CN108619243B - Method for synergistically extracting active ingredients in ramulus mori (leaves) by solid acid enhanced hydrothermal reaction - Google Patents
Method for synergistically extracting active ingredients in ramulus mori (leaves) by solid acid enhanced hydrothermal reaction Download PDFInfo
- Publication number
- CN108619243B CN108619243B CN201810501775.5A CN201810501775A CN108619243B CN 108619243 B CN108619243 B CN 108619243B CN 201810501775 A CN201810501775 A CN 201810501775A CN 108619243 B CN108619243 B CN 108619243B
- Authority
- CN
- China
- Prior art keywords
- mulberry leaf
- mulberry
- reaction kettle
- active ingredients
- hydrothermal reaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/60—Moraceae (Mulberry family), e.g. breadfruit or fig
- A61K36/605—Morus (mulberry)
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D11/00—Solvent extraction
- B01D11/02—Solvent extraction of solids
- B01D11/028—Flow sheets
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/17—Preparation or pretreatment of starting material involving drying, e.g. sun-drying or wilting
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Alternative & Traditional Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention belongs to the technical field of extraction of active ingredients in natural products, and particularly relates to a phosphotungstic heteropoly acid catalysis and hydrothermal reaction regulation and control method. The mulberry twig (leaf) has high-efficiency pharmacological action, is widely applied to the industries of food, medicine, skin care products and the like, has closely related pharmacological action with active ingredients thereof, improves the extraction efficiency and resource utilization value of the active ingredients in the mulberry twig (leaf), and has research significance and practical economic value. Aiming at the defects of the existing mulberry twig (leaf) active ingredient extraction method, the invention provides a phosphotungstic acid catalysis-hydrothermal regulation extraction method, wherein phosphotungstic acid catalysis and hydrothermal reaction are cooperated to degrade plant tissues, destroy hydrogen bonds between cellulose and hemicellulose, destroy a cell wall compact structure, reduce mass transfer barriers between cell walls and intercellular substances, facilitate the full dissolution of active ingredients, and realize the characteristics of few and simple extraction conditions, less environmental pollution, low production energy consumption, low cost and the like.
Description
Technical Field
The invention belongs to the technical field of extraction of active ingredients in natural products, and particularly relates to a phosphotungstic heteropoly acid catalysis and hydrothermal reaction regulation and control method.
Background
Folium Mori is rich in vitamins (100 g folium Mori dried product contains retinol 670 μ g, carotene 7440 μ g, and V)B10.59mg，VB21.35mg，VC31.6 mg) and amino acids (containing as many as 18, 10.10% by dry weight, with an essential amino acid content of 3.281%, the plant protein first limiting amino acid lysine of 0.454%, the semi-essential amino acid of 1.843%); the mulberry leaf extract has the advantages of good blood sugar and blood fat reduction, anti-inflammation, anti-aging, anti-filariasis and anti-tumor. The ramulus Mori contains polyhydroxy alkaloid, flavonoids, folium Mori polysaccharide, phytosterol, polyphenols, etc(ii) a Has pharmacological effects of inhibiting serum cholesterol increase, preventing atherosclerosis, and resisting bacteria and virus. Modern medical and nutritional researches prove that the high-efficiency pharmacological action of the mulberry twig (leaf) is closely related to the active ingredients contained in the mulberry twig (leaf). The mulberry leaves have the health-care pharmacological action, so that the mulberry leaves are favored by the food industry, and products such as mulberry tea, mulberry leaf surfaces, mulberry leaf biscuits, mulberry leaf wine, mulberry leaf vinegar, mulberry leaf sauce and the like are developed, so that the aims of preventing diseases and prolonging life are fulfilled by people through daily diet; with the continuous and deep research, the mulberry leaves are expected to become therapeutic drugs and health care products for some diseases, especially senile diseases and chronic diseases, and have wide prospects in the aspects of beauty, skin care, livestock raising and the like.
The existing extraction method of active ingredients of mulberry twigs (leaves) mainly comprises a reflux extraction method, an ultrasonic extraction method, an enzymatic extraction method, a supercritical fluid extraction method and the like. Wherein, the reflux method has long heating time of the extracting solution, the biological activity of the components in the raw materials is easy to be damaged due to long-time heating, and the solvent consumption is large, thereby limiting the production and application of the method; the enzymatic extraction reaction time is long, and the enzyme is protein, so that sugar liquor is difficult to filter; the ultrasonic extraction method is simple to operate, but sometimes the extraction efficiency is lower than that of other methods; the extraction efficiency of the supercritical fluid extraction is greatly improved, but the method requires complicated equipment, has narrow solvent selection range, needs a high-pressure container and a high-pressure pump, and has higher investment cost. These methods affect the quality of the extracted product more or less, and cause environmental pollution. Therefore, the research on the high-efficiency extraction method of the active ingredients of the mulberry leaves is developed, and the research significance and the practical economic value are realized for improving the extraction efficiency and the resource utilization value of the active ingredients in the mulberry twigs (leaves).
The invention of patent CN201610569828.8 published in 2016, 11, and 2 discloses a method for extracting tea polysaccharide by a hydrothermal method, which is to extract tea polysaccharide by a hydrothermal method in a high-temperature high-pressure reaction kettle, namely, the tea polysaccharide is quickly and completely dissolved by using a high-temperature high-pressure solution to extract the tea polysaccharide, so that the tea polysaccharide can be effectively extracted, but the following defects still exist: (1) the complex compact network structure of the plant cell wall is destroyed only by the pyrolysis effect of the hydrothermal reaction; (2) the invention only extracts one substance of tea polysaccharide; (3) water is used as an extracting agent, and other substances such as tea polyphenol, amino acid, protein and the like in the tea cannot be fully dissolved out, so that the maximization of resource utilization cannot be realized. The existing extraction method of active ingredients in ramulus mori (leaves), such as "extraction method of active substances in folium mori with publication number CN 201711354749.6" published in 2016, 1, 27, discloses the following methods: weighing dried folium Mori powder 300.0g, adding 6.0L petroleum ether at a ratio of 1: 20, refluxing at 70 deg.C for 4 hr, filtering, adding 6.0L 65% ethanol into the residue at a ratio of 1: 20, soaking for one day and night, filtering, and concentrating the filtrate under reduced pressure to obtain folium Mori extract; the mulberry leaf extract is separated and purified by resin to respectively obtain three crude extracts of alkaloid, flavone and polysaccharide. The active substance extracted by the method has stable chemical components and contents, but has the following defects: (1) the whole extraction process takes longer time; (2) the usage amounts of petroleum ether and ethanol are respectively 6.0L; (3) the flammability, explosiveness and chronic toxicity of petroleum ether threaten human health and environment, and increase the safe production cost.
Disclosure of Invention
The invention aims to provide a method for extracting active ingredients of ramulus mori (leaves) by catalysis-hydrothermal regulation of phosphotungstic heteropoly acid, aiming at the defects of the existing method for extracting the active ingredients of ramulus mori (leaves), and the method has the characteristics of few and simple extraction conditions and operation steps, small environmental pollution, low production energy consumption, low cost and the like.
The mechanism of the invention is as follows: the plant cell wall is a complex and compact network structure consisting of lignin, and the compact structure is destroyed under the action of external force, so that hydrogen bonds and covalent bonds can be broken, and the dissolution of the compound is accelerated. The beta-O-4 ether linkage connection mode of lignin is the most abundant and important structural unit in the lignin, and the bond energy is the smallest and the most easily broken, while the lignin can undergo hydrolysis reaction under hydrothermal conditions, and the main process is the breakage of the ether linkage. The phosphotungstic heteropoly acid is used for catalyzing and hydrothermal reaction to synergistically degrade plant tissues, and mainly utilizes acid sites with catalytic activity on the surface and heteropoly acid redox carriers (e and II)+) The diffusion of (A) and the oxidation of plant tissues in the biomass under hydrothermal conditions, waterThe synergistic effect of the hydrolysis reaction and the acid catalytic oxidation destroys hydrogen bonds between cellulose and hemicellulose, degrades plant fiber tissues, destroys cell wall compact structures, reduces mass transfer barriers between cell walls and intercellular substances, and is beneficial to fully dissolving out active ingredients.
The technical scheme for realizing the purpose of the invention is as follows: a method for extracting active ingredients from ramulus Mori (folium Mori) by solid acid reinforced hydrothermal reaction comprises subjecting folium Mori powder to catalytic degradation by phosphotungstic heteropoly acid in hydrothermal reaction kettle system, centrifuging, metering volume, and detecting and analyzing with high performance liquid. The method comprises the following specific steps:
(1) pretreatment of raw materials
The picked fresh mulberry leaves are dried for 4-6 hours in a drying oven at 50-70 ℃, the dried mulberry leaves are crushed by a crusher, sieved by a 60-100 mesh sieve and put into a sealing bag for later use.
(2) Solid acid reinforced hydrothermal reaction treatment raw material
After the step (1) is finished, weighing 2g of pretreated mulberry leaf powder in a hydrothermal reaction kettle, adding 4-6 per mill of phosphotungstic heteropoly acid based on the mass (g) of the mulberry leaves, and finally weighing 6-22 ml of absolute ethyl alcohol according to the ratio of the volume (ml) of the absolute ethyl alcohol to the mass (g) of the mulberry leaf powder of 3-11: 1 and pouring the absolute ethyl alcohol into the hydrothermal reaction kettle. And then putting the reaction kettle into an oven at 170-190 ℃ for reaction for 90-130 min.
(3) Assay for active ingredients
And (3) after the step (2) is finished, taking the reaction kettle out, cooling to room temperature, pouring the reaction kettle into a centrifuge tube, centrifuging for 10-20min at the rotating speed of 2000-4000 r/min, pouring out supernatant, and fixing the volume to a 25nl volumetric flask. Transferring 1ml of mulberry leaf extract into a sample bottle, and detecting under the conditions of mobile phase acetonitrile (ml) = 35: 65-50: 50, temperature of 25-40 ℃, sample injection amount of 5-20 mu L and flow rate of 0.6-1 ml/min.
(4) Active ingredient product extraction
Placing the centrifugal clear liquid collected in the step (3) in a rotary evaporator, and carrying out reduced pressure concentration to obtain a thick mulberry leaf extract crude product at the vacuum pressure of 0.4-0.6 Mpa and the temperature of 80-100 ℃; then, adding the mulberry leaf extract and an ethanol solution into an extraction container according to the ratio of the mass (g) of the mulberry leaf extract product to the volume (ml) of the ethanol of 1: 8-10, and mixing and dissolving uniformly; and finally, performing reduced pressure concentration for 2-4 times through a rotary evaporator under the vacuum pressure of 0.4-0.6 Mpa and the temperature of 80-100 ℃ to obtain the mulberry leaf extract product.
By adopting the technical scheme, the invention mainly has the following effects
(1) The invention carries out catalytic reaction by the phosphotungstic heteropoly acid, and has good catalytic degradation effect on mulberry leaves due to unique strong proton acidity, high hydrothermal stability, multiple functions (acid, oxygen and photocatalysis), easily modulated molecular structure and unique quasi-liquid phase behavior.
(2) The method adopts the hydrothermal reaction of ethanol instead of water to extract the active ingredients in the mulberry leaves, realizes the synchronous operation of the hydrothermal reaction and the alcohol extraction, only uses absolute ethanol in the extraction process, successfully avoids the use of other toxic and harmful solvents, increases the safety of the experimental process, is beneficial to environmental protection, and has simple operation.
(3) The invention adopts waste mulberry leaves as raw materials, after pretreatment, crude extracts are obtained by the method, and three active ingredients of iso-dendrophyllin, astragalin and chlorogenic acid are analyzed and detected. After the mulberry leaves are sieved by a 80-mesh sieve and dried in a 60-DEG C drying oven for 5 hours for pretreatment, 5 per thousand of heteropoly acid and 22ml of absolute ethyl alcohol are added, the reaction temperature is 180 ℃, the reaction time is 110min, and the extraction effect is the best, which shows that the heteropoly acid has important practical significance in the extraction of active ingredients in the mulberry leaves so as to realize the resource utilization of the heteropoly acid.
The present invention will be further described with reference to the following embodiments. The method for extracting the active ingredients in the mulberry twigs (leaves) by the cooperation of the hydrothermal reaction and the solid acid catalysis comprises the following specific steps:
example 1
(1) Pretreatment of raw materials
Drying picked fresh folium Mori in a 60 deg.C oven for 5 hr, pulverizing the dried folium Mori into powder by a pulverizer, sieving with 80 mesh sieve, and packaging into sealed bags.
(2) Solid acid reinforced hydrothermal reaction treatment raw material
After the step (1) is finished, firstly weighing 2g of the pretreated mulberry leaf powder in a hydrothermal reaction kettle, then adding 0.01g of phosphotungstic heteropoly acid which is 5 per thousand of the mass (g) of the mulberry leaf, and finally weighing 18ml of absolute ethyl alcohol according to the ratio of the volume (ml) of the absolute ethyl alcohol to the mass (g) of the mulberry leaf powder of 9: 1 and pouring the absolute ethyl alcohol into the hydrothermal reaction kettle. Then the reaction kettle is put into an oven at 180 ℃ for reaction for 110 min.
(3) Assay for active ingredients
And (3) after the step (2) is finished, taking the reaction kettle out, cooling to room temperature, pouring the reaction kettle into a centrifuge tube, centrifuging for 15min at the rotating speed of 4000r/min, pouring out supernatant, and fixing the volume to a 25nl volumetric flask. Transferring 1ml of folium Mori extract into a sample bottle, detecting with mobile phase acetonitrile (ml) = 40: 60, temperature 25 deg.C, sample amount 10 μ L, and flow rate 0.6ml/min, at which time active substance in folium Mori is fully dissolved out, and displaying peak area (mAU · s) reaching 23769.7 according to liquid phase detection result.
(4) Active ingredient product extraction
After the step (3), placing the collected centrifugal clear liquid in a rotary evaporator, and carrying out reduced pressure concentration to a viscous state under the vacuum pressure of 0.5Mpa and the temperature of 90 ℃ to obtain a crude mulberry leaf extract product; then, according to the mass ratio of the mulberry leaf extract product (g) to the volume (ml) of ethanol being 1: 9, adding the mulberry leaf extract and ethanol solution into an extraction container, and mixing and dissolving uniformly; and finally, performing reduced pressure concentration for 3 times through a rotary evaporator under the vacuum pressure of 0.5Mpa and the temperature of 90 ℃, and obtaining the mulberry leaf extract product.
Example 2
The same as example 1, a method for synergistically extracting active ingredients in mulberry twigs (leaves) by a solid acid enhanced hydrothermal reaction, wherein:
in the step (1), the temperature of the oven is 50 ℃, the drying time is 6h, and the mixture is sieved by a 60-mesh sieve.
In the step (2), the mass (g) of the phosphotungstic heteropoly acid is 4 per mill of the mass (g) of the mulberry leaves, the ratio of absolute ethyl alcohol (ml) to the mass (g) of the mulberry leaf powder is 7: 1, the reaction temperature is 170 ℃, and the reaction time is 130 min.
In the step (3), the rotating speed is 2000r/min, and the centrifugation time is 20 min. Acetonitrile (ml) to water (ml) ratio was 35: 65, temperature was 30 ℃, sample size was 5 μ L, flow rate was 0.8ml/min, peak area (mAu × s) was 19738.334.
In the step (4), the ratio of the mass (g) of the mulberry leaf extract product to the volume (ml) of ethanol is 1: 8 under vacuum pressure of 0.4MPa and at 80 deg.C, and the vacuum concentration is repeated 4 times.
Example 3
The same as example 1, a method for synergistically extracting active ingredients in mulberry twigs (leaves) by a solid acid enhanced hydrothermal reaction, wherein:
in the step (1), the temperature of the oven is 70 ℃, the drying time is 4h, and the mixture is sieved by a 100-mesh sieve.
In the step (2), the mass (g) of the phosphotungstic heteropoly acid is 6 per mill of the mass (g) of the mulberry leaves, the ratio of absolute ethyl alcohol (ml) to the mass (g) of the mulberry leaf powder is 11: 1, the reaction temperature is 190 ℃, and the reaction time is 90 min.
In the step (3), the rotating speed is 3000r/min, and the centrifugation time is 15 min; acetonitrile (ml) to water (ml) ratio 50: 50, temperature 40 ℃, sample size 20 μ L, flow rate 1ml/min, peak area (mAu · s) 18549.287.
In the step (4), the ratio of the mass (g) of the mulberry leaf extract product to the volume (ml) of ethanol is 1: 10 under vacuum pressure of 0.6MPa at 100 deg.C, and the concentration under reduced pressure is repeated 2 times.
Results of the experiment
1. The experimental results of extracting and preparing the mulberry leaf extract under different conditions are as follows:
TABLE 1-1 Effect of different temperatures on Mulberry leaf extract
| Serial number | Temperature (. degree.C.) | Amount of mulberry leaf (g) | Peak area |
| 1 | 150 | 2.0005 | 4952.58206 |
| 2 | 160 | 2.0002 | 5957.05733 |
| 3 | 170 | 2.0004 | 6633.87159 |
| 4 | 180 | 2.0005 | 7278.36817 |
| 5 | 190 | 2.0001 | 4767.52964 |
TABLE 1-2 influence of different solid-liquid ratios on mulberry leaf extract
| Serial number | Solid-to-liquid ratio | Amount of mulberry leaf (g) | Peak area |
| 1 | 1﹕3 | 2.0005 | 3994.49018 |
| 2 | 1﹕5 | 2.0002 | 4179.25257 |
| 3 | 1﹕7 | 2.0004 | 5305.81655 |
| 4 | 1﹕9 | 2.0005 | 9335.01751 |
| 5 | 1﹕11 | 2.0001 | 8241.60411 |
Tables 1-3 Effect of different times on Mulberry leaf extract
| Serial number | Time (min) | Amount of mulberry leaf (g) | Peak area |
| 1 | 70 | 2.0005 | 9197.33561 |
| 2 | 90 | 2.0002 | 9850.28037 |
| 3 | 110 | 2.0004 | 23769.7 |
| 4 | 130 | 2.0005 | 8074.78674 |
| 5 | 150 | 2.0001 | 7530.05743 |
From the above experiments, it can be seen that: according to the invention, solid acid is adopted to coordinate with a catalytic hydrothermal reaction to extract active ingredients in the waste mulberry leaves, after the mulberry leaves are pretreated by passing through a 80-mesh sieve and drying in a 60 ℃ oven for 5 hours, 5 thousandth of heteropolyacid and 18ml of absolute ethyl alcohol are added, the reaction temperature is 180 ℃, the reaction time is 110min, the extraction effect is the best, according to a high-efficiency liquid phase result, the peak area reaches 23769.7, the active ingredients in the mulberry leaves can be fully dissolved out, and three active ingredients of isoquercitrin, resveratrol and chlorogenic acid are analyzed and detected, so that the application of the active ingredients in the mulberry leaves in the extraction of the heteropolyacid to realize the resource utilization is shown to have very important practical significance. In the extraction process, only absolute ethyl alcohol is used, so that the use of other toxic and harmful solvents is successfully avoided, the safety performance is improved, the safety of the experimental process is improved, the environmental protection is facilitated, and the operation is simple.
Claims (3)
1. A method for extracting active ingredients from mulberry leaves by a hydrothermal reaction in cooperation with solid acid catalysis is characterized by comprising the following specific operation steps:
(1) pretreatment of raw materials
In the step (1), picked fresh mulberry leaves are dried in a drying oven at 60 ℃ for 5 hours, the dried mulberry leaves are crushed into powder by a crusher, sieved by a 80-mesh sieve and put into a sealed bag for later use;
(2) solid acid reinforced hydrothermal reaction treatment raw material
After the step (1), weighing 2g of pretreated mulberry leaf powder in a hydrothermal reaction kettle, adding 0.01g of phosphotungstic heteropoly acid which is 5 per thousand of the mass g of the mulberry leaves, weighing 18ml of absolute ethyl alcohol according to the ratio of the volume ml of the absolute ethyl alcohol to the mass g of the mulberry leaf powder being 9: 1, pouring the absolute ethyl alcohol into the hydrothermal reaction kettle, and putting the reaction kettle into an oven at 180 ℃ for reaction for 110 min;
(3) assay for active ingredients
After the step (2) is finished, taking out the reaction kettle, cooling to room temperature, pouring the reaction kettle into a centrifuge tube, centrifuging for 15min at the rotation speed of 4000r/min, pouring out a supernatant, fixing the volume to a 25ml volumetric flask, transferring 1ml of mulberry leaf extract into a sample bottle, and detecting under the conditions that the mobile phase acetonitrile ml, the water ml is 40: 60, the temperature is 25 ℃, the sample injection amount is 10 mu L, and the flow rate is 0.6ml/min, wherein active substances in the mulberry leaves are fully dissolved out, and the peak area mAu s reaches 23769.7 according to a liquid phase detection result;
(4) active ingredient product extraction
After the step (3), placing the collected centrifugal clear liquid in a rotary evaporator, and carrying out reduced pressure concentration to a viscous state under the vacuum pressure of 0.5Mpa and the temperature of 90 ℃ to obtain a crude mulberry leaf extract product; then, according to the ratio of the mass g of the mulberry leaf extract product to the volume ml of ethanol being 1: 9, adding the mulberry leaf extract and ethanol solution into an extraction container, and mixing and dissolving uniformly; and finally, performing reduced pressure concentration for 3 times through a rotary evaporator under the vacuum pressure of 0.5Mpa and the temperature of 90 ℃, and obtaining the mulberry leaf extract product.
2. A method for extracting active ingredients from mulberry leaves by a hydrothermal reaction in cooperation with solid acid catalysis is characterized by comprising the following specific operation steps:
(1) pretreatment of raw materials
In the step (1), picked fresh mulberry leaves are dried in a drying oven at 50 ℃ for 6 hours, the dried mulberry leaves are crushed into powder by a crusher, screened by a 60-mesh sieve and put into a sealed bag for later use;
(2) solid acid reinforced hydrothermal reaction treatment raw material
After the step (1), firstly weighing 2g of pretreated mulberry leaf powder in a hydrothermal reaction kettle, then adding 0.008g of phosphotungstic heteropoly acid which is 4 per thousand of the mass g of the mulberry leaves, finally weighing 18ml of absolute ethyl alcohol according to the ratio of the volume ml of the absolute ethyl alcohol to the mass g of the mulberry leaf powder being 7: 1, pouring the absolute ethyl alcohol into the hydrothermal reaction kettle, and then putting the reaction kettle into an oven at 170 ℃ for reaction for 130 min;
(3) assay for active ingredients
After the step (2) is finished, taking out the reaction kettle, cooling to room temperature, pouring the reaction kettle into a centrifuge tube, centrifuging at the rotating speed of 2000r/min for 20min, pouring out a supernatant, fixing the volume to a 25ml volumetric flask, transferring 1ml of mulberry leaf extract into a sample bottle, and detecting under the conditions that the mobile phase acetonitrile ml, the water ml is 35: 65, the temperature is 30 ℃, the sample injection amount is 5 mu L, and the flow rate is 0.8ml/min, wherein active substances in the mulberry leaves are fully dissolved out, and the peak area mAu s reaches 19738.334 according to a liquid phase detection result;
(4) active ingredient product extraction
After the step (3), placing the collected centrifugal clear liquid in a rotary evaporator, and carrying out reduced pressure concentration to a viscous state under the vacuum pressure of 0.4Mpa and the temperature of 80 ℃ to obtain a crude mulberry leaf extract product; then, according to the ratio of the mass g of the mulberry leaf extract product to the volume ml of ethanol being 1: 8, adding the mulberry leaf extract and ethanol solution into an extraction container, and mixing and dissolving uniformly; and finally, performing reduced pressure concentration on the mulberry leaves by a rotary evaporator under the vacuum pressure of 0.4Mpa and the temperature of 80 ℃ for 4 times to obtain a mulberry leaf extract product.
3. A method for extracting active ingredients from mulberry leaves by a hydrothermal reaction in cooperation with solid acid catalysis is characterized by comprising the following specific operation steps:
(1) pretreatment of raw materials
In the step (1), picked fresh mulberry leaves are dried in a drying oven at 70 ℃ for 4 hours, the dried mulberry leaves are crushed into powder by a crusher, screened by a 100-mesh sieve and put into a sealed bag for later use;
(2) solid acid reinforced hydrothermal reaction treatment raw material
After the step (1), weighing 2g of pretreated mulberry leaf powder in a hydrothermal reaction kettle, adding phosphotungstic acid which is 0.012g and 6 per thousand of the mass g of the mulberry leaf, finally weighing 18ml of absolute ethyl alcohol according to the ratio of the volume ml of the absolute ethyl alcohol to the mass g of the mulberry leaf powder being 11: 1, pouring the absolute ethyl alcohol into the hydrothermal reaction kettle, and then putting the reaction kettle into a 190 ℃ oven for reaction for 90 min;
(3) assay for active ingredients
After the step (2) is finished, taking out the reaction kettle, cooling to room temperature, pouring the reaction kettle into a centrifuge tube, centrifuging for 15min at the rotating speed of 3000r/min, pouring out a supernatant, fixing the volume to a 25ml volumetric flask, transferring 1ml of mulberry leaf extract into a sample bottle, and detecting under the conditions that the mobile phase acetonitrile ml, the water ml, the temperature of 40 ℃, the sample injection amount of 20 mu L and the flow rate of 1ml/min, wherein active substances in the mulberry leaves are fully dissolved out, and the peak area mAU s reaches 18549.287 according to a liquid phase detection result;
(4) active ingredient product extraction
After the step (3), placing the collected centrifugal clear liquid in a rotary evaporator, and carrying out reduced pressure concentration to a viscous state under the vacuum pressure of 0.6Mpa and the temperature of 100 ℃ to obtain a crude mulberry leaf extract product; then, according to the ratio of the mass g of the mulberry leaf extract product to the volume ml of ethanol being 1: 10, adding the mulberry leaf extract and ethanol solution into an extraction container, and mixing and dissolving uniformly; and finally, performing reduced pressure concentration for 2 times through a rotary evaporator under the vacuum pressure of 0.6Mpa and the temperature of 100 ℃ to obtain the mulberry leaf extract product.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810501775.5A CN108619243B (en) | 2018-05-23 | 2018-05-23 | Method for synergistically extracting active ingredients in ramulus mori (leaves) by solid acid enhanced hydrothermal reaction |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810501775.5A CN108619243B (en) | 2018-05-23 | 2018-05-23 | Method for synergistically extracting active ingredients in ramulus mori (leaves) by solid acid enhanced hydrothermal reaction |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108619243A CN108619243A (en) | 2018-10-09 |
| CN108619243B true CN108619243B (en) | 2021-04-23 |
Family
ID=63694065
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810501775.5A Active CN108619243B (en) | 2018-05-23 | 2018-05-23 | Method for synergistically extracting active ingredients in ramulus mori (leaves) by solid acid enhanced hydrothermal reaction |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108619243B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110551229B (en) * | 2019-09-19 | 2022-07-01 | 重庆工商大学 | Method for extracting natural product by catalyzing and activating mulberry twigs with heteropoly acid solution |
| CN114539132B (en) * | 2022-04-13 | 2023-09-12 | 重庆工商大学 | DNJ method for hydrothermally acid-controlled alcohol extraction of mulberry leaves |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101428069A (en) * | 2008-12-08 | 2009-05-13 | 广东省农业科学院蚕业与农产品加工研究所 | Process for producing mulberry leaf polyphenol, microcapsule and uses thereof |
| CN107282106A (en) * | 2017-06-28 | 2017-10-24 | 重庆工商大学 | A kind of weak water-soluble supported catalyst preparation method and application |
-
2018
- 2018-05-23 CN CN201810501775.5A patent/CN108619243B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101428069A (en) * | 2008-12-08 | 2009-05-13 | 广东省农业科学院蚕业与农产品加工研究所 | Process for producing mulberry leaf polyphenol, microcapsule and uses thereof |
| CN107282106A (en) * | 2017-06-28 | 2017-10-24 | 重庆工商大学 | A kind of weak water-soluble supported catalyst preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| 一测多评法测定新疆桑枝中槲皮素_绿原酸和紫云英苷的含量;郝晓婷;《中国卫生检验杂志》;20180228;第28卷(第4期);第405-408页 * |
| 水热法提取金银花中绿原酸的工艺研究;李杰等;《食品研究与开发》;20180131;第39卷(第2期);第62-67页，尤其是第66页左栏最后一段和右栏第1段 * |
| 紫苏梗催化强化降解及其天然活性成分提取工艺研究;赵莉娟;《紫苏梗催化强化降解及其天然活性成分提取工艺研究》;20180315(第3期);B016-50，尤其是第52页最后一段 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108619243A (en) | 2018-10-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104856015A (en) | A cordyceps sinensis enzyme and a preparation method thereof | |
| CN101961371B (en) | Method for extracting and separating ginsenoside, flavone and polysaccharide from sweet gynostemma pentaphylla | |
| CN102274246B (en) | Raw material pretreatment method for improving dissolution rate of active ingredients of medicinal plants | |
| CN109329500A (en) | A kind of Fu-brick tea polyphenol and the composite instant tea of polysaccharide and its preparation method and application | |
| CN101585885A (en) | Method for preparing polygonatum odoratum polysaccharide | |
| CN103865966B (en) | The method that a kind of microorganism substep solid fermentation prepares tea polysaccharide | |
| CN111821233B (en) | Plant whitening natural active compound composition and preparation method and application thereof | |
| CN110386860A (en) | A kind of highly effective extraction method of cannabidiol | |
| CN108619243B (en) | Method for synergistically extracting active ingredients in ramulus mori (leaves) by solid acid enhanced hydrothermal reaction | |
| CN106806160A (en) | Ultrasonic microwave aids in Enzymatic Extraction green tea polyphenols and its application in shower cream | |
| CN111135123B (en) | Preparation method and application of selaginella tamariscina fermentation extracting solution | |
| CN102697988A (en) | Method for extracting flavone from banana skin | |
| CN111096983A (en) | Phellinus igniarius phenolic extract with hypoglycemic activity and preparation and application thereof | |
| CN107213185A (en) | A kind of blueberry leaf polyphenol preparation method with antioxidation activity | |
| CN102321144A (en) | Method for extracting ursolic acid from red jujube | |
| CN106699916A (en) | Combined extraction method of polyphenols and polysaccharides from lotus roots | |
| CN109170885A (en) | A method of extracting ginseng effective component | |
| CN102940688A (en) | Extraction method for polysaccharides and total flavonoids in fig leaves | |
| CN104940280A (en) | Method for extracting total flavones from radix puerariae employing enzyme preparation | |
| CN111840349B (en) | Method for simultaneously extracting polysaccharide and flavonoid components from folium artemisiae argyi | |
| CN108567836A (en) | A method of combined extraction separation flavones and polysaccharide from hawthorn skin slag | |
| CN101830881B (en) | Enzyme-induced method for efficiently extracting toxifolin from larch processing leftover | |
| CN101619108B (en) | Method for extracting radiation resistant potentilla anserine polysaccharide oxide by ultrasonic waves | |
| WO2017185289A1 (en) | Method for preparing active extract of sea buckthorn | |
| CN109846932A (en) | A kind of extraction purification and technology of preparing of mulberries slag blood-sugar decreasing active |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |