CN116784475A - Compound functional food and preparation method - Google Patents

Abstract

The invention belongs to the field of food and discloses a compound functional food, which includes the following components by weight: 1.0-2.0 parts of nuciferine, 0.1-0.5 parts of quercetin, and 5.0-15.0 parts of chlorogenic acid. Charged nuciferine and quercetin-liposome solutions were prepared by encapsulating nuciferine and quercetin liposomes in chitosan/proanthocyanidin microgels through ion cross-linking method. Vitamin-liposomes and chlorogenic acid co-encapsulated microgels. It forms a hypolipidemic composite microgel co-loaded with nuciferine, quercetin-liposomes, and chlorogenic acid, which effectively protects nuciferine, quercetin, and chlorogenic acid from being destroyed by oral administration, thereby improving the performance of nuciferine. , bioavailability of quercetin and chlorogenic acid. This preparation has obvious synergistic effect and can also effectively reduce blood lipids and improve lipid metabolism.

Description

Compound functional food and preparation method

Technical field

The invention relates to the field of food, in particular to compound functional food containing nuciferine, chlorogenic acid and quercetin.

Background technique

Lipid metabolism disorder is a common type of lipid metabolic syndrome in today's society, and is the main cause of diseases such as atherosclerosis and fatty liver. At present, chemical drug intervention is mainly used in clinical practice, but the effect is not ideal and has significant toxic and side effects. Therefore, intervention from the perspective of daily diet is an important way to regulate lipid metabolism disorders in the body. At present, there are a large number of literatures reporting that plant-derived extracts have significant regulatory functions on lipid metabolism disorders. Their main functional components include flavones, flavonoids, saponins and alkaloids. Research mainly uses single components or single plant extracts for research. However, a single plant often has limited functions and is difficult to achieve the desired effect. Achieving functional synergy through the compounding of multiple medicinal and edible homologous ingredients has become a focus of attention in recent years. It is also in line with today's "whole food" and "compound" food" concept.

Lotus leaf (Nelumbinis folium), hawthorn (Crataegus pinnatifida Bunge) and Eucommia ulmoides Oliv (Eucommia ulmoides Oliv) are all important new food resources with homologous medicinal and food origins in my country. Lotus leaves are bitter and neutral in nature, and are related to the liver, spleen, stomach, and heart meridians. They have the functions of clearing away heat and reducing dampness, raising hair and clearing yang, cooling blood and stopping bleeding. Modern research shows that lotus leaves are rich in flavonoids and alkaloids. , Lotus leaf has high lipid-lowering effect and health care effect. Hawthorn is sour, sweet, and slightly warm in nature. It returns to the liver, spleen, and stomach meridians. It has the functions of digesting and strengthening the stomach, promoting qi and dissipating blood stasis, resolving turbidity, and lowering lipids. It contains a large amount of vitamins, organic acids, and flavonoids, mainly hypericin. , epicatechin, chlorogenic acid, maslinic acid, ursolic acid, etc. Eucommia ulmoides leaves are warm in nature and sweet in taste, and belong to the liver and kidney meridian. As a new resource food, it has the function of regulating hypertension, hyperlipidemia and hyperglycemia. Therefore, based on the advantages of food properties and tastes of lotus leaves, hawthorn and eucommia leaves, and studying their natural functional factor complex to synergistically regulate lipid metabolism function and its mechanism, it is necessary to study the homologous medicinal and food products of lotus leaves, hawthorn and eucommia leaves. Provide theoretical basis for functional food efficacy mechanism and new product development.

Contents of the invention

The technical problem to be solved by the present invention is that a single plant used for regulating lipid metabolism disorders has limited functions and is difficult to achieve the desired effect.

In order to solve the above technical problems, the present invention provides a new compound functional food including the following components by weight: 1.0-2.0 parts by weight of nuciferine, 0.1-0.5 parts by quercetin, and 5.0-15.0 parts by chlorogenic acid; Positively charged nuciferine and quercetin-liposome solutions were prepared by encapsulating nuciferine and quercetin liposomes in chitosan/proanthocyanidin microgels through ion cross-linking method. Quercetin-liposome and chlorogenic acid co-encapsulated microgel.

Preferably, it includes the following components by weight: 1.6 parts of nuciferine, 0.4 parts of quercetin, and 8 parts of chlorogenic acid.

The above-mentioned positively charged nuciferine and quercetin co-encapsulated liposomes include the following components by weight: 100-500 parts of neutral phospholipid, 1-100 parts of cholesterol, 1.0-2.0 parts of lotus leaf Alkali and quercetin The ratio of nuciferine to quercetin is 1:0.25.

The microgel is co-encapsulated with nuciferine, quercetin-liposome and chlorogenic acid, and also includes the following components by weight: 300-1000 parts of chitosan and 100-400 parts of proanthocyanidins.

Preferably, the Zeta potential of the positively charged nuciferine and quercetin-liposomes is 30-70mV;

Preferably, the particle size range of the positively charged nuciferine and quercetin-liposomes is 60-150nm;

Preferably, the Zeta potential of the microgel co-encapsulated by nuciferine, quercetin-liposome and chlorogenic acid is - (30-60) mV;

Preferably, the particle size range of the microgel co-encapsulated by nuciferine, quercetin-liposome and chlorogenic acid is 1-5um;

Preferably, the phospholipid is selected from soybean lecithin; the procyanidins are selected from proanthocyanidins with a purity of more than 95% extracted from grape seeds. A method for preparing a hypolipidemic composite chitosan/proanthocyanidin microgel co-encapsulated with nuciferine, quercetin-liposomes and chlorogenic acid, including the following steps:

1) Fully dissolve soy lecithin and cholesterol in a round-bottomed flask with an appropriate amount of volatile organic solvent, and stir evenly with a magnetic stirrer;

2) After fully dissolving nuciferine and quercetin in an appropriate amount of volatile organic solution, add it to the round-bottomed flask in step 1) above and continue stirring for 1 hour;

3) Rotary evaporate the solution in step 2) under reduced pressure at 30°C and 30 rpm, evaporate the organic solvent to dryness and burn a thin film on the round bottom;

4) Add buffer solution and bathe in 50°C water for 20 minutes to fully dissolve the film obtained in step 3);

5) Homogenize the fully dissolved solution in step 4) under an ice bath at 80% power and normal pressure for 1-5 minutes for 2-5 times to obtain a translucent nuciferine and quercetin-liposome solution;

6) Fully dissolve chitosan in a round-bottomed flask with glacial acetic acid, add the nuciferine and quercetin-liposome solution obtained in step 5), and stir magnetically for 0.5-2 hours until the mixture is complete;

7) After proanthocyanidins and chlorogenic acid are fully dissolved with an appropriate amount of water, add them to the round-bottomed flask in step 6), and continue magnetic stirring for 2-4 hours to obtain co-encapsulation of nuciferine, quercetin-liposomes, and chlorogenic acid. Anti-hyperlipidemic composite microgel.

The preparation method of the present invention adopts a thin film evaporation-ultrasonic homogenization-ionic cross-linking method. In the preparation method, an ultrasonic homogenization method is performed after thin film evaporation to obtain uniform and stable positively charged nuciferine and quercetin-liposomes. solution, and then encapsulate nuciferine and quercetin liposomes in chitosan/proanthocyanidin microgels through ion cross-linking method.

Preferably, the volatile organic solvent in step 1) is selected from chloroform, and the mass fraction of phospholipid-cholesterol and added chloroform is 1:10-1:30.

Preferably, the organic solvent in step 2) is methanol, and the mass fraction of nuciferine and quercetin to the added methanol is 1:0.25:50-1:0.25:100.

Preferably, the buffer in step 4) is 0.05M phosphate buffer with pH 7.2-7.4, and the added buffer volume is 40-80 ml.

Preferably, the concentration of glacial acetic acid added in step 6) is 1%.

The beneficial effects of the present invention are:

The present invention uses chitosan nanoparticles to encapsulate functional factors, and prepares a new type of multifunctional factor co-encapsulated with chitosan nanoparticles for use in the food industry. Therefore, screening and preparing microgels from natural polymers is of great significance for the development of modern new foods and the application of food additives. At the same time, exploring and elucidating the compound action mechanism, physical and chemical characteristics, gastrointestinal release and digestion rules of functional factors with different physical and chemical properties will have a profound impact on the protection and development of new food resources and food-borne natural active ingredients. Therefore, based on the preparation and characterization of chitosan nanoparticles co-loaded with phylline, chlorogenic acid, and quercetin, the present invention carries out research on the mechanism of multifunctional factors cooperatively regulating lipid metabolism, with a view to providing edible polymers and natural functional factors. It provides certain reference for the research and exploration of compounding mechanism.

The three different natural active factors nuciferine, chlorogenic acid and quercetin selected in the present invention target different action targets, thereby achieving a synergistic effect.

The present invention first encapsulates nuciferine and quercetin in liposomes, and then co-encapsulates the prepared co-encapsulated liposomes and chlorogenic acid into chitosan/proanthocyanidin microgel, which not only affects lotus leaves Alkali and quercetin play a dual protective and sustained-release role, and also play the role of non-contact and sequential release of the three functional factors to further strengthen the synergistic effect.

The present invention studies an effective hypolipidemic synergistic complex by studying nuciferine, quercetin and chlorogenic acid, which have clear hypolipidemic effects, and prepares them into nuciferine, quercetin-liposomes and chlorogenic acid. The co-loaded hypolipidemic composite microgel protects nuciferine, quercetin and chlorogenic acid from being destroyed by oral administration, thereby improving the bioavailability of nuciferine, quercetin and chlorogenic acid and their use in dosage forms flexible application. The synergistic effect of this preparation can also effectively lower blood lipids and improve lipid metabolism.

The invention protects nuciferine from being destroyed by gastric acid in the stomach, improves its oral bioavailability, and solves the problem of poor solubility of nuciferine and quercetin. They are almost insoluble in water, difficult to administer, and have a single dosage form. question, which is of great significance for the in vitro stability and in vivo application of the functional ingredients nuciferine and quercetin.

The invention provides an idea and method for the synergistic compounding of natural active factors, can realize sequential release and sustained release functions for the encapsulated material, and can be applied to the encapsulation and compounding of various natural factors.

Description of the drawings

Figure 1 shows the effect of nuciferine, quercetin-liposome, and chlorogenic acid co-encapsulated microgel on the body weight of rats;

Figure 2 shows the effect of nuciferine, quercetin-liposome, and chlorogenic acid co-encapsulated microgel on serum biochemical indicators in rats;

Figure 3 shows the effect of nuciferine, quercetin-liposome, and chlorogenic acid co-encapsulated microgel on the expression of related genes in rats.

Detailed ways

The present invention will be further described below in conjunction with the accompanying drawings and specific embodiments. It should be understood that these examples are only used to illustrate the invention and are not intended to limit the scope of the invention. In addition, it should be understood that after reading the content described in the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of this application.

The natural active factor for lowering blood lipids of the present invention is selected based on the main pathways of lipid metabolism. The specific instructions are as follows:

Nuciferine, Nuciferine (NF) extracted from lotus, is an isoquinoline alkaloid with a molecular formula of C19H21NO2 and a relative molecular weight of 295.4. It is not only an indicated component in lotus leaves, but also an important component in producing pharmacological effects. Quercetin and chlorogenic acid are the two most common components in natural plants. They belong to natural flavonoids and organic acids respectively. These two natural functional factors can significantly regulate lipid metabolism and achieve the effect of lowering blood lipids.

Through the compounding of the above three ingredients, it can basically target the main action pathways of lowering blood lipids, relieve inflammation, resist oxidation, regulate intestinal microecology, and inhibit lipid synthesis and absorption, thereby achieving a synergistic effect in lowering blood lipids. The present invention will be further described below with reference to specific embodiments, but is not limited thereto.

Example 1

Accurately weigh 0.1g soy lecithin into a 250ml round-bottomed flask, add 15ml chloroform, stir magnetically until dissolved; add 0.02g cholesterol after dissolution, then add 10ml chloroform, continue stirring until completely dissolved; accurately weigh 0.04g Dissolve nuciferine and 0.01g quercetin in 4 ml of methanol, add it to a round-bottomed flask and stir evenly; spin evaporate the organic solvent to dryness at 40 rpm and 30°C. After forming a light yellow film, add 50 ml of 0.05 mol/L, pH = 7.4 PBS solution, let stand for 10 minutes, dissolve the film, and bathe in 50°C water for 20 minutes. After the film is dissolved, ultrasonically crush it for 6 minutes under ice bath conditions to obtain a translucent liposome solution; weigh 400 mg of proanthocyanidins and 1600 mg of chlorogenic acid and dissolve it in 50 ml. In water, you can add no more than 1% ethanol to help dissolve, use a magnetic stirrer to stir until all is dissolved, use a 5ml syringe to filter the solution with a 0.45um inorganic filter membrane, and obtain a proanthocyanidin solution with a concentration of 8mg/ml for later use; weigh 400mg Dissolve chitosan in 50ml of 1% glacial acetic acid solution, stir with a magnetic stirrer until all is dissolved, and obtain a chitosan solution with a concentration of 8mg/ml; take 20ml of the prepared nuciferine and quercetin-liposome solution Add 15 ml of the chitosan solution prepared above, stir continuously for 1 hour with a magnetic stirrer until completely mixed and dissolved, let stand at room temperature for 1 hour, and eliminate foam. Add 5 ml of the above-mentioned proanthocyanidin solution (containing 160 mg of chlorogenic acid) to the mixed solution. ), and continued stirring for 2 hours in a magnetic stirrer to prepare chitosan/proanthocyanidin microgel co-encapsulated with nuciferine, quercetin-liposomes and chlorogenic acid.

The encapsulation rates of nuciferine, quercetin and chlorogenic acid by the nanocarriers were 90.16%, 88.98% and 83.25%, respectively. The chromatographic analysis conditions for nuciferine are: Shimadzu SHIMADAZU 20AT high performance liquid chromatography system, Zhejiang University Zhida N2000 chromatography workstation, the chromatographic column is Agilent ODS C18 column (5μm, 250mm×4.6mm), and the guard column ODS C18 (4.6mm ×12.5mm), the mobile phase is acetonitrile:0.1% triethylamine aqueous solution=60:40; the column temperature is 30°C, the wavelength is 270nm; the flow rate is 1.0mL·min-1; the injection volume is 20μL. The obtained standard curve is: y=13121x-25393 (R ² =0.9999), and the linear range is 2.0-500.0ug/ml.

The chromatographic analysis conditions of quercetin were: Shimadzu SHIMADAZU 20AT high performance liquid chromatography system, Zhejiang University Zhida N2000 chromatography workstation, the chromatographic column was Discovery ODS C18 column (5μm, 250mm×4.6mm), and the guard column ODS C18 (4.6mm× 12.5mm), the mobile phase is methanol:0.1% phosphoric acid solution=60:40; the column temperature is 30°C, the wavelength is 360nm; the flow rate is 1.0mL·min-1; the injection volume is 20μL. The obtained standard curve is: y=13121x-25624 (R ² =0.9999), and the linear range is 2.0-500.0ug/ml.

The chromatographic analysis conditions of chlorogenic acid were: Shimadzu SHIMADAZU 20AT high performance liquid chromatography system, Zhejiang University Zhida N2000 chromatography workstation, the chromatographic column was Discovery ODS C18 column (5 μm, 250 mm × 4.6 mm), and the guard column ODS C18 (4.6 mm × 12.5mm), the mobile phase is acetonitrile:0.1% phosphoric acid solution=20:80; the column temperature is 30°C, the wavelength is 327nm; the flow rate is 1.0mL·min-1; the injection volume is 20μL. The obtained standard curve is: y=3094.9x-505.75 (R ² =0.9999), and the linear range is 2.0-500.0ug/ml.

The effect experiment is as follows:

Serum biochemical index evaluation

The lipid-lowering compound chitosan/proanthocyanidin microgel co-encapsulated with nuciferine, quercetin-liposome, and chlorogenic acid was evaluated for its lipid-lowering effect in a hyperlipidemia rat model.

Thirty rats were randomly divided into 7 groups, respectively set as blank control group (NC), high-fat model group (HFD), nuciferine microgel + high-fat group (NF), quercetin microgel + high-fat Lipid group (HF), chlorogenic acid microgel + high fat group (LF), nuciferine + quercetin + chlorogenic acid co-encapsulated microgel + high fat group (NF+HF+LF), shell In the polysaccharide/proanthocyanidin microgel + high-fat group (MG), after one week of adaptive breeding in the laboratory, except for the normal control group, which continued to be given basic feed, the other five groups were given high-fat feed, and the high-fat feed formula was 78.8% basic. Feed, cholesterol 1%, lard 10%, egg yolk powder 10% and cholate 0.2%. The temperature of the animal room is 23±2°C and the relative humidity is 40%-60%. Drinking water and food are available freely. Body weight changes are measured weekly and the dosage is adjusted appropriately. The experimental animals were continuously gavaged and anesthetized with ether. Blood was collected from the choroid plexus vein of the eye, the serum was separated, and the changes in serum lipid composition at different time points were detected using corresponding kits. At the same time, after intragastric administration at the 8th week, all experimental animals were anesthetized and killed, and blood was collected from the heart, and the serum was separated for content detection of lipid components such as relevant biochemical indicators (TC, TG, LDL-C and HDL-C), and collected Animal liver tissue was stored at -80°C for molecular expression detection. The results of the changes in rat weight are shown in Figure 1, the results of serum lipid levels of rats in different treatment groups are shown in Figure 2, and the expression levels of liver lipid metabolism genes in rats in different treatment groups are shown in Figure 3.

The results in Figure 1 show that the blank microgel group, nuciferine microgel group, quercetin microgel group, chlorogenic acid microgel group, and nuciferine + quercetin + chlorogenic acid co-encapsulated Compared with the high-fat model group, the microgel group could significantly reduce the body weight of rats. However, the nuciferine + quercetin + chlorogenic acid co-encapsulated microgel group had the most obvious weight reduction. The results in Figure 2 show that the NF, HF, LF, NF+HF+LF, and MG groups can significantly reduce the levels of TC, TG, and LDL-C in rat serum compared with the HFD group, and nuciferine + quercetin The pigment + chlorogenic acid co-encapsulated microgel group had the most obvious weight loss, and the contents of TC, TG, and LDL-C were significantly different from the MG group. The results in Figure 3 show that the NF, HF, LF, NF+HF+LF groups can significantly reduce the expression levels of PPAR-α and CYP7A1 in rat livers compared with the HFD group, among which nuciferine + quercetin The +chlorogenic acid co-encapsulated microgel group reduced body weight most significantly;

The preparations made in this research are mainly used for the research, development and production of functional foods, and the formula of the present invention is pure natural active ingredients and pure natural extracts, does not contain any toxic ingredients, is safe and effective within the dosage range, and provides information for the development of health food. A new way of thinking and approach.

The above embodiments are preferred embodiments of the present invention, but the implementation of the present invention is not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, etc. may be made without departing from the spirit and principles of the present invention. All simplifications should be equivalent substitutions, and are all included in the protection scope of the present invention.

Finally, it should be noted that the above examples are only specific examples of the compound preparation composed of nuciferine, quercetin and chlorogenic acid in the present invention. Obviously, the present invention is not limited to the above embodiments, and many modifications are possible. All modifications that a person of ordinary skill in the art can directly derive or associate from the disclosure of the present invention should be considered to be within the protection scope of the present invention.

Claims (10)

1. The compound functional food is characterized by comprising the following components in parts by weight: 1.0-2.0 parts of nuciferine, 0.1-0.5 parts of quercetin and 5.0-15.0 parts of chlorogenic acid; the nuciferine and quercetin-liposome solution with positive charges are encapsulated in chitosan/procyanidine microgel by an ionic crosslinking method to prepare the nuciferine and quercetin-liposome and chlorogenic acid co-encapsulated microgel.

2. The compound functional food according to claim 1, which is characterized by comprising the following components in parts by weight: 1.6 parts of nuciferine, 0.4 part of quercetin and 8 parts of chlorogenic acid.

3. The compound functional food according to claim 1, wherein the positively charged nuciferine and quercetin-liposome comprise the following components in parts by weight: 100-500 parts of phospholipid, 1-100 parts of cholesterol and 1.0-2.0 parts of nuciferine and quercetin, wherein the ratio of nuciferine to quercetin is 1:0.25.

4. The compound functional food according to claim 1, wherein the compound chitosan/procyanidin microgel comprises the following components in parts by weight: 300-1000 parts of chitosan and 100-400 parts of procyanidine.

5. The compounded functional food according to claim 1, characterized in that: the Zeta potential of the nuciferine and quercetin-liposome with positive charges is 30-70mV.

6. The compounded functional food according to claim 1, characterized in that: the particle size of the nuciferine and quercetin-liposome with positive charges ranges from 60nm to 150nm.

7. The functional food according to claim 1, wherein the Zeta potential of the microgel co-entrapped by nuciferine and quercetin-liposome and chlorogenic acid is- (30-60) mV.

8. The functional food according to claim 1, wherein the particle size of the microgel co-entrapped by nuciferine and quercetin-liposome and chlorogenic acid is in the range of 1-5um.

9. The method for preparing a compound functional food according to any one of claims 1 to 8, comprising the steps of:

1) Fully dissolving soybean lecithin and cholesterol in a round bottom flask by using a proper amount of volatile organic solvent, and uniformly stirring by using a magnetic stirrer;

2) Fully dissolving nuciferine and quercetin in volatile organic solution, adding into the solution obtained in the step 1), and stirring uniformly;

3) Evaporating the solution obtained in the step 2) under reduced pressure at 30 ℃ and 30rpm to dryness, evaporating the organic solvent to dryness, and burning a layer of film at a round bottom;

4) Adding a buffer solution, and carrying out water bath at 50 ℃ for 20min, so as to fully dissolve the film obtained in the step 3);

5) Homogenizing the fully dissolved solution obtained in the step 4) for 1-5min under the ice bath with the power of 80% and the normal pressure for 2-5 times to obtain semitransparent nuciferine and quercetin-liposome solution;

6) Fully dissolving chitosan in a round-bottom flask by glacial acetic acid, adding nuciferine and quercetin-liposome solution obtained in the step 5), and magnetically stirring for 0.5-2h until the mixture is completely mixed;

7) After the procyanidine and the chlorogenic acid are fully dissolved by a proper amount of water, adding the mixture into the round-bottomed flask in the step 6), and continuing to magnetically stir for 2-4 hours to obtain the hypolipidemic composite microgel co-entrapped by nuciferine, quercetin-liposome and chlorogenic acid.

10. The method for preparing a compound functional food according to claim 9, wherein:

the volatile organic solvent in the step 1) is selected from chloroform, and the mass fraction of the phospholipid-cholesterol and the added chloroform is 1:10-1:30;

the organic solvent in the step 2) is methanol, and the mass fraction of nuciferine and quercetin to the added methanol is 1:0.25:50-1:0.25:100;

the buffer solution in the step 4) is 0.05M phosphate buffer solution with the pH of 7.2-7.4, and the volume of the buffer solution is 40-80ml;

the concentration of glacial acetic acid added in step 6) was 1%.