CN114029502A

CN114029502A - Method for synthesizing nano platinum by using artemisia apiacea extract and application

Info

Publication number: CN114029502A
Application number: CN202111341494.6A
Authority: CN
Inventors: 陈雪; 王军; 吴丽芳; 张义森
Original assignee: Hefei Institutes of Physical Science of CAS
Current assignee: Hefei Institutes of Physical Science of CAS
Priority date: 2021-11-12
Filing date: 2021-11-12
Publication date: 2022-02-11

Abstract

The invention discloses a method for synthesizing nano platinum by utilizing a sweet wormwood herb extract, which comprises the following steps of preparing raw materials of sweet wormwood herb and chloroplatinic acid; the preparation method comprises the following steps: adding the dried sweet wormwood powder into ultrapure water for soaking, filtering, concentrating and drying to obtain sweet wormwood extract, and mixing the sweet wormwood extract with chloroplatinic acid for reaction to obtain the nano platinum particles. The nano platinum of sweet wormwood prepared by the preparation method provided by the invention is safe and simple, has no pollution, is low in cost and has high biological activity. The nano platinum generated by reduction of the sweet wormwood herb has the oxidation resistance, can remove free radicals, promote the secretion of collagen, keep the skin elasticity, further achieve the anti-aging effect, inhibit the activity of tyrosinase, and achieve the whitening effect.

Description

Method for synthesizing nano platinum by using artemisia apiacea extract and application

Technical Field

The invention relates to the technical field of nano platinum preparation, in particular to a method for synthesizing nano platinum by utilizing a sweet wormwood herb extract and application thereof.

Background

In recent years, nanotechnology has received much attention due to its great advantages in the cosmetics industry and drug delivery. Nanotechnology refers to scientific technology on the nanometer scale (1 to 100 nanometers), and in recent 40 years, nanotechnology methods are also continuously entering the fields of cosmetics and other health products. To date, various nanoparticle delivery systems have been developed for dermal and transdermal delivery, and most studies have shown that Nanoparticles (NPs) enhance the penetration of drugs into and through the skin and increase the activity and tolerability of drugs. Skin aging not only affects appearance but also is associated with many skin diseases, and thus prevention and delay of skin aging are receiving much attention.

The nano platinum has very good oxidation resistance and plays a key role in resisting skin aging. At present, the synthesis of nano platinum is mostly based on physical and chemical methods, and reducing agents and stabilizing agents used in the synthesis are easy to pollute the environment.

For example, the Chinese patent application number is: CN202110366271.9 discloses "a nano platinum catalyst using carbon nanotube as carrier for fuel cell oxygen reduction electrochemical reaction and its preparation method".

The technical scheme disclosed in the patent discloses the following preparation method:

the carbon nano tube carrier is characterized in that the surface of the carbon nano tube carrier is covered with a manganese oxide hydrophilic functional layer which grows in situ, and platinum nano particles are uniformly dispersed on the manganese oxide hydrophilic layer and form a composite crystalline phase with the manganese oxide hydrophilic functional layer. The preparation method is characterized in that a manganese oxide functional layer covered on the surface of the carbon nano tube is utilized to cause the platinum ions in the liquid phase and the manganese with lower valence state to generate displacement reaction, thereby uniformly depositing the nano platinum atomic layer on the manganese oxide for forming. The preparation method has simple process, and the synthesized catalyst has high oxygen reduction performance and good industrial application prospect.

The method for synthesizing the nano platinum by utilizing the biological resources has more advantages than other two methods. The biosynthesis method has the characteristics of low cost, ecological friendliness, easiness in large-scale synthesis and the like, and is more suitable for being applied to the field of beauty and health care.

Sweet wormwood herb, known as "Chinese Shencao", has the functions of resisting virus, diminishing inflammation, clearing heat, relieving cough and asthma, lowering blood pressure, protecting liver and benefiting gallbladder. The artemisia apiacea extract is utilized to reduce the nano platinum, so that the method can be applied to the wider skin care field.

Disclosure of Invention

The invention aims to solve the technical problem of providing a method for synthesizing nano platinum by using a sweet wormwood herb extract.

The invention solves the technical problems through the following technical scheme:

a method for synthesizing nano platinum by using an artemisia apiacea extract comprises the following steps:

(1) mixing dried herba Artemisiae Annuae powder with ultrapure water at a mass ratio of 1:1-1:100 to obtain mixed solution, performing ultrasonic treatment at 0-100 deg.C for 0.5-6 hr, and standing at-20 deg.C for 24 hr;

(2) centrifuging at 15000r/min for 10-30min at 8000-;

(3) weighing the sweet wormwood herb extract obtained in the step (2), and mixing the materials according to the weight ratio of 1: 100-10: 100, dissolving in water, and filtering to obtain sweet wormwood extracting solution;

(4) adding 0.001-0.1mol of chloroplatinic acid into 1-4ml of extracting solution, and violently stirring for 30min-3 h to ensure that a reaction mixture turns from yellow to black to form nano platinum;

(5) and (4) centrifuging the nano platinum solution obtained in the step (4) for 10-30min at 8000-.

Preferably, in the step (1), the dried sweet wormwood powder is taken and mixed with ultrapure water according to the mass ratio of 1:10 to obtain a mixed solution, and ultrasonic treatment is carried out at 0 ℃ for 30 min;

in the step (2), centrifuging for 10min at 8000r/min by a centrifuge, taking the supernatant, and drying to obtain sweet wormwood herb extract dry powder;

in the step (3), the sweet wormwood herb extract obtained in the step (2) is weighed according to the weight ratio of 1:100, dissolving in water, and filtering to obtain sweet wormwood extracting solution;

in the step (4), taking 1ml of extracting solution, adding 0.001mol of chloroplatinic acid, violently stirring for 30min, and changing the reaction mixture from yellow to black to form nano platinum;

and (5) centrifuging the nano platinum solution obtained in the step (4) at 12000r/min for 20min, washing the precipitate twice with pure water, and dissolving the precipitate in the pure water to obtain the nano platinum solution.

Preferably, in the step (1), the dried sweet wormwood powder is taken and mixed with ultrapure water according to the mass ratio of 1:20 to obtain a mixed solution, and ultrasonic treatment is carried out at 30 ℃ for 2 hours;

(2) centrifuging at 10000r/min for 15min, collecting supernatant, and drying to obtain herba Artemisiae Annuae extract dry powder;

(3) weighing the sweet wormwood herb extract obtained in the step (2), and mixing the components in a ratio of 2: 100, dissolving in water, and filtering to obtain sweet wormwood extracting solution;

(4) adding 0.01mol of chloroplatinic acid into 2ml of extracting solution, and violently stirring for 1 hour until the reaction mixture turns from yellow to black to form nano platinum;

(5) and (4) centrifuging the nano platinum solution obtained in the step (4) at 12000r/min for 20min, washing the precipitate twice with pure water, and dissolving the precipitate in the pure water to obtain the nano platinum solution.

Preferably, in the step (1), the dried sweet wormwood powder is taken and mixed with ultrapure water according to the mass ratio of 1:50 to obtain a mixed solution, and ultrasonic treatment is carried out at 60 ℃ for 4 hours;

in the step (2), centrifuging for 20min at 12000r/min by a centrifuge, taking the supernatant, and drying to obtain sweet wormwood herb extract dry powder;

in the step (3), the sweet wormwood herb extract obtained in the step (2) is weighed, and the weight ratio is 5: 100, dissolving in water, and filtering to obtain sweet wormwood extracting solution;

in the step (4), 0.05mol of chloroplatinic acid is added into 3ml of extracting solution, and the mixture is stirred vigorously for 2 hours, so that the reaction mixture is changed from yellow to black to form nano platinum;

Preferably, in the step (1), the dried sweet wormwood powder is taken and mixed with ultrapure water according to the mass ratio of 1:100 to obtain a mixed solution, and the mixed solution is subjected to ultrasonic treatment at 100 ℃ for 6 hours;

in the step (2), centrifuging for 30min at 15000r/min by a centrifuge, taking the supernatant, and drying to obtain sweet wormwood herb extract dry powder;

in the step (3), the sweet wormwood herb extract obtained in the step (2) is weighed according to the weight ratio of 10: 100 (dissolving in water, filtering to obtain herba Artemisiae Annuae extractive solution;

in the step (4), 0.1mol of chloroplatinic acid is added into 4ml of extracting solution, and the mixture is stirred vigorously for 3 hours, so that the reaction mixture is changed from yellow to black to form nano platinum;

Preferably, the drying mode in the step (2) is freeze-drying.

Preferably, the ratio of the sweet wormwood herb extract to the water in the step (3) is a mass-to-volume ratio.

The invention also discloses the application of the nano platinum synthesized by the method for synthesizing the nano platinum by utilizing the sweet wormwood herb extract in preparing cosmetics.

Preferably, the cosmetic is any one of a mask, essence, cream, eye cream or toner.

Compared with the prior art, the invention has the following advantages:

the invention discloses a method for synthesizing nano platinum by utilizing a sweet wormwood herb extract, which has the following beneficial effects:

1. the method for synthesizing the nano platinum has the advantages of high content of secondary metabolites, no secondary pollution, cost saving and simplified production steps.

2. The synthesis method adopted by the invention has the advantages that the synthesized nano particles are stable, and substances such as polyphenol, flavone and the like contained in the sweet wormwood herb extract can be combined with the surfaces of the nano particles to prevent aggregation.

3. The southernwood nano platinum provided by the invention has high biological activity and has remarkable antioxidation and anti-aging effects.

4. The nano platinum solution provided by the invention can remove DPPH free radicals, has anti-aging capacity, can promote collagen synthesis, and is suitable for being applied to cosmetics.

Drawings

FIG. 1 is a diagram of the ultraviolet absorption spectrum of nano platinum prepared by the embodiment of the invention;

FIG. 2 is a transmission electron microscope image of nano platinum ions prepared by the embodiment of the invention;

FIG. 3 is a bar graph of DPPH radical scavenging effect of nano platinum in an embodiment of the present invention;

FIG. 4 is a graph showing the toxicity of nano-platinum on HFF-1 cells, which is obtained by the inventive example;

FIG. 5 is a graph showing the effect of nano-platinum prepared in the examples on type I collagen secretion in HFF-1 cells;

FIG. 6 is a graph showing toxicity analysis of nano-platinum prepared in the present invention on A375 cells;

FIG. 7 is a graph showing the effect of nano-platinum prepared by the example of the present invention on tyrosinase in A375 cells.

Detailed Description

The following examples are given for the detailed implementation and specific operation of the present invention, but the scope of the present invention is not limited to the following examples.

Example 1

(1) Taking dry sweet wormwood powder, uniformly mixing the dry sweet wormwood powder with ultrapure water according to the mass ratio of 1:10 to obtain a mixed solution, and carrying out ultrasonic treatment at 0 ℃ for 30 minutes;

(2) centrifuging at 8000 rpm for 10min, collecting supernatant, and lyophilizing to obtain herba Artemisiae Annuae extract powder;

(3) weighing the sweet wormwood herb extract obtained in the step (2), and mixing the materials according to the weight ratio of 1:100 (w: v) are dissolved in water and filtered to obtain the sweet wormwood herb extract.

(4) 0.001mol of chloroplatinic acid was added to 1ml of the extract, and the reaction mixture was vigorously stirred for 30 minutes to change from yellow to black, indicating the formation of nano-platinum.

(5) And (4) centrifuging the nano platinum solution obtained in the step (4) at 12000rpm for 20 minutes, washing the precipitate twice with pure water, and dissolving the precipitate in the pure water to obtain the nano platinum solution.

Example 2

(1) Mixing dried herba Artemisiae Annuae powder with ultrapure water at a mass ratio of 1:20 to obtain mixed solution, and performing ultrasonic treatment at 30 deg.C for 2 hr;

(2) centrifuging for 15 minutes at 10000 revolutions by a centrifuge, taking supernatant, and freeze-drying to obtain sweet wormwood herb extract dry powder;

(3) weighing the sweet wormwood herb extract obtained in the step (2), and mixing the components in a ratio of 2: 100(w: v) are dissolved in water and filtered to obtain the sweet wormwood herb extract.

(4) 0.01mol of chloroplatinic acid was added to 2ml of the extract, and the reaction mixture was vigorously stirred for 1 hour to change from yellow to black, indicating the formation of nano-platinum.

Example 3

(1) Mixing dried herba Artemisiae Annuae powder with ultrapure water at a mass ratio of 1:50 to obtain mixed solution, and performing ultrasonic treatment at 60 deg.C for 4 hr;

(2) centrifuging for 20 minutes at 12000rpm by a centrifuge, collecting supernatant, and lyophilizing to obtain herba Artemisiae Annuae extract dry powder;

(3) weighing the sweet wormwood herb extract obtained in the step (2), and mixing the materials according to the weight ratio of 5: 100(w: v) are dissolved in water and filtered to obtain sweet wormwood herb extracting solution;

(4) 0.05mol of chloroplatinic acid was added to 3ml of the extract, and the reaction mixture was vigorously stirred for 2 hours, changing from yellow to black, indicating the formation of nano-platinum.

Example 4

(1) Mixing dried herba Artemisiae Annuae powder with ultrapure water at a mass ratio of 1:100 to obtain mixed solution, and performing ultrasonic treatment at 100 deg.C for 6 hr;

(2) centrifuging for 30min at 15000 rpm, collecting supernatant, and lyophilizing to obtain herba Artemisiae Annuae extract powder;

(3) weighing the sweet wormwood herb extract obtained in the step (2), and mixing the components in a ratio of 10: 100(w: v) are dissolved in water and filtered to obtain the sweet wormwood herb extract.

(4) 0.1mol of chloroplatinic acid was added to 4ml of the extract, and the reaction mixture was vigorously stirred for 3 hours, whereby the color of the reaction mixture turned from yellow to black, indicating the formation of nano-platinum.

Comparative example

(3) and (3) dissolving the sweet wormwood herb extract dry powder obtained in the step (2) in pure water to obtain a comparative example solution.

Analytical experiments

Performing ultraviolet-visible photometer analysis on the Artemisia apiacea reduced nano platinum;

under ultraviolet visible light, chloroplatinic acid has an absorption peak at 262nm, the artemisia apiacea extract has an absorption peak at 283nm, the absorption peak of the chloroplatinic acid basically disappears along with the reduction of platinum, and the increase of the absorption intensity of the whole band in the ultraviolet visible region can be seen, as shown in figure 1.

Performing transmission electron microscope characterization on the Artemisia apiacea reduced nano platinum;

the shape and size of the nano particles can be observed more directly by using a transmission electron microscope, the nano platinum prepared in different embodiments is taken out for characterization, and the size of the nano particles prepared in the embodiments is less than 10nm as can be seen from an electron diffraction pattern, which is shown in figure 2.

Histogram of DPPH radical scavenging action of Artemisia annua nano platinum;

preparing 50 mu g/mL DPPH working solution by absolute ethyl alcohol, storing in dark, adding 100 mu L of southernwood nano platinum composition solution with the concentration of 10mg/L and 900 mu L of DPPH working solution (A1) into an experimental group, adding 100 mu L of southernwood nano platinum composition solution with the concentration of 10mg/L and 900 microliter of absolute ethyl alcohol solution (A2) into a blank group, adding 100 mu L of absolute ethyl alcohol solution and 900 microliter of DPPH working solution (A0) into a control group, mixing uniformly, standing for half an hour in a dark place at room temperature, centrifuging, removing supernatant, and measuring the light absorption value at 517 nm. The measurement results of examples 1 to 4 are shown in Table 1 and FIG. 3.

TABLE 1 ability of Artemisia annua Nanoplatin to scavenge DPPH free radicals

As can be seen from Table 1 and FIG. 3, the scavenging efficiency of DPPH is higher in the case of the four examples of Artemisia annua nano platinum, the scavenging efficiency of free radicals is 85.4% at a concentration of 10mg/L, and the scavenging effect is better than that of examples 1-3 and comparative example.

The sweet wormwood reduces the toxicity of nano platinum to HFF-1 cells;

taking human HFF-1 cells with the logarithmic growth phase and the concentration of 5000 cells/mL, and inoculating the cells into a 96-well cell culture plate, wherein each well of the cell sap is 100 mu L; the 96-well plate is placed in a cell culture box at 37 ℃ for adherent culture. Adding herba Artemisiae Annuae nano platinum with concentration of 0.25mg/L, 1mg/L and 2.5mg/L into 96-well cell culture plate, adding equal volume of cell culture solution into control group, and incubating for 24h at 37 deg.C in cell culture box containing 5% CO 2; then adding 10 mu L of CCK8 solution into each well, and incubating for 1-4h at 37 ℃ in a cell culture box containing 5% CO 2; absorbance at 450nm was measured with a microplate reader. The measurement results of examples 1 to 4 are shown in Table 1.

Table 2 shows the effect of nano platinum in Artemisia annua on human HFF-1 cell survival rate (100%)

As the survival rate of cells is higher than 80% which is the standard of no toxicity to human HFF-1 fibroblasts, it can be seen from Table 2 and FIG. 4 that the nano platinum Artemisia annua in examples 3 and 4 has no toxicity to HFF-1 fibroblasts at a concentration of not higher than 1 mg/mL.

5. The influence of the nano platinum prepared for the invention embodiment on the secretion of type I collagen of HFF-1 cells;

taking human HFF-1 cells with the logarithmic growth phase and the concentration of 5000 cells/mL, and inoculating the cells into a 96-well cell culture plate, wherein each well of the cell sap is 100 mu L; respectively adding 10 μ L of herba Artemisiae Annuae nano platinum with concentration of 0.25mg/L, 1mg/L, 2.5mg/L into experimental group, adding no sample into blank group, respectively incubating at 37 deg.C for 24 h; collecting cell culture supernatant, measuring light absorption value at 450nm by ELISA method and using ELISA reader, and calculating the relative content of type I collagen. The measurement results of examples 1 to 4 are shown in Table 3 and FIG. 5.

TABLE 3 Effect of Artemisia annua nanoplatinum on type I collagen secretion in HFF-1 cells

As can be seen from table 3 and fig. 5, after the artemisia apiacea nano platinum provided by the invention with the concentration of 1mg/L is added into HFF-1 cells, the content of type I collagen in the skin is 135% in the fourth example, which shows that the artemisia apiacea extract provided by the example 4 of the invention can remarkably promote the expression of type I collagen, and can be added into cosmetics as an anti-aging factor.

Analyzing the toxicity of the artemisia apiacea reduced nano platinum on the A375 cells;

taking human A375 cells with logarithmic growth phase and concentration of 3000/mL, inoculating the cells into a 96-well cell culture plate, wherein each well of cell sap is 100 mu L, and each sample is provided with 3 multiple wells; the 96-well plate is placed in a cell culture box at 37 ℃ for adherent culture. Adding herba Artemisiae Annuae nano platinum with concentration of 0.1mg/L, 1mg/mL and 10mg/mL into 96-well cell culture plate, adding equal volume of cell culture solution into control group, and incubating at 37 deg.C in cell culture box containing 5% CO2 for 24 h; then adding 10 mu L of CCK8 solution into each well, and incubating for 1-4h at 37 ℃ in a cell culture box containing 5% CO 2; absorbance at 450nm was measured with a microplate reader. The measurement results of examples 1 to 4 are shown in Table 4 and FIG. 6.

Table 4 shows the effect of nano-platinum in Artemisia annua on the survival rate of human A375 cells (100%)

As the cell survival rate is higher than 80% which is a non-toxic criterion for human A375 melanoma cells, it can be seen from Table 4 and FIG. 6 that the nano platinum Artemisia annua in examples 1-4 has no toxicity for A375 melanoma cells at a concentration of not higher than 1 mg/mL.

7. The nano platinum prepared by the embodiment of the invention has the effect of inhibiting tyrosinase in A375 cells;

taking human A375 cells with logarithmic growth phase and 106/mL concentration, inoculating 200 mu L of the cells into a 35mm cell culture dish with 1mL of culture solution, and placing the cell culture dish in a cell culture box with 37 ℃ and 5% CO2 for adherent culture; respectively adding 10 μ L of herba Artemisiae Annuae nano platinum with concentration of 0.1mg/L, 0.5mg/L, 1mg/L into experimental group, adding no sample into blank group, respectively incubating at 37 deg.C for 24 h; the cell culture solution was aspirated, washed 2 times with PBS, 100. mu.L of 1% Troton-X-100 cell lysate containing PMSF was added, rapidly frozen at-20 ℃ for 30 minutes, thawed at 4 ℃ and then the sample was collected and centrifuged at 4 ℃ for 15 minutes at 12000 rpm. Collecting supernatant, quantifying BCA, adding 100 microliters of 4mg/mL levodopa solution, carrying out water bath at 37 ℃ for 3 hours, measuring the light absorption value at 490nm by using an enzyme-labeling instrument, and calculating the inhibition effect of tyrosinase.

The measurement results of examples 1 to 4 are shown in Table 5 and FIG. 7.

TABLE 5 inhibitory Effect of Artemisia annua Nanoplatin on tyrosinase in A375 cells

As can be seen from table 5 and fig. 7, after the nano-platinum artemisia annua with the concentration of 0.1mg/L provided by the invention is added to the a375 cells, the inhibition rate of the nano-platinum artemisia annua in the a375 cells is 67.4% in example 2, which indicates that the nano-platinum artemisia annua provided by the invention can significantly inhibit the expression of tyrosinase and can be added to cosmetics as a whitening factor.

In conclusion, compared with the nano platinum prepared by the prior art, the nano platinum for sweet wormwood provided by the invention has good anti-aging and free radical removal capabilities, and can be used as a cosmetic additive to be added into cosmetics

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims

1. A method for synthesizing nano platinum by utilizing a sweet wormwood herb extract is characterized by comprising the following steps:

(2) centrifuging at 15000r/min for 10-30min at 8000-;

2. The method for synthesizing nano platinum by using the artemisia apiacea extract as claimed in claim 1, wherein in the step (1), the dried artemisia apiacea powder is uniformly mixed with ultrapure water according to a mass ratio of 1:10 to obtain a mixed solution, and ultrasonic treatment is carried out at 0 ℃ for 30 min;

3. The method for synthesizing nano platinum by using the artemisia apiacea extract as claimed in claim 1,

in the step (1), taking dry sweet wormwood powder, uniformly mixing the dry sweet wormwood powder with ultrapure water according to the mass ratio of 1:20 to obtain a mixed solution, and carrying out ultrasonic treatment at 30 ℃ for 2 hours;

4. The method for synthesizing nano platinum by using the artemisia apiacea extract as claimed in claim 1, wherein in the step (1), the dried artemisia apiacea powder is uniformly mixed with ultrapure water according to the mass ratio of 1:50 to obtain a mixed solution, and the mixed solution is subjected to ultrasonic treatment at 60 ℃ for 4 hours;

5. The method for synthesizing nano platinum by using the artemisia apiacea extract as claimed in claim 1, wherein in the step (1), the dried artemisia apiacea powder is uniformly mixed with ultrapure water according to a mass ratio of 1:100 to obtain a mixed solution, and the mixed solution is subjected to ultrasonic treatment at 100 ℃ for 6 hours;

6. The method for synthesizing nano platinum by using the artemisia apiacea extract as claimed in any one of claims 1 to 5, wherein the drying manner in the step (2) is freeze-drying.

7. The method for synthesizing nano platinum by using the artemisia apiacea extract as claimed in any one of claims 1 to 5, wherein the ratio between the artemisia apiacea extract and the water in the step (3) is a mass to volume ratio.

8. The use of the nano platinum synthesized by the method for synthesizing nano platinum by using the artemisia apiacea extract as claimed in any one of claims 1 to 5 in the preparation of cosmetics.

9. The application of the nano platinum in the preparation of cosmetics according to claim 8, wherein the cosmetics are any one of facial masks, essence, facial creams, eye creams or toner.