Method for extracting and separating aucubin, protein peptide and gutta-percha from folium cortex eucommiae and cortex eucommiae

Abstract

The invention discloses a method for extracting and separating aucubin, protein peptide and gutta-percha from folium cortex eucommiae and cortex eucommiae, and relates to the field of natural product extraction. S2, ultrasonic extraction; s3, filtering is carried out; s4, separating an extracting solution; s5, treatment of an eluent; s6, obtaining of aucubin; s7, treating the washing liquid; s8, obtaining of protein oligopeptide; s9, treating filter residues; s10, filtering the extract liquor; s11, obtaining eucommia ulmoides refined glue; according to the method for extracting and separating aucubin, protein peptide and gutta-percha from folium cortex eucommiae and cortex eucommiae, a continuous countercurrent ultrasonic extraction technology, a subcritical ultrasonic extraction technology and a gutta-percha refining group tank preparation technology are combined, so that various effective components in the folium cortex eucommiae and cortex eucommiae can be extracted, and maximum utilization of resources is realized; eucommia ulmoides leaves are fully utilized, and production benefits are improved.

Description

Technical Field

[0001] The invention relates to the technical field of natural product extraction, and in particular to a method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark. Background Art

[0002] Eucommia ulmoides is a plant of the Eucommia genus of the Eucommia family. It is a unique relict tree species in my country. As a traditional Chinese medicinal material, Eucommia ulmoides has the characteristics of no toxic side effects. Eucommia ulmoides contains chemical elements such as geniposide and aucubin, as well as various trace elements such as potassium. It has the effects of promoting laxative and urination, whitening and skin care. Eucommia ulmoides has the advantages of multiple energy efficiency and high utilization value.

[0003] At present, the extraction methods for producing aucubin in Eucommia ulmoides and Eucommia rubber include organic solvent extraction, acid-base water extraction, and enzymatic hydrolysis. Most of these methods directly use Eucommia ulmoides leaves, Eucommia ulmoides fruits, and Eucommia ulmoides bark for extraction. The raw materials are used once and cannot be reused after extraction, so they have the disadvantages of high extraction cost, large consumption of acid-base water, low extraction rate, and low product content. In addition, petroleum ether or n-hexane and other flammable and explosive organic solvents are traditionally used for extraction. Organic solvents such as petroleum ether and n-hexane have flammable and explosive properties, which increases the safety risk in the extraction process. During the operation, if improperly handled or encountering a fire source, it is very easy to cause a fire or explosion accident, posing a serious threat to the safety of personnel and equipment. In addition, this type of organic solvent may evaporate into the air during the extraction process, causing pollution to the atmospheric environment. At the same time, the treatment of waste liquid after extraction is also a difficult problem. If improperly handled, it may cause long-term impact on the environment. Summary of the invention

[0004] The purpose of the present invention is to provide a method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark, so as to solve the problem that the eucommia leaves, eucommia fruits and eucommia bark are directly used for extraction in the prior art, the raw materials are used once and cannot be reused after extraction, thus having the disadvantages of high extraction cost, large consumption of acid and alkaline water, low extraction rate, low product content, and organic solvents such as petroleum ether and n-hexane are flammable and explosive.

[0005] In order to achieve the above object, the present invention provides the following technical scheme: a method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark, which specifically comprises the following steps:

[0006] S1. Preparation of raw materials: first weigh 500 kg of Eucommia ulmoides leaves and skins as raw materials, then wash and dry the Eucommia ulmoides leaves and skins, and then crush the Eucommia ulmoides leaves and skins into Eucommia ulmoides leaf coarse powder;

[0007] S2, ultrasonic extraction: the prepared Eucommia leaf powder is then placed in a continuous countercurrent ultrasonic extractor, and then 4000 liters of deionized water is added to the continuous countercurrent ultrasonic extractor as an extracting liquid to extract the substances contained in the Eucommia leaf powder;

[0008] S3, filtration: then use a bag filter to filter to obtain the extract and filter residue;

[0009] S4, separation of extract: first pre-treat the macroporous resin, then uniformly load the pre-treated macroporous resin into the adsorption column, then add the extract into the adsorption column by a constant flow pump, then wash the macroporous resin column with water to obtain a washing solution, and then elute the resin column with ethanol to obtain an eluent;

[0010] S5, treatment of the eluate: the eluate is then concentrated using a reverse osmosis membrane to remove most of the ethanol and water, and the concentrated liquid is then concentrated using a low-temperature vacuum concentrator to obtain a concentrated viscous liquid;

[0011] S6. Obtaining aucubin: placing the concentrated viscous liquid into a low-temperature vacuum dryer for vacuum drying to obtain aucubin;

[0012] S7, treatment of the washing liquid: firstly, the washing liquid is hydrolyzed with 0.3% to 0.5% neutral protease or alkaline protease to obtain a hydrolyzate, and then the hydrolyzate is separated with a 3000D ultrafiltration membrane to obtain a filtrate;

[0013] S8. Obtaining protein oligopeptides: then placing the filtrate into a vacuum concentrator to concentrate under reduced pressure to obtain a concentrated solution, and then placing the concentrated solution into a low-temperature vacuum dryer to vacuum dry to obtain protein oligopeptides;

[0014] S9, filter residue treatment: firstly, the filter residue is dried and granulated to obtain granules, and then the granules are placed in a subcritical ultrasonic extraction device, and then R134a mixed solvent is added to the subcritical ultrasonic extraction device, and then the subcritical ultrasonic extraction device is turned on to perform subcritical ultrasonic extraction on the granules for 2 to 3 hours to obtain an extract;

[0015] S10, filtering the extract: then filtering the extract with a filter to obtain a separated liquid, and then pumping the separated liquid into a desorption tank to convert the R134a mixed solvent in the separated liquid into a gaseous state at a certain temperature and pressure to obtain a coarse viscous paste of Eucommia ulmoides;

[0016] S11. Obtaining Eucommia refined gum: Then put the Eucommia crude gum paste into the Eucommia gum refining tank and dissolve it with petroleum ether to obtain a dissolved solution, and then filter it through a filter to remove impurities, and then concentrate, crystallize, filter, purify and dry to obtain Eucommia refined gum.

[0017] Preferably, the continuous countercurrent ultrasonic extractor in S2 extracts the Eucommia leaf crude powder for 60 to 90 minutes.

[0018] Preferably, the macroporous resin in S4 is selected from LSA-21, D101 or AB-8.

[0019] Preferably, the specific steps of the macroporous resin pretreatment in S4 are:

[0020] First, the macroporous resin is soaked in 95% ethanol for 24 hours, and the resin is repeatedly washed with deionized water until the conductivity of the washing solution reaches the requirement, and then the macroporous resin is placed in deionized water for storage.

[0021] Preferably, the volume fraction of the ethanol eluent in S4 is 65% to 75%.

[0022] Preferably, the temperature for hydrolysis of S7 is 45-55° C., and the hydrolysis time is 2-4 hours.

[0023] Preferably, the S9 subcritical ultrasonic extraction temperature is 40-50° C. and the pressure is 0.4 MPa-2.0 MPa.

[0024] Preferably, the temperature in the S10 analysis tank is 10° C. to 50° C., and the pressure is 0.4 MPa to 2.0 MPa.

[0025] Compared with the prior art, the method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and peels provided by the invention adopts a combination of continuous countercurrent ultrasonic extraction technology, subcritical ultrasonic extraction technology and eucommia gum refining and assembly tank preparation technology, which can extract multiple effective ingredients from eucommia leaves and peels, realize the maximum utilization of resources, make full use of eucommia leaves, improve production efficiency, and solve the outstanding problems that the eucommia raw material extraction material is single, the extraction production cost is high, and the production efficiency of the enterprise is seriously affected.

[0026] When preparing Eucommia ulmoides fine rubber, R134a mixed solvent is used as the extracting liquid, and the extraction is carried out using a subcritical ultrasonic extraction device. Compared with the traditional use of flammable and explosive organic solvents such as petroleum ether or n-hexane, R134a mixed solvent has higher safety. Its non-flammable and non-explosive characteristics greatly reduce the risk of fire and explosion in the production process. Secondly, traditional solvents may evaporate into the air during the preparation process, causing certain pollution to the environment. R134a mixed solvent has low volatility and is easy to recycle and handle, thereby reducing pollution to the environment. Finally, the subcritical ultrasonic extraction device can maintain a low temperature and pressure during the extraction process, thereby avoiding the destruction of the active ingredients in Eucommia ulmoides by high temperature and high pressure, and maintaining the natural quality and biological activity of the product. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For ordinary technicians in this field, other drawings can also be obtained based on these drawings.

[0028] Figure 1 It is a flowchart of the Eucommia ulmoides extraction of the present invention. DETAILED DESCRIPTION

[0029] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.

[0030] Example:

[0031] See also Figure 1 The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark specifically comprises the following steps:

[0032] S1. Preparation of raw materials: first weigh 500 kg of Eucommia ulmoides leaves and skins as raw materials, then wash and dry the Eucommia ulmoides leaves and skins, and then crush the Eucommia ulmoides leaves and skins into Eucommia ulmoides leaf coarse powder;

[0033] S2, ultrasonic extraction: the prepared Eucommia leaf powder is then placed in a continuous countercurrent ultrasonic extractor, and then 4000 liters of deionized water is added to the continuous countercurrent ultrasonic extractor as an extracting liquid to extract the substances contained in the Eucommia leaf powder. The continuous countercurrent ultrasonic extractor extracts the Eucommia leaf powder for 60 to 90 minutes;

[0034] S3, filtration: then use a bag filter to filter to obtain the extract and filter residue;

[0035] S4, separation of extract: First, select a macroporous resin of model LSA-21, D101 or AB-8, and pretreat the macroporous resin. Specific steps:

[0036] First, the macroporous resin is soaked in 95% ethanol for 24 hours, and the resin is repeatedly washed with deionized water until the conductivity of the washing solution reaches the requirement, and then the macroporous resin is placed in deionized water for storage.

[0037] Then, the pretreated macroporous resin is uniformly loaded into the adsorption column, and the extract is added into the adsorption column by a constant flow pump, and then the macroporous resin column is washed with water to obtain a washing solution, and then the resin column is eluted with ethanol to obtain an eluent, and the volume fraction of the ethanol eluent is 65% to 75%;

[0038] S5, treatment of the eluate: the eluate is then concentrated using a reverse osmosis membrane to remove most of the ethanol and water, and the concentrated liquid is then concentrated using a low-temperature vacuum concentrator to obtain a concentrated viscous liquid;

[0039] S6. Obtaining aucubin: placing the concentrated viscous liquid into a low-temperature vacuum dryer for vacuum drying to obtain aucubin;

[0040] S7, treatment of the washing liquid: firstly, the washing liquid is hydrolyzed with 0.3% to 0.5% neutral protease or alkaline protease to obtain a hydrolyzate, the hydrolysis temperature is 45 to 55° C., the hydrolysis time is 2 to 4 hours, and then the hydrolyzate is separated with a 3000D ultrafiltration membrane to obtain a filtrate;

[0041] S8. Obtaining protein oligopeptides: then placing the filtrate into a vacuum concentrator to concentrate under reduced pressure to obtain a concentrated solution, and then placing the concentrated solution into a low-temperature vacuum dryer to vacuum dry to obtain protein oligopeptides;

[0042] S9, filter residue treatment: firstly, the filter residue is dried and granulated to obtain particles, and then the particles are placed in a subcritical ultrasonic extraction device, and then R134a mixed solvent is added to the subcritical ultrasonic extraction device, and then the subcritical ultrasonic extraction device is turned on to perform subcritical ultrasonic extraction on the particles for 2 to 3 hours to obtain an extract, wherein the subcritical ultrasonic extraction temperature is 40 to 50° C. and the pressure is 0.4 MPa to 2.0 MPa;

[0043] S10, filtering the extract: then filtering the extract with a filter to obtain a separated liquid, and then pumping the separated liquid into a desorption tank to convert the R134a mixed solvent in the separated liquid into a gaseous state at a certain temperature and pressure to obtain a coarse viscous paste of Eucommia ulmoides;

[0044] S11, obtaining Eucommia refined gum: then put the Eucommia crude gum paste into the Eucommia gum refining group tank and dissolve it with petroleum ether to obtain the dissolved liquid, then filter it through a filter to remove impurities, and then concentrate, crystallize, filter, purify and dry it to obtain Eucommia refined gum. The temperature in the S10 analysis tank is 10℃~50℃, and the pressure is 0.4MPa~2.0MPa. In step S2, a continuous countercurrent ultrasonic extraction device is used to extract aucubin from Eucommia leaves and skins. The continuous countercurrent ultrasonic extraction device can effectively extract the heat-sensitive component aucubin under low temperature conditions, and the product activity is maintained, the extraction efficiency is high, the degree of automation is good, and it saves labor, time and energy.

[0045] When preparing Eucommia ulmoides fine rubber, R134a mixed solvent is used as the extracting liquid, and the extraction is carried out using a subcritical ultrasonic extraction device. Compared with the traditional use of flammable and explosive organic solvents such as petroleum ether or n-hexane, R134a mixed solvent has higher safety. Its non-flammable and non-explosive characteristics greatly reduce the risk of fire and explosion in the production process. Secondly, traditional solvents may evaporate into the air during the preparation process, causing certain pollution to the environment. R134a mixed solvent has low volatility and is easy to recycle and handle, thereby reducing pollution to the environment. Finally, the subcritical ultrasonic extraction device can maintain a low temperature and pressure during the extraction process, thereby avoiding the destruction of the active ingredients in Eucommia ulmoides by high temperature and high pressure, and maintaining the natural quality and biological activity of the product.

[0046] Subcritical ultrasonic extraction technology is used. The pressure of subcritical ultrasonic extraction equipment is much lower than that of traditionally used supercritical extraction equipment. The equipment volume is greatly increased, making large-scale production of eucommia gum a reality. This solves the problem that supercritical CO2 extraction requires high pressure, resulting in strict equipment manufacturing requirements and small extraction equipment volume, which seriously affects large-scale production.

[0047] To sum up, the above steps adopt a combination of continuous countercurrent ultrasonic extraction technology, subcritical ultrasonic extraction technology and eucommia gum refined group tank preparation technology, which can extract a variety of effective ingredients in eucommia leaves and peels, realize the maximum utilization of resources, make full use of eucommia leaves, improve production efficiency, and solve the outstanding problems of single eucommia raw material extraction substance, high extraction production cost, and serious impact on the production efficiency of enterprises.

[0048] The above description is only by way of illustration of certain exemplary embodiments of the present invention. It is undoubted that those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the above drawings and descriptions are illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.

Claims

1. A method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark, characterized in that: The specific steps include: S1. Preparation of raw materials: first weigh 500 kg of Eucommia ulmoides leaves and skins as raw materials, then wash and dry the Eucommia ulmoides leaves and skins, and then crush the Eucommia ulmoides leaves and skins into Eucommia ulmoides leaf coarse powder; S2, ultrasonic extraction: the prepared Eucommia leaf powder is then placed in a continuous countercurrent ultrasonic extractor, and then 4000 liters of deionized water is added to the continuous countercurrent ultrasonic extractor as an extracting liquid to extract the substances contained in the Eucommia leaf powder; S3, filtration: then use a bag filter to filter to obtain the extract and filter residue; S4, separation of extract: first pre-treat the macroporous resin, then uniformly load the pre-treated macroporous resin into the adsorption column, then add the extract into the adsorption column by a constant flow pump, then wash the macroporous resin column with water to obtain a washing solution, and then elute the resin column with ethanol to obtain an eluent; S5, treatment of the eluate: the eluate is then concentrated using a reverse osmosis membrane to remove most of the ethanol and water, and the concentrated liquid is then concentrated using a low-temperature vacuum concentrator to obtain a concentrated viscous liquid; S6. Obtaining aucubin: placing the concentrated viscous liquid into a low-temperature vacuum dryer for vacuum drying to obtain aucubin; S7, treatment of the washing liquid: firstly, the washing liquid is hydrolyzed with 0.3% to 0.5% neutral protease or alkaline protease to obtain a hydrolyzate, and then the hydrolyzate is separated with a 3000D ultrafiltration membrane to obtain a filtrate; S8. Obtaining protein oligopeptides: then placing the filtrate into a vacuum concentrator to concentrate under reduced pressure to obtain a concentrated solution, and then placing the concentrated solution into a low-temperature vacuum dryer to vacuum dry to obtain protein oligopeptides; S9, filter residue treatment: firstly, the filter residue is dried and granulated to obtain granules, and then the granules are placed in a subcritical ultrasonic extraction device, and then R134a mixed solvent is added to the subcritical ultrasonic extraction device, and then the subcritical ultrasonic extraction device is turned on to perform subcritical ultrasonic extraction on the granules for 2 to 3 hours to obtain an extract; S10, filtering the extract: then filtering the extract with a filter to obtain a separated liquid, and then pumping the separated liquid into a desorption tank to convert the R134a mixed solvent in the separated liquid into a gaseous state at a certain temperature and pressure to obtain a coarse viscous paste of Eucommia ulmoides; S11. Obtaining Eucommia refined gum: Then put the Eucommia crude gum paste into the Eucommia gum refining tank and dissolve it with petroleum ether to obtain a dissolved solution, and then filter it through a filter to remove impurities, and then concentrate, crystallize, filter, purify and dry to obtain Eucommia refined gum.

2. The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark according to claim 1, characterized in that: The continuous countercurrent ultrasonic extractor in S2 extracts the Eucommia ulmoides leaf coarse powder for 60 to 90 minutes.

3. The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark according to claim 1, characterized in that: The macroporous resin in S4 is selected from LSA-21, D101 or AB-8.

4. The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark according to claim 1, characterized in that: The specific steps of macroporous resin pretreatment in S4 are as follows: First, the macroporous resin is soaked in 95% ethanol for 24 hours, and the resin is repeatedly washed with deionized water until the conductivity of the washing solution reaches the requirement, and then the macroporous resin is placed in deionized water for storage.

5. The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark according to claim 1, characterized in that: The volume fraction of the ethanol eluent in S4 is 65% to 75%.

6. The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark according to claim 1, characterized in that: The temperature of the S7 hydrolysis is 45-55° C., and the hydrolysis time is 2-4 hours.

7. The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark according to claim 1, characterized in that: The S9 subcritical ultrasonic extraction temperature is 40-50° C. and the pressure is 0.4 MPa-2.0 MPa.

8. The method for extracting and separating aucubin, protein peptide and eucommia gum from eucommia leaves and bark according to claim 1, characterized in that: The temperature in the S10 analysis tank is 10° C. to 50° C., and the pressure is 0.4 MPa to 2.0 MPa.

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