CN115400056A

CN115400056A - Method for improving efficacy of plant extract by using electrolysis method

Info

Publication number: CN115400056A
Application number: CN202211087898.1A
Authority: CN
Inventors: 王刚
Original assignee: Shanghai Ruidian Biotechnology Co ltd
Current assignee: Shanghai Ruidian Biotechnology Co ltd
Priority date: 2022-09-07
Filing date: 2022-09-07
Publication date: 2022-11-29

Abstract

The invention relates to the technical field of cosmetics, in particular to the field of IPC A61K8, and more particularly relates to a method for improving the efficacy of a plant extract by using an electrolytic method. The extraction method comprises the following steps: s1, drying and crushing plants, and extracting with an ethanol solution to obtain a pretreatment solution; s2, carrying out negative pressure rotary evaporation on the pretreatment solution, and filtering to obtain a primary concentrated solution; s3, adding alkali to enable the primary concentrated solution to be alkalescent, and obtaining a secondary concentrated solution; s4, adding the secondary concentrated solution into an electrolytic cell, inserting an ion exchange membrane into the electrolytic cell, and electrolyzing; and S5, enriching at the anode after electrolysis to obtain the plant extract. The amino acid is enriched by using an electrolysis method, so that the efficacy of the plant extract can be improved, and the active substances can be efficiently enriched by using the electrolysis method while the active substances are not damaged.

Description

Method for improving efficacy of plant extract by using electrolysis method

Technical Field

The invention relates to the technical field of cosmetics, in particular to the field of IPC A61K8, and more particularly relates to a method for improving the efficacy of a plant extract by using an electrolytic method.

Background

With the improvement of living standard, people increasingly have increased consciousness of pursuing nature, pursuing green, pursuing health and safety, cosmetics become indispensable components in the life of people, and with the wider application of plant extracts in cosmetics and more applied varieties, since the execution of the evaluation specification of cosmetic efficacy declaration in 2021, the plant extracts are used as raw materials of cosmetics, and the efficacy and the extraction process of the plant extracts become one of the most important items of various cosmetic companies. How to improve the efficacy of plant extracts has become a topic of well-known food in the current cosmetic industry, and the current methods for extracting plant extracts are generally divided into several categories, namely: ultrasonic extraction, enzyme extraction, microwave-assisted extraction, solvent extraction, and supercritical fluid extraction.

CN201611210177 provides a cosmetic with plant extract, which is prepared by mixing plant components according to weight percentage, drying, pulverizing, performing carbon dioxide supercritical extraction on the medicinal powder, extracting with anhydrous ethanol, concentrating under reduced pressure, and recovering ethanol to obtain ethanol extractive concentrate; the method comprises the steps of taking ethanol solution with the concentration of 10% -70% as an extraction solvent for dregs after carbon dioxide supercritical extraction, heating, extracting and concentrating, enriching a concentrated solution through a macroporous resin column, eluting the concentrated solution for multiple times by using 30% -90% ethanol, dissolving an eluent by using 1.3 butanediol after concentrating the eluent under reduced pressure, filtering, and finally mixing the eluent with the solution obtained by carbon dioxide supercritical extraction to obtain the plant extracting solution.

CN200410072587.3 discloses a prinsepia utilis royle cosmetic. The weight percentage of the mixed liquid of the natural plant extracts is 1.0 to 3.0 percent. The Prinsepia utilis royle cosmetic provided by the invention is characterized in that Prinsepia utilis royle extract prepared by low-temperature extraction at 35-40 ℃ is added into conventional cosmetic components together with mixed liquid of various natural plant extracts such as tea, lucid ganoderma, ginkgo and the like, wherein pure natural bioactive components are reserved by the Prinsepia utilis royle extract extracted at 35-40 ℃, and the fatty acid structure contained in the Prinsepia utilis royle extract is very close to human body lipid, so that the skin permeability of the skin is particularly good, and the skin of a human body is very easy to absorb.

Disclosure of Invention

On the basis of the invention, the extracted plant extract has low solid content or impurities cannot be effectively removed, and the invention adopts an electrolytic method to improve the concentrations of amino acid, enzyme and the like in the unit volume of the plant extract, thereby improving the efficacy of the plant extract. In order to achieve the above objects, the present invention provides a method for improving the efficacy of a plant extract by using an electrolysis method, the extraction method comprising the steps of:

s1, crushing plants, and extracting with an ethanol solution to obtain a pretreatment solution;

s2, carrying out negative pressure rotary evaporation on the pretreatment liquid, and filtering to obtain a primary concentrated solution;

s3, adding alkali to enable the primary concentrated solution to be alkalescent, and obtaining a secondary concentrated solution;

s4, adding the secondary concentrated solution into an electrolytic cell, inserting an ion exchange membrane into the electrolytic cell, and electrolyzing;

and S5, enriching at the anode after electrolysis to obtain the plant extract.

The plant is at least one of radix et caulis Opuntiae Dillenii (Opuntiastascta), herba Sophorae Alopecuroidis (Sophora Alopecuroides L), herba Patriniae (Patrinia Bisaosa), tea Tree (Camellia sinensis), aloe (Aloe vera), chinese juniper (Juniperus chinensis L), and folium Artemisiae Argyi (Artemisia argyi Levl. Et Van). One or more of chamomile (matricaria L), purslane (Portulaca oleracea L), olive (Oleaeuropaea L.), centella asiatica (centellaasia L Urban), white willow bark (Salix alba L), horse chestnut (Aesculus chinensis Bunge), and witch hazel (Hamamelismollis Oliver).

Preferably, the plant is one or more of patrinia (patrinia bifosa folia), tea tree (Camellia sinensis), aloe (Aloe vera), centella asiatica (centella asiatica L Urban), horse chestnut (Aesculuschinensis Bunge), witch hazel (Hamamelismollis Oliver).

Further preferably, the plant is tea tree (Camellia sinensis).

The drying in the step S1 comprises one or more of hot air drying, vacuum drying, spray drying and freeze drying.

Preferably, the drying in step S1 is freeze drying at-20-0 ℃ for 6-12h.

Further preferably, the drying in step S1 is freeze-drying at-20 ℃ for 12h.

In the step S1, a wall breaking machine or a pulverizer is used in the pulverizing process, and the rotating speed is 800-10000r/min.

Preferably, the crushing process in step S1 uses a crusher, and the model is: IKA 10, the rotating speed is set to be 5000r/min.

Preferably, the diameter of the filtration mesh in step S2 is 0.22 to 0.45. Mu.m.

More preferably, the filter mesh diameter in the step S2 is 0.35. Mu.m.

Preferably, the solid content of the secondary concentrated solution in the step S3 is 10-15%.

Further preferably, the solid content of the secondary concentrated solution in the step S3 is 12%.

Preferably, the alkali in step S3 is one or more of sodium hydroxide, potassium hydroxide and sodium bicarbonate.

Further preferably, the base in step S3 is sodium hydroxide (CAS: 1310-73-2) at a concentration of 1mol/L.

Preferably, the amount of the secondary concentrated solution added in step S4 is 300-800g.

More preferably, the amount of the second concentrated solution added in step S4 is 500g.

When the mass fraction of the ethanol solution is 65-72%, not only the extraction efficiency of amino acid can be improved, but also the dissolution of water-soluble substances such as saccharides and the like can be reduced, and the purity of the plant extract can be improved. When the concentration of the ethanol is low, a large amount of water-soluble substances such as saccharides and proteins are dissolved out, and meanwhile, the amino acid is easy to generate hydrolysis reaction due to excessive water content, and although the amino acid is not changed essentially, the amino acid is changed into an ionic state to a certain extent, so that the amino acid and the foreign protein in the plant are easy to react. When the ethanol concentration is too high, a large amount of fat-soluble substances are dissolved out, the components compete with amino acid substances and are combined with an ethanol-water molecular system to influence the dissolution of amino acid, and further research shows that when an ethanol solution with the mass fraction of 70% is selected, the extraction efficiency of alpha-amino acid in plants is further improved, and the polarity of the ethanol solution with the concentration is presumed to be similar to that of the alpha-amino acid in plants, so that the alpha-amino acid can be fully dissolved in a solvent and extracted.

Preferably, the mass fraction of the ethanol solution is 65-72%.

More preferably, the ethanol solution is 70% by mass.

The weight ratio (liquid-material ratio) of the ethanol solution to the plants is 15-25:1, the extraction efficiency of amino acids in plants is optimal, in the mass transfer process of plant extraction, the internal diffusion of solutes has a large influence on extraction, the solutes can diffuse into plant tissues through cell walls gradually under the action of concentration gradients inside and outside plant cells, when the liquid-material ratio is too low, the concentration gradients inside and outside the plant cells are small, the extraction amount and efficiency are too slow, the solubility of active substances such as amino acids in the plant cells is improved along with the increase of the liquid-material ratio, most of the amino acids are dissolved in ethanol, the effect of continuously increasing the amount of an extraction solvent on the increase of the extraction rate is not large, and the production period and the production cost in practical production application are increased.

Preferably, the weight ratio of the ethanol solution to the plants is (15-25): 1

Further preferably, the weight ratio of the ethanol solution to the plants is 20:1

Further research shows that when the extraction time is controlled to be 2.5-3.5h, the extraction efficiency can be further improved, probably because the osmotic pressure generated by the concentration gradient inside and outside the plant cell can be maintained at a higher level.

Preferably, the extraction time is 2.5-3.5h.

More preferably, the extraction time is 3h.

When the rotary evaporation temperature is 45-55 ℃, the stability of amino acid in the extract can be improved to a greater extent, the concentration efficiency is continuously improved along with the increase of the temperature within a certain range, when the temperature is higher than 55 ℃, the high temperature can influence the hydrogen bond of carboxyl in the amino acid, irreversible damage such as hydrogen bond breakage and the like can be caused, but when the rotary evaporation temperature is lower, the boiling point is far lower than 78 ℃ of ethanol, and the improvement of the extraction efficiency of the amino acid is limited.

Preferably, the rotary evaporation temperature is 45-55 ℃.

More preferably, the rotary evaporation temperature is 50 ℃.

Further research shows that the ethanol can reach the saturated vapor pressure at 45-55 deg.c when the pressure is 1.5-2.5KPa. Further improving the efficiency of concentration.

Preferably, the negative pressure is 1.5-2.5KPa.

Further preferably, the negative pressure is 1.8KPa.

When the pH value of the system is 7.5-8.1, the effect of enriching the obtained extract can be improved. The superoxide dismutase is an active substance in the plant extract, has the effects of resisting oxidation and aging, has the highest activity value when the pH is 7.5-8.1, and improves the efficacy of the plant extract, and further research finds that amino acids such as arginine, lysine and the like can be enriched at the anode to the greatest extent in the electrolysis process when the pH is 7.7-7.9, and possibly that when the amino acids migrate in the form of anions, a plurality of amino acids reach the maximum migration number value when the pH is 7.7-7.9, and the anode enrichment rate is improved.

Preferably, the pH of the system is 7.5 to 8.1.

It is further preferred that the pH of the system is from 7.7 to 7.9.

Even more preferably, the system pH is 7.8.

The method for enriching the amino acid by using the electrolysis method can improve the efficacy of the plant extract, the electrolysis method can efficiently enrich the active substances without destroying the active substances, and the applicant finds that the superoxide dismutase and the amino acid can be efficiently enriched simultaneously under the alkaline condition, and the synergistic effect is supposed to be generated under the alkaline condition of the two active substances. Further research shows that the ion exchange membrane has the best enrichment effect when the exchange capacity is 0.9-2.3meq/g, and the active groups in the ion exchange membrane are generally hydrophilic, so that the ion exchange efficiency is higher when the active groups are high, but the water content and the swelling degree in the membrane are increased when the exchange capacity is too large, so that the membrane body structure is too loose, and the selective permeability of the membrane is reduced.

Preferably, the ion exchange membrane has an exchange capacity of 0.9 to 2.3mmol/g.

Further preferably, the ion exchange membrane has an exchange capacity of 1.2 to 2.1mmol/g.

Still more preferably, the anion exchange membrane is selected to have an exchange capacity of 2.02meq/g, available from sandinow technologies, inc: FAA-3-50. The cation exchange membrane is selected to have an exchange capacity of 1.3meq/g, purchased from Suzhou Cheng Er Nuo science and technology Co., ltd, model number: FKS-50.

The preferred electrode for use in the cell is a graphite electrode.

Preferably, the voltage of the electrolytic cell is 6-12V, and the electrolytic time is 0.5-3h.

More preferably, the voltage of the electrolytic cell is 9V, and the electrolysis time is 1h.

Has the beneficial effects that:

1. when the mass fraction of the ethanol solution is 65-72%, not only the extraction efficiency of amino acid can be improved, but also the dissolution of water-soluble substances such as saccharides and the like can be reduced, and the purity of the plant extract can be improved.

2. The weight ratio of the ethanol solution to the plants (liquid-material ratio) is 15-25:1, the extraction efficiency of amino acids in plants is best.

3. The rotary evaporation temperature is 45-55 ℃, the pressure is 1.5-2.5KPa, and the concentration efficiency is improved.

4. The electrolysis method can enrich the active substances efficiently without destroying the active substances.

5. When the pH value of the system is 7.5-8.1, the effect of enriching to obtain the extract can be improved, and the anode enrichment rate is improved.

Detailed Description

Example 1

A method for improving the efficacy of a plant extract by using an electrolytic method comprises the following steps:

s1, drying and crushing plants, and extracting with an ethanol solution to obtain a pretreatment solution;

s2, carrying out negative pressure rotary evaporation on the pretreatment solution, and filtering to obtain a primary concentrated solution;

and S5, enriching at the anode after electrolysis to obtain the plant extract.

The plant in step S1 is tea tree (Camellia sinensis); the drying is freeze drying at-20 ℃ for 12h; the crushing process uses the rubbing crusher, and the model is: IKA 10, the rotating speed is set to be 5000r/min; the mass fraction of the ethanol solution is 70%; the weight ratio of the ethanol solution to the plants is 20:1; the extraction time is 3h.

The aperture of the filter screen for filtering in the step S2 is 0.35 mu m; the rotary evaporation temperature is 50 ℃; the pressure of the negative pressure is 1.8KPa.

In the step S3, the alkali is sodium hydroxide (CAS: 1310-73-2) with the concentration of 1mol/L; the solid content of the secondary concentrated solution is 12 percent; the system pH was 7.8.

The electrode used in the electrolytic cell in the step S4 is a graphite electrode; the voltage of the electrolytic cell is 9V, and the electrolytic time is 1h; anion exchange membranes are available at 2.02meq/g exchange capacity from sandinow technologies ltd, suzhou: FAA-3-50. The cation exchange membrane has exchange capacity of 1.3meq/g, is purchased from Suzhou Cheng Er Nuo science and technology Limited company, and has the model number: FKS-50; the amount of the second concentrate added was 500g.

Example 2

A method for improving the efficacy of a plant extract by using an electrolysis method comprises the following steps:

and S5, enriching at the anode after electrolysis to obtain the plant extract.

The plant in step S1 is witch hazel (Hamamelismollis Oliver); the drying is freeze drying for 8 hours at the temperature of minus 20 ℃; the crushing process uses the rubbing crusher, and the model is: IKA A10, the rotating speed is set to be 6000r/min; the mass fraction of the ethanol solution is 70 percent; the weight ratio of the ethanol solution to the plants is 22:1; the extraction time is 2.5h.

The aperture of the filter screen for filtering in the step S2 is 0.30 mu m; the rotary evaporation temperature is 50 ℃; the pressure of the negative pressure is 1.8KPa.

In the step S3, the alkali is sodium hydroxide (CAS: 1310-73-2) with the concentration of 1mol/L; the solid content of the secondary concentrated solution is 10 percent; the system pH was 7.8.

The electrode used in the electrolytic cell in the step S4 is a graphite electrode; the voltage of the electrolytic cell is 9V, and the electrolytic time is 1h; the anion exchange membrane has exchange capacity of 2.02meq/g, and is purchased from Suzhou Cheng Er Nuo science and technology Limited company, model number: FAA-3-50. The cation exchange membrane has exchange capacity of 1.3meq/g, is purchased from Suzhou Cheng Er Nuo science and technology Co., ltd, and has the model number: FKS-50; the amount of the second concentrate added was 400g.

Example 3

and S5, enriching at the anode after electrolysis to obtain the plant extract.

The plant in step S1 is centella asiatica (Centellaasiatica L Urban); the drying is freeze drying for 10 hours at the temperature of minus 20 ℃; the crushing process uses rubbing crusher, and the model is: IKA 10, the rotating speed is set to be 4000r/min; the mass fraction of the ethanol solution is 70 percent; the weight ratio of the ethanol solution to the plants is 18:1; the extraction time is 3h.

The aperture of the filter screen for filtering in the step S2 is 0.30 mu m; the rotary evaporation temperature is 50 ℃; the negative pressure is 1.8KPa.

In the step S3, the alkali is sodium hydroxide (CAS: 1310-73-2) with the concentration of 1mol/L; the solid content of the secondary concentrated solution is 15 percent; the system pH was 7.7.

The electrode used in the electrolytic cell in the step S4 is a graphite electrode; the voltage of the electrolytic cell is 9V, and the electrolytic time is 1h; anion exchange membranes are available at 2.02meq/g exchange capacity from sandinow technologies ltd, suzhou: FAA-3-50. The cation exchange membrane has exchange capacity of 1.3meq/g, is purchased from Suzhou Cheng Er Nuo science and technology Co., ltd, and has the model number: FKS-50; the amount of the second concentrate added was 500g.

Comparative example 1

s2, carrying out negative pressure rotary evaporation on the pretreatment liquid, and filtering to obtain a plant extract;

in the step S1, the plant is tea tree (Camellia sinensis); the drying is freeze drying at-20 ℃ for 12h; the crushing process uses the rubbing crusher, and the model is: IKA 10, the rotating speed is set to be 5000r/min; the mass fraction of the ethanol solution is 70 percent; the weight ratio of the ethanol solution to the plants is 20:1; the extraction time is 3h.

The diameter of the filter screen for filtering in the step S2 is 0.35 mu m; the rotary evaporation temperature is 50 ℃; the negative pressure is 1.8KPa.

Comparative example 2

The specific implementation manner is the same as that of example 1; except that the weight ratio of the ethanol solution of the step S1 to the plants in the comparative example 2 was 10:1; the mass fraction of the ethanol solution is 60 percent.

Comparative example 3

The specific implementation manner is the same as that of example 1; except that the system pH in step S3 described in comparative example 3 was 8.5.

Comparative example 4

The specific implementation manner is the same as that of example 1; except that in comparative example 4 the rotary evaporation temperature was 78 ℃ and the pressure was 101.3KPa.

Performance test method

Determination of the Total amount of amino acids

1. 0.6g of recrystallized ninhydrin was weighed into a beaker, and 15ml of n-propanol, 30ml of n-butanol, 60ml of ethylene glycol and 9ml of acetate buffer solution with pH 4.54 were added and mixed well.

2. Preparing an amino acid standard solution: weighing 23.4mg of leucine dried at 80 ℃, dissolving the leucine into 50ml (containing 50ug/ml of nitrogen) by using 10% of isopropanol solution by mass fraction, taking 5ml of the solution, and adding water to the volume of 50ml, wherein the volume of the solution is 5ug/ml of working solution containing the nitrogen.

3. Preparing an ascorbic acid solution with the mass concentration of 1 percent: 0.1g of ascorbic acid is weighed up to 100ml.

4. Taking 6 20ml test tubes, adding the following agents according to the following table, uniformly mixing the solutions of the test tubes, sealing, heating in boiling water for 15min, taking out, immediately shaking and cooling with cold water, fixing the volume to 20ml with ethanol with the mass fraction of 60%, shaking uniformly, and testing the light absorption value with lambda =570 nm.

Reagent	1	2	3	4	5	6
							Leucine Standard solution (ml)	0	0.2	0.4	0.6	0.8	1.0
Ammonia-free distilled water (ml)	2.0	1.8	1.6	1.4	1.2	1.0
							Ninhydrin (ml)	3.0	3.0	3.0	3.0	3.0	3.0
Ascorbic acid (ml)	0.1	0.1	0.1	0.1	0.1	0.1
							Amino nitrogen content (ug/tube)	0	1.0	2.0	3.0	4.0	5.0
Lambda =570nm absorbance	0	0.025	0.055	0.099	0.146	0.186

5. Equation of standard curve

And (3) taking the absorbance as a ordinate and the amino nitrogen amount/ug number as a abscissa to obtain a standard curve equation: y =0.0382x-0.01032 (R) ² ＝0.98820.15)

6. Determination of a sample to be tested

Taking 20ml test tube, taking 1ml of extract, adding distilled water lml, ninhydrin 3.0ml and ascorbic acid 0.1ml, mixing uniformly and sealing. Heating in boiling water bath for 1.5 min, shaking in cold water for cooling, dissolving in 60% ethanol to 20ml, shaking, and measuring absorbance A at 570mn wavelength.

7. And (3) calculating the result:

and (3) measuring absorbance, substituting the absorbance into a formula to calculate: a =0.0382m-0.0103, m is the amount of amino nitrogen.

Performance test data

	Absorbance of the solution	Amino nitrogen content (ug/tube)
			Example 1	0.452	12.1
Example 2	0.391	10.5
			Example 3	0.402	10.8
Comparative example 1	0.093	2.7
			Comparative example 2	0.154	4.3
Comparative example 3	0.207	5.7
			Comparative example 4	0.112	3.2

Claims

1. A method for improving the efficacy of a plant extract by using an electrolysis method is characterized by comprising the following steps:

and S5, enriching at the anode after electrolysis to obtain the plant extract.

2. The method for improving the efficacy of plant extracts by using an electrolysis method as claimed in claim 1, wherein the weight ratio of the ethanol solution to the plants in the step S1 is (15-25): 1.

3. the method for improving the efficacy of plant extracts by using an electrolysis method as claimed in claim 2, wherein the ethanol solution in step S1 is 65-72% by weight.

4. The method for improving efficacy of plant extracts through the electrolysis according to claim 1, wherein the rotary evaporation temperature in step S2 is 45-55 ℃.

5. The method for improving the efficacy of plant extracts by using an electrolysis method as claimed in claim 4, wherein the negative pressure in step S2 is 1.5-2.5KPa.

6. The method for improving the efficacy of plant extracts by using an electrolysis method as claimed in claim 5, wherein the pore size of the filter screen for filtration in step S2 is 0.22-0.45 μm.

7. The method for improving the efficacy of a plant extract by an electrolysis process as claimed in claim 1, wherein the pH of weak base in step S3 is 7.5-8.1.

8. The method for improving the efficacy of plant extracts by using an electrolysis method as claimed in claim 1, wherein the solid content of the secondary concentrated solution in the step S3 is between 10% and 15%.

9. The method for improving efficacy of plant extracts by using the electrolysis method as claimed in claim 1, wherein the ion exchange membranes in step S4 are anion exchange membranes and cation exchange membranes.

10. The method for improving efficacy of plant extracts by electrolysis as claimed in claim 9, wherein the ion exchange membrane in step S4 has an exchange capacity of 0.9-2.3meq/g.