Bioactive substance extraction method

Abstract

The invention discloses a method for extracting a bioactive substance, and particularly relates to the field of substance extraction, and the bioactive substance is a polyphenol substance. The extraction of the polyphenols specifically comprises the following steps: S1, pretreatment: putting fruits / vegetables rich in the polyphenols into a low-temperature vacuum machine, and carrying out low-temperature vacuum microwave pretreatment in a vacuum environment with the vacuum degree of-0.1 MPa to-0.06 MPa to obtain a prefabricated raw material; s2, crushing: performing airflow crushing on the prefabricated raw material obtained in the step S1 at the air pressure of 0.5-1.5 MPa, wherein the airflow crushing environment temperature is set to be-5 DEG C to 0 DEG C or below. According to the invention, through the research on a novel extraction method for pretreating, crushing and extracting different fruits and vegetables, the step setting of vacuum low-temperature treatment and airflow crushing is utilized, and the crushed powder is extracted by using methanol and ethanol aqueous solution, so that effective substances of polyphenols in the raw materials can be effectively extracted; and the yield of effective substances in the raw materials is greatly improved.

Description

Technical Field

[0001] The present invention relates to the technical field of substance extraction, and more specifically, the present invention relates to a method for extracting bioactive substances. Background Art

[0002] Natural bioactive substances have physiological activities such as anti-inflammatory, anti-cancer, and antioxidant, and are widely distributed in various animals and plants, marine organisms, and microorganisms. Fruits and vegetables are an important part of daily diet. Currently, the main functional factors that have been confirmed include dietary fiber, fructooligosaccharides, phytosterols, carotenoids, flavonoids, diallyl disulfide, resveratrol, isothiocyanates, limonene, etc. For example, apples are the main source of flavanols and flavonols in daily diet and have the effects of enhancing immunity and antioxidant; longan polysaccharide has health care functions such as regulating and enhancing immunity, memory, antioxidant, and anti-tumor; resveratrol contained in grape fruits and seeds has the effect of inhibiting lipid oxidation. The main bioactive substances in vegetables include carotenoids, organosulfur compounds, polysaccharides, and lycopene, anthocyanins, etc. in flavonoids, which can significantly promote human health and prevent diseases. For example, pumpkin polysaccharide has various health care functions such as diet therapy for preventing diseases; the active chemical components such as triterpenoids, steroids, alkaloids, proteins, organic acids, and sugars in bitter gourd have functions such as lowering blood sugar and blood lipids, regulating enzyme activity, antiviral, and anti-tumor.

[0003] Polyphenolic substances have strong antioxidant effects. Polyphenolic substances are called "the seventh nutrient". Its main active ingredient is polyphenolic substances. Polyphenolic compounds refer to the general term of plant components with several phenolic hydroxyl groups in the molecular structure, including flavonoids, tannins, phenolic acids, and anthocyanins, etc. Phenols are compounds found in plant-based foods that have potential health-promoting effects. It exists in some common plant-based foods, such as cocoa beans, tea, soybeans, vegetables, and fruits.

[0004] When extracting and preparing existing polyphenolic substances, common drawbacks are mostly situations such as low extraction rate or excessive production cost. How to prepare polyphenolic substances environmentally friendly and economically is an urgent research topic. Summary of the Invention

[0005] In order to overcome the above-mentioned defects of the prior art, an embodiment of the present invention provides a method for extracting bioactive substances. The technical problem to be solved by the present invention is: how to extract polyphenolic substances in natural plants economically and inexpensively.

[0006] To achieve the above object, the present invention provides the following technical solution: A method for extracting bioactive substances, wherein the bioactive substance is polyphenolic substances;

[0007] The extraction of the polyphenolic substances specifically adopts the following steps:

[0008] S1. Pretreatment: Put fruits / vegetables rich in polyphenols into a low-temperature vacuum machine and conduct low-temperature vacuum microwave pretreatment in a vacuum environment with a vacuum degree of -0.1 MPa to -0.06 MPa to obtain prefabricated raw materials;

[0009] S2. Crushing: Conduct pneumatic crushing on the prefabricated raw materials obtained in step S1 at a gas pressure of 0.5 - 1.5 MPa. Set the temperature of the pneumatic crushing environment to below -5°C to -0°C, and set the pneumatic crushing time to 20 - 25 min to obtain crushed materials;

[0010] S3. Extraction;

[0011] S3.1: Select an ethanol solvent with a purity index of 70% and prepare a mixed solution with the crushed materials according to a weight ratio of 1 - 1.5:1. Mix the mixed solution, methanol, and pure water according to a molar ratio of 1:2 - 3:2 - 3, and react at 25°C - 30°C for 0.5 h under nitrogen protection to conduct an extraction reaction;

[0012] S3.2: Cool the reaction solution after extraction in step S3.1, then centrifuge and filter to remove residues to obtain a supernatant of polyphenol substances;

[0013] S3.3: Put the supernatant of polyphenol substances prepared in step S3.2 into a short-path molecular distillation unit and concentrate it at a temperature of 45 - 60°C to obtain a concentrated solution of polyphenol substances.

[0014] In a preferred embodiment, the fruits / vegetables rich in polyphenols in step S1 include but are not limited to apples, strawberries, blueberries, cherries, and plums; among them, the low-temperature environment of the low-temperature vacuum machine is set between -10°C and 20°C. In a vacuum state, the internal pressure of the container is low, which helps with the drying, degassing, impurity removal, etc. of substances, only increasing the brittleness of the cell walls of fruits / vegetables and not directly breaking the cell walls of fruits / vegetables. Conduct microwave treatment for 10 - 30 minutes, and the low-temperature environment keeps the nutrients from being lost. The vibration of water molecules breaks the cellulose of the cell walls, thereby increasing the brittleness of the cells. Conducting microwave treatment in a vacuum environment can avoid the influence of active factors in the air (such as oxygen) on the nutritional components in the cells.

[0015] In a preferred embodiment, the methanol selected in step S3.1 is methanol with a mass fraction concentration of 95%.

[0016] In a preferred embodiment, a low-speed centrifuge is selected for centrifugation in step S3.2, and it rotates at a speed of 3000 - 3500 rpm for 10 - 15 min; the filter screen for filtration is a 6000-mesh fiberglass filter screen.

[0017] In a preferred embodiment, the molecular distillation in step S3.3 is carried out in a vacuum environment, and the internal vacuum of the short-path molecular distillation is set to 0.1 - 10 Pa.

[0018] Technical effects and advantages of the present invention:

[0019] Through the research on a new extraction method of pre-treating, crushing and extracting different fruits and vegetables, by setting steps of vacuum low-temperature treatment and airflow pulverization, and extracting the crushed powder with ethanol, methanol and pure water, the effective polyphenolic substances in the raw materials can be effectively extracted, greatly improving the recovery rate of the effective substances in the raw materials. Specific embodiments

[0020] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0021] Example 1:

[0022] The present invention provides a method for extracting bioactive substances, and the bioactive substances are polyphenolic substances;

[0023] The extraction of the polyphenolic substances specifically adopts the following steps:

[0024] S1. Pretreatment: Put apples rich in polyphenolic substances into a low-temperature vacuum machine, and carry out low-temperature vacuum microwave pretreatment in a vacuum environment with a vacuum degree of -0.1 MPa and a temperature of -10 °C to obtain prefabricated raw materials. In a vacuum state, the internal pressure of the container is low, which helps with the drying, degassing, and impurity removal of substances, only increasing the brittleness of the fruit / vegetable cell wall without directly breaking the fruit / vegetable cells. Microwave treatment is carried out for 10 minutes, and the low-temperature environment keeps the nutrients from being lost. The vibration of water molecules breaks the cellulose of the cell wall, thereby increasing the brittleness of the cells. Microwave treatment in a vacuum environment can avoid the influence of active factors in the air (such as oxygen) on the nutrients in the cells;

[0025] S2. Crushing: Carry out airflow pulverization on the prefabricated raw materials obtained in step S1 at an air pressure of 0.5 MPa. The temperature of the airflow pulverization environment is set below -5 °C, and the airflow pulverization time is set to 20 min to obtain pulverized materials;

[0026] S3. Extraction;

[0027] S3.1: Select an ethanol solvent with a purity index of 70% and the crushed material to prepare a mixed solution according to a weight ratio of 1:1, and mix the mixed solution, methanol with a mass fraction concentration of 95%, and pure water in a molar ratio of 1:2:2, and react at 25°C for 0.5 h under nitrogen protection to carry out an extraction reaction;

[0028] S3.2: Cool the reaction solution after extraction in step S3.1, then centrifuge and filter to remove residues, and centrifuge at a speed of 3000 rpm for 10 min; The filter screen for filtration is a 6000-mesh fiberglass filter screen to obtain the supernatant of polyphenol substances;

[0029] S3.3: Put the supernatant of polyphenol substances prepared in step S3.2 into a short-path molecular distillation device and concentrate it at a vacuum degree of 0.1 Pa and a temperature of 45°C to obtain a concentrated solution of polyphenol substances.

[0030] Example 2:

[0031] The present invention provides a method for extracting bioactive substances, and the bioactive substances are polyphenol substances;

[0032] The extraction of the polyphenol substances specifically adopts the following steps:

[0033] S1. Pretreatment: Put blueberries rich in polyphenol substances into a low-temperature vacuum machine, and carry out low-temperature vacuum microwave pretreatment in a vacuum environment with a vacuum degree of -0.08 MPa and a temperature of 0°C to obtain a prefabricated raw material. In a vacuum state, the internal pressure of the container is low, which helps with the drying, degassing, and impurity removal of substances, only increasing the brittleness of the fruit / vegetable cell wall without directly breaking the fruit / vegetable cells. Microwave treatment is carried out for 20 minutes, and the low-temperature environment keeps the nutrients from being lost. The vibration of water molecules breaks the cellulose of the cell wall, thereby increasing the brittleness of the cells. Microwave treatment in a vacuum environment can avoid the influence of active factors in the air (such as oxygen) on the nutrients in the cells;

[0034] S2. Crushing: Carry out air flow crushing on the prefabricated raw material obtained in step S1 at a gas pressure of 1 MPa. The air flow crushing environment temperature is set below -2°C, and the air flow crushing time is set to 22 min to obtain a crushed material;

[0035] S3. Extraction;

[0036] S3.1: Select an ethanol solvent with a purity index of 70% and the crushed material to prepare a mixed solution according to a weight ratio of 1.25:1, and mix the mixed solution, methanol with a mass fraction concentration of 95%, and pure water in a molar ratio of 1:3:3, and react at 30°C for 0.5 h under nitrogen protection to carry out an extraction reaction;

[0037] S3.2: Cool the reaction solution after extraction in step S3.1, then centrifuge and filter to remove residues. Centrifuge at a speed of 3200 rpm for 12 min. Select a 6000-mesh fiberglass filter screen for filtration to obtain the supernatant of polyphenolic substances.

[0038] S3.3: Put the supernatant of polyphenolic substances obtained in step S3.2 into a short-path molecular distillation unit and concentrate it under a vacuum of 5 Pa and a temperature of 50 °C to obtain a concentrated solution of polyphenolic substances.

[0039] Example 3:

[0040] The present invention provides a method for extracting bioactive substances, and the bioactive substances are polyphenolic substances.

[0041] The extraction of the polyphenolic substances specifically adopts the following steps:

[0042] S1. Pretreatment: Put plums rich in polyphenolic substances into a low-temperature vacuum machine and perform low-temperature vacuum microwave pretreatment in a vacuum environment with a vacuum degree of -0.06 MPa and a temperature of 20 °C to obtain prefabricated raw materials. In a vacuum state, the internal pressure of the container is low, which helps with the drying, degassing, and impurity removal of substances. It only increases the brittleness of the fruit / vegetable cell walls and does not directly break the fruit / vegetable cells. Microwave treatment is carried out for 30 minutes, and the low-temperature environment keeps the nutrients from being lost. The vibration of water molecules breaks the cellulose of the cell walls, thereby increasing the brittleness of the cells. Microwave treatment in a vacuum environment can avoid the influence of active factors in the air (such as oxygen) on the nutrients in the cells.

[0043] S2. Crushing: Subject the prefabricated raw materials obtained in step S1 to airflow crushing at an air pressure of 1.5 MPa. Set the temperature of the airflow crushing environment to below 0 °C and the airflow crushing time to 25 min to obtain crushed materials.

[0044] S3. Extraction;

[0045] S3.1: Select an ethanol solvent with a purity index of 70% and the crushed materials to prepare a mixed solution according to a weight ratio of 1.5:1. Then mix the mixed solution, methanol with a mass fraction concentration of 95%, and pure water according to a molar ratio of 1:3:3, and carry out an extraction reaction at 30 °C for 0.5 h under nitrogen protection.

[0046] S3.2: Cool the reaction solution after extraction in step S3.1, then centrifuge and filter to remove residues. Centrifuge at a speed of 3500 rpm for 15 min. Select a 6000-mesh fiberglass filter screen for filtration to obtain the supernatant of polyphenolic substances.

[0047] S3.3: Put the supernatant of polyphenolic substances obtained in step S3.2 into a short-path molecular distillation unit and concentrate it at a vacuum degree of 10 Pa and a temperature of 65 °C to obtain a concentrated solution of polyphenolic substances.

[0048] Example 4:

[0049] Respectively take the polyphenolic substances prepared in the above Examples 1-3 for purity detection, and the following data are obtained:

[0050]

[0051] As can be seen from the above table, the concentration of polyphenolic substances in the concentrated solutions prepared by the extraction methods in Examples 1-3 is above 89%, which can well ensure a great increase in the recovery rate of raw materials, and is applicable to the extraction of active substances in fruits and vegetables with different polyphenolic substance contents, and has remarkable effects compared with traditional extraction methods.

[0052] Finally: The above are only the preferred embodiments of the present invention and are not used to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for extracting biologically active substances, characterized in that: The biologically active substances are polyphenols; The extraction of the polyphenols specifically adopts the following steps: S1. Pretreatment: putting fruits / vegetables rich in polyphenols into a low-temperature vacuum machine, and performing low-temperature vacuum microwave pretreatment in a vacuum environment with a vacuum degree of -0.1 MPa to -0.06 MPa to obtain prefabricated raw materials; S2, crushing: the prefabricated raw material obtained in step S1 is subjected to air flow crushing at an air pressure of 0.5-1.5 MPa, the air flow crushing environment temperature is set to below -5°C-0°C, and the air flow crushing time is set to 20-25 min to obtain crushed material; S3, extraction; S3.1: Select an ethanol solvent with a purity index of 70% and a crushed material in a weight ratio of 1-1.5:1 to prepare a mixed solution, and mix the mixed solution, methanol and pure water in a molar ratio of 1:2-3:2-3, react at 25°C-30°C for 0.5h under nitrogen protection to perform an extraction reaction; S3.2: cooling the reaction solution extracted in step S3.1, centrifuging it, filtering and removing residues, and obtaining a polyphenol supernatant; S3.3: The polyphenol supernatant obtained in step S3.2 is placed in a short-range molecular distillation and concentrated at a temperature of 45-65° C. to obtain a polyphenol concentrated solution.

2. A method for extracting biologically active substances according to claim 1, characterized in that: The fruits / vegetables rich in polyphenols in step S1 include but are not limited to apples, strawberries, blueberries, cherries, and plums; wherein the low temperature environment of the low temperature vacuum machine is set to between -10°C and 20°C, and the microwave treatment is performed for 10 to 30 minutes.

3. A method for extracting biologically active substances according to claim 1, characterized in that: In the step S3.1, the methanol is selected to have a mass fraction concentration of 95%.

4. A method for extracting biologically active substances according to claim 1, characterized in that: In step S3.2, a low-speed centrifuge is used for centrifugation at a speed of 3000-3500 rpm for 10-15 min; a 6000-mesh glass fiber filter is used for the filtration.

5. A method for extracting biologically active substances according to claim 1, characterized in that: In step S3.3, the molecular distillation is carried out in a vacuum environment, and the vacuum degree in the short-range molecular distillation is set to 0.1-10Pa.

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