阅读说明：本技术 一种含京尼平、绿原酸活性成分的杜仲提取物制备方法 (Preparation method of eucommia ulmoides extract containing genipin and chlorogenic acid active ingredients ) 是由 赵德刚 雷跃 宋莉 李民学 赵懿琛 于 2020-05-28 设计创作，主要内容包括：本发明公开一种含京尼平、绿原酸活性成分的杜仲提取物制备方法,其特征在于：包括下列步骤：1)杜仲鲜叶烘干,粉碎过筛制成杜仲叶超微粉；2)取杜仲叶超微粉加入蒸馏水浸泡后水浴提取,提取物离心收集上清液,滤渣重复提取多次；3)取上清液进行干燥后即获得杜仲提取物；4)取杜仲提取物适量,通过气相色谱-质谱技术测定其活性成分。(The invention discloses a preparation method of an eucommia ulmoides extract containing active ingredients of genipin and chlorogenic acid, which is characterized by comprising the following steps of: comprises the following steps: 1) drying fresh folium Eucommiae, pulverizing, and sieving to obtain folium Eucommiae micropowder; 2) soaking folium Eucommiae micropowder in distilled water, extracting in water bath, centrifuging the extract, collecting supernatant, and repeatedly extracting the residue for several times; 3) drying the supernatant to obtain cortex Eucommiae extract; 4) taking a proper amount of eucommia ulmoides extract, and measuring active ingredients of the eucommia ulmoides extract by a gas chromatography-mass spectrometry technology.)

1. A preparation method of an eucommia ulmoides extract containing active ingredients of genipin and chlorogenic acid is characterized by comprising the following steps: comprises the following steps:

1) drying fresh folium Eucommiae, pulverizing, and sieving to obtain folium Eucommiae micropowder;

2) soaking folium Eucommiae micropowder in distilled water, extracting in water bath, centrifuging the extract, collecting supernatant, and repeatedly extracting the residue for several times;

3) drying the supernatant to obtain cortex Eucommiae extract;

4) taking a proper amount of eucommia ulmoides extract, and measuring active ingredients of the eucommia ulmoides extract by a gas chromatography-mass spectrometry technology.

2. The method for preparing eucommia ulmoides extract containing genipin and chlorogenic acid as active ingredients according to claim 1, is characterized in that: picking the fresh eucommia leaves in the step 1) in 4 months, drying at the temperature of 80-90 ℃ for 4-6h, crushing, and sieving with 80 meshes.

3. The method for preparing eucommia ulmoides extract containing genipin and chlorogenic acid as active ingredients according to claim 1, is characterized in that: the volume of the solvent in the step 2) is 32 times of that of the eucommia leaf powder, the water bath extraction time is 2 hours, and the water bath temperature is 80-90 ℃.

4. The method for preparing eucommia ulmoides extract containing genipin and chlorogenic acid as active ingredients according to claim 1, is characterized in that: and 2) the rotation number of a centrifuge is 10000rpm, the centrifugation time is 5min, the repeated extraction times are 5 times, and the filtrates obtained by 1-5 times of extraction are combined.

5. The method for preparing eucommia ulmoides extract containing genipin and chlorogenic acid as active ingredients according to claim 1, is characterized in that: the volume of the supernatant in the step 3) is 30-40ml, the drying temperature is 50-70 ℃, and the drying time is 48 h.

6. The method for preparing eucommia ulmoides extract containing genipin and chlorogenic acid as active ingredients according to claim 1, is characterized in that: step 4), the chromatographic conditions are as follows: column size C18(1.8 μm, 2.1 × 100 mm); the column temperature was 35 ℃; the flow rate is 0.3 ml/min; the sample amount is 5 mul; the mobile phase was eluted using a gradient of acetonitrile-0.1% aqueous formic acid.

7. The method for preparing eucommia ulmoides extract containing genipin and chlorogenic acid as active ingredients according to claim 1, is characterized in that: step 4) mass spectrum conditions are as follows: the ionization mode is an electrospray positive ion mode, the temperature of an ion source is 600 ℃, the atomizing gas is nitrogen, the scanning range of primary mass spectrum parent ions is 50-1000, and the scanning range of secondary ions is 50-1000; the ionization mode is an electrospray negative ion mode, the temperature of an ion source is 500 ℃, the atomizing gas is nitrogen, the scanning range of primary mass spectrum parent ions is 50-1000, and the scanning range of secondary ions is 50-1000.

Technical Field

The invention belongs to the field of traditional Chinese medicine extracts, and particularly relates to a preparation method of an eucommia ulmoides extract containing active ingredients of genipin and chlorogenic acid.

Background

Eucommia ulmoides (eucommia ulmoides)Eucommia ulmoedesoliver) also known as Sec.kaempferi, Siberian cottonwood, Yusimian bark, etc., is a unique and rare medicinal tree species in China. Cortex Eucommiae and folium Eucommiae contain lignans and glycosides, iridoids (including genipin), phenylpropanoids (including chlorogenic acid), flavonoids, polysaccharides, gutta percha, alkaloids, vitamins, microelements, amino acids, and polysaccharides, and has antiinflammatory, antibacterial, antioxidant, blood glucose reducing, immunity enhancing, and antifatigue effects. Eucommia bark is used as a Chinese medicinal material for bark, and leaves are not used, so that resource waste is caused. Establishes a rapid, simple and effective separation and preparation technology of the medicinal active ingredients stored in the eucommia ulmoides leaf resources, and has important significance for the development of the eucommia ulmoides industry chain.

Disclosure of Invention

The purpose of the invention is: compared with the existing extraction method, the method is simple and easy to implement, low in cost and free of organic solvent pollution, the content of genipin and chlorogenic acid in the prepared eucommia ulmoides extract is high, and the eucommia ulmoides leaf extract prepared by the method has high medicinal value due to the effects of oxidation resistance, inflammation resistance, antibiosis, immunity regulation and the like.

The invention is realized by the following steps: a method for preparing eucommia ulmoides extract containing genipin and chlorogenic acid active ingredients comprises the following steps:

1) drying fresh folium Eucommiae, pulverizing, and sieving to obtain folium Eucommiae micropowder;

2) soaking folium Eucommiae micropowder in distilled water, extracting in water bath, centrifuging the extract, collecting supernatant, and repeatedly extracting the residue for several times;

3) drying the supernatant to obtain cortex Eucommiae extract;

4) taking a proper amount of eucommia ulmoides extract, and measuring active ingredients of the eucommia ulmoides extract by a gas chromatography-mass spectrometry technology.

Picking fresh eucommia leaves in the step 1) for 5 months, drying in an oven, wherein the stacking thickness is 2cm, the drying temperature is 80-90 ℃, the drying time is 5h, and sieving with 80 meshes after crushing;

the volume of the solvent in the step 2) is 32 times of that of the eucommia leaf powder, the water bath extraction time is 2 hours, and the water bath temperature is 80-90 ℃;

the revolution of the centrifuge in the step 3) is 10000rpm, the centrifugation time is 5min, the repeated extraction times are 5 times, and the filtrates obtained by 1-5 times of extraction are combined;

in the step 4), the volume of the supernatant is 30-40ml, the drying temperature is 50-70 ℃, and the drying time is 48 h;

step 5) chromatographic conditions are as follows: the chromatographic instrument is as follows: shimadzu LC-30A; column size C18(1.8 μm, 2.1 × 100 mm); the column temperature was 35 ℃; the flow rate is 0.3 ml/min; the sample amount is 5 mul; the mobile phase adopts a gradient elution program of acetonitrile-0.1 percent formic acid water solution;

step 6) the mass spectrum conditions are as follows: the ionization mode is an electrospray positive ion mode, the temperature of an ion source is 600 ℃, the atomizing gas is nitrogen, the scanning range of primary mass spectrum parent ions is 50-1000, and the scanning range of secondary ions is 50-1000; the ionization mode is an electrospray negative ion mode, the temperature of an ion source is 500 ℃, the atomizing gas is nitrogen, the scanning range of primary mass spectrum parent ions is 50-1000, and the scanning range of secondary ions is 50-1000.

The invention has the beneficial effects that: due to the adoption of the technical scheme, compared with the prior art, the invention uses the superfine powder as an extraction material and uses water as a solvent to extract active ingredients in the eucommia ulmoides leaves, and the prepared eucommia ulmoides extract has higher contents of genipin and chlorogenic acid. The method is simple and feasible, efficient, convenient and low in cost, and the superfine powder can be prepared for standby after being harvested in season and is not limited by seasons.

Follow-up processing procedure is loaded down with trivial details than this patent in this patent of concrete CN200910147928.1, and probably has organic solvent's pollution, and this patent adopts the water for extracting the solvent, and water is a fine solvent, and most material all dissolves in water in the eucommia ulmoides, and this patent extraction method is simple, does not have organic solvent pollution, and the residue after the extraction need not handle, can directly abandon, can not produce the pollution to the environment.

Drawings

FIG. 1 shows that the prepared Eucommiae cortex extract is brown powder after being pulverized and sieved with a 80-mesh sieve;

FIG. 2 is a total ion flow graph in sample extract and blank positive ion mode;

FIG. 3 is a total ion flow graph of sample extracts and blank anion mode.

Detailed Description