| Lycopus lucidus. is a kind of perennial herb of the genus Lycopus L.from the Labiatae and widely distributed in our country. The stems and leaves of Zeeland is a Chinese herbal medicines which has been recorded in Chinese Materia Medica and The dictionary of Chinese Herbal Medicine for its effects of blood circulation. several essential nutrient content, Total phenols content and total tlavonoids content in ground leaves and rhizomes of Lycopus lucidus from Henan, Chongqing, Yunnan, Xizang were measured and compared in this study at the background of less research literature of Lycopus lucidus..Many kinds of triterpenoids acid such as oleanolic acid, ursolic acid, betulinic acid has been found in Lycopus lucidus.. It is of great significance to extract and separate the triterpenic acid for its low content in plants and animals. In this stuy, the reaction conditions and extraction process of triterpenic acid of Lycopus lucidus. was optimized. It studied the influence of added amounts of Vanillin-glacial acetic acid and perchloric acid, the reaction temperature and reaction time on the measurement results of triterpenic acid of Lycopus lucidus.; then studied the influence of ethanol concentration, heating time, heating temperature, solid-liquid ratio on extraction rate of triterpenic acid of Lycopus lucidus., and obtained the optimum extraction process of triterpenic acid of Lycopus lucidus. by response surface experiments.In this study, macroporous resin was used to purify triterpenic acid of Lycopus lucidus.. It studied the influence of the type of resin, sample rate, eluent concentration, elution rate, the volume of the eluent on purification effect triterpenic acid of Lycopus lucidus., which indicated with macroporous resin’s adsorption, absorption rate, elution rate and purity of triterpenic acid. Then purified and measured its content of triterpenic acid of Lycopus lucidus. from different areas with the above experimental conditions.Based on the above background and experiment procedure, the main results were summarized as following:(1) The experimental results of nutrients content in Lycopus lucidus. showed that the protein content is 5%～6%, fat content is 2%～4%, the total sugar content is 13%～60%, reducing sugar content is 2%～4%, starch content is 2%～16%, an ash content is about 4%～6.5%, potassium content is about 1000～3000 mg/100 g, the sodium content is about 40～60 mg/100 g, the calcium content is about 70～200 mg/100 g, the magnesium content is about 90～500 mg/100 g, the zinc content is about 5～12 mg/100 g, the iron content is about 15～35 mg/100 g. the copper content is 1～3 mg/100 g, the manganese content is 2～8 mg/100 g, total phenols content is 1%～3%, total tlavonoids content is 5%～12%, triterpenic acid content is 1%～3%.(2) The various ingredients content were significant in Lycopus lucidus. from different areas and different parts, but comparing different origin, different parts had greater effect on the ingredients content in Lycopus lucidus.. There were 55% of total sugar content and 13% of starch in the root and about 5 times than the content in leaves of Lycopus lucidus., but the content of total phenols, total tlavonoids and triterpenic acid in the root wre about 3 times than the content in leaves of Lycopus lucidus.. In addition, the fatty acid mainly consisted of palmitic acid, oleic acid, linoleic acid, linolenic fatty acid and eicosanoids in ground leaves and rhizomes of Lycopus lucidus..(3) The experimental results showed that the absorbance values of Lycopus lucidus’ s triterpene acid reached the maximum when the added amounts of Vanillin-glacial acetic acidand perchloric acid was respectively 0.4 m L and 1 m L, the reaction temperature and reaction time was respectively 65℃ and 20 min and the best measurement wavelength was 540 nm. The extraction rate of triterpenic acid of Lycopus lucidus. reached the maximum when the ethanol concentration, heating time, heating temperature, solid-liquid ratio was respectively 85%, 85℃, 1 h, 1:25 in the Single-factor experiment. Analysis of response surface experimental results showed that data model fitted well with the actual experimental results, the total regression equation in ANOVA was extremely significant(P <0.0001), the lack of fit items was not significant(P=0.0662)and the coefficient of determination R2=0.9962. While the influence level of individual factors on yield of triterpenes were ethanol concentration > heating time > heating temperature. The optimum extraction process of triterpenic acid of Lycopus lucidus. optimized by response surface experiments are the ethanol concentration was 80%, the heating time was 1 h, the heating temperature was 83℃, the solid-liquid ratio was 1:25, and the extract value of triterpenic acid of Lycopus lucidus. was 3.21%.(4) Experimental results showed that X-5 resin was selected to purify triterpenic acid of Lycopus lucidus. for its adsorption rate of 74.24%, desorption rate of 85.86% and decolorization rate of 86%, which significantly higher than others’. In the adsorption experiment, according to the static adsorption experiments resins graph and isothermal experiments graph, which in line with Langmuir adsorption theory and it can be speculated that triterpenic acid of Lycopus lucidus. adsorbed by X-5 belongs to a single molecule adsorption. It can be speculated that the optimum sample concentration is 2 BV/h in resin dynamic adsorption process from the leak graph macroporous resin.(5) In the experiment of dynamic desorption, it can be speculated that the best eluent was 85% ethanol and the optimum elution rate was 2 BV/h from the graph of effection of ethanol concentration on desorption rates and the graph of effection of different elution rates respectively. It can be speculated that when the eluent volume was 7 BV, the elution complete almostly. It showed that triterpenic acid content in roots and leaves of Lycopus lucidus. were about 1% and 2.5%. The purity of triterpenic acid of Lycopus lucidus. was about 74% after purification.In conclusion, there were rich primary nutrients and secondary metabolic components in Lycopus lucidus.. There were much carbohydrate and secondary metabolic components in root and leaves of Lycopus lucidus. respectively. It used ethanol as the extraction solvent and combined with reflux extraction method to Extract the triterpenic acid of Lycopus lucidus., and used macroporous resin to further separation and purification, combined with optimization and control a variety of conditions in separation and purification process, it can be showed that X-5 macroporous resin has good purification effect on triterpenic acid of Lycopus lucidus.. |
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