CN111657415A - Curcumin nanoparticle and preparation method and application thereof - Google Patents

Abstract

The invention provides a curcumin nanoparticle, wherein the nanoparticle uses curcumin as a core material, and gum arabic and zein as a wall material; the mass ratio of the core material to the wall material is 5.5-7.5:100; The mass ratio of the gum arabic and zein is 1-5:5. The present invention also includes a preparation method of the curcumin nanoparticles and a beverage capable of delaying aging prepared from the curcumin nanoparticles. The preparation method of curcumin nano-particles of the present invention improves the problem of photo-instability in solution, so that it can be added to functional beverages as functional pigments to prepare anti-aging functional beverages. The curcumin beverage of the invention has a sweet and sour taste, and has the smell of curcumin, can improve the activity of SOD enzyme, reduce the content of malondialdehyde (MDA), and has anti-oxidation and anti-aging effects.

Description

A kind of curcumin nanoparticle and its preparation method and application

technical field

The invention relates to the technical field of functional food production technology, in particular to a curcumin nanoparticle and a preparation method and application thereof.

Background technique

Curcumin (curcumin, cur) is a polyphenolic compound proposed from the rhizome of the ginger plant, turmeric. It has a symmetrical molecular structure composed of beta-diketone heptadiene and two ortho-methylated phenols. The powder is orange. Yellow crystalline, slightly bitter taste. It has strong tinting power and low toxicity. It has been widely used in food, textile, cosmetics and other fields as a common natural pigment for a long time. In recent years, many scholars have shown that curcumin has anti-inflammatory, antioxidant, anti-tumor, anti-depressant, anti-viral, anti-viral, cardiovascular protection, repairing brain damage, relieving Parkinson's symptoms and other pharmacological and health care effects. However, because curcumin is insoluble in water, soluble in organic solvents such as ethanol, acetic acid, acetone, and chloroform, it is easily affected by external factors such as light, temperature, metal ions, and pH value, and it is easy to lose its color rendering ability, especially its high photosensitivity. Wang Xuemei et al. studied the photostability of curcumin compounds, and used high performance liquid chromatography to analyze the solutions of curcumin compounds before and after illumination, and found that under outdoor illumination, the solution of curcumin was extremely unstable, while the solid curcumin Powder is more stable.

In recent years, there have been many reports on the research on nanometerization of curcumin. Liu Yanmin et al. prepared the solid dispersion of curcumin through the method of melting rapid cooling and room temperature cooling, and found that the in vitro dissolution rate of curcumin solid dispersion is much higher than Pure Curcumin. The lyophilized powder of curcumin nanoparticles prepared by anti-solvent method can improve the water solubility of curcumin, which is beneficial to improve the bioavailability of curcumin. In addition, there are preparation methods for curcumin nanoparticles such as liposomes, micelles, and microemulsions.

Curcumin has a variety of pharmacological and health-care effects. People hope to use it as a functional pigment in the development of functional foods. However, because curcumin is insoluble in water, it is easily affected by external factors such as light, temperature, metal ions, and pH value. , it is easy to lose its color rendering ability. Especially under the outdoor light, the curcumin aqueous solution is unstable, undergoes a degradation reaction, and loses its original pharmacological effect and color, which limits the application field of curcumin, especially in the industrial development and application of liquid food. At present, there are many research reports on improving the solubility and bioavailability of curcumin. For example, after it is made into nanoliposomes, its water solubility and bioavailability can be significantly improved; , pigment molecular structure modification, improvement of natural pigment processing and storage conditions, etc. to improve the stability of natural pigments, but the research on improving its photosensitivity is rarely reported. With the rapid development of social economy and the rapid progress of science and technology, when people's food and clothing problems are solved, people's demand for beverages is no longer just for the pursuit of taste and vision, but now people pay more attention to the nutrition and health of the beverage itself. It has gone from the most primitive thirst quenching function to the current pursuit of nutrition, health and wellness. In recent years, there have been many functional beverages with nutrition, health and other functions in the market, which have various functions such as anti-aging and beauty, and are gradually favored by consumers. According to a large amount of data, the total output and sales of health-care functional beverages of domestic beverage companies have increased year by year in recent years. From the long-term development trend, health-care functional beverages will have a good development trend in my country in the future. If the photosensitivity problem of curcumin can be solved and applied in beverages, it will enrich the nutritional health beverage market and have high economic value.

SUMMARY OF THE INVENTION

The first object of the present invention is to provide a curcumin nanoparticle and a preparation method thereof in order to solve the shortcomings and deficiencies of the above-mentioned prior art.

The present invention is achieved through the following technical solutions:

A curcumin nanoparticle uses curcumin as a core material, gum arabic and zein as wall materials; the mass ratio of the core material to the wall material is 5.5-7.5:100; the gum arabic and zein are soluble in The mass ratio of protein is 1-5:5.

The preparation method of described curcumin nanoparticle, comprises the following steps:

1) Preparation of alcohol phase: Dissolve zein in 85% ethanol, stir magnetically for 1 h, remove insoluble matter by centrifugation as a zein solution, add curcumin and stir in the zein solution for 30 min to obtain an alcohol phase ;

2) Preparation of water phase: Dissolve gum arabic in 0.3g/L zinc sulfate solution (dissolve by heating in a water bath at 60°C), stir until completely dissolved, and obtain a water phase, and the water phase and the alcohol phase in step 1) are mixed. The volume ratio is 1.5-3.5:1;

3) anti-solvent co-precipitation: adding the alcohol phase obtained in step 1) into the water phase obtained in step 2) in a fine flow state and stirring for 30 min to obtain a curcumin nanoparticle dispersion;

4) rotary evaporation: the curcumin nanoparticle dispersion liquid obtained in step 3) is removed by rotary evaporation to obtain ethanol and water to obtain a curcumin nanoparticle concentrate;

5) drying: the curcumin nano-particles are obtained after freeze-drying the curcumin nano-particle concentrate obtained in step 4);

The addition amounts of the curcumin, gum arabic and zein are added according to the described mass ratio.

Further, the volume ratio of the water phase described in step 2) to the alcohol phase described in step 1) is 2:1; the mass ratio of the gum arabic to zein is 4:5; the core material and the wall material are The mass ratio is 7.5:100.

The second object of the present invention is to provide a curcumin beverage for delaying aging and a preparation method thereof.

The present invention is achieved through the following technical solutions:

An anti-aging curcumin beverage, in mass percentage, the beverage comprises: 15% of emperor orange concentrate, 0.05%-0.25% of xanthan gum, 0.2%-1% of curcumin nanoparticles, octenyl succinic acid 0.2%-1% of sodium starch, 0.05%-0.25% of sodium carboxymethyl cellulose, 0.1% of citric acid, 10% of xylitol, and the rest are water; the curcumin nanoparticles are obtained by the above preparation method.

Further, in terms of mass percentage, the beverage comprises: emperor mandarin orange concentrate 15%, xanthan gum 0.15%, curcumin nanoparticles 0.8%, sodium starch octenyl succinate 0.4%, sodium carboxymethyl cellulose 0.15% %, citric acid 0.1% and xylitol 10%, and the rest is water.

The preparation method of described anti-aging curcumin beverage, comprises the following steps:

1) Preparation of the emperor mandarin concentrate: after the emperor mandarin orange is peeled and cored, the juice is squeezed with a juicer and filtered twice with double-layer gauze, and then the juice is concentrated to 1/2 of the original volume to obtain the emperor mandarin concentrate;

2) mixing: adding the curcumin nanoparticles and sodium starch octenyl succinate of the mass percent under high-speed stirring, and mixing with the emperor orange concentrate;

3) Homogenization: pour the mixed solution prepared in step 2) into a high pressure homogenizer for homogenization;

4) Allocate: in the product of step 3), add the sodium carboxymethyl cellulose, xanthan gum, citric acid and xylitol of the mass percentage;

5) Secondary homogenization: pour the mixed solution prepared in step 4) into a high-pressure homogenizer for homogenization;

6) degassing: utilize water bath heating to degas the product of step 5);

7) Filling and sterilization: filling and sterilizing the product of step 6) while it is still hot to obtain the curcumin beverage for delaying aging.

Further, the pressure of the homogenization in step 3) is 25Mpa.

Further, the pressure of the secondary homogenization in step 5) is 5Mpa.

Further, the sterilization conditions in step 7) are sterilization at 121° C. for 10 minutes.

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

1. The preparation method of curcumin nanoparticles of the present invention improves the problem of photo-instability in solution, so that it can be added to functional beverages as functional pigments to prepare anti-aging functional beverages.

2. The curcumin beverage of the present invention has a sweet and sour taste, and has the smell of curcumin, which can improve the activity of SOD enzyme, reduce the content of malondialdehyde and MDA, and has anti-oxidation and anti-aging effects.

For better understanding and implementation, the present invention is described in detail below with reference to the accompanying drawings.

Description of drawings

Figure 1 is a curcumin absorption curve.

Figure 2 is a curcumin standard curve.

Figure 3 is the screening of the optimum concentration of zinc ions.

Figure 4 is the volume ratio of water phase to alcohol phase.

Figure 5 is the ratio of gum arabic to zein mass.

Figure 6 is the mass ratio of curcumin to zein-gum arabic.

Figure 7 is the curcumin retention rate of curcumin nanoparticles and pure curcumin under natural light.

Figure 8 is a picture taken by a 100-fold microscope under the condition of Experiment 1.

Figure 9 is a picture taken by a 100-fold microscope under the condition of Experiment 2.

Figure 10 is a picture taken by a 100x microscope under the condition of Experiment 3.

Figure 11 is the effect of CMC-Na addition on the free radical scavenging rate.

Figure 12 shows the effect of CMC-Na addition on curcumin beverages.

Figure 13 is the effect of xanthan gum addition on the free radical scavenging rate.

Figure 14 shows the effect of xanthan gum addition on curcumin beverages.

Figure 15 is the effect of the addition amount of sodium starch octenyl succinate on the free radical scavenging rate.

Figure 16 is the effect of the addition amount of sodium starch octenyl succinate on curcumin beverages.

Figure 17 is the effect of curcumin freeze-dried powder addition on free radical scavenging rate.

Figure 18 is the effect of curcumin freeze-dried powder addition on the quality of curcumin beverage.

Detailed ways

The materials and reagents used in the present invention are shown in Table 1.

Table 1 Materials and reagents used in the test

The equipment adopted in the present invention is as shown in Table 2:

Table 2 List of equipment used in the test

The data processing software used in the present invention are GraphPad Prism 7.04, Origin 9, EZ OMNIC and Orthogonal Design Assistant respectively.

All materials, reagents and instruments selected in the present invention are well known in the art, but do not limit the implementation of the present invention, and some other reagents and equipment well known in the art are applicable to the implementation of the following embodiments of the present invention.

Example 1

Test method for preparation of curcumin nanoparticles

1. Analysis of curcumin raw materials

1.1 Curcumin maximum absorption wavelength and regression equation

Maximum absorption wavelength: Accurately weigh 0.01g of curcumin powder sample into a 100ml volumetric flask, dilute to volume with 95% ethanol to obtain a curcumin solution with a concentration of 0.1mg/ml, pipette 500ul from it and dilute to volume with 95% ethanol solution To 100ml, obtain a curcumin solution with a concentration of 0.5ug/ml, adjust the wavelength to measure the absorbance of curcumin at different wavelengths, take the wavelength as the abscissa and the absorbance as the ordinate to determine the maximum absorption wavelength.

Regression equation: accurately weigh 25mg of curcumin standard in a 50mL volumetric flask, dilute to 0, 15, 30, 45, 60, 75ug/mL standard solution with 95% absolute ethanol, and measure at 425nm Absorbance, linear regression of absorbance Y with concentration X.

It can be seen from Figure 1 and Figure 2 that the maximum absorption wavelength of curcumin is 425nm, the regression equation of the standard curve is Y=0.3168x+0.6349, R ² =0.9974, and the curcumin concentration is in the range of 0～75μg/mL, and the linear relationship is good .

1.2 Analysis of mineral element content of curcumin

Microwave digestion method: refer to "GB 5009.268-2016 Determination of Multi-elements in Food"; Weigh 0.3g of curcumin solid (actually weighed 0.2824g) in the microwave digestion inner tank, add 8mL of nitric acid, put it on for 1h or overnight, Tighten the jar lid and digest according to the standard operating procedure of the microwave digester. After the digestion is completed, take it out after cooling, slowly open the tank cover to exhaust, rinse the inner cover with a small amount of water, put the digestion tank on a temperature-controlled electric hot plate to drain acid until it is nearly dry, and then use ultrapure water to make up the volume in 25mL volumetric flask, mix well for later use, and do the same number of blank tests at the same time.

It can be seen from Table 3 that the chemical composition of curcumin contains less zinc element, which has a certain influence on the stability of curcumin.

Table 3 Curcumin mineral element content

2. Screening of optimum concentration of zinc ion for stabilizer

2.1 Preparation of sample solution

Preparation of curcumin mother liquor: 0.175 g of curcumin was weighed and dissolved in 280 ml of absolute ethanol, and stirred until the curcumin powder was completely dissolved.

Preparation of different zinc ion concentration stock solutions: Weigh 0.03g, 0.06g, 0.09g, 0.12g, 0.15g, 0.18g of zinc sulfate and dissolve them in 100ml of distilled water, stir evenly, and configure into 0.3g/L, 0.6g/L L, 0.9g/L, 1.2g/L, 1.5g/L, 1.8g/L zinc sulfate stock solutions, save them for later use.

2.2 Screening test of optimum concentration of zinc ions

Divide the curcumin mother liquid into 7 samples of 40ml, and add 10ml, 0.0g/L, 0.3g/L, 0.6g/L, 0.9g/L, 1.2g/L to No. 1-7 sample solutions respectively. , 1.5g/L, 1.8g/L zinc sulfate stock solution, stir evenly to obtain zinc sulfate concentrations of 0.0g/L, 0.06g/L, 0.12g/L, 0.18g/L, 0.24g/L, 0.3g respectively /L, 0.36g/L sample solution, placed at room temperature. The absorbance was measured with a microplate reader at a wavelength of 425 nm every 12 hours, and the average value was obtained in parallel three times.

2.3 Analysis of results

It can be seen from Figure 3 that when different concentrations of zinc ions are added to the curcumin solution within 0 to 48 hours, the curcumin is fading, but it is more stable than that without zinc ions. From 48 to 60 hours, the absorbance of adding different concentrations of zinc ion curcumin solution increased. After 60 hours, the slope of zinc ion concentration of 0.3g/L was the smallest and the absorbance was relatively stable. Although the absorbance of 0.36g/L zinc ion concentration was relatively stable, The overall trend is still declining. Therefore, 0.3g/L zinc ion concentration has the best effect on improving the photostability of curcumin.

3. The preparation process of curcumin nanoparticles:

1) Preparation of alcohol phase: Dissolve zein in 85% ethanol, stir magnetically for 1 h, remove insoluble matter by centrifugation as a zein solution, add curcumin and stir in the zein solution for 30 min to obtain an alcohol phase ;

2) Preparation of water phase: Dissolve gum arabic in 0.3g/L zinc sulfate solution (dissolve by heating in a water bath at 60°C), stir until completely dissolved, and obtain a water phase, and the water phase and the alcohol phase in step 1) are mixed. The volume ratio is 1.5-3.5:1;

3) anti-solvent co-precipitation: adding the alcohol phase obtained in step 1) into the water phase obtained in step 2) in a fine flow state and stirring for 30 min to obtain a curcumin nanoparticle dispersion;

4) rotary evaporation: the curcumin nanoparticle dispersion liquid obtained in step 3) is removed by rotary evaporation to obtain ethanol and water to obtain a curcumin nanoparticle concentrate;

5) Drying: freeze-drying the curcumin nanoparticle concentrate obtained in step 4) to obtain curcumin nanoparticles.

3.1 Single factor experiment of preparation of curcumin nanoparticles

Taking the entrapment rate as the investigation index, the influences of the volume ratio of water phase to alcohol phase, the mass ratio of gum arabic and zein, and the mass ratio of curcumin and zein-gum arabic on the preparation of curcumin nanoparticles were determined to determine The optimum dosage of the three factors.

3.1.1 Volume ratio of water phase to alcohol phase

Zein 1g, curcumin 0.02g, gum arabic 1g, 85% ethanol 50ml, zinc sulfate 0.3g/L (calculated by total volume), the volume ratio of distilled water to ethanol is 1.5:1, 2:1, 2.5:1 , 3:1, 3.5:1, prepare curcumin nanoparticles according to the above process operation, and determine the optimum ratio of the volume of the water phase to the alcohol phase according to the entrapment rate.

It can be seen from Figure 4 that when the volume ratio of the water phase to the alcohol phase is from 1.5:1 to 2.5:1, the encapsulation rate increases from 85.4% to 93.2%, and when the volume ratio increases from 2.5:1 to 3.5:1, the encapsulation rate increases. The rate dropped from 93.2% to 84.6%. It may be that as the volume ratio of the water phase to the alcohol phase increases, the concentration of the alcohol phase decreases, and the dissolution of curcumin reaches a supersaturated state. During the deposition process, the precipitation of curcumin leads to the increase of free and the decrease of the entrapment rate. Determine the optimum volume ratio of water phase to alcohol phase to be 2.5:1.

3.1.2 The mass ratio of gum arabic and zein

Zein 1g, curcumin 0.02g, distilled water 75ml, 85% ethanol 50ml, zinc sulfate 0.3g/L (calculated by total volume), the mass ratio of gum arabic to zein is 2:10, 4:10, At 6:10, 8:10, and 10:10, the curcumin nanoparticles were prepared according to the above-mentioned technological operations, and the optimum ratio of gum arabic and zein was determined according to the encapsulation rate.

It can be seen from Figure 5 that as the mass ratio of gum arabic and zein increases, the encapsulation first increases and then decreases. When the mass ratio is 6:10, the encapsulation rate is the largest, which is 87.1%. The charges carried by gliadin were completely neutralized, and the entrapment rate was the largest, and the subsequent increase in the ratio resulted in charge imbalance and the entrapment rate decreased. The optimum mass ratio of gum arabic to zein was determined to be 6:10.

3.1.3 Total mass ratio of curcumin to zein-Gum Arabic

Zein 1g, gum arabic 1g, 85% ethanol 50ml, zinc sulfate 0.3g/L (calculated by total volume), distilled water 75ml, curcumin and zein-acacia total mass ratio 5.5:100, 6: 100, 6.5:100, 7:100, 7.5:100, prepare curcumin nanoparticles according to the above process operation, and determine the optimum ratio of curcumin and zein-acacia total mass according to the entrapment rate.

As can be seen from Figure 6, when the mass ratio of curcumin to zein-gum arabic from 5.5:100 to 6.5:100, the encapsulation rate increased from 97.1% to 97.6%, and when the ratio was from 6.5:100 to 7.5:100 , the embedding rate decreased. It may be that when the mass ratio of curcumin to zein-gum arabic was 6.5:100, the loading of the wall material reached the maximum loading, and then the increase of the ratio resulted in an increase in the amount of free curcumin and a decrease in the encapsulation rate. The optimum mass ratio of curcumin to zein-gum arabic was determined to be 6.5:100.

3.2 Orthogonal experiment of preparation of curcumin nanoparticles

3.2.1 Orthogonal experimental method

On the basis of the single factor test, the L9(3 ⁴ ) orthogonal table was used to compare the volume ratio of water phase to alcohol phase, the mass ratio of wall material components (mass ratio of gum arabic and zein), and the mass ratio of core wall (turmeric). The three factors were optimized, as shown in the table below; the encapsulation rate, photostability score and particle size were divided into the investigation indicators, and the multi-index comprehensive balance method was selected. Light stability score=(1-(A7-A1)/A7), particle size score=(1-particle size), the larger the value, the better.

Table 4 Factor level design table of L9(3 ⁴ ) orthogonal experiment for preparation of curcumin nanoparticles

3.2.1.1 Determination of the entrapment rate of curcumin nanoparticles

Pipette 1 mL of curcumin-loaded nanoparticles and 4 mL of 95% ethanol into a beaker, extract free curcumin by ultrasonic for 15 min, filter with a 0.22 μm organic membrane, and repeat the extraction 3 times. Combine the extracts, use 95% ethanol as a blank control, and measure the absorbance of the sample at 425 nm. According to the standard curve equation of curcumin in 95% ethanol, after calculating the free curcumin, calculate the entrapment rate and drug loading according to the following formula. quantity:

3.2.1.2 Particle size determination of curcumin nanoparticles

Dilute the sample by a certain factor, place it in the sample cell, and measure its particle size using a laser particle size analyzer.

3.2.1.3 Observation of photostability of curcumin nanoparticles

The samples were diluted 4 times, placed at room temperature, and the absorbance value was measured by a microplate reader every 24 hours to observe the effect of microcapsule technology on the photosensitivity of curcumin. Taking the reaction time t as the abscissa and the retention rate ln(At*/A0*) as the ordinate, plot the retention rates of the (Zein-Gum Arabic-Curcumin) nanosystem and unembedded curcumin, respectively , to observe the effect of curcumin nanoparticles on the photosensitivity of curcumin.

Sample grouping situation: sample 1 is the process condition of water-alcohol volume ratio (water phase to alcohol phase volume ratio) 2:1, wall mass ratio (acacia and zein mass ratio) 4:10, core wall mass ratio (curcumin and zein-gum arabic total mass ratio) 6:100; sample 2 is the process condition of hydroalcoholic volume ratio 2:1, wall mass ratio 6:10, core wall mass ratio 6.5:100; Sample 3 is a process condition of water-alcohol volume ratio of 2:1, wall material mass ratio of 8:10, core wall mass ratio of 7:100; sample 4 is a process condition of water-alcohol volume ratio of 2.5:1, wall material mass ratio of 4: 10. The mass ratio of core to wall is 7:100; for sample 5, the process condition is that the volume ratio of water to alcohol is 2.5:1, the mass ratio of wall material is 6:10, and the mass ratio of core wall is 6:100; sample 6 is that the process condition is the volume ratio of water to alcohol. 2.5:1, the mass ratio of wall material is 8:10, and the mass ratio of core wall is 6.5:100; for sample 7, the process conditions are that the volume ratio of water to alcohol is 3:1, the mass ratio of wall material is 4:10, and the mass ratio of core wall is 6.5:100; Sample 8 is a process condition of water-alcohol volume ratio of 3:1, wall material mass ratio of 6:10, core wall mass ratio of 7:100; sample 9 is a process condition of water-alcohol volume ratio of 3:1, wall material mass ratio of 8: 10. The mass ratio of core to wall is 6:100; the blank is pure curcumin 0.02g.

It can be seen from Figure 7 that, in general, after 48h, the retention rates of all samples tend to be stable, and the curcumin retention rates of curcumin nanoparticles are higher than those of unembedded curcumin nanoparticles. The photolysis reaction of curcumin nanoparticle-encapsulated is slower than that of unencapsulated curcumin. The volume ratio of water to alcohol (volume ratio of water phase to alcohol phase) is 3:1, the mass ratio of wall material (mass ratio of gum arabic to zein) is 4:10, and the mass ratio of core wall (curcumin to gum arabic-corn) The sample 7 prepared with the total mass ratio of gliadin) of 6.5:100 had the highest curcumin retention rate and the slowest curcumin photolysis reaction after 144h.

It can be seen from the above that the photolysis reaction of curcumin nanoparticles is slower than that of unembedded curcumin, and the stability of curcumin to light is improved after curcumin is embedded.

3.2.2 The orthogonal experiment results are as follows:

Table 5 Orthogonal test results of preparation process L ₉ (3 ⁴ ) of curcumin nanoparticles

3.2.3 The results of the variance analysis of the inspection indicators are as follows:

Table 6 The results of variance analysis of the investigation index based on the embedding rate

Table 7 The results of variance analysis with particle size as the investigation index

Table 8 The results of variance analysis of the indicators under investigation based on the absorbance degradation rate

Note: * means significant P<0.05

Table 9 Comparison table of optimal combination of different indicators

Available from Table 5, Table 6, Table 7, Table 8, Table 9, factor A has a highly significant influence on the embedding rate, according to the R value, preferably A ₁ when the embedding rate is an index; factor B affects the embedding rate There is a highly significant effect, so B ₃ is selected according to the R value, and factor C has no significant effect on the three indicators, but because the nanoparticles are prepared, the particle size needs to be smaller, and C ₃ is selected according to the R value. Through the comprehensive equilibrium method, the optimal level of curcumin nanoparticles preparation is A ₁ B ₃ C ₃ , that is, the volume ratio of water phase to alcohol phase is 2:1, the mass ratio of gum arabic to zein is 8:10, The mass ratio of curcumin to zein-gum arabic is 7.5:100, and the verification test shows that under the optimal action conditions, the encapsulation rate of curcumin nanoparticles is 96.730%, the drug load is 62mg/g, and the particle size is 0.940um , the slope of absorbance decline is 0.73.

Example 2

Test method for development of curcumin beverage for delaying aging

1. The preparation process of curcumin beverage:

1) Preparation of the emperor mandarin concentrate: after the emperor mandarin orange is peeled and cored, the juice is squeezed with a juicer and filtered twice with double-layer gauze, and then the juice is concentrated to 1/2 of the original volume to obtain the emperor mandarin concentrate;

2) mixing: adding the curcumin nanoparticles and sodium starch octenyl succinate of the mass percent under high-speed stirring, and mixing with the emperor orange concentrate; the curcumin nanoparticles are obtained from Example 1;

3) Homogenization: pour the mixed solution prepared in step 2) into a high pressure homogenizer for homogenization;

4) Allocate: in the product of step 3), add the sodium carboxymethyl cellulose, xanthan gum, citric acid and xylitol of the mass percentage;

5) Secondary homogenization: pour the mixed solution prepared in step 4) into a high-pressure homogenizer for homogenization;

6) degassing: utilize water bath heating to degas the product of step 5);

7) Filling and sterilization: filling and sterilizing the product of step 6) while it is still hot to obtain the anti-aging curcumin beverage.

1.1 Determination of homogenization pressure

The number of times of homogenization is twice, the beverages are homogenized under the homogenization pressure of 25MPa for the first time, 5MPa for the second homogenization, and the first homogenization pressure is 5MPa and the secondary homogenization pressure is 25MPa. Beverage was used as a control to investigate the effect of homogenization pressure on particle size. The following table shows the results of homogenization pressure.

Table 10 Homogenization pressure results

It can be seen from the particle size that Experiment 3>2>1, combined with the photomicrographs (Fig. 8-10), when the primary pressure is 25MPa, and the secondary homogenization pressure is 5MPa, the large particles are more fully broken into small particles. Molecular substances are beneficial to the storage stability of beverages. When the primary pressure is 5 MPa and the secondary pressure is 25 MPa, large substances can be broken, but the effect is not as good as the homogeneous condition of experiment 1 but better than the non-homogeneous effect. Therefore, the primary homogenization pressure is 25MPa, and the secondary homogenization pressure is 5MPa.

1.2 Determination of sterilization method

The pH of curcumin beverage is 6, which is a low-acid food. It can be sterilized by high temperature and high pressure, and sterilized at 121 °C for 10 minutes. You can also add citric acid to acidify the beverage, adjust its pH to 4.6, and pasteurize it at 80. Sterilize at ℃ for 20 min, and use cold sterilization to filter with 0.22 μm organic membrane, and use unsterilized beverage as a control. Taking DPPH scavenging free radical ability as an index, the sterilization conditions were determined.

1.3 Preparation of samples for analysis

Beverage sample solution: take curcumin nanoparticle freeze-dried powder and sodium starch octenyl succinate as raw materials, xanthan gum and CMC-Na as stabilizers, and prepare curcumin beverage according to the above-mentioned process. The curcumin content in this beverage is: 100 mg/mL. Then diluted with distilled water to the concentration of 20, 40, 60, 80, 100 μg/mL;

Taking the ascorbic acid solution as the positive control, 2, 4, 6, 8 and 10 mg of ascorbic acid were weighed and dissolved in distilled water and made up to 100 mL. The concentration of the ascorbic acid solution was 20, 40, 60, 80, and 100 μg/mL.

Table 11 Sterilization conditions

It can be seen from Table 11 that the sample IC ₅₀ value is 2>3>1>4, and the Vc positive control IC ₅₀ value is 2>1>3>4. The lower the _IC50 value, the higher the DPPH·free radical scavenging rate. The better the oxidation resistance of the sample, so the oxidation resistance of sample 1 is the highest. After sterilization, the IC ₅₀ values of the three sterilization methods all increased, while the sample 2 was acidified, the pH was 4.6, and the acid anion had an inhibitory effect on superoxide anion, so its IC ₅₀ value was the highest, and the DPPH·free radical scavenging rate was the lowest. Antioxidant is minimal. According to GB7101-2015 National Food Safety Standard (Beverage), the total bacterial counts of samples 1, 2 and 3 after sterilization all meet the national standard, while the total bacterial count of the unsterilized samples does not meet the national safety standard. Based on the above results, 121°C/10min was used as the sterilization condition in this experiment.

2. Evaluation method of curcumin beverage quality

2.1 Sensory scoring criteria

The sensory evaluation of this experiment consisted of 8 members, and the sensory tasting scores were carried out on various indicators of curcumin beverages.

Table 12 Sensory scoring criteria

2.2 Light absorption stability

Weigh a certain amount of sample, add water to dilute it 20 times, mix the diluent evenly, put it into a centrifuge at 4000r/min for 15min. Measure the absorbance values Ao and A before and after centrifugation at the maximum absorption wavelength of the sample. The light absorption stability is expressed by A/Ao.

2.3 Centrifugal sedimentation rate

Weigh the sample and the total weight of the centrifuge tube and the weight of the centrifuge tube respectively, centrifuge at 4000r/min for 15min, discard the supernatant, and accurately weigh the total amount of the centrifuge tube and the remaining precipitate.

The calculation formula is: sedimentation rate=precipitation mass/sample liquid mass×100%

2.4 Determination of DPPH·free radical scavenging ability

(1) Sample preparation, same as 1.3.

(2) Determination of DPPH·free radical scavenging ability

100 μL of 0.2 mmol/L DPPH ethanol solution was added to the 96-well plate. Then add 100 μL of curcumin beverage sample solutions of different concentration gradients, mix well, react at room temperature for 30 min in the dark place, measure the absorbance value A1 at the wavelength of 517 nm, and replace the DPPH ethanol solution (A2) with an equal volume of absolute ethanol in the control group. In the control group, the sample solution (A0) was replaced by an equal volume of distilled water, and ascorbic acid was used as a positive control.

The calculation formula is: clearance %=[1-(A1-A2)/A0]×100

where:

A0: The absorbance of a mixed solution of 100 μL distilled water and 100 μL LDPPH ethanol;

A1: The absorbance of 100 μL sample solution and 100 μL LDPPH ethanol mixture;

A2: The absorbance of 100 μL sample solution and 100 μL absolute ethanol mixture.

2.5 Determination of ABTS ⁺ · free radical scavenging ability

(1) Sample preparation, same as 1.3.

(2) Determination of ABTS ⁺ · free radical scavenging ability

Measure 5ml of 2.6mmol/L K2S2O8 solution and 10ml of 7.4mmol/L ABTS+ solution, mix them well, store them for 12h at room temperature and avoid light, dilute 40 to 50 times, and use phosphate buffer ( pH 6.6) Dilute the ABTS+ solution to the absorbance of 0.70±0.02 to obtain the working solution. Take 40 μL of curcumin anti-aging beverage sample solution in a 96-well plate, add 160 μL of ABTS ⁺ · working solution, mix and shake well and let stand in the dark for 6 min, then measure the absorbance at a wavelength of 734 nm. The scavenging rate of each sample to be tested to ABTS ⁺ · radical was calculated according to the following formula. Using distilled water as a blank control, measure the absorbance A0. Ascorbic acid was used as a positive control.

The calculation formula is: clearance rate (%)=[1-(A1-A2)/A0]×100%

where:

A0 is the absorbance value after the reaction of 40 μL of distilled water and 160 μL of ABTS+ solution;

A1 is the absorbance value after 40 μL of sample solution reacted with 160 μL of LBTS+ solution;

A2 is the absorbance value after mixing 40 μL of sample solution with 160 μL of absolute ethanol.

2.6 Comprehensive scoring method

A comprehensive scoring method was used to determine the formulation of curcumin anti-aging beverage. In this experiment, sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH·free radical scavenging ability, and ABTS ⁺ · free radical scavenging ability are determined, and these five are evaluation indicators. Total score = sensory evaluation score x sensory evaluation weight + light absorption stability score x light absorption stability weight + centrifugal sedimentation rate score x centrifugal sedimentation rate weight + DPPH·radical scavenging rate score x DPPH·radical scavenging rate weight+ ABTS ⁺ · + radical scavenging rate score x ABTS ⁺ · + radical scavenging rate weight. See Tables 1, 2, and 3 for the rank assignments, ranking scores and weight assignments below.

Table 13 Grade Allocation Table

Table 14 Ranking score table

Table 15 Weight distribution table

2.7 Detection of curcumin content

2.7.1 Standard solution configuration

Accurately weigh 5 mg of curcumin standard, dilute to 100 mL with methanol, and prepare a standard solution of 0.05 mg/mL respectively. Accurately measure 0.2, 0.4, 0.8, 1, and 2ml of the standard solution into a 10ml volumetric flask, then dilute to volume with methanol, filter through a microporous membrane, and store the filtrate in a 5°C refrigerator away from light.

2.7.2 Sample preparation

Put 5mL of sample in a 50mL centrifuge tube, add 25mL of 90% ethanol, extract with ultrasonic for 15min, then centrifuge at 4000r/min for 15min, take the supernatant into a 50mL volumetric flask, dilute to volume with 90% ethanol, and use 0.45μm The filter membrane filters the sample to be tested for analysis.

2.7.3 Chromatographic conditions

Chromatographic column: Hypersil ODS C18 (150nm×4.6nm, 5μm); Mobile phase: A: methanol, B: 5% glacial acetic acid buffer (A:B=65:35V/V); flow rate: 1mL/min; detection wavelength : 420 nm; the column temperature was 30 °C, and the injection volume was 20 μL.

2.7.4 HPLC analysis

According to the chromatographic conditions in 2.7.3, carry out HPLC analysis, take the mass concentration of curcumin as the abscissa and the peak area value as the ordinate to create a linear equation, draw a standard curve, and bring the sample peak area into the standard curve to obtain turmeric element content.

3. Single factor test of curcumin beverage formula

3.1 Effect of CMC-Na addition on the quality of curcumin beverages

Add 0.05%, 0.1%, 0.15%, 0.2%, 0.25% of CMC-Na in turn, the addition of curcumin nanoparticles is 0.4%, the addition of sodium starch octenyl succinate is 0.4%, and the addition of xanthan gum is 0.1%, the addition amount of xylitol is 10%, and the addition amount of citric acid is 0.1%. According to the above preparation process, sensory evaluation, light absorption stability, centrifugal sedimentation rate, scavenging DPPH · free radical capacity, ABTS ⁺ · free radical The scavenging ability is the investigation index, and the optimal addition amount of CMC-Na is determined. The results are shown in the following table.

Table 16 Effect of CMC-Na addition on the quality of curcumin beverages

It can be seen from Table 16 that with the increase of the addition amount of CMC-Na, the sensory score shows a trend of first increasing and then decreasing. When the addition amount of CMC-Na is 0.1%, the sensory score is the highest. This is because the added amount of CMC-Na is 0.1%, the beverage has no stratification, no precipitation, and has the best taste. As the amount thereof increases, the taste gradually deteriorates. And when the addition amount of CMC-Na is 0.1%, the solution system is in a stable state, the absorption stability coefficient is the highest and the centrifugal sedimentation rate is the lowest. The sedimentation rate increases. It can be seen from Fig. 11 that the addition amount of CMC-Na has little effect on DPPH·radical scavenging rate% and ABTS ⁺ ·radical scavenging rate%. Combining Table 16 and Figures 11 and 12, when the addition amount of CMC-Na is 0.1%, the comprehensive score is the highest, so it is determined that the optimum addition amount of CMC-Na is 0.1%.

3.2 The effect of xanthan gum addition on the quality of curcumin beverages

Add 0.05%, 0.1%, 0.15%, 0.2%, 0.25% of xanthan gum in sequence, the amount of curcumin nanoparticles is 0.4%, the amount of sodium starch octenyl succinate is 0.4%, and the amount of CMC-Na is 0.1%, the addition amount of xylitol is 10%, and the addition amount of citric acid is 0.1%. According to the above preparation process, sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH free radical scavenging ability, ABTS free radical scavenging The ability is the inspection index, and the optimal amount of xanthan gum added is determined. The results are shown in the following table.

Table 17 Effect of xanthan gum addition on the quality of curcumin beverages

It can be seen from Table 17 that with the increase of the addition amount of xanthan gum, the sensory score shows a trend of first increasing and then decreasing. When the addition amount of xanthan gum is 0.15%, the sensory score is the highest. This is because the addition of xanthan gum is 0.15%, the beverage has no stratification, no precipitation, and has the best taste. As the addition amount increases, the beverage becomes viscous and affects the taste. And when the addition amount of xanthan gum is 0.15%, the solution system is in a stable state, the absorption stability coefficient reaches the highest, and the centrifugal sedimentation rate is the lowest. The sedimentation rate increases. It can be seen from FIG. 13 that the addition amount of xanthan gum has little effect on DPPH·free radical scavenging rate% and ABTS ⁺ ·radical scavenging rate %. Combining Table 17, Figure 13 and Figure 14, when the addition amount of xanthan gum is 0.15%, the comprehensive score is the highest, thus it is determined that the optimum addition amount of xanthan gum is 0.15%.

3.3 Effect of sodium starch octenyl succinate addition on curcumin beverage quality

The additions were set to five levels of 0.2%, 0.4%, 0.6%, 0.8%, and 1.0% respectively, the addition of curcumin nanoparticles was 0.4%, the addition of CMC was 0.1%, the addition of xanthan gum was 0.1%, and the addition of wood was 0.4%. The addition amount of sugar alcohol is 10%, and the addition amount of citric acid is 0.1%. According to the above preparation process, sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH·free radical scavenging ability, and ABTS ⁺ · free radical scavenging ability are investigated. index to determine the optimal addition amount of sodium starch octenyl succinate, the results are shown in the following table.

Table 18 The effect of adding sodium starch octenyl succinate on curcumin beverage

It can be seen from Table 18 that with the increase of the addition amount of sodium starch octenyl succinate, the sensory score shows a trend of first increasing and then decreasing. When the addition amount of sodium starch octenyl succinate is 0.4%, the sensory score is the highest. This is because when the sodium starch octenyl succinate is 0.4%, the odor of sodium starch alkenyl succinate and curcumin are the most abundant in the beverage. When the amount of sodium starch octenyl succinate continued to increase, the smell of sodium starch octenyl succinate was too strong, covered the smell of curcumin, and the taste decreased. Therefore, the sensory score decreased. The highest absorption stability coefficient and centrifugal sedimentation rate decreased and increased with the addition of sodium starch octenyl succinate, respectively. It can be seen from FIG. 15 that the addition amount of sodium starch octenyl succinate slightly increases DPPH·free radical scavenging rate %, but has little effect on ABTS ⁺ · free radical scavenging rate %. Combining Table 18, Figure 15 and Figure 16, when the addition amount of starch sodium octenyl succinate is 0.4%, the comprehensive score is the highest, so it is determined that the optimum addition amount of starch sodium octenyl succinate is 0.4%.

3.4 Effect of curcumin nanoparticles addition on the quality of curcumin beverages

Curcumin nanoparticles were set to five levels of 0.2%, 0.4%, 0.6%, 0.8% and 1.0%, the addition of sodium starch octenylsuccinate was 0.4%, the addition of CMC was 0.1%, and the addition of xanthan gum It is 0.1%, the addition amount of xylitol is 10%, and the addition amount of citric acid is 0.1%. According to the above preparation, sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH free radical scavenging ability, ABTS ⁺ free radical The scavenging ability is the investigation index, and the optimal addition amount of curcumin nanoparticles is determined. The results are shown in the following table.

Table 19 Effect of curcumin nanoparticle addition on the quality of curcumin beverages

It can be seen from Table 19 that with the increase of the addition amount of curcumin nanoparticles, the sensory score showed a trend of first increasing and then decreasing. When the addition amount of curcumin nanoparticles was 0.8%, the sensory score was the highest. This is because the odor of curcumin in the beverage is the strongest when the addition of curcumin nanoparticles is 0.8%. When the addition amount exceeds 0.8%, the smell of curcumin is too strong, which affects the sensory evaluation. The highest absorption stability coefficient and centrifugal sedimentation rate decreased and increased with the addition of curcumin nanoparticles, respectively. It can be seen from FIG. 17 that DPPH·free radical scavenging rate % and ABTS ⁺ · free radical scavenging rate % increase with the increase of the addition amount of curcumin nanoparticles. Combining Table 19, Figure 17 and Figure 18, when the addition amount of curcumin nanoparticles is 0.8%, the comprehensive score is the highest, thus it is determined that the optimum addition amount of curcumin nanoparticles is 0.8%.

4. Orthogonal optimization experiment of curcumin beverage formula

4.1 Orthogonal experimental method

Based on the single-factor experimental results, the addition amount of sodium carboxymethyl cellulose (A), the addition amount of xanthan gum (B), the addition amount of curcumin freeze-dried powder (C), the addition amount of sodium starch octenyl succinate (D) is the experimental factor, taking sensory evaluation, light absorption stability coefficient, centrifugal sedimentation rate, DPPH free radical scavenging ability, and ABTS ⁺ free radical scavenging ability as the inspection indicators, the comprehensive scoring method is adopted, and L ₉ (3 ⁴ ) positive The formula of curcumin beverage for anti-aging was optimized by cross-experiment, and the design table is shown below.

Table 20 Level design table of orthogonal experimental factors for curcumin beverage formula L ₉ (3 ⁴ )

Table 21. Scores of the inspection indicators in the experimental group

4.2 Orthogonal experimental results and analysis

Table 22 L ₉ (3 ⁴ ) orthogonal test results

The orthogonal experimental design was carried out with the comprehensive scoring method. The results are shown in Table 22. From the R value, it can be seen that the primary and secondary order of the influence of each factor on the curcumin anti-aging beverage is D>C>B>A, that is, curcumin nanoparticles The addition amount of freeze-dried powder was the main influencing factor, followed by the addition amount of sodium starch octenyl succinate, and again the addition amount of xanthan gum, and the addition amount of CMC-Na had the least effect. From the K value, it can be seen that the best solution is A ₃ B ₂ C ₂ D ₂ , that is, the addition amount of CMC-Na is 0.15%, the addition amount of xanthan gum is 0.15%, the addition amount of sodium starch octenyl succinate is 0.4%, The addition amount of curcumin nanoparticles is 0.8%.

The variance analysis of the orthogonal test results is shown in the table below.

Table 23 Orthogonal test analysis of variance

The results are shown in Table 23. The results show that: curcumin nanoparticles and sodium starch octenyl succinate are significant factors affecting the curcumin anti-aging drink, and the curcumin nanoparticles are more significant than sodium starch octenyl succinate, However, CMC-Na and xanthan gum were not significant factors.

4.3 Verification experiment

The orthogonal optimal group is A ₃ B ₂ C ₂ D ₂ . The verification test was repeated three times. The results are shown in Table 24. The comprehensive score of curcumin anti-aging beverage is 98.64 points, and the test results are stable and reliable, so A ₃ B ₂ C ₂ D ₂ is an excellent combination with a high comprehensive score.

Table 24 Verification test

In conclusion, the optimal formula of curcumin beverage is: 0.15% xanthan gum, 0.15% CMC-Na, 0.4% sodium starch octenyl succinate, 0.8% curcumin nanoparticle freeze-dried powder, 0.1% citric acid addition amount and 10% xylitol, under this condition, the product has the highest comprehensive evaluation score, which is 98.64±0.05; sensory score is 93.33±0.24, light absorption stability coefficient is 0.518±0.005, centrifugal sedimentation rate is 37.8%±0.4%, curcumin The scavenging rate of DPPH·free radicals was 82.86%±0.00%, and the scavenging rate of ABTS+·free radicals was 85.33%±0.00%. Curcumin beverages had certain antioxidant properties. The curcumin content in the product is 4mg/mL. The drink is orange-yellow, sweet and sour, and has the characteristic smell of curcumin.

4.4 Comparative test

Preparation and orthogonal experiments used curcumin combined with the same concentration of zinc ions, nano-particle curcumin and pure curcumin, and used it as a control. Compared with the beverage made in the orthogonal optimal group (A ₃ B ₂ C ₂ D ₂ ), the sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH·free radical scavenging ability, and ABTS ⁺ · free radical scavenging ability were compared. . Compare the changes of various indicators after making curcumin beverages. The results are shown in the table below.

Table 25 Comparative test

It can be seen from Table 25 that the pure curcumin solution, combined with Zn ²⁺ curcumin solution, curcumin nanoemulsion, and anti-aging curcumin beverage, the DPPH free radical scavenging rate % and ABTS ⁺ free radical scavenging rate % decreased in turn, while The stability increased in turn, while the pure curcumin solution, Zn ²⁺ curcumin solution, and curcumin nanoparticle solution were precipitated due to the insolubility of curcumin in water, which affected the tissue morphology, and the curcumin smell was too strong, affecting the sensory score, because curcumin The solubility of nanoparticles in aqueous solution is greatly enhanced, which is beneficial to the sensory evaluation of beverages. Therefore, the comprehensive score of curcumin beverage for anti-aging is the highest, which is 98.64 points.

5. Research on the efficacy of anti-aging curcumin drink

Taking Drosophila as a model, the survival test indicators of Drosophila, such as half days of death, average lifespan and average maximum lifespan, were used to reflect the efficacy of curcumin beverages in prolonging the lifespan of Drosophila; by analyzing the changes in the levels of biochemical indicators SOD and MDA in Drosophila, curcumin was preliminarily explored. The mechanism of action of beverages to delay aging.

5.1 Preparation of Drosophila Medium

Basic medium: Liquid A: 20 g of sucrose, 3 g of agar, 300 mL of distilled water, fully stir to dissolve, heat and boil until the agar is completely melted. Liquid B: 33 g of corn flour, 21.24 g of glucose, 4.5 g of soybean flour, and 200 mL of distilled water, fully stir and mix; slowly pour liquid B into liquid A, and keep stirring, boil to a paste, stop heating, and use Dissolve 0.125g of paraben in 1.25mL of absolute ethanol; when the temperature of the mixture of A and B drops to about 75°C, add 12.5g of yeast powder, stir well and add 3.5mL of propionic acid, stir well Immediately pour it into a clean culture tube, the height of each tube is 1.5-2mL, and after it is completely solidified, the culture medium can be inoculated into the fruit fly after 24 hours.

5.2 Curcumin Beverage Dose Gradient Selection

According to the model required by the State Food and Drug Administration for health food, the recommended dose of human (0.0033g/kg·bw·d), the daily amount of food and drinking water for a person weighing 60kg is set as 3000g, to calculate the experimental intermediate concentration , 1 to 2 concentration groups were set up and down by 3 times the group distance above and below this concentration, namely four dose groups, and a blank control group was set up.

5.3 Drosophila survival test method

Collect 300 male and female Drosophila newly emerged within 8 hours, distinguish male and female with ether anesthesia, and randomly divide them into 5 groups, 60 males and 60 males in each dose group. In a constant temperature incubator with a relative humidity of 45% to 75% and a temperature of 25±1°C, the medium was changed every four days to prevent the flies from dying due to food adhesion. The survival activity and the number of dead flies were observed every day until all the flies died. And calculate the average lifespan, the average maximum lifespan (calculated by the lifespan of the last 10 fruit flies that died) and the half number of dead days and other indicators for statistics.

5.4 Data Analysis

Statistical analysis was performed on the data using SPSS17 statistical analysis software. P<0.01 and P<0.05 were considered to be statistically significant.

5.5 Analysis of experimental results

5.5.1 The effect of curcumin drink on the lifespan of Drosophila

The efficacy of curcumin beverages in prolonging the lifespan of Drosophila was reflected by indicators such as the half number of dead days, the average lifespan and the average maximum lifespan. The results are shown in Table 26.

Table 26 Effects of curcumin drink on the life span of Drosophila

*. The significance level of the mean difference is 0.05, and the extremely significant level is 0.01.

It can be seen from the results in Table 26 that there are significant differences in the average lifespan and the average maximum lifespan between the blank group and the dose group. When the addition amount of curcumin drink was 0.0201%, both the average lifespan and the average maximum lifespan of male and female fruit flies reached the highest value.

Among female flies, the average lifespan and average maximum lifespan of the 0.0201% dose group were 43.87±7.97 and 52.01±1.23, respectively, which were higher than those in the blank group, and significantly higher than those in the other groups, and their lifespan extension rates reached 13.95% and 8.81%, respectively. , 0.0022%, 0.0067% and 0.0603% of the three groups, the average maximum lifespan were higher than the blank group; in male Drosophila, the average lifespan and the average maximum lifespan of the 0.0067% and 0.0201% dose groups were 38.83±9.77 and 48.20±1.29, respectively were higher than the blank group, with an average life extension rate of 14.17% and 9.10% and 16.67% and 10.80%. The average maximum life expectancy of the 4 dose groups was significantly higher than that of the blank group. There were also significant differences in maximum lifespan. Therefore, the curcumin beverage described in this example has the effect of prolonging the lifespan of Drosophila.

5.5.2 The effect of curcumin beverage on the activity of Drosophila antioxidant enzymes

Table 27 Effects of curcumin drink on the activity of Drosophila antioxidant enzymes

*. The significance level of the mean difference is 0.05, and the extremely significant level is 0.01.

It can be seen from the results in Table 27 that there are differences in SOD activity and MDA content between the dose group and the blank group. When the concentration of curcumin drink increased, the SOD activity in male and female flies also increased. When the concentration of curcumin drink was 0.0201%, the SOD activity in male and female flies reached the highest value. The MDA content in male and female flies decreased with the increase in the concentration of curcumin beverage in the medium. When the concentration of curcumin drink was 0.0201%, the MDA content in male and female flies reached the minimum value. Therefore, in female Drosophila, the SOD activity in the 0.0201% dose group was significantly higher than that in other groups, while the MDA content was significantly lower than that in other dose groups. In male Drosophila, the SOD activity and MDA content in the 0.0201% dose group were significantly different from those in the blank group.

In the fly Drosophila survival experiment, for male and female Drosophila, their average lifespan, average maximum lifespan, and half-death time were prolonged with the increase of curcumin drink concentration, reaching the maximum in the 0.0201% dose group, and higher than other dose groups and blank group, curcumin beverage can prolong the lifespan of Drosophila and has anti-aging effect; with the increase of curcumin beverage addition, SOD enzyme activity increases, and MDA content decreases, when the addition amount of curcumin beverage is 0.0201%, SOD enzyme activity the highest and the lowest MDA content. It indicated that curcumin beverage played an anti-aging effect through the internal mechanism of increasing SOD enzyme activity and reducing malondialdehyde MDA content.

The present invention is not limited to the above-mentioned embodiments. If various changes or deformations of the present invention do not depart from the spirit and scope of the present invention, and if these changes and deformations belong to the claims of the present invention and the equivalent technical scope, then the present invention is also It is intended that these modifications and variations are included.

Claims (9)

1. The curcumin nanoparticle is characterized in that the nanoparticle takes curcumin as a core material and Arabic gum and zein as wall materials; the mass ratio of the core material to the wall material is 5.5-7.5: 100, respectively; the mass ratio of the Arabic gum to the zein is 1-5: 5.

2. The method for preparing curcumin nanoparticles as claimed in claim 1, comprising the steps of:

1) preparation of an alcohol phase: dissolving zein in 85% ethanol, magnetically stirring for 1h, centrifuging to remove insoluble substances to obtain zein solution, adding curcumin in the zein solution, and stirring for 30min to obtain alcohol phase;

2) preparation of the aqueous phase: dissolving Arabic gum in 0.3g/L zinc sulfate solution, stirring at 60 ℃ until the Arabic gum is completely dissolved to obtain a water phase, wherein the volume ratio of the water phase to the alcohol phase in the step 1) is 1.5-3.5: 1;

3) anti-solvent coprecipitation: adding the alcohol phase obtained in the step 1) into the water phase obtained in the step 2) in a fine stream state, and stirring for 30min to obtain a curcumin nanoparticle dispersion liquid;

4) rotary evaporation: removing ethanol and water from the curcumin nanoparticle dispersion liquid obtained in the step 3) through rotary evaporation to obtain a curcumin nanoparticle concentrated solution;

5) and (3) drying: freeze-drying the curcumin nanoparticle concentrated solution obtained in the step 4) to obtain curcumin nanoparticles;

the added amounts of curcumin, acacia and zein are added according to the mass ratio of claim 1.

3. The method for preparing curcumin nanoparticles as claimed in claim 2, wherein the volume ratio of the aqueous phase in step 2) to the alcohol phase in step 1) is 2: 1; the mass ratio of the Arabic gum to the zein is 4: 5; the mass ratio of the core material to the wall material is 7.5: 100.

4. a curcumin beverage for delaying aging is characterized by comprising the following components in percentage by mass: 15% of emperor orange concentrated solution, 0.05-0.25% of xanthan gum, 0.2-1% of curcumin nanoparticles, 0.2-1% of sodium starch octenylsuccinate, 0.05-0.25% of sodium carboxymethylcellulose, 0.1% of citric acid, 10% of xylitol and the balance of water; the curcumin nanoparticles are obtained by the preparation method of claim 3.

5. A curcumin beverage for delaying aging as claimed in claim 4, wherein said beverage comprises, in mass percent: 15% of emperor orange concentrated solution, 0.15% of xanthan gum, 0.8% of curcumin nanoparticles, 0.4% of sodium starch octenyl succinate, 0.15% of sodium carboxymethyl cellulose, 0.1% of citric acid, 10% of xylitol and the balance of water.

6. A method for preparing a curcumin beverage for delaying aging as set forth in claim 4, comprising the steps of:

1) preparing an emperor orange concentrated solution: peeling and removing kernels of the emperor oranges, juicing by using a juicer, filtering twice by using double-layer gauze, and concentrating the juice to 1/2 of the original volume to obtain an emperor orange concentrated solution;

2) mixing: adding the curcumin nanoparticles and sodium starch octenyl succinate in percentage by mass into the mixture under high-speed stirring, and mixing the mixture with the citrus emperor concentrated solution;

3) homogenizing: pouring the mixed solution prepared in the step 2) into a high-pressure homogenizer for homogenization;

4) blending: adding the sodium carboxymethylcellulose, the xanthan gum, the citric acid and the xylitol into the product obtained in the step 3) in percentage by mass;

5) and (3) secondary homogenization: pouring the mixed solution prepared in the step 4) into a high-pressure homogenizer for homogenization;

6) degassing, namely heating the product obtained in the step 5) by using a water bath to perform degassing treatment;

7) filling and sterilizing: filling and sterilizing the product obtained in the step 6) while the product is hot to obtain the curcumin beverage for delaying aging.

7. The method for preparing a curcumin beverage for delaying aging as set forth in claim 6, wherein the homogenizing pressure in the step 3) is 25 Mpa.

8. The method for preparing a curcumin beverage for delaying aging as set forth in claim 6, wherein the pressure of the secondary homogenization in the step 5) is 5 Mpa.

9. The method for preparing a curcumin beverage for delaying aging as set forth in claim 6, wherein said sterilization condition in the step 7) is sterilization at 121 ℃ for 10 min.

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