Studies On The Preparation Of Effective Fractions Of Compound Yibazhen And Their Quality Standards

Hypertensive Renal Disease is nephrosclerosis and malignant arteriolar nephrosclerosis, which is caused by essential hypertension. Hypertensive kidney disease is one of the major complications of the hypertension, and the morbidity rate is about 42%, which is only lower than that of heart complications. The long-term use of chemicals tends to cause adverse reactions and tolerance, as well as many contraindications to these chemicals. Thus, chemicals have limitations for this disease.The prescription of Compound Yibazhen comes from Tang Shu-hua's experience prescription, which is proved effective and has been used for many years in the treatment of Hypertensive Renal Disease. The recipe is made up of wine-prepared Fructus corni, prepared radix polygoni Multiflori and Bidens biPinnata L.. In this paper, the selection, preparation, quality standards of the effective fractions of Compound Yibazhen were studied.The main research processes are as follows:1. The confirmation of effective fractionsThe effective fractions of Compound Yibazhen were confirmed according to pharmacology experiments and literature, which including iridoid glycoside of wine-prepared Fructus corni, stilbene glycosides of prepared radix polygoni multiflori, total flavonoids of Bidens biPinnata L2. Quality control of the botanical raw materials of Compound Yibazhen(1) The method was established to determine the content of loganin and morroniside, stilbene glycoside, total flavonoids.(2) Each active constituent in botanical raw materials was determined. It was confirmed that the contents of loganin and morroniside of wine-prepared Fructus corni were in the range of 0.50%～0.90%,0.90%～1.60%; stilbene glycoside of prepared radix polygoni multiflori was in the range of 0.90%～2.00%; total flavonoids of Bidens biPinnata L.. was in the range of 2.50%～4.50%.3. Studies on the preparation technology of each effective fractions(1) Using the translation rate (content) of the loganin and morroniside as the index, extracting method, alcohol precipitation process, separation and purification of iridoid glycoside have been optimized:wine-prepared Fructus corni was extracted 3 times, using 10-fold volume of water,80 mins each time. Water extracts should be concentrated until relative density of the extraction is 1.20(60℃), then alcohol solution was added to the extraction until its alcohol concentration is 60%, precipitated for 12 hours. the concentration of the extracted solution was diluted to 0.5 g raw material per 1 mL before adsorbing, wet resin vs crude herbs was 1:0.75 (weight ratio), current velocity was 2 BV/h, the volume of distilled water was 5 BV, the eluting reagent was 15% ethanol and the volume of 15% ethanol was 9 BV, then concentrate the eluting reagent to 1.05～1.15(60℃), the temperature is 60℃.(2) Using the Translation rate of the stilbene glycoside as the index, method of extracting and purification were optimized:prepared radix polygoni multiflori was dissolved with 8 times of water for three successive extraction and 75 mins per time, water extracts should be concentrated until elative density of the extraction is 1.10 (60℃), then extracted the solution with same volume of ethyl acetate four times. The layer of ethyl acetate was evaporated in vacuum and the residue was desiccated.(3) To get the optimized preparation technology, method and conditions of extraction and purification were studied on the contents of total flavonoids. The optimized extraction conditions were:13 times as much as the volume 70% ethanol for three successive extraction and 1 hour per time. D101 resin was chosed to purify total flavonoids from Bidens biPinnata L.. Technological conditions were as the follows:the concentration of the extracted solution was diluted to 0.5 g raw material per 1 mL before adsorbing, crude herbs vs wet resin was 1:0.5, current velocity was 3 BV/h, the volume of distilled water was 6 BV, the eluting reagent was 50% ethanol and the volume of 50% ethanol was 9 BV, Drying temperature is 80℃.(4) Pilot-scale experiments have been carried out using the studied preparation process.4. Quality standards of effective fractions(1) The identification of Bidens biPinnata L. and each effective fractions were established by TLC method.(2) The content of loganin, morroniside, and stilbene glycoside were determined by HPLC; total flavonoids were determined by UV; and the standard contents of these constituents were set more than 50%,55%,50%, respectively.(3) The moisture, ignition residue and heavy metal of every effective fractions, and ethyl acetate residue of stilbene glycoside were investigated and established the suitable content limitation.