WO2020215756A1 - Method for extracting active substance extraction solution and application - Google Patents

Abstract

A method for extracting an active substance extraction solution and an application. The method comprises the steps of: preparing an extract; wherein the extract comprises at least an eutectic solvent; adding an animal and plant or microbial sample to the extract for mechanochemical extraction; and centrifuging, and removing a supernatant to obtain an active substance extraction solution. The active ingredient in an animal and plant or microbial sample can be extracted efficiently without pollution, and there is no need for high temperature and no consumption of a toxic organic solvent during an extraction process, thereby reducing environmental pollutions. There is no need to remove the solvent during the process of making a daily chemical product from the extract and the extract can be used directly. The method features simple operation, short time-consuming, low costs and high extraction efficiency.

Description

An extraction method and application of active substance extract Technical field

The invention relates to an extraction method and application of an active substance extract.

Background technique

In recent years, biologically active natural products have attracted more and more attention in the world in pharmacology and biology. These products have the effects of curing diseases, improving physical fitness and skin care. The active ingredients in animals, plants and microorganisms have attracted more and more attention due to their safety, reliability and remarkable activity. The biologically active substances contained in them have anti-oxidation, elimination of free radicals, anti-inflammatory and other physiological activities. Therefore, its active ingredients are increasingly used in the field of daily chemicals. However, in many cases, these active ingredients need to be extracted with organic solvents, and most organic solvents are usually toxic and volatile, which can cause environmental pollution. The obtained active ingredients need to be completely removed from the toxic solvent before being added to the daily chemical products to prevent the residual solvent from causing harm to the human body. This process also increases the production cost. Therefore, the use of new green solvents to extract active ingredients is beneficial to reduce environmental pollution and reduce the production cost of daily chemicals.

The eutectic solvent is synthesized from two or more natural, inexpensive and biodegradable components, and has the advantages of simple preparation, low cost, low toxicity, good solubility, non-volatile, and biodegradable. Therefore, it can replace traditional organic solvents for extracting biologically active substances from animal, plant or microbial materials.

Summary of the invention

The technical problem to be solved by the present invention is to overcome the defects of the prior art and provide a method for extracting active substance extracts, which can extract the active ingredients in animal, plant or microbial samples efficiently and without pollution, and during the extraction process There is no need for high temperature, no consumption of toxic organic solvents, and environmental pollution is reduced. The extraction liquid does not need to be removed during the process of making daily chemical products, and can be used directly. This method has simple operation, short time-consuming, low cost and high extraction efficiency specialty.

In order to solve the above technical problems, the present invention provides a method for extracting an active substance extract. The steps of the method include:

Preparing an extract; wherein the extract includes at least a eutectic solvent;

Add animal, plant or microbial samples to the extraction solution for mechanochemical extraction;

Centrifuge, and take the supernatant, which is the active substance extract.

Further, the method of preparing the extract includes:

Mix hydrogen bond acceptor and hydrogen bond donor;

Heat and stir to a homogeneous phase, that is, the eutectic solvent;

The eutectic solvent is mixed with water.

Further, the mole fraction ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:4 to 4:1.

Further, the hydrogen bond acceptor is at least one of quaternary ammonium salts, zwitterions, amino acids, sugars, and carboxylic acids;

Further, the hydrogen bond donor is at least one of urea, thiourea, carboxylic acid, polyol, amino acid, sugar, and amide.

Further, the heating temperature is 60-120°C.

Further, the mass content of the eutectic solvent in the extract is 50-100%.

Further, the solid-liquid ratio of the animal, plant or microbial sample and the extract is 0.01-0.20 g/mL.

Further, the mechanochemical extraction is suitable for using a cell lysis instrument, a cell homogenizer, or a ball mill;

The extraction time is 0.1-120min.

In another aspect, the present invention also provides an application of the active substance extract extracted by the extraction method, which is applied to the processing and production of daily chemicals.

The daily chemicals are facial cleanser, cleansing foam, facial mask, shampoo, shower gel, air freshener, anti-cracking oil, essence, whitening cream, sunscreen, hand cream, moisturizing lotion, moisturizing cream, softening lotion , Astringent, firming lotion, washing powder, conditioner, hair cream, sunscreen, shampoo, aftershave, rouge, mascara, toothpaste.

After adopting the above technical scheme, the method of the present invention uses a eutectic solvent as the extraction solvent and a mechanochemical extraction method as an extraction method to extract the active ingredients in animal, plant or microbial samples. It is necessary to operate at high temperature to reduce their viscosity, and high temperature is easy to destroy and decompose biologically active substances. The mechanical action in mechanochemical extraction can effectively overcome the shortcomings of high viscosity. The mechanical collision and low co-existence of grinding beads or grinding balls The melting solvent destroys the cell wall and cell membrane, achieving the synergy of mechanical force and chemical reaction. Therefore, this application combines the eutectic solvent with the mechanical action in an efficient organic combination to extract the active ingredients in animal, plant and microbial samples efficiently and without pollution. Compared with other extraction methods, there is no need in the extraction process. High temperature, no need to consume toxic organic solvents, reducing environmental pollution. The extraction liquid does not need to remove the solvent during the process of making daily chemicals, and can be used directly. The method has the characteristics of simple operation, short time-consuming, low cost and high extraction efficiency.

Description of the drawings

Fig. 1 is a graph showing the influence of different eutectic solvent contents on the extraction amount in Example 1 of the present invention.

Fig. 2 is a graph showing the influence of different solid-liquid ratios on the extraction amount in Example 1 of the present invention.

FIG. 3 is a chromatogram of tea extract by mechanochemical extraction method at a wavelength of 231 nm (a) in Example 1 of the present invention;

Fig. 4 is a chromatogram of tea extract by mechanochemical extraction method at a wavelength of 274 nm (b) in Example 1 of the present invention.

Fig. 5 is a chromatogram of tea leaf extract by mechanochemical extraction at a wavelength of 365 nm (c) in Example 1 of the present invention.

Detailed ways

In order to make the content of the present invention easier to be understood clearly, the following further describes the present invention in detail based on specific embodiments in conjunction with the accompanying drawings.

Example one

The eutectic solvent combined with mechanochemical extraction method is used to extract alkaloids, flavonoids and tea polyphenols in tea.

(1) Target extract

Caffeine (CAF), Theobromine (TBM), Kaempferol (KAE), (-)-Epicatechin (EC), (-)-Epicatechin gallate (ECg), (-)- Epigallocatechin (EGC) and (-)-Epigallocatechin (EGCg).

(2) Preparation of eutectic solvent

The proline and glycerin with a molar ratio of 1:1-1:4 are heated and stirred at 60-120°C for 30 minutes to 2 hours to a homogeneous phase, and then the eutectic solvent is diluted with water to 10-90% (mass percentage ).

Of course, the eutectic solvent of this embodiment can also be obtained by mixing other hydrogen bond acceptors and other hydrogen bond donors and then heating them. Other hydrogen bond acceptors can be quaternary ammonium salts, zwitterions, amino acids, sugars, and carboxylic acids. At least one of

Other hydrogen bond donors may be at least one of urea, thiourea, carboxylic acid, polyol, amino acid, sugar, and amide.

(3) Extraction of alkaloids, flavonoids and tea polyphenols in tea

Take 0.5-5.0g of tea powder mixed with 5-50ml of eutectic solvent aqueous solution and add it to a cell homogenizer containing grinding beads or grinding balls for mechanochemical extraction. After extraction for 10-80 seconds, centrifuge to get the supernatant. Quantification is carried out by high performance liquid chromatography; of course, the mechanochemical extraction in this embodiment can also be carried out by using a cell lysator or a ball mill.

(4) The influence of water content in eutectic solvent on extraction rate

Because the viscosity of the eutectic solvent is high, it will affect the extraction effect, so it needs to be diluted with water. The present invention explores the extraction effect of the eutectic solvent with a ratio of 10-90%. It can be seen from Figure 1 that when the ratio is 50%, the extraction effect is the best. This may be that the addition of water to the eutectic solvent reduces the extraction liquid Viscosity and changed polarity. According to the principle of similar compatibility, the target has high solubility under this condition, so 50% eutectic solvent aqueous solution is selected as the extractant.

(5) Influence of solid-liquid ratio

In the range of the solid-liquid ratio of 0.01-0.20g/mL, the effect of the solid-liquid ratio on the extraction of anthraquinone by the eutectic solvent was studied. It can be seen from Figure 2 that the extraction rate does not change much, and the solid-liquid ratio can be selected at will within this range.

(6) Influence of vibration speed

The vibration speed of the cell homogenizer is very important for grinding and breaking tea cells. The higher the vibration speed, the better the extraction effect. Choose vibration speeds of 4m/s, 4.5m/s, 5m/s, 5.5m/s and 6m/s, but the vibration speed is important for the extraction of seven biologically active compounds in tea. There is no significant impact. Considering energy saving and keeping the ceramic balls intact, 4m/s is used for the remaining extraction experiments.

(7) Effect of extraction time

The time is set to 10s, 20s, 40s, 60s and 80s. From 10s to 20s, the amount of extraction increases with the increase of time, but almost unchanged over 20s. Therefore, we choose 20s for future experiments, which also shows another advantage of this experiment: the extraction time is short, and the biologically active substances are not easy. transsexual.

(8) Stability of extracted biologically active compounds

Biologically active compounds are generally sensitive to oxygen and temperature. In order to evaluate the stability of the method, the method extracts seven biologically active compounds under selected experimental conditions. The extract was placed in the air at 20°C and analyzed by UPLC-MS every 24h. As shown in Figures 3, 4, and 5, the extraction amount of biologically active compounds did not change significantly after 96 hours.

Example two

The eutectic solvent extract is used to process liquid soap 1.

(1) Soap paste production

Coconut oil 85g, palm kernel oil 10g, cocoa butter 5g, potassium hydroxide 18g, and water 55g are mixed and heated to 70-90 degrees to completely dissolve, and then continue to heat for 2-3 hours until it appears transparent.

(2) Soap paste dilution

30g of soap paste, 65g of water, and 5g of eutectic solvent extract are mixed and poured into the pot until completely dissolved, and then used in a bottle.

Example three

The eutectic solvent extract is used to process liquid soap2.

(1) Soap paste production

Coconut oil 75g, olive oil 10g, castor oil 15g, potassium hydroxide 24g, and water 71g are mixed and heated to 70-90 degrees to completely dissolve, and then continue to heat for 2-3 hours until transparent.

(2) Soap paste dilution

Mix 57g of soap paste, 38g of water, and 5g of eutectic solvent extract and pour it into a pot until it is completely dissolved, then bottling for use.

Example three

The eutectic solvent extract is used to process moisturizing emulsions.

Paraffin wax 1%, beeswax 2%, lanolin 2%, liquid paraffin 20%, isotaricon 10%, sorbitan sesquioleate 4%, sorbitan monooleate 1%, low total Mix and dissolve 7% of the molten solvent (glycerin is replaced by propylene glycol) extract and 53% of refined water at 70°C, pour the water phase into the oil phase, emulsify with a homogenizer, and cool to room temperature.

In summary, the present invention combines eutectic solvents and mechanochemical extraction technology with high efficiency, and can extract active ingredients in animal, plant and microbial samples efficiently and without pollution. The invention has obvious advantages in the sample extraction stage, short time required for the entire process, high extraction rate, no additional pollution to the environment, safety and greenness, low cost, simple operation, and broad development prospects in extraction. At the same time, due to the non-toxic or low-toxicity of the eutectic solvent, the extract can be directly added to the daily chemical products, reducing the processing flow.

The specific embodiments described above further describe the technical problems, technical solutions, and beneficial effects solved by the present invention in further detail. It should be understood that the above descriptions are only specific embodiments of the present invention and are not intended to limit the present invention. The invention, any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

In the description of the present invention, it should be understood that the terms indicating the orientation or the positional relationship are based on the orientation or the positional relationship shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying what is meant. The device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation of the present invention.

In the present invention, unless otherwise clearly defined and defined, the terms "installed", "connected", "connected", "fixed" and other terms should be understood in a broad sense, for example, it can be a fixed connection or a detachable connection. , Or integrated; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, and it can be the internal communication of two components or the interaction relationship between two components. For those of ordinary skill in the art, the specific meaning of the above-mentioned terms in the present invention can be understood according to specific circumstances.

In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. The indicated position or position relationship is based on the position or position relationship shown in the drawings, or the position or position relationship usually placed when the product of the invention is used. It is only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying The referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation of the present invention. In addition, the terms "first", "second", "third", etc. are only used for distinguishing description, and cannot be understood as indicating or implying relative importance.

In addition, the terms "horizontal", "vertical", "overhanging" and other terms do not mean that the component is required to be absolutely horizontal or overhanging, but may be slightly inclined. For example, “horizontal” only means that its direction is more horizontal than “vertical”, it does not mean that the structure must be completely horizontal, but can be slightly inclined.

In the present invention, unless otherwise clearly defined and defined, the first feature above or below the second feature may include the first and second features in direct contact, or may include the first and second features not in direct contact but It is through the other characteristic contact between them. Moreover, the first feature being above, above, and above the second feature includes the first feature being directly above and obliquely above the second feature, or it simply means that the level of the first feature is higher than the second feature. The first feature below, below, and below the second feature includes the first feature directly below and obliquely below the second feature, or it simply means that the level of the first feature is smaller than the second feature.

Claims (10)

1. A method for extracting an active substance extract, which is characterized in that the steps of the method include:

   Preparing an extract; wherein the extract includes at least a eutectic solvent;

   Add animal, plant or microbial samples to the extraction solution for mechanochemical extraction;

   Centrifuge, and take the supernatant, which is the active substance extract.
2. The extraction method according to claim 1, wherein:

   Methods of preparing the extract include:

   Mix hydrogen bond acceptor and hydrogen bond donor;

   Heat and stir to a homogeneous phase, that is, the eutectic solvent;

   The eutectic solvent is mixed with water.
3. The extraction method according to claim 2, wherein:

   The mole fraction ratio of the hydrogen bond acceptor to the hydrogen bond donor is 1:4 to 4:1.
4. The extraction method according to claim 2, wherein:

   The hydrogen bond acceptor is at least one of quaternary ammonium salts, zwitterions, amino acids, sugars, and carboxylic acids;

   The hydrogen bond donor is at least one of urea, thiourea, carboxylic acid, polyol, amino acid, sugar, and amide.
5. The extraction method according to claim 2, wherein:

   The heating temperature is 60-120°C.
6. The extraction method according to claim 1, wherein:

   The mass content of the eutectic solvent in the extract is 50-100%.
7. The extraction method according to claim 1, wherein:

   The solid-liquid ratio of the animal, plant or microorganism sample to the extract is 0.01-0.20 g/mL.
8. The extraction method according to claim 1, wherein:

   The mechanochemical extraction is suitable for using a cell lysis instrument, a cell homogenizer, or a ball mill;

   The extraction time is 0.1-120min.
9. An application of the active substance extract extracted by the extraction method according to any one of claims 1 to 8, characterized in that:

   It is used in the production of daily chemicals.
10. The application according to claim 9, characterized in that,

    The daily chemicals are facial cleanser, cleansing foam, facial mask, shampoo, shower gel, air freshener, anti-cracking oil, essence, whitening cream, sunscreen, hand cream, moisturizing lotion, moisturizing cream, softening lotion , Astringent, firming lotion, washing powder, conditioner, hair cream, sunscreen, shampoo, aftershave, rouge, mascara, toothpaste.

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