CN119564769B

CN119564769B - A preparation method and application of DES four yellow extract

Info

Publication number: CN119564769B
Application number: CN202411798027.XA
Authority: CN
Inventors: 詹海莺; 陈美怡; 曹华; 刘想; 王星月; 顾凯迪; 丰元昊
Original assignee: Guangdong Pharmaceutical University
Current assignee: Guangdong Pharmaceutical University
Priority date: 2024-12-09
Filing date: 2024-12-09
Publication date: 2025-09-23
Anticipated expiration: 2044-12-09
Also published as: CN119564769A

Abstract

The invention provides a preparation method and application of a DES four-yellow extract, and relates to the technical field of traditional Chinese medicine extraction. A process for preparing the DES four-yellow extract includes such steps as preparing eutectic solvent from amino acid as HBA and alcohol or carboxylic acid as HBD, heating while stirring to obtain DES, preparing aqueous solution of DES, mixing it with water to obtain aqueous solution of DES, extracting from phellodendron bark, coptis root, rhubarb and scutellaria root, pulverizing, sieving, adding aqueous solution of DES prepared from S2, ultrasonic treating at 55-65 deg.C for 30-60min, cooling to room temp., centrifugal separation, and drying supernatant. The invention is a novel solvent method based on green and nontoxic, and the obtained DES four-yellow extract has stable components and good solubility.

Description

Preparation method and application of DES four-yellow extract

Technical Field

The invention relates to the technical field of traditional Chinese medicine extraction, in particular to a preparation method and application of a DES four-yellow extract.

Background

Acne, comedo and pimple are commonly called acne in medicine, are common chronic diseases of sebaceous glands, have various types of existing acne removing methods, are various acne removing cosmetics, acne removing external medicines and acne removing internal medicines, and also comprise various chemical skin changing and physical treatment methods. However, the existing acne-removing product has poor effect, strong dependence and great side effect. The traditional Chinese medicine extract is mild and has no stimulation, and is widely concerned.

Flavonoid traditional Chinese medicines such as radix scutellariae, cortex phellodendri, rhizoma coptidis, rheum officinale and the like have the effects of clearing heat and detoxicating, resisting inflammation and bacteria, resisting oxidization and the like, and have the effects of inhibiting acne, removing acnes, repairing and relieving skin. The Sanhuang Xiexin decoction composed of 3 Chinese medicinal herbs including coptis root, baikal skullcap root and rhubarb is recorded in the 'jin Kui Yao Lloyd' of Han dynasty, and is a classical recipe for treating internal heat, stasis heat and internal accumulation, etc. and purging fire and detoxicating. In addition, the Huanglian Jiedu Tang from the Jie Fang Bei Jiu (elbow-treating preparation) is prepared by decocting three or two coptis chinensis, two or two phellodendron bark and two or four baikal skullcap root respectively, six liters of water, two liters of decoction and separate administration. Has the functions of purging fire and detoxicating, is mainly used for treating all excessive heat fire toxin, and has obvious curative effect in surgical treatment of carbuncle and furuncle. According to the ancient prescription, the four-yellow extract prepared by mixing the phellodendron bark, the coptis chinensis, the rheum officinale and the scutellaria baicalensis according to a certain proportion can improve the acne removing effect to a certain extent, and forms unique and innovative combination of Chinese herbal medicines and cosmetics.

Chinese patent CN 106420490A discloses a traditional Chinese medicine composition with acne-removing effect, and a preparation method and application thereof, wherein the composition comprises, by weight, 10-20% of pulsatilla chinensis, 10-20% of gentian, 5-10% of kuh-seng, 5-10% of garden balsam stem, 5-10% of wrinkled gianthyssop herb, 5-10% of dandelion, 5-10% of virgate wormwood herb, 5-10% of rhubarb, 5-10% of amur corktree bark, 1-5% of thorn coptis root and 1-5% of baical skullcap root. Chinese patent CN 105232725A relates to an acne-removing formula which comprises a plurality of medicines of 1.5-4.5% of liquorice, 2-5% of white mulberry root-bark, 1-5% of chamomile, 0.5-5% of red sage root, 1-5% of baikal skullcap root, 1.5-5% of amur corktree bark, 1-7% of lightyellow sophora root, 0.5-3.5% of honeysuckle, 1.5-6% of weeping forsythiae capsule, 1-4% of coptis chinensis, 1-3% of dyers woad leaf, 0.5-5% of rhubarb and 3-5% of purslane and solvent.

However, in the conventional method for extracting the medicinal materials used as raw materials of cosmetics, water or organic solvents such as methanol and ethanol are generally used, as in the above-mentioned chinese patent. The water is used as the extraction solvent, which often causes the problems of more water-soluble impurities, difficult separation and purification, and the like, and the organic solvent has the defects of inflammability, easy volatilization, toxicity, more solvent consumption, easy residue, and the like.

The eutectic solvent (DES) is a binary or ternary liquid eutectic mixture formed by combining Hydrogen Bond Donors (HBD) such as carboxylic acid and alcohol and Hydrogen Bond Acceptors (HBA) such as quaternary ammonium salt according to a certain molar ratio. The DES has the characteristics of low vapor pressure, high thermal stability, structural adjustability, high solubility to a plurality of solutes and the like, and has the advantages of low price, biodegradability, good compatibility, low toxicity and the like. These features and advantages make DES a green solvent of great interest. Notably, water can form hydrogen bonds with the DES component, and increasing the water content can reduce DES viscosity, so that DES can better penetrate into plant tissues, enhance DES surface tension, and increase extraction efficiency. In addition, it has been found that DES can be used as an adjuvant drug to improve the stability, oral bioavailability and skin permeability of some traditional Chinese medicines, thereby enhancing the therapeutic effect of these drugs.

Based on the above, an anti-acne raw material based on four traditional Chinese medicine components of radix scutellariae, cortex phellodendri, coptis chinensis and rheum officinale is developed, and the extraction mode is optimized. Solves the problems of the prior art of the traditional extraction and is a problem to be solved by researchers in the field.

Disclosure of Invention

Aiming at the problems, the invention provides a DES four-yellow extract and a preparation method thereof, which are characterized in that a ternary DES system formed by specific amino acid, alcohol/carboxylic acid and water is used for extracting baical skullcap root, amur corktree bark, golden thread and rhubarb and optimizing the formula ratio of the ternary DES system. The extraction rate of the effective components of the four traditional Chinese medicines is high under the selected DES system. Compared with water extraction and alcohol extraction, the method has the advantage of natural non-toxicity. In addition, the DES tetrayellow extract also has good antioxidant activity, solubility and anti-acne bacillus capability.

In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:

In one aspect, the invention provides a method for preparing a DES tetrayellow extract, comprising the steps of:

S1, preparing a eutectic solvent, namely taking amino acid as HBA, taking alcohol or carboxylic acid as HBD, heating and stirring to obtain DES;

s2, preparing a DES aqueous solution, namely mixing the DES prepared in the step S1 with water to obtain the DES aqueous solution;

S3, extracting, namely taking phellodendron, coptis chinensis, rheum officinale and scutellaria baicalensis, crushing, sieving, adding the DES aqueous solution prepared in the step S2, carrying out ultrasonic treatment for 30-60min at 55-65 ℃, cooling to room temperature, centrifuging, taking supernatant and drying to obtain the DES four-yellow extract.

Preferably, in S1, the amino acid is selected from at least one of lysine and arginine, and further preferably, in S1, the amino acid is selected from lysine or arginine.

Preferably, in S1, the alcohol is selected from at least one of xylitol and 1, 3-propanediol, and further preferably, in S1, the alcohol is selected from at least one of xylitol and 1, 3-propanediol.

Preferably, in S2, the water is selected from at least one of ultrapure water, deionized water and distilled water, and further preferably, in S2, the water is selected from ultrapure water.

Preferably, in S1, the molar ratio of the HBA to the HBD is 1:2-4. Further preferably, in S1, the molar ratio of the HBA to the HBD is 1:2 or 1:4.

Preferably, in S2, the mass of the water is 35% -65% of the total mass of the DES aqueous solution. Further preferably, in S2, the mass of the water is 40% -60% of the total mass of the DES aqueous solution. More preferably, in S2, the mass of water is 40% -50% of the total mass of DES aqueous solution.

Preferably, in S1, the heating and stirring is stirring at 65-75 ℃.

Preferably, in S2, the mixing is stirring at 55-65 ℃.

Preferably, in S3, the DES aqueous solution is transferred after being cooled to room temperature and stored in a dark place.

Preferably, in S3, the mesh number of the screen is 60-80 mesh, and further preferably, in S3, the mesh number of the screen is 70 mesh.

Preferably, in S3, the mass ratio of the phellodendron, the coptis chinensis, the rheum officinale and the scutellaria baicalensis is 1-2:1-2:1-2, further preferably, in S3, the mass ratio of the phellodendron, the coptis chinensis, the rheum officinale and the scutellaria baicalensis is 1:1:1-2, and further preferably, in S3, the mass ratio of the phellodendron, the coptis chinensis, the rheum officinale and the scutellaria baicalensis is 1:1:1:2.

Preferably, in S3, the solid-to-liquid ratio of the phellodendron, coptis, rheum officinale and scutellaria baicalensis to the DES aqueous solution is 1:20-40, and further preferably, in S3, the solid-to-liquid ratio of the phellodendron, coptis, rheum officinale and scutellaria baicalensis to the DES aqueous solution is 1:20 or 1:40.

Preferably, in S3, the temperature of the ultrasonic wave is 55-60 ℃ and the time is 30-40min.

Preferably, in S3, the speed of centrifugation is 5500-6500r/min for 15-25min, and further preferably, in S3, the speed of centrifugation is 5500-6000r/min for 15-20min.

Preferably, in the step S3, the specific parameters of the drying are that the temperature is 55-65 ℃, the vacuum degree is minus 0.3 to minus 1MPa, and the time is 10-14 hours. Further preferably, in S3, the specific parameters of the drying are that the temperature is 55-60 ℃, the vacuum degree is minus 0.5 to minus 1MPa, and the time is 10-12 hours.

The invention provides the DES tetrayellow extract prepared by the preparation method.

Compared with the prior art, the invention has the following beneficial effects:

The invention is a novel solvent method based on green and harmless extraction of radix scutellariae, cortex phellodendri, rheum officinale and coptis chinensis, and the obtained DES four-yellow extract has stable components, good solubility and certain acne bacillus resistance. The invention can solve the problems of low bioavailability, use of harmful organic solvents and the like faced by the traditional Chinese medicine, so that the invention has important significance in the field of Chinese medicine extraction.

Drawings

FIG. 1 is a graph showing the comparison of the extraction rates of Scutellariae radix in examples 1-4.

FIG. 2 is a graph showing the comparison of the extraction rates of phellodendron bark in examples 5-8.

FIG. 3 is a graph showing the comparison of extraction rates of Coptidis rhizoma in examples 9-12.

FIG. 4 is a graph showing comparison of extraction rates of rheum officinale of examples 13-16.

Fig. 5 is a DES extract graph of example 2, example 5, example 11 and example 16, wherein:

(a) Transparent homogeneous DES aqueous solutions for example 2, example 5, example 11 and example 16;

(b) DES extracts of examples 2, 5, 11 and 16;

(c) DES extracts of examples 2, 5, 11 and 16.

Fig. 6 is a DES extract temperature-viscosity diagram of examples 2,5, 11 and 16.

FIG. 7 is a graph showing the comparison of antioxidant effect of DES tetrayellow extracts prepared in examples 17-20.

FIG. 8 is a dissolution chart of DES tetrayellow extracts prepared in example 20 and comparative examples 1-2.

Detailed Description

In order to make the technical means, the creation features, the achievement of the purpose and the effect of the present invention easy to understand, the present invention will be further elucidated with reference to the specific embodiments, but the following embodiments are only preferred embodiments of the present invention, not all of them. Based on the examples in the embodiments, those skilled in the art can obtain other examples without making any inventive effort, which fall within the scope of the invention. It is to be noted that the raw materials used in the present invention are all common commercial products, and the sources thereof are not particularly limited. Technical and scientific terms used in the examples have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

The reagents, methods and equipment adopted by the invention are conventional reagents, methods and equipment in the technical field.

Example 1:

DES baical skullcap root extract (lysine: citric acid molar ratio is 1:2, water content is 50%)

Weighing 0.1000g of lysine and 0.2628g of citric acid in a 10mL centrifuge tube, heating and stirring at 70 ℃ in a water bath at 700-900rpm until clear and stable DES is presented. 0.3744g of ultra pure water was weighed into the centrifuge tube and stirred in a 60℃water bath at 700-900rpm until a clear and homogeneous DES aqueous solution was formed. 0.0370g of baical skullcap root powder is weighed in DES aqueous solution (solid-liquid ratio 1:20) and uniformly shaken. Ultrasonic extraction is carried out for 40min at 60 ℃. And (3) cooling the DES solution to room temperature, centrifuging at 6000r/min for 20min, and taking the supernatant to obtain the corresponding baical skullcap root DES extract. Removing water in vacuum for 12 hr to obtain DES Scutellariae radix extract.

Example 2:

DES baical skullcap root extract (lysine: citric acid molar ratio is 1:4, water content is 50%)

Lysine 0.1008g and citric acid 0.5251g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.6272g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0625g of baical skullcap root powder is weighed into the DES aqueous solution (solid-liquid ratio 1:20) and uniformly shaken. The subsequent procedure is as in example 1. Namely the DES baical skullcap root extract.

Example 3:

DES baical skullcap root extract (lysine: citric acid molar ratio is 1:4, water content is 60%)

Weighing 0.1003g of lysine and 0.5255g of citric acid in a 10mL centrifuge tube, heating and stirring in a 70 ℃ water bath at 700-900rpm until clear and stable DES is presented. 0.9386g of ultrapure water was weighed into the centrifuge tube, stirred in a 60 ℃ water bath at 700-900rpm until a transparent uniform DES aqueous solution was formed, and then 0.0784g of baical skullcap root powder was weighed into the DES aqueous solution (solid-liquid ratio 1:20) and shaken uniformly. The subsequent procedure is as in example 1. Namely the DES baical skullcap root extract.

Example 4:

Weighing 0.1002g of lysine and 0.5257g of citric acid in a 10mL centrifuge tube, heating and stirring in a 70 ℃ water bath at 700-900rpm until clear and stable DES is presented. 0.6252g of ultrapure water was weighed into the centrifuge tube, stirred in a 60 ℃ water bath at 700-900rpm until a transparent uniform DES aqueous solution was formed, and then 0.0311g of baikal skullcap root powder was weighed into the DES aqueous solution (solid-liquid ratio 1:40) and uniformly shaken. The subsequent procedure is as in example 1. Namely the DES baical skullcap root extract.

Example 5:

DES cortex Phellodendri extract (lysine: xylitol molar ratio of 1:2, water content of 50%)

Weighing 0.1005g of lysine and 0.2081g of xylitol in a 10mL centrifuge tube, heating and stirring at 70 ℃ in a water bath at 700-900rpm until clear and stable DES is presented. 0.3483g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, 0.0328g of phellodendron amurense powder is weighed into the DES aqueous solution (solid-liquid ratio is 1:20), and the phellodendron amurense powder is uniformly shaken. The subsequent procedure is as in example 1. The DES cortex phellodendri extract is obtained.

Example 6:

DES cortex Phellodendri extract (lysine: xylitol molar ratio of 1:4, water content of 50%)

Weighing 0.1005g of lysine and 0.4164g of xylitol in a 10mL centrifuge tube, heating and stirring at 70 ℃ in a water bath at 700-900rpm until clear and stable DES is presented. 0.5175g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0515g of phellodendron amurense powder is weighed into the DES aqueous solution (solid-liquid ratio is 1:20) and uniformly shaken. The subsequent procedure is as in example 1. The DES cortex phellodendri extract is obtained.

Example 7:

DES cortex Phellodendri extract (lysine: xylitol molar ratio is 1:2, water content is 60%)

Weighing 0.1001g of lysine and 0.2082g of xylitol in a 10mL centrifuge tube, heating and stirring in a 70 ℃ water bath at 700-900rpm until clear and stable DES is presented. 0.4620g of ultrapure water was weighed into the centrifuge tube, stirred in a 60 ℃ water bath at 700-900rpm until a transparent uniform DES aqueous solution was formed, and then 0.0384g of phellodendron amurense powder was weighed into the DES aqueous solution (solid-liquid ratio 1:20) and uniformly shaken. The subsequent procedure is as in example 1. The DES cortex phellodendri extract is obtained.

Example 8:

Weighing 0.1002g of lysine and 0.2083g of xylitol in a 10mL centrifuge tube, heating and stirring in a 70 ℃ water bath at 700-900rpm until clear and stable DES is presented. 0.3107g of ultrapure water is weighed into the centrifuge tube, stirred in a 60 ℃ water bath at 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0154g of phellodendron amurense powder is weighed into the DES aqueous solution (solid-liquid ratio 1:40) and uniformly shaken. The subsequent procedure is as in example 1. The DES cortex phellodendri extract is obtained.

Example 9:

DES Coptidis rhizoma extract (arginine: malic acid molar ratio is 1:2, water content is 50%)

Arginine 0.1006g and malic acid 0.1540g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.2973g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0280g of coptis powder is weighed into the DES aqueous solution (solid-liquid ratio is 1:20) and uniformly shaken. The subsequent procedure is as in example 1. The DES coptis extract is obtained.

Example 10:

DES Coptidis rhizoma extract (arginine: malic acid molar ratio is 1:4, water content is 50%)

Arginine 0.1004g and malic acid 0.3076g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.4308g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, 0.0413g of coptis powder is weighed into the DES aqueous solution (solid-to-liquid ratio is 1:20), and the mixture is uniformly shaken. The subsequent procedure is as in example 1. The DES coptis extract is obtained.

Example 11:

DES Coptidis rhizoma extract (arginine: malic acid molar ratio is 1:4, water content is 40%)

Arginine 0.1002g and malic acid 0.3078g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.6115g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, 0.0509g of coptis powder is weighed into the DES aqueous solution, (solid-to-liquid ratio is 1:20), and the mixture is uniformly shaken. The subsequent procedure is as in example 1. The DES coptis extract is obtained.

Example 12:

Arginine 0.1006g and malic acid 0.3077g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.4096g of ultrapure water is weighed into the centrifuge tube, stirred in a 60 ℃ water bath at 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0204g of coptis powder is weighed into the DES aqueous solution (solid-liquid ratio 1:40) and uniformly shaken. The subsequent procedure is as in example 1. The DES coptis extract is obtained.

Example 13:

DES radix et rhizoma Rhei extract (arginine: 1, 3-propylene glycol molar ratio of 1:2, water content of 50%)

Arginine 0.1006g, 1, 3-propanediol 0.0889g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.1922g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0191g of rheum officinale powder is weighed into the DES aqueous solution (solid-liquid ratio 1:20) and uniformly shaken. The subsequent procedure is as in example 1. Namely the DES rheum officinale extract.

Example 14:

DES radix et rhizoma Rhei extract (arginine: 1, 3-propylene glycol molar ratio of 1:4, water content of 50%)

Arginine 0.1001g, 1, 3-propanediol 0.1797g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.2786g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0280g of rheum officinale powder is weighed into the DES aqueous solution (solid-liquid ratio is 1:20) and uniformly shaken. The subsequent procedure is as in example 1. Namely the DES rheum officinale extract.

Example 15:

DES radix et rhizoma Rhei extract (arginine: 1, 3-propylene glycol molar ratio of 1:2, water content 60%)

Arginine 0.1003g, 1, 3-propanediol 0.0881g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.2817g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0234g of rheum officinale powder is weighed into the DES aqueous solution (solid-liquid ratio is 1:20) and uniformly shaken. The subsequent procedure is as in example 1. Namely the DES rheum officinale extract.

Example 16:

Arginine 0.1006g, 1, 3-propanediol 0.0874g were weighed into a 10mL centrifuge tube and heated in a 70 ℃ water bath with stirring at 700-900rpm until clear and stable DES was exhibited. 0.1922g of ultrapure water is weighed into the centrifuge tube, stirred in a water bath at 60 ℃ for 700-900rpm until a transparent uniform DES aqueous solution is formed, and then 0.0191g of rheum officinale powder is weighed into the DES aqueous solution (solid-liquid ratio 1:40) and uniformly shaken. The subsequent procedure is as in example 1. Namely the DES rheum officinale extract.

Example 17:

DES four-yellow extract compounded by cortex phellodendri, coptis chinensis, rheum officinale and radix scutellariae

Weighing 1g of DES cortex Phellodendri extract, 0.5g of DES Coptidis rhizoma extract, 0.5g of DES radix et rhizoma Rhei extract and 0.5g of DES Scutellariae radix extract respectively, and placing into a beaker. 60% ethanol/water was dissolved and transferred to a 25mL volumetric flask to give 100mg/mL (compound weight ratio 2:1:1:1) of DES tetrayellow extract.

Example 18:

The other operations were the same as in example 17 except that extracts of DES cortex Phellodendri, coptidis rhizoma, radix et rhizoma Rhei and Scutellariae radix were weighed respectively (compound weight ratio 1:2:1:1).

Example 19:

The DES cortex Phellodendri, coptidis rhizoma, radix et rhizoma Rhei and Scutellariae radix extracts (compound weight ratio 1:1:2:1) were weighed respectively, and the rest was the same as in example 17.

Example 20:

The procedure of example 17 was repeated except that extracts of DES cortex Phellodendri, coptidis rhizoma, radix et rhizoma Rhei and Scutellariae radix (compound weight ratio 1:1:1:2) were weighed separately.

Comparative example 1:

a tetrandra and radix Et rhizoma Rhei composition, and water extract

Weighing 10g of coarse powder of cortex phellodendri, rhizoma coptidis, rheum officinale and radix scutellariae in a certain proportion (weight ratio of 1:1:1:2) into a 250mL three-neck flask, adding 100mL distilled water (liquid-material ratio is 10:1 mg/mL), reflux-extracting for 1 hour, extracting for 2 times, filtering with gauze, and filtering the filtrate for later use. The 2 filtrates were combined, centrifuged at 1000rpm for 10min and filtered. Pouring the filtered sample into a culture dish, pre-cooling in a refrigerator at-24 ℃, vacuum freeze-drying, and weighing after drying.

Comparative example 2:

a tetrayellow composition is prepared by extracting with 60wt% ethanol

Weighing 10g of four-yellow coarse powder with a certain proportion (phellodendron, coptis chinensis, rheum officinale and scutellaria baicalensis according to the weight ratio of 1:1:1:2), adding 100mL of 60% ethanol (the liquid-material ratio is 10:1 mg/mL) into a three-neck flask with 250mL, carrying out reflux extraction for 1 hour, carrying out total extraction for 2 times, filtering with gauze, and filtering the filtrate for later use. The 2 filtrates were combined, centrifuged at 1000rpm for 10min and filtered. Pouring the filtered sample into a culture dish, pre-cooling in a refrigerator at-24 ℃, vacuum freeze-drying, and weighing after drying.

Test example 1:

1. examples 1-4 extraction ratio of scutellaria baicalensis extract of DES:

Baicalin is used as analysis control. 5mg of baicalin analysis reference substance is weighed, the concentration of 0.0025mg/mL-0.015mg/mL is prepared, and the absorbance is measured at 278 nm. The linear equation was found to be y=50.35 x-0.00905 (y: absorbance; x: concentration mg/mL) from the linear fit, and R ² =0.9990.

The supernatants of example 1, example 2, example 3 and example 4 were weighed, the corresponding absorbance was measured at 278nm, and the corresponding extraction rate was calculated by substituting into a linear equation. As shown in FIG. 1, the extraction rate of the radix scutellariae in example 2 is highest under the conditions of a molar ratio of 1:4, a water content of 50% and a solid-to-liquid ratio of 1:20 based on a lysine and citric acid DES system.

2. Examples 5-8 phellodendron bark extraction yield of phellodendron bark extract:

The analysis control of cortex Phellodendri is berberine hydrochloride. Weighing 2.2mg berberine hydrochloride analysis control substance, and preparing into 0.0022mg/mL-0.0132mg/mL concentration. Absorbance was measured at 350 nm. The linear equation was found from the linear fit to be y=78.14x-0.01754 (y: absorbance; x: concentration mg/mL), and R ² =0.9997.

The supernatants of examples 5,6,7 and 8 were weighed, the corresponding absorbance was measured at 350nm, and the linear equation was substituted to calculate the corresponding extraction rate. As shown in FIG. 2, the extraction rate of cortex Phellodendri of example 5 is highest based on lysine and xylitol DES system at a molar ratio of 1:2, water content of 50% and solid-liquid ratio of 1:20.

3. Examples 9-12 coptis extraction yield of DES coptis extract:

The analysis and control of Coptidis rhizoma is berberine hydrochloride. Weighing 2.2mg berberine hydrochloride analysis control substance, and preparing into 0.0022mg/mL-0.0132mg/mL concentration. Absorbance was measured at 350 nm. The linear equation was found from the linear fit to be y=78.14x-0.01754 (y: absorbance; x: concentration mg/mL), and R ² =0.9997.

The supernatants of examples 9, 10, 11 and 12 were weighed, absorbance was measured under a 350nm ultraviolet spectrometer, and the corresponding extraction ratios were calculated by substituting into a linear equation. As shown in FIG. 3, the extraction rate of rhizoma Coptidis of example 11 was highest based on arginine and malic acid DES system at a molar ratio of 1:4, a water content of 40% and a solid-liquid ratio of 1:20.

4. Examples 13-16 extraction yield of rheum officinale of DES rheum officinale extract:

Rhubarb uses 1, 8-dihydroxyanthraquinone as analysis control. 5mg of 1, 8-dihydroxyanthraquinone analysis reference substance is weighed and the concentration of 0.0025mg/mL-0.0125mg/mL is prepared. Absorbance was measured at 252 nm. The linear equation was found to be y=83.8x-0.0009 (y: absorbance; x: concentration mg/mL) and R ² =0.9990 according to the linear fit.

The supernatants of examples 13, 14, 15 and 16 were weighed, absorbance was measured under a 252nm ultraviolet spectrometer, and the corresponding extraction ratios were calculated by substituting the absorbance into a linear equation. As shown in FIG. 4, the extraction rate of the rheum officinale of example 16 is highest under the conditions that the mol ratio is 1:4, the water content is 50% and the solid-liquid ratio is 1:40 based on the arginine and the 1, 3-propanediol DES system.

Thus, the preferred embodiments are example 2, example 5, example 11 and example 16, and the subsequent test is represented by the four preferred embodiments described above.

(b) DES extracts of examples 2, 5, 11 and 16;

(c) DES extracts of examples 2, 5, 11 and 16.

Test example 2:

the viscosity of the DES scutellaria baicalensis, phellodendron amurense, coptis chinensis and rheum officinale extracts prepared in example 2, example 5, example 11 and example 16 was measured using a rheometer, and the temperature was measured by heating to 60 ℃ at 2 ℃ per 30s in a temperature programmed manner.

As a result, as shown in fig. 6, the viscosity of DES extracts prepared in each example decreased with increasing temperature, and the rate of decrease in the low temperature region was faster and gradually slowed down with increasing temperature. This is because stable hydrogen bonding and van der waals forces are formed between DES systems and DES extracts prepared in the examples form stable liquid DES forms.

Test example 3:

Antioxidant activity as measured by 1, 1-diphenyl-2-picrylhydrazine (1, 1-Diphenyl-2-picryl-hydrazyl, DPPH) free radical scavenging method. The DPPH is weighed to prepare a DPPH solution with the concentration of 0.1mg/mL, and the solution is stored in a dark place and is prepared for use. DES four yellow extract was prepared according to examples 17-20 and dissolved in 60% ethanol to prepare a stock solution of 0.2mg/mL-0.7 mg/mL. 2mL of the stock solution was placed in a 10mL centrifuge tube, 2mL of DPPH solution was added thereto, and after shaking sufficiently, the reaction was carried out at room temperature for 30 minutes in the absence of light. Absorbance a _i was measured at a wavelength of 517nm, wherein the absorbance measured after 2mL of 60% absolute ethanol solution and 2mL of DPPH solution were mixed was a ₀. The clearance of DPPH was calculated by the following formula.

Wherein A ₀ is absorbance of a blank control group, and A _i is absorbance of samples with different concentrations after DPPH reaction.

Specific data are shown in Table 1 and in FIG. 7, sample data are fitted linearly, trend lines are made and linear regression equations are obtained, IC ₅₀ for each sample group is calculated according to the equation to be 0.52mg/mL for IC ₅₀ for example 17 (weight ratio of four colors of 2:1:1), 0.48mg/mL for IC ₅₀ for example 18 (weight ratio of four colors of 1:2:1), 0.50mg/mL for IC ₅₀ for example 19 (weight ratio of four colors of 1:1:2:1), and 0.50mg/mL for IC ₅₀ for example 20 (weight ratio of four colors of 1:1:1:2). According to the results, examples 17-20 all had good oxidation resistance, with example 18 being more resistant to oxidation.

TABLE 1 clearance of examples 17-20

Test example 4:

The DES tetrayellow extracts and tetrayellow extracts prepared in examples 17 to 20 and comparative examples 1 to 2 were prepared as 200mg/mL mother liquor, respectively. 200. Mu.L of the mother solution was added to column 1 of the 96-well plate, and the working concentrations of 100, 50, 25, 12.5, 6.25, 3.125, 1.5625 and 0.78125mg/mL were prepared by a medium half-dilution method, whereby the concentration of the acne bacteria was 1.5X10. 10 ⁵. Wherein, 100. Mu.L of culture medium is added to the control group, 100. Mu.L of bacterial liquid is added to the blank group, and 200. Mu.L of culture medium is added to the blank group. After the gradient concentration was prepared, the 96-well plate was placed in a 37℃incubator for 12 hours, and the bacterial growth was observed. The results are shown in tables 2-3, example 20 being the best ratio of acnes with a minimum MIC of 3.125mg/mL. In combination with the antioxidant results, example 20 was excellent in antioxidant effect and anti-acne bacillus ability. Therefore, the ratio of DES four yellow extracts to be compounded for the following experiments is 1:1:1:2.

TABLE 2 MIC comparative tables of examples 17-20

(Note: "-" indicates sterile formation at this concentration, successful inhibition of bacterial growth; "+": indicates bacterial growth at this concentration.)

TABLE 3 MIC comparison Table for Water extraction, DES extraction, 60% absolute ethanol extraction

As shown in Table 3, the anti-acne bacillus MIC of the DES tetrayellow extract prepared in example 20, the aqueous extract of comparative example 1 and the 260% alcoholic extract of comparative example were 3.125mg/mL, 25mg/mL and 25mg/mL, respectively, and thus, the antibacterial ability of the DES tetrayellow extract was significantly higher than that of the aqueous extract and 60% alcoholic extract.

Test example 5:

the DES tetrayellow extract and tetrayellow extract prepared in example 20 and comparative examples 1-2 were diluted to the same concentration of 2mg/mL, and then dissolved in water and 60wt% ethanol solution, respectively, and the dissolution was observed.

As a result (see FIG. 8), the aqueous extraction or the alcoholic extraction was poor in solubility and high in impurity content, whereas the DES extraction was good in dissolution capacity and free from precipitation and impurities.

The DES extract formed by respectively extracting phellodendron, coptis chinensis, rheum officinale and scutellaria baicalensis by using specific amino acid, carboxylic acid/alcohol substances and water has the advantages of greenness and no toxicity compared with water extraction and alcohol extraction, and simultaneously has good antioxidant activity, solubility and acne bacillus resistance.

Finally, it should be noted that the above description is only for illustrating the technical solution of the present invention, and not for limiting the scope of the present invention, and that the simple modification and equivalent substitution of the technical solution of the present invention can be made by those skilled in the art without departing from the spirit and scope of the technical solution of the present invention.

Claims

1. A method for preparing DES tetrayellow extract, comprising the steps of:

when the DES baical skullcap root extract is prepared, lysine is used as HBA, citric acid is used as HBD, and the molar ratio of lysine to citric acid is 1:4;

When the DES cortex phellodendri extract is prepared, lysine is used as HBA, xylitol is used as HBD, and the molar ratio of lysine to xylitol is 1:2;

when the DES coptis extract is prepared, arginine is used as HBA, malic acid is used as HBD, and the molar ratio of arginine to malic acid is 1:4;

When the DES rheum officinale extract is prepared, arginine is used as HBA, 1, 3-propylene glycol is used as HBD, and the mol ratio of arginine to 1, 3-propylene glycol is 1:2;

when the DES baical skullcap root extract is prepared, the mass of the water is 50% of the total mass of the DES aqueous solution, and the solid-to-liquid ratio of the baical skullcap root to the DES aqueous solution is 1:20;

when the DES cortex phellodendri extract is prepared, the mass of water is 50% of the total mass of the DES aqueous solution, and the solid-to-liquid ratio of the cortex phellodendri to the DES aqueous solution is 1:20;

When the DES coptis chinensis extract is prepared, the mass of the water is 40% of the total mass of the DES aqueous solution, and the solid-to-liquid ratio of the coptis chinensis to the DES aqueous solution is 1:20;

When the DES rheum officinale extract is prepared, the mass of the water is 50% of the total mass of the DES aqueous solution, and the solid-to-liquid ratio of rheum officinale to the DES aqueous solution is 1:40;

S3, extracting, namely taking phellodendron, coptis chinensis, rheum officinale and scutellaria baicalensis, crushing, sieving, respectively adding the DES aqueous solution prepared in the step S2, carrying out ultrasonic treatment for 30-60min at 55-65 ℃, cooling to room temperature, centrifuging, taking supernatant, and drying to obtain a DES four-yellow extract, wherein the weight ratio of the DES four-yellow extract to the DES coptis chinensis extract to the DES rheum officinale extract to the DES scutellaria baicalensis extract is 1:1:1:2.

2. The process according to claim 1, wherein in S3,

The temperature of the ultrasonic wave is 55-60 ℃ and the time is 30-40min;

the centrifugal speed is 5500-6500r/min, and the time is 15-25min;

The specific parameters of the drying are that the temperature is 55-65 ℃, the vacuum degree is minus 0.3 to minus 1MPa, and the time is 10-14h.

3. The DES tetrayellow extract prepared by the method of any one of claims 1-2.

4. Use of DES tetrayellow extract prepared by the preparation method of any one of claims 1-2 in preparing an anti-acne product.