CN111657415A - Curcumin nanoparticle and preparation method and application thereof - Google Patents
Curcumin nanoparticle and preparation method and application thereof Download PDFInfo
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- CN111657415A CN111657415A CN202010501007.7A CN202010501007A CN111657415A CN 111657415 A CN111657415 A CN 111657415A CN 202010501007 A CN202010501007 A CN 202010501007A CN 111657415 A CN111657415 A CN 111657415A
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- curcumin
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- zein
- nanoparticles
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract
The invention provides a curcumin nanoparticle, which 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. The invention also comprises a preparation method of the curcumin nanoparticles and a beverage prepared from the curcumin nanoparticles and capable of delaying aging. The preparation method of the curcumin nano-particles improves the problem of light instability in the solution, so that the curcumin nano-particles can be used as functional pigment to be added into functional beverage to prepare the anti-aging functional beverage. The curcumin beverage provided by the invention is palatable in sour and sweet taste, has curcumin smell, can improve SOD enzyme activity, reduces malonaldehyde MDA content, and has antioxidant and anti-aging effects.
Description
Technical Field
The invention relates to the technical field of functional food production processes, and particularly relates to curcumin nanoparticles and a preparation method and application thereof.
Background
Curcumin (curcumin, cur) is a polyphenol compound extracted from rhizome of Curcuma longa of Zingiberaceae, has a symmetrical molecular structure formed by connecting beta-diketone heptadiene and two o-methylated phenols, and has orange yellow crystal powder and bitter taste. Has strong tinting strength and low toxicity, and is widely applied to the fields of food, textile, cosmetics and the like as a common natural pigment for a long time. In recent years, researches of a plurality of scholars show that curcumin has a plurality of pharmacological and health-care effects of resisting inflammation, oxidation, tumors, depression, viruses, cardiovascular protection, brain injury repair, Parkinson symptom relief and the like. However, curcumin is insoluble in water, soluble in organic solvents such as ethanol, acetic acid, acetone, chloroform and the like, and is easily influenced by external factors such as light, temperature, metal ions, pH value and the like, so that the curcumin easily loses the color development capability, and particularly has high photosensitivity. The King Chimonanthus nitens and the like research the light stability of the curcumin compounds, and analyze the curcumin compound solution before and after illumination by adopting a high performance liquid chromatography, and find that the curcumin solution is extremely unstable under outdoor illumination, and the curcumin solid powder is relatively stable.
In recent years, much reports have been made on curcumin nanocrystallization research, Liuyanmin and the like prepare curcumin solid dispersions by a method of melting rapid cooling and room temperature cooling, and the in-vitro dissolution rate of the curcumin solid dispersions is found to be far higher than that of pure curcumin. The lyophilized powder of curcumin nanoparticles prepared by Gekko Swinhonis et al by anti-solvent method can improve curcumin water solubility, and is beneficial for improving curcumin bioavailability. In addition, the preparation method of curcumin nanoparticles such as liposome, micelle and microemulsion is also provided.
Curcumin has various pharmacological and health-care effects, people hope to apply curcumin as a functional pigment in functional food development, but curcumin is difficult to dissolve in water and is easily influenced by external factors such as light, temperature, metal ions, pH value and the like, so that the color development capability of curcumin is easily lost. Particularly, under outdoor illumination, the curcumin aqueous solution is unstable, has degradation reaction and loses the original pharmacological action and color, so that the application field of curcumin is limited, and the curcumin aqueous solution is especially suitable for industrial development and application in liquid food. At present, there are many research reports aiming at improving the solubility and bioavailability of curcumin, for example, after the curcumin is prepared into a nano liposome, the water solubility of the curcumin can be obviously improved, and the bioavailability is improved; the stability of the natural pigment is improved by adding a stabilizer, microencapsulation, pigment molecular structure modification, improvement of the processing and storage conditions of the natural pigment and the like, and the research on improving the photosensitivity of the natural pigment is rarely reported. With the rapid development of socioeconomic and rapid progress of science and technology, after the problem of satiety is solved, people's demand for beverages is no longer just for pursuing taste and vision, but now pay more attention to the nutrition and health of the beverages themselves, from the most original function of quenching thirst to the now pursuing nutrition, health and health. In recent years, a plurality of functional beverages with the characteristics of nutrition, health and the like, and multiple efficacies of anti-aging, beautifying and the like appear on the market, and are gradually favored by consumers. According to a large amount of data, the total yield and sales of health-care functional beverages of domestic beverage enterprises are increased year by year in recent years, and the development trend of the health-care functional beverages in China in the future is good from the long-term development trend. If the problem of photosensitivity of curcumin can be solved and the curcumin can be applied to beverages, the market of nutritional health-care beverages can be enriched, and the curcumin has high economic value.
Disclosure of Invention
A first object of the present invention is to solve the above-mentioned drawbacks and disadvantages of the prior art, and to provide curcumin nanoparticles and a method for preparing the same.
The invention is realized by the following technical scheme:
a curcumin nanoparticle comprises curcumin as core material, acacia and zein as wall material; 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.
The preparation method of the curcumin nanoparticles comprises the following steps:
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 (heated in water bath at 60 ℃) and stirring 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 addition amounts of the curcumin, the Arabic gum and the zein are added according to the mass ratio.
Further, the volume ratio of the aqueous phase of step 2) to the alcohol phase of 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.
the second purpose of the invention is to provide a curcumin beverage for delaying aging and a preparation method thereof.
The invention is realized by the following technical scheme:
a curcumin beverage for delaying senescence, which comprises 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 nano-particles are prepared by the preparation method.
Further, the beverage comprises the following components in percentage by mass: 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.
The preparation method of the curcumin beverage for delaying aging comprises the following steps:
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.
Further, the homogenizing pressure in the step 3) is 25 MPa.
Further, the pressure of the secondary homogenization in the step 5) is 5 MPa.
Further, the sterilization condition in the step 7) is sterilization at 121 ℃ for 10 min.
Compared with the prior art, the invention has the following advantages:
1. the preparation method of the curcumin nano-particles improves the problem of light instability in the solution, so that the curcumin nano-particles can be used as functional pigment to be added into functional beverage to prepare the anti-aging functional beverage.
2. The curcumin beverage provided by the invention is palatable in sour and sweet taste, has curcumin smell, can improve SOD enzyme activity, reduces malonaldehyde MDA content, and has antioxidant and anti-aging effects.
For a better understanding and practice, the invention is described in detail below with reference to the accompanying drawings.
Drawings
Fig. 1 is a curcumin absorption curve.
Fig. 2 is a curcumin standard curve.
FIG. 3 is a screening of the optimum concentration of zinc ions.
FIG. 4 is the volume ratio of the aqueous phase to the alcohol phase.
FIG. 5 is the mass ratio of acacia gum to zein.
Fig. 6 is the curcumin to zein-gum arabic mass ratio.
Fig. 7 shows curcumin retention of curcumin nanoparticles and pure curcumin in natural light.
FIG. 8 is a photograph taken by 100-fold microscopy under the condition of experiment 1.
FIG. 9 is a photograph taken under the conditions of experiment 2 by 100 times microscope.
FIG. 10 is a photograph taken under the conditions of experiment 3 by 100 times microscope.
FIG. 11 is a graph showing the effect of the addition of CMC-Na on the radical scavenging rate.
FIG. 12 is the effect of CMC-Na addition on curcumin beverages.
FIG. 13 is a graph of the effect of xanthan gum addition on radical scavenging.
Fig. 14 is a graph of the effect of xanthan gum addition on curcumin beverages.
FIG. 15 is a graph of the effect of sodium starch octenylsuccinate addition on radical clearance.
Fig. 16 is a graph of the effect of sodium starch octenyl succinate addition on curcumin beverages.
Fig. 17 is the effect of curcumin lyophilized powder addition on free radical clearance.
FIG. 18 shows the effect of the addition of lyophilized curcumin powder on the quality of curcumin beverages.
Detailed Description
The materials and reagents used in the present invention are shown in table 1.
TABLE 1 materials and reagents used for the assay
The equipment used in the present invention is shown in table 2:
TABLE 2 summary of the equipment used in the tests
The data processing software adopted by the invention is GraphPad Prism 7.04, Origin 9, EZ OMNIC and orthogonal design assistant respectively.
All materials, reagents and equipment selected for use in the present invention are well known in the art, but do not limit the practice of the invention, and other reagents and equipment well known in the art may be suitable for use in the practice of the following embodiments of the invention.
Example 1
Preparation test method of curcumin nanoparticles
1. Curcumin feedstock analysis
1.1 curcumin maximum absorption wavelength and regression equation
Maximum absorption wavelength: accurately weighing 0.01g of curcumin powder sample in a 100ml volumetric flask, fixing the volume with 95% ethanol to obtain a curcumin solution with the concentration of 0.1mg/ml, transferring 500ul of the curcumin solution from the curcumin solution to fix the volume to 100ml with the 95% ethanol solution to obtain the curcumin solution with the concentration of 0.5ug/ml, adjusting the wavelength to measure the light absorption value of the curcumin under different wavelengths, and determining the maximum absorption wavelength by taking the wavelength as the abscissa and the absorbance as the ordinate.
The regression equation: accurately weighing 25mg of curcumin standard substance into a 50mL volumetric flask, fixing the volume with 95% absolute ethyl alcohol, diluting to 0, 15, 30, 45, 60 and 75ug/mL standard solutions respectively, measuring absorbance at 425nm, and performing linear regression on absorbance Y by using concentration X.
As can be seen from fig. 1 and 2: the maximum absorption wavelength of curcumin is 425nm, the regression equation of the standard curve is that Y is 0.3168x +0.6349, R2The curcumin concentration is 0.9974, and the linear relation is good, wherein the curcumin concentration is in a range of 0-75 mu g/mL.
1.2 curcumin mineral element content analysis
A microwave digestion method comprises the following steps: refer to GB 5009.268-2016 determination of multiple elements in food; weighing 0.3g curcumin solid (0.2824 g is actually weighed) into a microwave digestion inner tank, adding 8mL nitric acid, covering and placing for 1h or overnight, screwing a tank cover, and digesting according to the standard operation steps of a microwave digestion instrument. After digestion, taking out after cooling, slowly opening the tank cover to exhaust, washing the inner cover with a small amount of water, placing the digestion tank on a temperature-control electric heating plate to discharge acid until the digestion tank is nearly dry, then adding ultrapure water to a constant volume in a 25mL volumetric flask, uniformly mixing for later use, and simultaneously performing blank tests with the same quantity.
As can be seen from table 3, the chemical composition of curcumin contains less zinc element, which has a certain effect on the stability of curcumin.
TABLE 3 curcumin mineral element content
2. Screening of optimum concentration of stabilizer zinc ion
2.1 preparation of the sample solution
Preparing a curcumin mother solution: 0.175g of curcumin is weighed and dissolved in 280 ml of absolute ethyl alcohol, and stirred until curcumin powder is completely dissolved.
Preparing stock solutions with different zinc ion concentrations: 0.03g, 0.06g, 0.09g, 0.12g, 0.15g and 0.18g of zinc sulfate are respectively weighed and dissolved in 100ml of distilled water, and are uniformly stirred to prepare zinc sulfate stock solutions of 0.3g/L, 0.6g/L, 0.9g/L, 1.2g/L, 1.5g/L and 1.8g/L, and the zinc sulfate stock solutions are well preserved for later use.
2.2 optimum concentration screening test for Zinc ion
The curcumin mother liquor is divided into 7 sample solutions of 40ml, 10ml,0.0g/L, 0.3g/L, 0.6g/L, 0.9g/L, 1.2g/L, 1.5g/L and 1.8g/L zinc sulfate stock solutions are respectively added into No. 1 to No. 7 sample solutions, the mixture is uniformly stirred to obtain the sample solutions with the zinc sulfate concentrations of 0.0g/L, 0.06g/L, 0.12g/L, 0.18g/L, 0.24g/L, 0.3g/L and 0.36g/L respectively, and the sample solutions are placed at room temperature. The absorbance was measured every 12 hours at a wavelength of 425nm with a microplate reader, and the average value was taken in triplicate.
2.3 analysis of results
As can be seen from FIG. 3, the curcumin solutions containing different concentrations of zinc ions all discolored within 0 to 48 hours, but were more stable than those without zinc ions. After 48 to 60 hours, the absorbance of the curcumin solution added with zinc ions with different concentrations is increased, the gradient of the zinc ion concentration of 0.3g/L is minimum and the absorbance is stable after 60 hours, and the zinc ion concentration of 0.36g/L is stable, but the overall trend still shows a decline. Therefore, the effect of improving the light stability of the curcumin is best when the zinc ion concentration is 0.3 g/L.
3. The curcumin nanoparticle preparation process comprises the following steps:
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 (heated in water bath at 60 ℃) and stirring 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.
3.1 Single-factor experiment for curcumin nanoparticle preparation
The embedding rate is used as an investigation index, and the influence of the volume ratio of the water phase to the alcohol phase, the mass ratio of the Arabic gum to the zein and the mass ratio of the curcumin to the zein-Arabic gum on the preparation of the curcumin nano-particles is analyzed, so that the optimal addition amount of the three factors is determined.
3.1.1 volume ratio of aqueous phase to alcohol phase
1g of zein, 0.02g of curcumin, 1g of Arabic gum, 50ml of 85% ethanol, 0.3g/L of zinc sulfate (calculated by total volume), and the volume ratio of distilled water to ethanol is 1.5: 1. 2: 1. 2.5: 1. 3: 1. 3.5:1, preparing curcumin nanoparticles according to the process operation, and determining the optimal proportion of the volume of the water phase and the alcohol phase according to the embedding rate.
As can be seen from fig. 4, the volume ratio of the aqueous phase to the alcohol phase is from 1.5:1 to 2.5: at 1, the embedding rate increased from 85.4% to 93.2%, and when the volume ratio increased from 2.5:1 to 3.5:1, the embedding rate decreased from 93.2% to 84.6%. Probably, the concentration of the alcohol phase is reduced along with the increase of the volume ratio of the water phase to the alcohol phase, the curcumin is dissolved to reach a supersaturated state, and during the deposition process, the curcumin is separated out to cause the increase of the liberation and the reduction of the embedding rate. The volume ratio of the optimum aqueous phase to the alcohol phase was determined to be 2.5: 1.
3.1.2 weight ratio of Arabic gum to zein
1g of zein, 0.02g of curcumin, 75ml of distilled water, 50ml of 85% ethanol and 0.3g/L of zinc sulfate (calculated by total volume), wherein the mass ratio of the Arabic gum to the zein is 2:10, 4:10, 6:10, 8:10 and 10:10, curcumin nanoparticles are prepared by the process, and the optimal mass ratio of the Arabic gum to the zein is determined according to the embedding rate.
As can be seen from fig. 5, the embedding rate increased first and then decreased as the mass ratio of acacia gum to zein increased, and when the mass ratio was 6:10, the embedding rate was the largest and 87.1%, and at this time, the charges of acacia gum and zein were completely neutralized, and the embedding rate was the largest, and then the increase in the ratio resulted in charge imbalance and the embedding rate decreased. The optimal mass ratio of the Arabic gum to the zein is determined to be 6: 10.
3.1.3 Total Mass ratio of curcumin to zein-Arabic gum
1g of zein, 1g of Arabic gum, 50ml of 85% ethanol, 0.3g/L of zinc sulfate (calculated by total accumulation), 75ml of distilled water, and the total mass ratio of the curcumin to the zein-Arabic gum is 5.5:100, 6:100, 6.5:100, 7:100 and 7.5:100, preparing curcumin nanoparticles according to the process, and determining the optimal proportion of the curcumin to the total mass of the zein-Arabic gum according to the embedding rate.
As can be seen from fig. 6, when the mass ratio of curcumin to zein-gum arabic is from 5.5:100 to 6.5:100, the embedding rate is increased from 97.1% to 97.6%, and when the ratio is from 6.5:100 to 7.5: at 100, the embedding rate decreases. It is likely that when the mass ratio of curcumin to zein-gum arabic is 6.5:100, the loading of the wall material reaches the maximum load, and then the increase in the ratio leads to an increase in the amount of free curcumin and a decrease in the encapsulation rate. The optimal mass ratio of curcumin to zein-Arabic gum is determined to be 6.5: 100.
3.2 Quadrature experiments for curcumin nanoparticles preparation
3.2.1 orthogonal Experimental methods
Based on single-factor experimental investigation, L9 (3) is adopted4) The orthogonal table optimizes three factors of the volume ratio of the water phase to the alcohol phase, the mass ratio of wall material components (the mass ratio of the Arabic gum to the zein) and the mass ratio of the core wall (the mass ratio of the curcumin to the total mass of the zein and the Arabic gum), and is shown in the following table; the embedding rate, the light stability and the grain diameter are divided into investigation indexes, and a multi-index comprehensive balance method is selected. The larger the light stability (1- (a7-a1)/a7) and the larger the particle size (1-particle size), the better.
Table 4 curcumin nanoparticles preparation L9 (3)4) Orthogonal test factor level design table
3.2.1.1 determination of curcumin nanoparticle embedding rate
Transferring 1mL of curcumin-loaded nanoparticles and 4mL of 95% ethanol, placing in a beaker, performing ultrasonic extraction for 15min to obtain free curcumin, filtering with a 0.22 μm organic membrane, and repeatedly extracting for 3 times. Combining the extracts, taking 95% ethanol as a blank control, measuring the absorbance of the sample at 425nm, calculating the free curcumin according to a standard curve equation of the curcumin in the 95% ethanol, and calculating the embedding rate and the drug loading rate according to the following formula:
3.2.1.2 curcumin nanoparticle size determination
The sample is diluted by a certain factor, placed in a sample cell, and the particle size is measured using a laser particle sizer.
3.2.1.3 observations of light stability of curcumin nanoparticles
Diluting the sample by 4 times, standing at room temperature, measuring the absorbance value once every 24 hours by using an enzyme-labeling instrument, and observing the influence of the microcapsule technology on curcumin photosensitivity. And respectively drawing retention rate graphs of a (zein-Arabic gum-curcumin) nano system and non-embedded curcumin by taking the reaction time t as an abscissa and the retention rate ln (At/A0) as an ordinate, and observing the influence of the curcumin nanoparticles on curcumin photosensitivity.
Sample grouping conditions: sample 1 was prepared by the process conditions of 2:1 water-alcohol volume ratio (water phase to alcohol phase volume ratio), 4:10 wall material mass ratio (acacia to zein mass ratio), 6:100 core wall mass ratio (curcumin to zein-acacia total mass ratio); the sample 2 has the process conditions that the water-alcohol volume ratio is 2:1, the wall material mass ratio is 6:10, and the core wall mass ratio is 6.5: 100; the sample 3 has the process conditions of the hydroalcoholic volume ratio of 2:1, the wall material mass ratio of 8:10 and the core wall mass ratio of 7: 100; sample 4 was prepared by the process conditions of 2.5:1 hydroalcoholic volume ratio, 4:10 wall material mass ratio, 7:100 core wall mass ratio; sample 5 was prepared by the process conditions of 2.5:1 hydroalcoholic volume ratio, 6:10 wall material mass ratio, 6:100 core wall mass ratio; sample 6 was prepared by the process conditions of a hydroalcoholic volume ratio of 2.5:1, a wall material mass ratio of 8:10, and a core wall mass ratio of 6.5: 100; sample 7 was prepared by a process with a hydroalcoholic volume ratio of 3:1, a wall material mass ratio of 4:10, and a core wall mass ratio of 6.5: 100; the sample 8 has the process conditions of the hydroalcoholic volume ratio of 3:1, the wall material mass ratio of 6:10 and the core wall mass ratio of 7: 100; sample 9 was prepared by the process conditions of a hydroalcoholic volume ratio of 3:1, a wall material mass ratio of 8:10, and a core wall mass ratio of 6: 100; blank is 0.02g of pure curcumin.
As can be seen from fig. 7, in samples 1 to 9, after 48 hours, the retention rates of all samples tend to be stable, and the retention rate of curcumin of the curcumin nanoparticles is greater than that of non-embedded samples, so that the photolysis reaction of curcumin embedded by the curcumin nanoparticles is slower than that of non-embedded samples. Sample 7, which had a water-alcohol volume ratio (water phase to alcohol phase volume ratio) of 3:1, a wall material mass ratio (acacia to zein mass ratio) of 4:10, and a core wall mass ratio (curcumin to total mass ratio of acacia to zein) of 6.5:100, had the greatest curcumin retention rate after 144h, and the curcumin photolysis reaction was the slowest.
From the above, the photolysis reaction of curcumin nanoparticles is slower than that of non-embedded curcumin, and the stability of curcumin embedded to light is improved.
3.2.2 orthogonal experimental results are as follows:
table 5 curcumin nanoparticles preparation process L9(34) Table of results of orthogonal tests
3.2.3 survey index ANOVA results are as follows:
TABLE 6 ANOVA result Table with embedding rate as investigation index
TABLE 7 ANOVA RESULT TABLE USING PARTICULAR SIZE AS A TEST INDICATOR
TABLE 8 ANOVA result Table with absorbance degradation rate as investigation index
Note: significance P <0.05
TABLE 9 optimal combination comparison table for different indexes
As can be seen from tables 5, 6, 7, 8 and 9, the factor A has a highly significant influence on the embedding rate, and according to the R value, A is preferred when the embedding rate is used as an index1(ii) a Factor B has high significant influence on the embedding rate, and B is selected according to the R value3The factor C has no obvious influence on the three indexes, but the prepared nano particles have smaller particle size, and C is selected according to the R value3. The optimal level of curcumin nanoparticle preparation is A through comprehensive equilibrium analysis1B3C3The volume ratio of the water phase to the alcohol phase is 2:1, the mass ratio of the Arabic gum to the zein is 8:10, and the mass ratio of the curcumin to the zein-Arabic gum is 7.5:100, and the curcumin nanoparticle embedding rate is 96.730%, the drug loading rate is 62mg/g, the particle size is 0.940um and the absorbance decrease slope is 0.73 under the optimal action condition obtained by an empirical test.
Example 2
Curcumin beverage development and test method for delaying aging
1. The preparation process of the curcumin beverage comprises the following steps:
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; the curcumin nanoparticles were prepared from example 1;
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.
1.1 determination of the homogenization pressure
The number of homogenization times was two, and the beverages were homogenized at a homogenization pressure of 25MPa, a secondary homogenization at 5MPa, and a primary homogenization pressure of 5MPa and a secondary homogenization pressure of 25MPa, respectively, and the influence of the homogenization pressure on the particle size was examined using the non-homogenized beverage as a control, and the results of the homogenization pressure are shown in the table below.
TABLE 10 homogeneous pressure results
According to the particle size, the experiment 3 is more than 2 and more than 1, and the micro-photographing picture is combined (shown in the attached figure 8-10), when the primary pressure is 25MPa and the secondary homogenizing pressure is 5MPa, large particles are more fully crushed into small molecular substances, the storage stability of the beverage is facilitated, when the primary pressure is 5MPa and the secondary pressure is 25MPa, the large substances can be crushed, but the effect is not the same as the homogenizing condition of the experiment 1 but is better than the effect of non-homogenizing. Therefore, the primary homogenizing pressure is 25MPa, and the secondary homogenizing pressure is 5 MPa.
1.2 determination of Sterilization mode
The curcumin beverage has pH of 6, is low acidic food, and can be sterilized at high temperature and high pressure for 10min at 121 deg.C, or added with citric acid to acidify the beverage, adjust its pH to 4.6, pasteurize at 80 deg.C for 20min, and cold sterilize by filtering with 0.22 μm organic membrane, and using unsterilized beverage as control. The sterilization conditions were determined using DPPH ability to scavenge free radicals as an index.
1.3 preparation of analytical samples
Beverage sample liquid: the curcumin beverage is prepared by taking curcumin nanoparticle freeze-dried powder and sodium starch octenyl succinate as raw materials and xanthan gum and CMC-Na as stabilizers according to the process, wherein the curcumin content in the beverage is 100 mg/mL. Then diluting with distilled water to the concentration of 20, 40, 60, 80 and 100 mu g/mL;
weighing 2, 4, 6, 8 and 10mg of ascorbic acid by taking an ascorbic acid solution as a positive control, dissolving the ascorbic acid with distilled water and fixing the volume to 100mL, wherein the concentration of the ascorbic acid solution is 20, 40, 60, 80 and 100 mu g/mL.
TABLE 11 Sterilization conditions
From Table 11, sample IC50The value is 2 > 3 > 1 > 4, Vc positive control IC50The value is 2 > 1 > 3 > 4, IC50The lower the value, the higher the DPPH.radical scavenging ratio, and the better the oxidation resistance of the sample, so that the oxidation resistance of sample 1 was the highest. IC with three sterilization modes after sterilization50The values are all rising, while sample 2 has IC because of acidification, pH 4.6, and inhibition of superoxide anion by acid radical ion50The highest value, the lowest DPPH-free radical clearance rate and the lowest inoxidizability. According to the national food safety standard (beverage) of GB7101-2015, the total number of bacterial colonies of samples 1, 2 and 3 after sterilization all accord with the national standard, and the total number of bacterial colonies of samples without sterilization do not accord with the national safety standard. The above results were combined to obtain the experimentAdopts 121 ℃/10min as the sterilization condition.
2. Evaluation method of curcumin beverage quality
2.1 sensory Scoring Standard
The sensory evaluation of the experiment consists of 8 members, and all indexes of the curcumin beverage are subjected to sensory taste scoring.
TABLE 12 sensory Scoring standards
2.2 light absorption stability
Weighing a certain amount of sample, adding water to dilute by 20 times, mixing the diluent uniformly, putting the mixture into a centrifuge to centrifuge for 15min at 4000r/min, measuring the light absorption values Ao and A before and after centrifugation of the sample at the maximum absorption wavelength of the sample, and expressing the light absorption stability of the sample to be measured by A/Ao.
2.3 centrifugal sedimentation Rate
Respectively weighing the total weight of the sample, the centrifuge tube and the centrifuge tube, centrifuging for 15min at 4000r/min, discarding the supernatant, and accurately weighing the total weight of the centrifuge tube and the residual precipitate.
The calculation formula is as follows: the precipitation rate is equal to the mass of the precipitate/the mass of the sample solution multiplied by 100 percent
2.4 measurement of DPPH.radical scavenging ability
(1) Sample preparation, same as 1.3.
(2) Measurement of DPPH radical scavenging ability
100. mu.L of 0.2mmol/L ethanol DPPH was added to each 96-well plate. Adding 100 μ L curcumin beverage sample solutions with different concentration gradients, mixing, reacting at room temperature in the lacquer black for 30min, measuring absorbance A1 at 517nm, replacing DPPH ethanol solution (A2) with equal volume of anhydrous ethanol as control group, replacing sample solution (A0) with equal volume of distilled water as control group, and using ascorbic acid as positive control.
The calculation formula is as follows: clearance [% 1- (A1-A2)/A0] x 100
In the formula:
a0: absorbance of a mixed solution of 100. mu.L of distilled water and 100. mu.L of ethanol LLDPPH;
a1: absorbance of a mixture of 100. mu.L of the sample solution and 100. mu.L of the ethanol solution of LDPPH;
a2: absorbance of a mixture of 100. mu.L of the sample solution and 100. mu.L of absolute ethanol.
2.5 ABTS+Determination of the radical scavenging ability
(1) Sample preparation, same as 1.3.
(2)ABTS+Determination of the radical scavenging ability
Measuring 5ml of 2.6mmol/L K2S2O8 solution and 10ml of 7.4mmol/L ABTS +. solution, mixing uniformly, placing at room temperature under the condition of keeping out of the sun for reaction for 12 hours, diluting by 40-50 times, and diluting the ABTS +. solution by phosphate buffer solution (pH6.6) until the absorbance is 0.70 +/-0.02 to obtain working solution. Adding 40 μ L curcumin aging delaying beverage sample liquid into 96-well plate, adding ABTS +160. mu.L of the working solution was mixed, shaken well, and left standing for 6min in the dark, and then the absorbance was measured at a wavelength of 734 nm. ABTS of each sample pair to be tested was calculated according to the following formula+Clearance of free radicals. The absorbance A0 was measured using distilled water as a blank. Ascorbic acid was used as a positive control.
The calculation formula is as follows: the clearance (%) [1- (a1-a2)/a0] x 100%
In the formula:
a0 is the absorbance value after 40. mu.L of distilled water reacts with 160. mu.L of ABTS +. solution;
a1 is the absorbance value after 40. mu.L of sample solution reacts with 160. mu.L of LABTS +. solution;
a2 is the absorbance value of 40. mu.L of sample mixed with 160. mu.L of absolute ethanol.
2.6 comprehensive scoring method
The formula of the curcumin beverage for delaying aging is determined by adopting a comprehensive grading method. In this experiment, sensory evaluation, light absorption stability, centrifugal precipitation rate, DPPH-free radical scavenging ability, ABTS, were determined+The radical scavenging ability, and these five are evaluation indices. 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 clearance score xDPPH-radical clearance weight + ABTS+' free radical cleanerDivide ratio score x ABTS+· + radical clearance weight. The following are the rank assignments, rank scores and weight assignments in tables 1, 2 and 3.
TABLE 13 grade Allocation Table
TABLE 14 ranking score Table
Table 15 weight assignment table
2.7 curcumin content detection
2.7.1 Standard solution preparation
Accurately weighing 5mg curcumin standard substance, diluting to 100mL with methanol, and respectively preparing into 0.05mg/mL standard solution. Accurately weighing standard solutions 0.2 ml, 0.4 ml, 0.8 ml, 1ml and 2ml in a 10ml volumetric flask, then fixing the volume with methanol, filtering with a microporous membrane, collecting filtrate, and storing in a refrigerator at 5 ℃ in a dark place.
2.7.2 sample preparation
Putting 5mL of sample into a 50mL centrifuge tube, adding 25mL of 90% ethanol, extracting for 15min by ultrasonic waves, centrifuging for 15min at 4000r/min, taking supernatant into a 50mL volumetric flask, fixing the volume by 90% ethanol, and filtering the sample to be tested by a filter membrane of 0.45 mu m for analysis.
2.7.3 chromatographic conditions
A chromatographic column: hypersil ODS C18(150 nm. times.4.6 nm, 5 μm); mobile phase: a: methanol, B: 5% glacial acetic acid buffer (a: B ═ 65: 35V/V); flow rate: 1 mL/min; detection wavelength: 420 nm; the column temperature was 30 ℃ and the amount of sample was 20. mu.L.
2.7.4 HPLC analysis
And (3) performing HPLC analysis according to the chromatographic conditions in 2.7.3, establishing a linear equation by taking the mass concentration of the curcumin as a horizontal coordinate and the peak area value as a vertical coordinate, drawing a standard curve graph, and substituting the peak area of the sample into the standard curve to obtain the curcumin content.
3. Curcumin beverage formula single factor test
3.1 Effect of CMC-Na addition on curcumin beverage quality
Sequentially adding 0.05%, 0.1%, 0.15%, 0.2% and 0.25% of CMC-Na, wherein the addition amount of curcumin nanoparticles is 0.4%, the addition amount of sodium starch octenylsuccinate is 0.4%, the addition amount of xanthan gum is 0.1%, the addition amount of xylitol is 10%, and the addition amount of citric acid is 0.1%, and performing the preparation process according to the preparation process to evaluate the appearance, the light absorption stability, the centrifugal sedimentation rate, the DPPH-free radical scavenging capacity and ABTS+The radical scavenging ability was used as an index for investigation, and the optimum CMC-Na addition amount was determined, and the results are shown in the following table.
TABLE 16 influence of CMC-Na addition on curcumin beverage quality
As is clear from Table 16, the sensory score tended to increase and decrease with increasing CMC-Na addition, and the sensory score was the highest at 0.1% CMC-Na addition. This is because the addition amount of CMC-Na is 0.1%, and the beverage has no demixing and precipitation, and has the best taste. As the amount thereof increases, the taste becomes gradually poor. And when the addition amount of the CMC-Na is 0.1%, the solution system is in a stable state, the light absorption stability coefficient is highest and the centrifugal sedimentation rate is lowest, and as the addition amount of the CMC-Na is continuously increased, the stable state of the solution is destroyed, the light absorption stability coefficient is reduced and the centrifugal sedimentation rate is increased. As can be seen from FIG. 11, the amount of CMC-Na added was found to have a degree of DPPH.radical scavenging% and ABTS+The% radical clearance had little effect. When the CMC-Na addition amount is 0.1%, the comprehensive score is the highest by combining the table 16, the graph 11 and the graph 12, thereby determining that the optimal addition amount of the CMC-Na is 0.1%.
3.2 Effect of Xanthan Gum addition on curcumin beverage quality
0.05%, 0.1%, 0.15%, 0.2% and 0.25% of xanthan gum is sequentially added, the addition amount of curcumin nanoparticles is 0.4%, the addition amount of sodium starch octenylsuccinate is 0.4%, the addition amount of CMC-Na is 0.1%, the addition amount of xylitol is 10%, the addition amount of citric acid is 0.1%, the preparation process is carried out according to the above, sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH-free radical scavenging capacity and ABTS-free radical scavenging capacity are considered indexes, the optimal xanthan gum addition amount is determined, and the results are shown in the following table.
TABLE 17 influence of Xanthan Gum addition on curcumin beverage quality
As can be seen from table 17, as the amount of added xanthan gum increased, the sensory score tended to increase first and then decrease, and the sensory score was the highest at 0.15% added xanthan gum. This is because the addition amount of xanthan gum is 0.15%, the beverage is free from separation and sedimentation, and the taste is excellent, and as the addition amount increases, the beverage becomes viscous, which affects the taste. And when the addition amount of the xanthan gum is 0.15%, the solution system is in a stable state, the light absorption stability coefficient reaches the highest, and the centrifugal sedimentation rate is the lowest. As can be seen from FIG. 13, the amount of xanthan gum added was found to have a% DPPH.radical scavenging rate and ABTS+The% radical clearance had little effect. From the above table 17, fig. 13 and fig. 14, the overall score was highest when the xanthan gum was added at 0.15%, thereby determining that the optimum amount of xanthan gum was added at 0.15%.
3.3 Effect of sodium starch octenylsuccinate addition on curcumin beverage quality
The addition is respectively set to five levels of 0.2%, 0.4%, 0.6%, 0.8% and 1.0%, the addition amount of curcumin nanoparticles is 0.4%, the addition amount of CMC is 0.1%, the addition amount of xanthan gum is 0.1%, the addition amount of xylitol is 10% and the addition amount of citric acid is 0.1%, the preparation process is carried out according to the above preparation process,sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH & free radical scavenging ability, ABTS+The free radical scavenging ability was used as an index for investigation to determine the optimum amount of starch sodium octenylsuccinate to be added, and the results are shown in the following table.
TABLE 18 Effect of sodium starch octenyl succinate addition on curcumin beverages
As can be seen from table 18, as the addition amount of starch sodium octenylsuccinate increased, the sensory score tended to increase and then decrease, and the sensory score was the highest when the addition amount of starch sodium octenylsuccinate was 0.4%. This is because the odor of sodium starch octenyl succinate and the odor of curcumin are the most abundant in the beverage when the sodium starch octenyl succinate is 0.4%. When the amount of starch sodium octenyl succinate continues to increase, the starch sodium octenyl succinate has an excessively strong odor, masks the curcumin odor, and has a reduced mouthfeel. So sensory score is reduced. And the highest light absorption stability coefficient and the centrifugal sedimentation rate respectively decrease and increase along with the increase of the addition amount of the starch sodium octenyl succinate. As can be seen from FIG. 15, the amount of sodium starch octenylsuccinate added was slightly increased in DPPH.radical scavenging percentage and in ABTS+The% radical clearance had little effect. In combination with table 18, fig. 15 and fig. 16, the best overall score was obtained when the addition amount of starch sodium octenylsuccinate was 0.4%, thereby determining that the optimum addition amount of starch sodium octenylsuccinate was 0.4%.
3.4 influence of curcumin nanoparticle addition on curcumin beverage quality
Curcumin nanoparticles are set to five levels of 0.2%, 0.4%, 0.6%, 0.8% and 1.0% in sequence, the addition amount of starch sodium octenylsuccinate is 0.4%, the addition amount of CMC is 0.1%, the addition amount of xanthan gum is 0.1%, the addition amount of xylitol is 10%, the addition amount of citric acid is 0.1%, the preparation is carried out according to the preparation method, and sensory evaluation is carried outValence, light absorption stability, centrifugal sedimentation rate, DPPH & free radical scavenging ability, ABTS+The addition amount of the optimal curcumin nanoparticles was determined by using the radical scavenging ability as an index, and the results are shown in the following table.
TABLE 19 influence of curcumin nanoparticle addition on curcumin beverage quality
As can be seen from table 19, as the added amount of curcumin nanoparticles increases, the sensory score tends to increase first and then decrease, and when the added amount of curcumin nanoparticles is 0.8%, the sensory score is the highest. This is because curcumin smell is the strongest in the beverage when the added amount of curcumin nanoparticles is 0.8%. When the addition amount exceeds 0.8%, the curcumin has excessively strong odor, which affects sensory evaluation. And the highest light absorption stability coefficient and the centrifugal sedimentation rate respectively decrease and increase along with the increase of the addition amount of the curcumin nanoparticles. FIG. 17 shows the% DPPH.radical scavenging and ABTS+The% radical clearance increased with increasing curcumin nanoparticle addition. By combining table 19, fig. 17 and fig. 18, the composite score was highest when the curcumin nanoparticle addition amount was 0.8%, thereby determining that the optimum curcumin nanoparticle addition amount was 0.8%.
4. Quadrature optimization experiment for curcumin beverage formula
4.1 orthogonal Experimental method
Based on the results of the single-factor experiment, the additive amounts of sodium carboxymethylcellulose (A), xanthan gum (B), curcumin lyophilized powder (C) and sodium starch octenylsuccinate (D) are used as experimental factors, and sensory evaluation, light absorption stability coefficient, centrifugal sedimentation rate, DPPH-free radical scavenging capacity, ABTS are carried out+The radical scavenging ability is taken as an investigation index, a comprehensive grading method is adopted, and L is selected9(34) The formula of the curcumin beverage for delaying aging is optimized through orthogonal experiments, and the design table is shown as follows.
TABLE 20 curcumin beverage formula L9(34) Orthogonal experiment factor level design table
TABLE 21 test group survey index scores
4.2 orthogonal Experimental results and analysis
TABLE 22L9(34) Results of orthogonal experiments
The results of the orthogonal experimental design by the comprehensive scoring method are shown in Table 22, and the R values show that the main and secondary sequence of the influence degree of each factor on the curcumin aging-delaying beverage is D>C>B>A, namely the addition amount of curcumin nanoparticle freeze-dried powder is a main influence factor, the addition amount of octenyl succinic acid starch sodium is the addition amount of xanthan gum, and the influence of the addition amount of CMC-Na is the minimum. From the K value, the best solution is A3B2C2D2Namely, the addition amount of CMC-Na is 0.15 percent, the addition amount of xanthan gum is 0.15 percent, the addition amount of starch sodium octenyl succinate is 0.4 percent, and the addition amount of curcumin nano-particles is 0.8 percent.
The results of the cross-over experiments were analyzed for variance as shown in the following table.
TABLE 23 analysis of variance of orthogonal test
The results are shown in table 23 and show that: the curcumin nanoparticles and sodium starch octenyl succinate are significant influencing factors for the curcumin aging-delaying beverage, the curcumin nanoparticles are greater than the sodium starch octenyl succinate in significance, and CMC-Na and xanthan gum are insignificant factors.
4.3 validation experiments
The orthogonal optimal group is A3B2C2D2And the verification test is repeated for 3 times, the results are shown in Table 24, the comprehensive score of the curcumin aging-delaying beverage is 98.64 points, the test result is stable and reliable, and therefore, A3B2C2D2Is a better combination with high comprehensive score.
TABLE 24 verification test
In conclusion, the optimal formula of the curcumin beverage is as follows: 0.15% of xanthan gum, 0.15% of CMC-Na, 0.4% of starch sodium octenyl succinate, 0.8% of curcumin nanoparticle freeze-dried powder, 0.1% of citric acid addition and 10% of xylitol, wherein the product comprehensive evaluation score is the highest and is 98.64 +/-0.05 under the condition; the sensory score is 93.33 +/-0.24, the light absorption stability coefficient is 0.518 +/-0.005, the centrifugal sedimentation rate is 37.8% +/-0.4%, the DPPH-free radical clearance rate of the curcumin beverage is 82.86% +/-0.00%, the ABTS + -. free radical clearance rate is 85.33% +/-0.00%, and the curcumin beverage has certain oxidation resistance. The curcumin content in the product is 4 mg/mL. The beverage is orange yellow, has sour and sweet palatability, and has characteristic flavor of curcumin.
4.4 comparative test
Curcumin, nanoparticulate curcumin, pure curcumin combined with zinc ions at the same concentration were prepared for control in the orthogonal experiments. And orthogonal optimum group (A)3B2C2D2) The beverage is used as control, and evaluated from sensory evaluation, light absorption stability, centrifugal sedimentation rate, DPPH & free radical scavenging ability, ABTS+Comparison in radical scavenging ability. Comparing the changes of various indexes after preparing curcumin beverage. The results are shown in the following table.
TABLE 25 comparative test
As can be seen from Table 25, pure curcumin solution, combined with Zn2+Curcumin solution, curcumin nanoemulsion, curcumin beverage for delaying aging, DPPH-free radical clearance percent and ABTS+The% radical clearance decreases in turn and the stability increases in turn, while the pure curcumin solution, Zn2+The curcumin solution and the curcumin nanoparticle solution are separated out due to insolubility of curcumin in water, so that the tissue morphology is influenced, the curcumin smell is too strong, and the sensory evaluation is influenced, because the solubility of the curcumin nanoparticles in the water solution is greatly enhanced, the sensory evaluation of the beverage is facilitated. Therefore, the comprehensive score of the curcumin beverage for delaying senescence is the highest and is 98.64.
5. Efficacy research of aging-delaying curcumin beverage
The fruit fly is taken as a model, and the effect of prolonging the life of the fruit fly of the curcumin beverage is reflected by fruit fly survival experiment indexes such as half death days, average life, average maximum life and the like; the aging delaying action mechanism of the curcumin beverage is preliminarily researched by analyzing the level changes of biochemical indexes SOD and MDA in the drosophila.
5.1 preparation of Drosophila Medium
Basic culture medium: solution A: 20g of sucrose, 3g of agar and 300mL of distilled water are fully stirred and dissolved, and the mixture is heated and boiled until the agar is completely melted. And B, liquid B: 33g of corn flour, 21.24g of glucose, 4.5g of soybean meal and 200mL of distilled water, and fully stirring and mixing; slowly pouring the solution B into the solution A, continuously stirring, boiling to paste, stopping heating, and dissolving and adding 0.125g of p-hydroxybenzoate by using 1.25mL of absolute ethyl alcohol; when the temperature of the A, B mixed solution is reduced to about 75 ℃, adding 12.5g of yeast powder, fully stirring, adding 3.5mL of propionic acid, uniformly stirring, immediately pouring into a clean culture tube, wherein the height of each tube of culture medium is 1.5-2mL, completely solidifying, and inversely placing the culture medium in an operation platform for 24 hours to be inoculated with fruit flies.
5.2 curcumin beverage dose gradient selection
According to a model required by health food of the State food and drug administration, the recommended dosage of a person (0.0033g/kg · bw · d) and the daily food and drinking water amount of a person with the weight of 60kg are determined to be 3000g to calculate the experimental intermediate concentration, 1-2 concentration groups, namely four dosage groups, are respectively set according to 3 times of group distance above and below the concentration, and 1 blank control group is set.
5.3 Drosophila survival test method
Collecting 300 fruit fly males and females which are newly emerged within 8h, distinguishing the males and females by ether anesthesia, randomly dividing into 5 groups, 60 females in each dose group, subpackaging in test tubes of 2.5cm multiplied by 20cm, 20 females in each tube, placing in a constant temperature incubator with the relative humidity of 45-75% and the temperature of 25 +/-1 ℃, and replacing the culture medium once every four days to prevent fruit flies from being dead due to food adhesion. The survival activity and death number of the fruit flies are observed every day until all the fruit flies die. And calculating indexes such as average life, average maximum life (calculated by the life of 10 fruit flies dead at last), half death days and the like, and carrying out statistics.
5.4 data analysis
The data were statistically analyzed using SPSS17 statistical analysis software, and differences were judged to be statistically significant with P < 0.01 and P < 0.05.
5.5 analysis of the results of the experiment
5.5.1 Effect of curcumin beverages on Drosophila longevity
The effect of curcumin beverages on prolonging the life of fruit flies was reflected by the indexes of half the number of days to death, average life and average maximum life, and the results are shown in table 26.
TABLE 26 Effect of curcumin beverages on Drosophila longevity
*. mean difference was at a significance level of 0.05, and a very significant level of 0.01.
As can be seen from the results in table 26, there was a significant difference in the average lifetime and the average maximum lifetime between the blank and dose groups. When the added amount of the curcumin beverage is 0.0201%, the average life and the average maximum life of the male and female fruit flies reach the highest value.
In female fruit flies, the average life span and the average maximum life span of the fruit flies in 0.0201% of the dose group are respectively 43.87 +/-7.97 and 52.01 +/-1.23 which are higher than those in the blank group and are obviously higher than those in other groups, the life prolonging rate respectively reaches 13.95% and 8.81%, and the average maximum life spans of the three groups of 0.0022%, 0.0067% and 0.0603% are higher than those in the blank group; in male drosophila melanogaster, the average life span and the average maximum life span of the 0.0067% dose group and the 0.0201% dose group are respectively 38.83 +/-9.77 and 48.20 +/-1.29, higher than those of the blank group, the average life span rate is 14.17% and 9.10% and 16.67% and 10.80%, the average maximum life span of the 4 dose groups is obviously higher than that of the blank group, and the average maximum life span of the 0.0201% dose group is also obviously different from that of other groups. Therefore, the curcumin beverage described in the present example has the effect of prolonging the life of fruit flies.
5.5.2 Effect of curcumin beverages on Drosophila antioxidant enzyme Activity
TABLE 27 Effect of curcumin beverages on Drosophila antioxidant enzyme Activity
*. mean difference was at a significance level of 0.05, and a very significant level of 0.01.
As can be seen from the results in Table 27, the SOD activity and MDA content were different between the dose group and the blank group. The SOD activity in the female and male fruit flies is increased when the concentration of the curcumin beverage is increased, and the SOD activity in the female and male fruit flies reaches the highest value when the concentration of the curcumin beverage is 0.0201%. The MDA content in the male and female drosophila also decreases with the increase of the concentration of curcumin beverage in the culture medium. When the curcumin beverage concentration is 0.0201%, the MDA content in the male and female drosophila bodies reaches the minimum value. Therefore, in female drosophila, the SOD activity in 0.0201% of the dose groups was significantly higher than the other groups, while the MDA content was significantly lower than the other dose groups. In the male drosophila melanogaster, the SOD activity and MDA content in 0.0201% of the dose group are obviously different from those in the blank group.
In a survival experiment of drosophila melanogaster, for male and female drosophila melanogaster, the average life, the average maximum life and half death time of the male and female drosophila melanogaster are prolonged along with the increase of the concentration of the curcumin beverage, the curcumin beverage reaches the maximum in 0.0201% dosage group and is higher than other dosage groups and blank groups, the curcumin beverage can prolong the life of drosophila melanogaster and has the effect of delaying aging; along with the increase of the addition amount of the curcumin beverage, the SOD enzyme activity is improved, the MDA content is reduced, and when the addition amount of the curcumin beverage is 0.0201%, the SOD enzyme activity is the highest, and the MDA content is the lowest. The curcumin beverage plays a role in delaying aging through an internal mechanism of improving the SOD enzyme activity and reducing the MDA content.
The present invention is not limited to the above-described embodiments, and various modifications and variations of the present invention are intended to be included within the scope of the claims and the equivalent technology of the present invention if they do not depart from the spirit and scope of the present invention.
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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