Dynamic Accumulation Of The Synthesis And Content Difference Of The Of Bioactive Constituents In Isatis Indigotica Fort

Isatis indigotica is a traditional medicinal plant, belonging to the family of Cruciferae. Its roots can be prepared as the crude drug, radix isatidis, and the leaves can either be dried as another crude drug folium isatidis or be processed to the third crude drug, indigo naturalis. Indigo and indirubin both are bioactive constituents in thses three crude drugs. Three methods are systematically studied to measure these two bioactive constituents to probe their advantages and disadvantages, i.e., high performance liquid chromatography (HPLC), thin layer chromatographic scanning (TLCS) and spectrophotometry with double wavelength (DWSP) measurement. Different preparing methods are compared while processing folium isatidis and radix isatidis. The influence of the temperature of water bath on the contents of Indigo and indirubin was studied while measuring their contest according to the method described by "The Pharmacopoeia of the People's Republic of China". Rules of the dynamic accumulation of both index constituents in different climate zones and different growth periods and different locations are investigated. The results are as follows. 1. Based on the methodological study of three analysis methods, HPLC, TLCS, DWSP, it has been found that there exist advantages and disadvantages among them. They can be used within different scope of precision. Comparison of three methods on stabilities, precisions, respectabilities and recover rates show that HPLC is the best method. DWSP gets higher analytical results due to the interference of other impurities. TLCS is suitable to analyze indigo and indirubin contained in folium isatidis and indigo naturalis because of the higher contents, not suitable to radix isatidis for its low contents of the two constituents. 2. Different drying methods for processing radix isatidis and folium isatidis show that drying raw materials at the temperature of 60℃in an oven is the best method for its lowest losses of bioactive constituents. Drying raw materials at higher temperatures can result the losses of bioactive constituents as high as 40%~60% for the preparation of radix isatidis and 30%~60% for folium isatidis. Considering the appearance of the crude drugs, contents of bioactive contents, the suitable methods for processing crude drugs are drying at temperatures between 50~80℃or sun drying. 3. To determine the contents of indigo and indirubin in crude drugs by soxhlet extraction methods according to the pharmacopoeia, the suitable temperature of water bath is 80℃. 4. The contents of bioactive constituents are closely related to growth duration. The contents are higher in folium isatidis harvested in the first growth year than those in the second year, and reversely in radix isatidis. 5. The peak periods of the accumulation of bioactive constituents in the roots appear in July, October to the middle of November and April in the next year; in the leaves however, appear in July, and October, August shows the lowest contents. The peak periods of the accumulation of bioactive constituents are oppositely related to the growth of the plant. 6. The peak of dynamic accumulation of bioactive constituents in northern semitropical zone advances 2 weeks compared to northern temperate zone. The peak periods in the former appear in September to October; and in the middle of October to the middle of November. 7. Roots used for radix isatidis should be harvested when the contents of bioactive constituents are higher and while the growth of the plant is stable, and the appearance of the root can meet the standard. The harvest time should be varied in different climate zones. 8. The contents of bioactive constituents vary with locations where the medicinal plant grow, indicating it plays an important role for the quality of crude drugs.