Extraction method of eucommia ulmoides extract

Abstract

The invention discloses an extraction method of a eucommia ulmoides extract, and relates to the technical field of eucommia ulmoides extracts. The preparation method specifically comprises the following steps: step 1, taking eucommia ulmoides leaves, drying, and grinding and crushing with a grinding pestle to obtain eucommia ulmoides leaf powder; 2, adding an ethanol solution into the eucommia ulmoides leaf powder to obtain a eucommia ulmoides leaf powder mixture; extracting the folium cortex eucommiae powder mixture through an ultrasonic extraction method to obtain a folium cortex eucommiae powder extracted mixture; according to the extraction method of the eucommia ulmoides leaf extract, the eucommia ulmoides leaf powder is subjected to ultrasonic water bath, oscillation adsorption and other measures through the ethanol solution, extraction of the eucommia ulmoides leaf extract is achieved in a normal-temperature state, the integrity of the eucommia ulmoides leaf extract is fully guaranteed, the extraction effect of the eucommia ulmoides leaf extract is improved, and further utilization of the eucommia ulmoides leaf extract is facilitated; meanwhile, experiments prove that the eucommia ulmoides extract extracted by the method can obviously inhibit damage to cell membranes caused by increase of lipid peroxide free radicals.

Description

Technical Field

[0001] The invention relates to the technical field of eucommia ulmoides extracts, and in particular to a method for extracting eucommia ulmoides extracts. Background Art

[0002] Muscular dystrophy is a chronic, progressive myopathy that is relatively rare and difficult to treat in clinical practice. The cause is still unknown. Many cases have a family history or genetic history, and some people believe that it is caused by a disorder of the amino acid metabolism of the muscles themselves. The main clinical features are progressive skeletal muscle atrophy and weakness, which leads to obstacles in walking and other aspects.

[0003] Eucommia ulmoides is a traditional economic tree species in my country. According to the Chinese Pharmacopoeia, its bark is used as medicine. It is recorded in Shennong's Herbal Classic that Eucommia ulmoides can be used as a medicine for strengthening tendons and bones, replenishing qi, and calming the fetus. It is also recorded in Li Shizhen's Compendium of Materia Medica. Eucommia ulmoides leaves contain many functional ingredients, such as pinoresinol diglucoside, a lignin-based substance, which has anti-tumor and antihypertensive effects.

[0004] After searching, the Chinese patent application No. 01136208.1 discloses a method for extracting eucommia leaf extract, which comprises taking a certain amount of eucommia leaf, adding 10 times of water, boiling for half an hour, filtering out the water, adding 5 times of water, boiling for half an hour, filtering out the water; mixing the two filtrates, boiling and concentrating until the ratio of effective ingredients to water is 1:1, refrigerating for more than 12 hours, and then centrifuging at 2500 rpm for 30 minutes to make it into a paste; then drying under reduced pressure at -0.09Mpa and 60°C to obtain a dry paste; and then crushing it into powder according to conventional methods.

[0005] The extraction method of Eucommia leaf extract in the above patent has the following shortcomings: the extraction method is relatively rough, and the effective ingredients in Eucommia leaves are directly processed by decoction, and the extraction effect of various components in Eucommia leaves is relatively poor, which is not conducive to further research and further utilization of Eucommia leaf extract. Summary of the invention

[0006] The purpose of the invention is to solve the shortcomings in the prior art and to propose a method for extracting eucommia extract.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A method for extracting eucommia extract comprises the following steps:

[0009] Step 1: Take the Eucommia leaves, dry them, and grind them with a pestle to obtain Eucommia leaf powder;

[0010] Step 2: adding ethanol solution to eucommia leaf powder to obtain an eucommia leaf powder mixture; extracting the eucommia leaf powder mixture by ultrasonic extraction to obtain an eucommia leaf powder extraction mixture; then subjecting the eucommia leaf powder extraction mixture to sedimentation centrifugation treatment by a centrifuge, collecting the supernatant, and obtaining a crude eucommia leaf alcohol extract;

[0011] Step 3: Put the macroporous resin into the crude Eucommia leaf extract, add MBPS solution to dilute, and shake the Eucommia leaf alcohol extract by shaking on a shaker; after adsorption, use deionized water to filter; after filtering, immerse the macroporous resin adsorbing the Eucommia leaf alcohol extract in an ethanol solution, and desorb by shaking on a shaker;

[0012] Step 4: gradient eluting the ethanol solution containing the oleophila alcohol extract, concentrating the ethanol solution to obtain an extract, and vacuum drying the ethanol solution to obtain a powder;

[0013] Step 5: Use ethanol to elute the product to obtain the separated and purified Eucommia ulmoides leaf alcohol extract.

[0014] Preferably: the eucommia leaves in step 1 are set to 20 grams, the eucommia leaves are cleaned by ultrasound, and dried by low-temperature airflow; the eucommia leaf powder completely passes through a 100-mesh filter screen.

[0015] Further: the ethanol solution in step 2 is set to 100 ml, and the concentration is set to 80%; the equipment in the ultrasonic extraction method is an ultrasonic water bath pot, the working power of the ultrasonic water bath pot is 100 watts, and the extraction time is 3 hours; the centrifuge is set to a flat plate sedimentation centrifuge.

[0016] As a preferred embodiment of the present invention: the macroporous resin in step three is a polymer porous microsphere, the MBPS solution is set to 80 ml, and the concentration is set to 0.5 mg / ml; the working temperature of the shaker in the oscillating adsorption is set to 27 degrees Celsius, the working speed is 100 revolutions per minute, and the working time is 24 hours.

[0017] On the basis of the above scheme: the ethanol solution in step three is set to 40 ml and the concentration is set to 80%; the working temperature of the shaker in the oscillating desorption is set to 27 degrees Celsius, the working speed is 100 revolutions per minute, and the working time is 12 hours.

[0018] Preferably: in the step 4, the ethanol gradient elution uses 20%, 40%, 60% and 80% ethanol solutions in sequence to elute the macroporous resin adsorbing the eucommia leaf alcohol extract, and the four elution products are concentrated into an extract, and the extract is dried into a powder in a vacuum drying oven, and the powder completely passes through a 200-mesh filter screen.

[0019] Furthermore: the ethanol elution product in step 5 is a 40% ethanol solution elution product.

[0020] The beneficial effects of the present invention are:

[0021] The eucommia extract extraction method uses an ethanol solution to subject eucommia leaf powder to ultrasonic water bath, oscillation adsorption and other measures to extract the eucommia leaf extract at room temperature, fully ensuring the integrity of the eucommia leaf extract, improving the extraction effect of the eucommia leaf extract, and facilitating further use of the eucommia leaf extract; and experiments have shown that the eucommia extract extracted by the method can significantly inhibit the damage to cell membranes caused by increased lipid peroxide free radicals. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The schematic diagram of the process structure of the method for extracting the Eucommia ulmoides extract of the present invention;

[0023] Figure 2 This is a statistical table of the gripping strength of mice before treatment at different periods of the present invention;

[0024] Figure 3 The figure is a statistical table of the gripping strength and changes of mice after treatment at different stages of the present invention. DETAILED DESCRIPTION

[0025] The technical solution of the present invention is further described in detail below in conjunction with specific implementation methods.

[0026] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are only used to explain the present invention, and cannot be understood as limiting the present invention.

[0027] A method for extracting eucommia extract, such as Figure 1 As shown, the following steps are included:

[0028] Step 1: Take 20 grams of Eucommia ulmoides leaves, first clean them by ultrasonic, then dry them by low-temperature airflow, and finally grind them with a grinding pestle to obtain Eucommia ulmoides leaf powder; the Eucommia ulmoides leaf powder completely passes through a 100-mesh filter screen;

[0029] Step 2: adding 100 ml of 80% ethanol solution to the eucommia leaf powder to obtain an eucommia leaf powder mixture; extracting the eucommia leaf powder mixture by an ultrasonic water bath (ultrasonic extraction method), the working power of the ultrasonic water bath is 100 watts, the extraction time is 3 hours, and the eucommia leaf powder extraction mixture is obtained; then the eucommia leaf powder extraction mixture is subjected to sedimentation centrifugation treatment by a flat plate sedimentation centrifuge, and the supernatant is collected to obtain a crude eucommia leaf alcohol extract, the working speed of the flat plate sedimentation centrifuge is 8000 rpm, and the working time is 60 minutes;

[0030] Step 3: placing a macroporous resin (polymer porous microsphere) into a crude Eucommia leaf extract, and adding 80 ml of MBPS solution to dilute the crude Eucommia leaf extract to obtain a dilution, wherein the concentration of the MBPS solution is set to 0.5 mg / ml; shaking the dilution of the Eucommia leaf alcohol extract by shaking a shaker, wherein the operating temperature of the shaker is set to 27 degrees Celsius, the operating speed is 100 revolutions per minute, and the working time is 24 hours; after adsorption is completed, filtering with deionized water; after filtering, immersing the macroporous resin adsorbing the Eucommia leaf alcohol extract in 40 ml of an ethanol solution with a concentration of 80%, and desorbing by shaking on a shaker, wherein the operating temperature of the shaker is set to 27 degrees Celsius, the operating speed is 100 revolutions per minute, and the working time is 12 hours;

[0031] Step 4: gradient elution is performed on the ethanol solution in which the oleophyte alcohol extract is dissolved, wherein the ethanol gradient elution uses 20%, 40%, 60% and 80% ethanol solutions in sequence to concentrate the extract, and the extract is dried into powder in a vacuum drying oven, and the powder completely passes through a 200-mesh filter screen;

[0032] Step 5: Using 40% ethanol solution to elute the product, to obtain the separated and purified eucommia leaf alcohol extract.

[0033] Using mouse upper limb grip strength as the detection index, and normal mice and muscular dystrophy mice as experimental subjects, the advantages of the new Eucommia extraction method were demonstrated from the therapeutic effect on mice.

[0034] The specific control experiment method is as follows:

[0035] 1. Grip strength of mice before treatment at different stages

[0036] Two groups of mice: normal controls and muscular dystrophy mice;

[0037] Data were obtained at different time points: 8 to 11 weeks;

[0038] Detection index: Mouse upper limb grip strength (unit: grip strength (mN / g));

[0039] The experimental data refer to the instruction manual. Figure 2 .

[0040] 2. Grip strength and changes of mice after treatment at different stages

[0041] Two groups of mice: normal controls were given 0.1% DMSO weekly and muscular dystrophy mice were given Eucommia extract;

[0042] Data were obtained at different time points: 8 to 11 weeks;

[0043] Detection index: Mouse upper limb grip strength (unit: grip strength (mN / g));

[0044] The experimental data refer to the instruction manual. Figure 3 .

[0045] The specific results are as follows:

[0046] When aging model mice were gavage-fed with water extract of Eucommia leaves, it was found that the activities of oxidase dismutase and glutathione peroxidase in the mouse red blood cells were significantly reduced, proving that Eucommia extract can significantly inhibit the increase of lipid peroxide free radicals and the damage to cell membranes; the active ingredient in Eucommia leaves, cyclopentane glycosides, can promote the synthesis of antioxidant substances in the body and enhance the scavenging effect of SOD on superoxide anions: increase the activity of superoxide dismutase and reduce the level of lipid peroxidation caused by D-galactose; strengthen the reduction reaction of glutathione peroxidase to HO, protect the biological membrane from injury, and thus maintain the integrity of the structure and function.

[0047] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A method for extracting Eucommia ulmoides extract, characterized in that: The steps include: Step 1: Take the Eucommia leaves, dry them, and grind them with a pestle to obtain Eucommia leaf powder; Step 2: adding ethanol solution to eucommia leaf powder to obtain an eucommia leaf powder mixture; extracting the eucommia leaf powder mixture by ultrasonic extraction to obtain an eucommia leaf powder extraction mixture; then subjecting the eucommia leaf powder extraction mixture to sedimentation centrifugation treatment by a centrifuge, collecting the supernatant, and obtaining a crude eucommia leaf alcohol extract; Step 3: Put the macroporous resin into the crude Eucommia leaf extract, add MBPS solution to dilute, and shake the Eucommia leaf alcohol extract by shaking on a shaker; after adsorption, use deionized water to filter; after filtering, immerse the macroporous resin adsorbing the Eucommia leaf alcohol extract in an ethanol solution, and desorb by shaking on a shaker; Step 4: gradient elution of the ethanol solution containing the oleophila alcohol extract, concentration to an extract, and vacuum drying to a powder; Step 5: Using ethanol to elute the product to obtain the separated and purified eucommia leaf alcohol extract.

2. The method for extracting the Eucommia ulmoides extract according to claim 1, characterized in that: The eucommia leaves in step 1 are set to 20 grams, the eucommia leaves are cleaned by ultrasound, and dried by low-temperature air flow; the eucommia leaf powder completely passes through a 100-mesh filter screen.

3. The method for extracting the Eucommia ulmoides extract according to claim 1, characterized in that: The ethanol solution in step 2 is set to 100 ml and the concentration is set to 80%; the equipment in the ultrasonic extraction method is an ultrasonic water bath pot, the working power of the ultrasonic water bath pot is 100 watts, and the extraction time is 3 hours; the centrifuge is set to a flat plate sedimentation centrifuge.

4. The method for extracting the Eucommia ulmoides extract according to claim 1, characterized in that: The macroporous resin in step three is a polymer porous microsphere, the MBPS solution is set to 80 ml, and the concentration is set to 0.5 mg / ml; the working temperature of the shaker in the oscillating adsorption is set to 27 degrees Celsius, the working speed is 100 revolutions per minute, and the working time is 24 hours.

5. The method for extracting the Eucommia ulmoides extract according to claim 1, characterized in that: The ethanol solution in step three is set to 40 ml and the concentration is set to 80%; the working temperature of the shaker in the oscillating desorption is set to 27 degrees Celsius, the working speed is 100 revolutions per minute, and the working time is 12 hours.

6. The method for extracting the Eucommia ulmoides extract according to claim 1, characterized in that: In the step 4, the ethanol gradient elution uses 20%, 40%, 60% and 80% ethanol solutions in sequence to elute the macroporous resin adsorbing the eucommia leaf alcohol extract, and the four elution products are concentrated into an extract, and the extract is dried into powder in a vacuum drying oven, and the powder completely passes through a 200-mesh filter screen.

7. The method for extracting the Eucommia ulmoides extract according to claim 1, characterized in that: The ethanol elution product in step 5 is a 40% ethanol solution elution product.

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