatographic fingerprint of total flavones, respectively. The HPLC chromatogram of total flavones, showing 9 characteristic peaks, was established 10 batch of Herba Desmodll Styracifolii samples from Cultivation Basement. Moreover, comparison of the TLC and HPLC profile of samples from different producing area and its confusion breed. The results indicated that some of them have distinct differences . the FTIR method can distinguish sample different harvestry time and its confusion breed.4. examine experimentWe determinated moisture content, total ash, acid -insoluble ash, alcohol-soluble extractive, water-soluble extractive, heavy metals residue level and pesticide residue respectively according to the methods of supplement of China Pharmacopeia. The result showed that water-soluble extractive exceed than the request of China Pharmacopeia.5. Study on quantitative analysisThe method of determining total flavones content, amylose content and isovitexin content was found. We use UV-VIS method determined total flavones and amylose content., and use HPLC determined isovitexin content. Result shows that the sample's content of total flavones, amylose and isovitexin is stabilization from Cultivation Basement, but the sample's content is different obviously from different producing area .6. Experimental Study on diuresis effect of Herba Desmodll StyracifoliiEffect of diuresis by from Herba Desmodll Styracifolii were studied. The rat was divided into two part. One is Pathologic model group (the rat's abdomen injected by water) ,the other is unsettled group, each part was divide into control group, low dose group, middle dose group and high dose group. Result shows: There was no significant difference of 24 hour's amount of urine between the unsettled groups, but has significant difference in the concentration and content of Cl"(p<0.05). There was significant difference of amount of urine between three and six hours after feed by amylase and the concentration and content of Cl" between Pathologic model group and control group.Based on the research, the quality standard of Herba Desmodll Styracifolii was established.According to the Quality Control Standard on Production of Medicinal Herb(Draft), A Standardized Operation Practice(SOP) of Desmodium styracifolium (Osb.) Merr. was established on the basis of our study and planting practice. The SOP can be divided into the following aspects:(1) Natural conditions of planting field(2) Species and its breeds(3) Seeding cultivation and planting(4) Planting density(5) Field management(6) Prevention and treatment of diseases and insects(7) Collecting and processing(8) Seeding preservation(9) Quality criteria and control(10) Package, storage and transportation...