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Primary Study on the Extraction and Purification of Folium Eucommiae and the Aucubin Accumulation in Seeds of Eucommiae

Author: SunYanChao
Tutor: LiQin；DuHongYan
School: Henan University
Course: Of Pharmacy
Keywords: Eucommia leaf Chlorogenic acid Extraction and purification Aucubin Accumulation content
CLC: R284
Type: Master's thesis
Year: 2010
Downloads: 250
Quote: 0
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Abstract

Eucommia Eucommia Eucommia plants Eucommia ulmoides Oliv. Dried bark, only contained in the \(2005 edition a) collection contains varieties, Eucommia leaf and warm, slightly acrid taste, liver, kidney, with liver and kidney, strong bones role. This thesis is on the skin and leaves of Eucommia chemical composition, pharmacological effects and clinical applications, Eucommia leaf chlorogenic acid and other active ingredients, based on review of Eucommia leaf chlorogenic acid extraction and purification process was optimized by single factor test and positive post test, optimize Enzymatic preliminary extraction of chlorogenic acid in Eucommia leaf extract process parameters, combined ultrasonic extraction residue on Eucommia leaf extract chlorogenic acid efficiently determine the appropriate Eucommia leaf chlorogenic acid optimum extraction process : cellulase enzyme dosage 2 mg / g Du mid herbs, pH4.0, 16 times the amount of water, 45 ℃ water bath, hydrolysis 1.5 h, extracted once, pH4.0 residues plus 16 times the amount of water, sonication (power 180 W, frequency 40 kHz) 40 min, and the optimum verified. To chlorogenic acid eluate extraction rate indicators, NKA-Ⅱ macroporous resin enrichment and purification of chlorogenic acid Eucommia leaf process were investigated systematically studied on a sample solution pH, maximum adsorption capacity, washing except miscellaneous dosage, eluent ethanol and other key process parameters, and determine the best conditions for the enrichment and purification: leaf extract was concentrated to about 0.09 g crude drug / mL of the sample solution, adjust the sample solution pH3 ~ 4, according to the Sample solution: dry resin ratio = 24:1 on NKA-Ⅱ macroporous resin column, 1.0 mL / min dynamic adsorption, first with 10 times the amount of dry resin column column washed with distilled water, then 14 times the amount of 70% of the dry resin ethanol eluate was concentrated under reduced pressure at 40 ℃ drying, the leaves dry extract Eucommia chlorogenic acid crude. Chlorogenic acid content as an indicator of silica gel column chromatography purification Eucommia leaf dry extract chlorogenic acid crude process parameters were investigated, and to determine the final process: dry extract chlorogenic acid crude: Silicone = 1:40 proportion of the sample, ethyl acetate - formic acid - water (12:1:1) 400 mL elution, TLC TLC, the same merger chlorogenic acid fraction, 40 ℃ vacuum dried up, that was purified chlorogenic acid products. A HPLC method for the determination Eucommia Eucommia leaves and leaf dry extract content of chlorogenic acid in approach and methodology were carried out visits to chlorogenic acid as an indicator to measure Eucommia chlorogenic acid content in the range of 2.46 ~ 2.51 %, with an average content of 2.48%; determination of chlorogenic acid Eucommia leaf dry extract content of chlorogenic acid in the range of 27.63 ~ 27.89%, the average content of 27.52%. Chlorogenic acid refined products through average purity by HPLC purity of 93.13% over the average, the average recovery was 78.42%. Establish a RP-HPLC method for the determination Eucommia seed kernel content Aucubin approach and methodology for the detailed investigation of different origin and different varieties of seeds and peel Eucommia Aucubin content were determined. Eucommia peel Aucubin were not detected, the different varieties of seeds Eucommia Aucubin content results in a larger difference, the same species in different areas Eucommia Seeds Aucubin content have different degrees of difference, the same harvest time in different areas Eucommia Seeds Aucubin content differences, including Henan Shangqiu China Eucommia Seeds Chung on the 6th Aucubin highest content of up to 10.93%; while the same origin Shangqiu, different harvest Time Eucommia Seeds content Aucubin there are significant differences, including September 25, 2007 - October 25 Eucommia Seeds collected Aucubin content change is 10.94% ~ 11.90%, remained relatively high level, from the extraction and use of Aucubin point of view, September 25 - October 25 as Gutta suitable collection period. Establish a RP-HPLC method for simultaneous determination of Eucommia leaves geniposidic acid, chlorogenic acid, geniposide content in the same place, different acquisition time of Eucommia leaves three active ingredient content were determined. The results showed that the same origin, different acquisition time of Eucommia leaves three active ingredient content are different, May 5, 2009 - June 20 Eucommia leaves three active ingredient content to maintain a high level, which June 5 Eucommia leaves three active ingredient content of the highest three active ingredients Eucommia leaves the average levels were as high as 4.58%, 2.23%, 0.13%, but this acquisition will seriously affect Eucommia leaves of the tree growth. Later, as the leaf senescence, leaves three active ingredient content continues to decrease in mid-July to August chlorogenic acid decreased slowly, August 20, after three active ingredients rapidly decreased to deciduous ago (October the 20th) leaves the lowest three active ingredients. Taken together, mid-July to August more appropriate Eucommia leaf collection.

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