CN110559352B - A kind of hawthorn flavonoid microcapsule and its preparation method and application - Google Patents
A kind of hawthorn flavonoid microcapsule and its preparation method and application Download PDFInfo
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- CN110559352B CN110559352B CN201911013922.5A CN201911013922A CN110559352B CN 110559352 B CN110559352 B CN 110559352B CN 201911013922 A CN201911013922 A CN 201911013922A CN 110559352 B CN110559352 B CN 110559352B
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- hawthorn
- hawthorn flavone
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Abstract
本发明提供一种山楂黄酮微胶囊及其制备方法和应用，所述芯材包括山楂黄酮提取物和植物硒多肽，所述壁材包括酪蛋白磷酸肽、氯化钙、明胶和羧甲基纤维素钠。本发明在壁材中加入酪蛋白磷酸肽、明胶和羧甲基纤维素钠与氯化钙固化剂进行配合，提高了山楂黄酮的包埋率和缓释效果，同时还能协同促进山楂黄酮降血压、降血脂等生物活性，提高山楂黄酮的生物利用度；芯材选择植物硒多肽与山楂黄酮进行复配，不但提高了产品的应用价值，而且，植物硒多肽还能协同促进山楂黄酮的生物活性，从而进一步提高微胶囊降血脂、抗氧化等功效。同时，山楂黄酮微胶囊的制备工艺简单，条件温和，制备得到的微胶囊的尺寸较小，稳定性好，成本低廉，适用于企业大规模生产。
The invention provides hawthorn flavonoid microcapsules and a preparation method and application thereof. The core material includes hawthorn flavonoid extract and plant selenium polypeptide, and the wall material includes casein phosphopeptide, calcium chloride, gelatin and carboxymethyl fiber Sodium. In the invention, casein phosphopeptide, gelatin and sodium carboxymethyl cellulose are added to the wall material to cooperate with calcium chloride solidifying agent, so as to improve the embedding rate and slow-release effect of hawthorn flavonoids, and at the same time, it can synergistically promote the reduction of hawthorn flavonoids. Blood pressure, lowering blood lipids and other biological activities, improve the bioavailability of hawthorn flavonoids; the core material selects plant selenium polypeptides and hawthorn flavonoids for compounding, which not only improves the application value of the product, but also synergistically promotes hawthorn flavonoids. Activity, thereby further improving the blood lipid lowering, antioxidant and other effects of microcapsules. Meanwhile, the preparation process of hawthorn flavonoid microcapsules is simple, the conditions are mild, the prepared microcapsules are small in size, good in stability and low in cost, and are suitable for large-scale production in enterprises.
Description
Technical Field
The invention relates to the technical field of microcapsule preparation, in particular to a hawthorn flavone microcapsule and a preparation method and application thereof.
Background
The hawthorn fruit contains abundant flavonoids, has the effects of resisting oxidation, reducing blood pressure and blood fat, relieving pain and healing wounds by inhibiting the secretion of inflammatory biological enzymes and the like, and can be used as a functional factor to be added into food or medicines. However, the flavonoid compounds are not stable in nature and are susceptible to factors such as oxygen in the air, light in the storage environment, temperature and the like, so that the activity of the flavonoid compounds is reduced and even lost. The hawthorn flavone is embedded by microencapsulation technology, which is not only beneficial to the preservation of flavonoid substances, but also enables the flavonoid substances to stably play a role in organisms. Microencapsulation is a technique in which particles, droplets, bubbles, or the like are encapsulated into heterogeneous fine particles using a polymeric film-forming material. The technology can achieve the aims of protecting the physicochemical property of the microcapsule core material, keeping the functional activity of the microcapsule core material and achieving sustainable release. However, the existing preparation process of the hawthorn flavone microcapsules is complex, and the prepared microcapsules have incomplete structures, so that the stability of the microcapsules is poor, a large amount of hawthorn flavone is difficult to reach a specified absorption part after entering an organism, the organism cannot efficiently utilize the hawthorn flavone, and the bioavailability of the hawthorn flavone is to be further improved.
Disclosure of Invention
The invention provides a hawthorn flavone microcapsule and a preparation method and application thereof, aiming at the problems that the existing hawthorn flavone microcapsule is complex in preparation process, incomplete in prepared microcapsule structure and low in hawthorn flavone bioavailability.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
a hawthorn flavone microcapsule comprises a core material and a wall material, wherein the core material comprises hawthorn flavone extract and plant selenium polypeptide, and the wall material comprises casein phosphopeptide, calcium chloride, gelatin and sodium carboxymethylcellulose.
The existing microcapsule generally adopts gelatin, chitosan or sodium alginate and the like as wall materials, the prepared microcapsule has an incomplete structure and small strength of the wall, and the structure of the microcapsule is quickly destroyed after entering a body, so that hawthorn flavone is exposed under the conditions of gastric acid, pepsin and the like of the body, the activity of the hawthorn flavone is seriously influenced, and the hawthorn flavone is difficult to reach a designated absorption part in a large amount, and the oral bioavailability of the hawthorn flavone is low. According to the invention, casein phosphopeptide, gelatin, sodium carboxymethylcellulose and a calcium chloride curing agent are added into the wall material to be matched, so that the embedding rate and the slow-release effect of hawthorn flavone are improved, and the bioactivity of the hawthorn flavone, such as blood pressure reduction, blood fat reduction and the like, can be synergistically promoted, and the bioavailability of the hawthorn flavone is improved; the core material is compounded by the plant selenium polypeptide and the hawthorn flavone, so that the application value of the product is improved, and the plant selenium polypeptide can also synergistically promote the biological activity of the hawthorn flavone, so that the effects of reducing blood fat, resisting oxidation and the like of the microcapsule are further improved.
The casein phosphopeptide can obviously improve the binding rate of calcium ions and wall materials, improve the integrity and strength of a microcapsule capsule wall structure, simultaneously, the casein phosphopeptide structure contains a large number of phosphoserine residues, can prevent the further action of digestive enzymes in the alkalescent environment of intestinal pH, improve the stability of the hawthorn flavone microcapsule in the intestinal tract of a human body and improve the slow release effect in the intestinal tract. More importantly, the casein phosphopeptide can also interact with hawthorn flavone, so that the biological activity and water solubility of the hawthorn flavone compound are improved, the bioavailability of the hawthorn flavone is improved, and the effects of lowering blood pressure, reducing blood fat and the like of the hawthorn flavone are fully exerted.
The plant selenium polypeptide has the effects of resisting oxidation and cancer, enhancing immunity, regulating vitamin absorption and utilization, promoting protein synthesis and the like, is compounded with hawthorn flavone as a core material, not only improves the nutritional value of the microcapsule, but also promotes the synthesis of selenium protein in a human body, regulates the activity of immunocytes, regulates intestinal tracts and synergistically promotes the absorption and utilization of the hawthorn flavone by the intestinal tracts, thereby further improving the bioavailability of the hawthorn flavone.
Preferably, the mass ratio of the casein phosphopeptide to the calcium chloride to the gelatin to the sodium carboxymethyl cellulose is 4-6:4-6:75-85: 8-12.
Compared with the prior art, the casein phosphopeptide, the calcium chloride, the gelatin and the sodium carboxymethyl cellulose in a specific proportion are selected as wall materials of the microcapsule, and the wall materials and the calcium chloride can form a capsule wall with high polymerization degree and good stability after being solidified, so that the capsule wall has better compactness and integrity, the core material can be better protected, the core material is prevented from leaking, and the bioavailability of the hawthorn flavone is improved.
Preferably, the mass ratio of the hawthorn flavone extract to the plant selenium polypeptide is 200-400: 1.
The optimized mass ratio of the hawthorn flavone extract to the plant selenium polypeptide can enable the hawthorn flavone extract and the plant selenium polypeptide to synergistically exert the best biological activity, obviously promote the synthesis of matrix selenoprotein, improve the biological activity of the hawthorn flavone, promote the body to absorb and utilize the hawthorn flavone and improve the bioavailability of the hawthorn flavone.
Preferably, the mass ratio of the core material to the wall material is 1: 6-8.
The preferable mass ratio of the wall material to the core material can ensure that the microcapsule has better integrity and slow release performance and controllable release performance.
Preferably, the preparation method of the hawthorn flavone extract comprises the following steps: freezing fructus crataegi with water content of 5-15%, pulverizing, and sieving to obtain fructus crataegi powder; adding 30-40 times of anhydrous ethanol, ultrasonic extracting at 50-60 deg.C for 30-40min, and filtering to obtain residue and filtrate; adding 15-20 times of anhydrous ethanol into the residue, ultrasonic extracting at 50-60 deg.C for 30-40min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone extract.
The preferable preparation method of the hawthorn flavone can dissolve the hawthorn flavone in the raw materials into the extracting solution more, avoids the hawthorn flavone from being damaged due to high temperature, and simultaneously avoids the contradiction problem of poor quality of the extracted product caused by excessive entrainment of ineffective substances.
The invention also provides a preparation method of the hawthorn flavone microcapsule, which comprises the following steps:
weighing the components according to a designed ratio, and respectively dissolving the weighed casein phosphopeptide, calcium chloride, gelatin and sodium carboxymethylcellulose in water to respectively obtain a casein phosphopeptide solution, a calcium chloride solution, a gelatin solution and a sodium carboxymethylcellulose solution;
dissolving the weighed plant selenium polypeptide in water to obtain a plant selenium polypeptide aqueous solution;
step four, uniformly mixing the hawthorn flavone solution and the plant selenium polypeptide aqueous solution, adjusting the pH value to 4.8-5.2, and stirring at 50-70 ℃ for 2-4h to obtain a core material;
and step five, adding an emulsifier into the gelatin solution, uniformly mixing to obtain a gelatin emulsion, adding the core material into the gelatin emulsion, uniformly mixing, adding the sodium carboxymethylcellulose solution, uniformly mixing, adjusting the pH to 4.3-4.7, adding the casein phosphopeptide solution and the calcium chloride solution at the temperature of 0-5 ℃, adjusting the pH to 8-9, curing for 50-70min, centrifuging, and freeze-drying to obtain the hawthorn flavone microcapsule.
The microcapsule prepared by the preparation method has the particle size centralized distribution of about 2 μm, and the embedding rate of hawthorn flavone can reach more than 90%. The process for preparing the hawthorn flavone microcapsules is simple, the conditions are mild, the prepared microcapsules are small in size, high in separation degree, good in stability and low in cost, and the hawthorn flavone microcapsules are suitable for large-scale production of enterprises.
Preferably, the concentration of the casein phosphopeptide solution is 15-25mg/mL, the concentration of the calcium chloride solution is 15-25mg/mL, the concentration of the gelatin solution is 12-18mg/mL, and the concentration of the sodium carboxymethylcellulose solution is 1.2-1.8 mg/mL.
The optimized concentration of each wall material raw material is beneficial to the full and uniform mixing of the raw materials, and the microencapsulation efficiency is improved.
Preferably, the concentration of the hawthorn flavone solution is 35-45mg/mL, and the concentration of the plant selenium polypeptide aqueous solution is 180-220 mu g/mL.
The optimized concentration of the hawthorn flavone solution and the plant selenium polypeptide aqueous solution is beneficial to the full emulsification of the core material, and the concentration of active substances in the core material of the microcapsule can be improved, so that the biological activity of the prepared microcapsule is improved.
Preferably, the emulsifier is tween 80.
Preferably, the addition amount of the emulsifier is 0.8-1.2% of the mass of the gelatin solution.
The optimized dosage of the emulsifier can ensure that the hawthorn flavone is fully emulsified, achieve more excellent embedding effect and be beneficial to forming a stable microcapsule structure.
Optionally, in the fourth step, the stirring speed is 100-300 rpm/min.
The optimized stirring speed can ensure that the hawthorn flavone extract and the plant selenium polypeptide are fully and uniformly mixed, thereby being beneficial to subsequent emulsification to obtain emulsion with uniform size.
The invention also provides application of the hawthorn flavone microcapsule in antioxidant and blood fat reducing medicines or health-care foods.
The hawthorn flavone microcapsule prepared by the invention has good structural stability and strength, can resist the damage of gastrointestinal environment to hawthorn flavone activity, has good slow release effect, can improve the activity and solubility of the hawthorn flavone after inclusion, is easy to be absorbed by human body, further improves the bioavailability of the hawthorn flavone by selecting the plant selenium polypeptide and the hawthorn flavone for compounding in the core material, and has wide application prospect in the fields of antioxidant and blood fat reducing medicines or health-care foods.
Drawings
FIG. 1 is a scanning electron microscope image of the hawthorn flavone microcapsule prepared in example 1 of the present invention;
fig. 2 is a graph showing the sustained release effect of the hawthorn flavone microcapsules prepared in example 1 of the present invention in simulated gastrointestinal fluids.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
A preparation method of hawthorn flavone microcapsules comprises the following steps:
weighing gelatin, sodium carboxymethylcellulose, casein phosphopeptide and calcium chloride according to a mass ratio of 80:10:5:5, respectively dissolving the components in water to respectively obtain a casein phosphopeptide solution with a mass concentration of 20mg/mL, a calcium chloride solution with a mass concentration of 20mg/mL, a gelatin solution with a mass concentration of 16mg/mL and a sodium carboxymethylcellulose solution with a mass concentration of 1.6 mg/mL;
weighing hawthorn flavone extract and plant selenium polypeptide according to the mass ratio of the core material to the wall material of 1:7 and the mass ratio of the hawthorn flavone extract to the plant selenium polypeptide of 300:1, dissolving the weighed hawthorn flavone extract in 90% ethanol solution by mass concentration, and preparing hawthorn flavone solution with the concentration of 40 mg/mL; dissolving the weighed plant selenium polypeptide in water to obtain a plant selenium polypeptide water solution with the mass concentration of 200 mug/mL;
step three, uniformly mixing the hawthorn flavone solution and the plant selenium polypeptide aqueous solution, adjusting the pH value to 5.0, and stirring for 3 hours at the temperature of 60 ℃ and the stirring speed of 200rpm/min to obtain a core material;
and step four, adding tween 80 accounting for 1 wt% of the mass of the gelatin solution into the gelatin solution, uniformly mixing to obtain a gelatin emulsion, adding the core material into the gelatin emulsion, uniformly mixing, adding the sodium carboxymethylcellulose solution, uniformly mixing, adjusting the pH to 4.5, adding the casein phosphopeptide solution and the calcium chloride solution at the temperature of 0 ℃, adjusting the pH to 8.5, curing for 60min, carrying out centrifugal separation, and carrying out freeze drying for 12h to obtain the hawthorn flavone microcapsule.
The preparation method of the hawthorn flavone extract comprises the following steps: freezing and pulverizing fructus crataegi with water content of 5-15%, and sieving with 80 mesh sieve to obtain fructus crataegi powder; adding anhydrous ethanol 35 times the weight of the fructus crataegi powder, ultrasonic extracting at 55 deg.C for 35min, and filtering to obtain filter residue and filtrate; adding 18 times of anhydrous ethanol into the filter residue, performing ultrasonic extraction at 55 deg.C for 35min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone extract.
The scanning electron microscope picture of the hawthorn flavone microcapsule product prepared in the embodiment 1 is shown in fig. 1, and it can be seen from the picture that submicron-sized microcapsules with good shapes can be obtained by adopting the preparation method of the hawthorn flavone microcapsule provided by the invention, the particle size is mainly distributed in 2-5 μm, the separation degree of the microcapsules is high, and the encapsulation effect is good.
The embedding rate of the hawthorn flavone microcapsules prepared in the embodiment is 91.9%.
Example 2
A preparation method of hawthorn flavone microcapsules comprises the following steps:
step one, weighing gelatin, sodium carboxymethylcellulose, casein phosphopeptide and calcium chloride according to a mass ratio of 85:12:4:6, respectively dissolving the components in water to respectively obtain a casein phosphopeptide solution with a mass concentration of 15mg/mL, a calcium chloride solution with a mass concentration of 25mg/mL, a gelatin solution with a mass concentration of 12mg/mL and a sodium carboxymethylcellulose solution with a mass concentration of 1.8 mg/mL;
weighing the hawthorn flavone extract and the plant selenium polypeptide according to the mass ratio of the core material to the wall material of 1:6 and the mass ratio of the hawthorn flavone extract to the plant selenium polypeptide of 400:1, and dissolving the weighed hawthorn flavone extract in an ethanol solution with the mass concentration of 80% to prepare a hawthorn flavone solution with the concentration of 35 mg/mL; dissolving the weighed plant selenium polypeptide in water to obtain a plant selenium polypeptide water solution with the mass concentration of 180 mug/mL;
step three, uniformly mixing the hawthorn flavone solution and the plant selenium polypeptide aqueous solution, adjusting the pH value to 4.8, and stirring for 2 hours at the temperature of 70 ℃ and the stirring speed of 300rpm/min to obtain a core material;
and step four, adding Tween 80 with the mass of 0.8 wt% into the gelatin solution, uniformly mixing to obtain a gelatin emulsion, adding the core material into the gelatin emulsion, uniformly mixing, adding the sodium carboxymethylcellulose solution, uniformly mixing, adjusting the pH to 4.7, adding the casein phosphopeptide solution and the calcium chloride solution at the temperature of 5 ℃, adjusting the pH to 8.5, solidifying for 60min, carrying out centrifugal separation, and carrying out freeze drying for 12h to obtain the hawthorn flavone microcapsule.
The preparation method of the hawthorn flavone extract comprises the following steps: freezing and pulverizing fructus crataegi with water content of 5-15%, and sieving with 80 mesh sieve to obtain fructus crataegi powder; adding 30 times of anhydrous ethanol, ultrasonically extracting at 60 deg.C for 30min, and filtering to obtain filter residue and filtrate; adding 20 times of anhydrous ethanol into the residue, ultrasonic extracting at 60 deg.C for 30min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone extract.
The embedding rate of the hawthorn flavone microcapsules prepared in the embodiment is 91.3%.
Example 3
A preparation method of hawthorn flavone microcapsules comprises the following steps:
step one, weighing gelatin, sodium carboxymethylcellulose, casein phosphopeptide and calcium chloride according to a mass ratio of 75:8:6:4, respectively dissolving the components in water to respectively obtain a casein phosphopeptide solution with a mass concentration of 25mg/mL, a calcium chloride solution with a mass concentration of 15mg/mL, a gelatin solution with a mass concentration of 18mg/mL and a sodium carboxymethylcellulose solution with a mass concentration of 1.2 mg/mL;
weighing the hawthorn flavone extract and the plant selenium polypeptide according to the mass ratio of the core material to the wall material of 1:8 and the mass ratio of the hawthorn flavone extract to the plant selenium polypeptide of 200:1, and dissolving the weighed hawthorn flavone extract in an ethanol solution with the mass concentration of 70% to prepare a hawthorn flavone solution with the concentration of 45 mg/mL; dissolving the weighed plant selenium polypeptide in water to obtain a plant selenium polypeptide water solution with the mass concentration of 220 mug/mL;
step three, uniformly mixing the hawthorn flavone solution and the plant selenium polypeptide aqueous solution, adjusting the pH value to 5.2, and stirring for 4 hours at the temperature of 50 ℃ and the stirring speed of 100rpm/min to obtain a core material;
and step four, adding Tween 80 with the mass of 1.2 wt% of that of the gelatin solution into the gelatin solution, uniformly mixing to obtain a gelatin emulsion, adding the core material into the gelatin emulsion, uniformly mixing, adding the sodium carboxymethylcellulose solution, uniformly mixing, adjusting the pH to 4.3, adding the casein phosphopeptide solution and the calcium chloride solution at the temperature of 2 ℃, adjusting the pH to 8.0, curing for 70min, carrying out centrifugal separation, and freeze-drying for 12h to obtain the hawthorn flavone microcapsule.
The preparation method of the hawthorn flavone extract comprises the following steps: freezing and pulverizing fructus crataegi with water content of 5-15%, and sieving with 80 mesh sieve to obtain fructus crataegi powder; adding anhydrous ethanol 40 times the weight of the fructus crataegi powder, ultrasonic extracting at 50 deg.C for 40min, and filtering to obtain filter residue and filtrate; adding 15 times of anhydrous ethanol into the filter residue, performing ultrasonic extraction at 50 deg.C for 40min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone extract.
The embedding rate of the hawthorn flavone microcapsules prepared in the embodiment is 90.8%.
Comparative example 1
A preparation method of hawthorn flavone microcapsules comprises the following steps:
weighing gelatin, sodium carboxymethylcellulose, casein phosphopeptide and calcium chloride according to a mass ratio of 80:10:5:5, respectively dissolving the components in water to respectively obtain a casein phosphopeptide solution with a mass concentration of 20mg/mL, a calcium chloride solution with a mass concentration of 20mg/mL, a gelatin solution with a mass concentration of 16mg/mL and a sodium carboxymethylcellulose solution with a mass concentration of 1.6 mg/mL;
weighing a hawthorn flavone extract according to the mass ratio of the core material to the wall material of 1:7, and dissolving the weighed hawthorn flavone extract in an ethanol solution with the mass concentration of 90% to prepare a hawthorn flavone solution with the concentration of 40 mg/mL;
adding tween 80 accounting for 1 wt% of the mass of the gelatin solution into the gelatin solution, uniformly mixing to obtain a gelatin emulsion, adding the hawthorn flavone solution into the gelatin emulsion, uniformly mixing, adding the sodium carboxymethylcellulose solution, uniformly mixing, adjusting the pH to 4.5, adding the casein phosphopeptide solution and the calcium chloride solution at the temperature of 0 ℃, adjusting the pH to 8.5, solidifying for 60min, carrying out centrifugal separation, and carrying out freeze drying for 12h to obtain the hawthorn flavone microcapsule.
The embedding rate of the hawthorn flavone microcapsules prepared in the comparative example is 90.7%.
Comparative example 2
A preparation method of hawthorn flavone microcapsules comprises the following steps:
step one, weighing gelatin, sodium carboxymethylcellulose and calcium chloride according to a mass ratio of 80:10:5, and respectively dissolving the components in water to respectively obtain a calcium chloride solution with a mass concentration of 20mg/mL, a gelatin solution with a mass concentration of 16mg/mL and a sodium carboxymethylcellulose solution with a mass concentration of 1.6 mg/mL;
weighing a hawthorn flavone extract according to the mass ratio of the core material to the wall material of 1:7, and dissolving the weighed hawthorn flavone extract in an ethanol solution with the mass concentration of 90% to prepare a hawthorn flavone solution with the concentration of 40 mg/mL;
adding tween 80 accounting for 1 wt% of the mass of the gelatin solution into the gelatin solution, uniformly mixing to obtain a gelatin emulsion, adding the hawthorn flavone solution into the gelatin emulsion, uniformly mixing, adding the sodium carboxymethylcellulose solution, uniformly mixing, adjusting the pH to 4.5, adding the casein phosphopeptide solution and the calcium chloride solution at the temperature of 0 ℃, adjusting the pH to 9.0, solidifying for 50min, carrying out centrifugal separation, and carrying out freeze drying for 12h to obtain the hawthorn flavone microcapsules.
The embedding rate of the hawthorn flavone microcapsules prepared by the comparative example is 89.5%.
In vitro sustained Release assay
Preparing simulated gastric juice and simulated intestinal juice, respectively placing the hawthorn flavone microcapsules prepared in example 1 in the simulated gastric juice and the simulated intestinal juice, and then placing the hawthorn flavone microcapsules in a constant temperature incubator at 37 ℃ for in-vitro slow release test, wherein the results are shown in figure 2.
As can be seen from the figure, in simulated gastric fluid, the cumulative release amount of the hawthorn flavone microcapsules can reach 29.09% after 4 hours, the release speed is gradually reduced after 4-8 hours, and the cumulative release amount reaches 37.26% after 8 hours. In simulated intestinal juice, the accumulative release amount of the hawthorn flavone microcapsules in 1 hour is larger than that in gastric juice, and can reach 97.78 percent, and then the release speed is reduced, so that the hawthorn flavone microcapsules are in a stable state. The hawthorn flavone microcapsules prepared by the invention are relatively stable in gastric juice, and after reaching the small intestine of the main absorption part, the release is accelerated, the release rule of the hawthorn flavone microcapsules prepared by the invention accords with the characteristics of human nutrition digestion and absorption, and the hawthorn flavone microcapsules have good application value in protecting and utilizing the bioactivity of hawthorn flavone.
Research on blood fat reducing function
72 Wistar rats were randomly divided into 8 groups of 9 rats each, which were a blank group, a model group, a hawthorn flavone extract group, a comparative example 1 group, a comparative example 2 group, a comparative example 3 group, a comparative example 4 group and an example 1 group, and all rats were fed for one week before the experiment to adapt to the environment. According to the method in the technical Specification for health food inspection and evaluation (2003), gavage is performed while the feed is taken freely.
Wherein, the blank group had free intake of basal diet, and the other 4 groups had free intake of high fat diet. The blank group is subjected to intragastric perfusion with distilled water, and the single administration dose is 1mL/100g of body weight; model group, drenching stomach with distilled water, and single administration dosage is 1mL/100g body weight; the hawthorn flavone extract group is used for gastric lavage by using the hawthorn flavone extract extracted in the example 1, and the single administration dosage is 100mg/Kg of body weight; group of comparative example 1, the hawthorn flavone microcapsules prepared in comparative example 1 were used for intragastric administration, and the single administration dose was 50mg/Kg body weight; group of comparative example 2, the hawthorn flavone microcapsules prepared in comparative example 2 were used for intragastric administration, and the single administration dose was 50mg/Kg body weight; comparative example 3 group, the administration of casein phosphopeptide for intragastric administration was 50mg/Kg of body weight for a single administration; the mixture of hawthorn flavone extract and casein phosphopeptide extracted in the group of comparative example 4 and the group of example 1 is used for gastric lavage, the mass ratio of the hawthorn flavone extract to the casein phosphopeptide is the same as that in the group of example 1, and the single administration dosage is 50mg/Kg body weight; example 1 group, the hawthorn flavone microcapsules prepared in example 1 are used for gastric lavage, and the single administration dose is 50mg/Kg body weight; each group of rats is administrated 1 time a day, and after continuous administration for 5 weeks, various physiological and biochemical indexes of the rats are measured. The specific experimental design is shown in table 1.
The formula of the high-fat feed comprises the following components: 78.8% of basic feed, 10% of lard, 10% of egg yolk powder, 1% of cholesterol and 0.2% of cholate.
Table 1 experimental design protocol
After fasting for 12h on day 35, the femoral artery of the rat is bled, and serum is separated by centrifugation at 3000r/min for 15min and is used for measuring the blood fat index and the antioxidant index. Dissecting rat, quickly taking out brain, heart, spleen and kidney, removing fat membrane on its surface, washing with normal saline, removing water with filter paper, weighing, sealing in clean plastic box, and storing serum and organs in-80 deg.C refrigerator.
Detection indexes are as follows:
(1) body weight and feed intake: animal body weight and feed intake were recorded weekly.
(2) And (3) measuring various biochemical indexes in serum: the following biochemical indicators were detected from each group of rat sera following the protocol specification of each kit: serum Total Cholesterol (TC), Triglycerides (TG), high density lipoproteins (HDL-C) and low density lipoproteins (LDL-C), and an Arteriosclerosis Index (AI) value can be calculated according to the formula: AI-TC-HDL-C/HDL-C, the results are shown in Table 2.
(3) And (3) detecting an antioxidant index: the Malondialdehyde (MDA) content, Catalase (CAT) activity, superoxide dismutase (SOD) activity and glutathione peroxidase (GSH-Px) activity in serum and liver tissues were measured according to the procedures of the kit instructions, and the results are shown in tables 3 and 4.
(4) Organ index calculation
The organ index (g/100g) is the organ mass × 100/rat body weight, and the results are shown in table 5.
(5) Fat wet weight and fat coefficient: after blood was taken from the animals, perirenal and perigenital adipose tissues were dissected and peeled off, and the fat weight was measured to calculate the fat factor, and the results are shown in table 6.
Fat coefficient ═ fat wet weight (g) × 100%/body weight (g)
The experimental results are as follows:
TABLE 2 results of biochemical indicators in serum of rats of each experimental group
Notes comparison of P with blank group<0.05，**P<0.01; comparison with model group#P<0.05，##P<0.01。
As can be seen from the above table, after the drug withdrawal for 12h, compared with the blank group, TC, TG and LDL-C of the model group are all significantly improved (P is less than 0.01), and HDL-C is significantly reduced (P is less than 0.01), which indicates that the model building of the rat in the high-fat model group is successful. Compared with the model group, the rats of the flavone extract group, the comparative example 1 group, the comparative example 2 group, the comparative example 4 group and the example 1 group all have obviously reduced TC and TG, the comparative example 1 group and the example 1 group have extremely obviously reduced TC (P <0.01), the comparative example 2 group and the comparative example 4 group have obviously reduced TC (P <0.05), and the comparative example 4 group and the example 1 group have obvious difference of TC (P < 0.05).
Compared with the model group, the HDL-C content of the comparative example 1 group and the HDL-C content of the example 1 group are obviously increased (P <0.05), and the flavone extract group and the comparative example 4 group have an increased trend, but the difference has no statistical significance (P > 0.05); the contents of LDL-C in the comparative example 1 group and the example 1 group were significantly reduced, and the contents of the flavone extract group and the comparative example 4 group tended to be reduced, but the difference was not statistically significant (P > 0.05).
The AI mean value of each group of rats changed differently with the change of the blood lipid of the rats. The AI value of the model group is as high as 4.38 +/-1.51, the risk of atherosclerosis is higher, and the AI values of rats of the comparative example 1 group and the example 1 group are respectively reduced by 41.6 percent and 43.8 percent after the treatment by administration. It is shown that the group of example 1 has a more significant effect of reducing the risk of atherosclerosis in the experimental rats than the group of comparative example 1 and the group of comparative example 4. Meanwhile, TC, TG and HDL-C in the comparative example 4 group were significantly different (P <0.05) from those in example 1 and the flavone extract group.
TABLE 3 anti-oxidation index of liver in each experimental group
| Group of | CAT(U/mg) | GSH-PX(U/mg) | MDA(nmol/mg) | SOD(U/mg) |
| Blank group | 60.66±9.57# | 253.48±29.55# | 1.41±0.37# | 146.12±34.40 |
| Model set | 43.38±4.83* | 199.16±0.90* | 2.12±0.49* | 127.66±9.65 |
| Flavone extract group | 55.42±10.30 | 226.50±42.32 | 1.64±0.35 | 131.06±18.37 |
| Comparative example 1 group | 68.48±6.42# | 238.64±26.81# | 1.48±0.28 | 140.89±15.30 |
| Comparative example 2 group | 56.2±9.37 | 229.50±42.32 | 1.62±0.31 | 136.53±12.38 |
| Comparative example 3 group | 43.36±4.77* | 199.02±0.85* | 2.15±0.44* | 127.53±9.73 |
| Comparative example 4 group | 64.42±10.30 | 232.59±23.03# | 1.52±0.35 | 139.06±18.37 |
| EXAMPLE 1 group | 74.18±8.27## | 241.87±30.05# | 1.35±0.18# | 147.25±32.40 |
Note: comparison with blank group<0.05; comparison with model group#P<0.05。
As can be seen from the above table, the MDA content in the liver of the rat in the model group is significantly higher than that in the blank group (P <0.05), and the CAT activity and the GSH-Px activity are significantly lower than those in the blank group (P < 0.05). Compared with the model group, the CAT activity of the group in the example 1 is obviously higher than that of the rest groups, which shows that compared with the group in the comparative example 1 and the group in the comparative example 4, the CAT activity of the organism can be obviously improved and the oxidation resistance of the organism can be improved in the example 1. Compared with the group of the comparative example 4, the group of the comparative example 1 can obviously improve the CAT activity of organisms. Compared with the flavone extraction group, the group 4 can improve the CAT activity of the organism.
TABLE 4 serum antioxidant index of each experimental group
Note comparison with blank group*P<0.05，**P<0.01; comparison with model group#P<0.05，##P<0.01。
As can be seen from the above table, the CAT activity in the serum of the rats in the model group is significantly lower than that in the blank group (P <0.05), and the SOD activity is significantly lower than that in the blank group (P < 0.05). Compared with the model group, the CAT activity, the GPX activity and the SOD activity of the group in the example 1 are obviously improved, the MDA content is obviously reduced, and only the CAT activity and the SOD activity of the group in the comparative example 1 are obviously improved. This shows that example 1 has a better effect of improving the antioxidant ability of the body than the group of comparative example 1. Only CAT activity in the comparative example 4 group is remarkably improved, which shows that the comparative example 1 group can improve the oxidation resistance of organisms better than the comparative example 4 group.
TABLE 5 results of organ coefficient of rat in each experimental group
| Group of | Liver coefficient/%) | Spleen coefficient/% | Heart coefficient/%) | Renal coefficient/% |
| Blank group | 2.46±0.06# | 0.20±0.03## | 0.33±0.02 | 0.53±0.04 |
| Model set | 2.70±0.19* | 0.24±0.06** | 0.32±0.04 | 0.52±0.03 |
| Flavone extract group | 2.53±0.14 | 0.21±0.01# | 0.32±0.04 | 0.51±0.02 |
| Comparative example 1 group | 2.44±0.12# | 0.19±0.01## | 0.31±0.02 | 0.51±0.04 |
| Comparative example 2 group | 2.63±0.19 | 0.20±0.02# | 0.32±0.03 | 0.52±0.03 |
| Comparative example 3 group | 2.69±0.17* | 0.25±0.04** | 0.33±0.05 | 0.51±0.04 |
| Comparative example 4 group | 2.48±0.22 | 0.20±0.01# | 0.31±0.04 | 0.51±0.07 |
| EXAMPLE 1 group | 2.46±0.09# | 0.20±0.02## | 0.32±0.03 | 0.53±0.03 |
Note P <0.05, P <0.01 compared to blank; p <0.05, P <0.01 compared to model group.
The liver and spleen coefficients increased, indicating that feeding high-fat feed can cause liver injury and spleen injury in rats. As can be seen from the above table, the liver coefficient of the model group is increased compared with that of the blank group, and has significant difference (P < 0.05); compared with the model group, the liver coefficients of the comparative example 1 group and the example 1 group have significant difference (P <0.05), and are respectively reduced by 9.6% and 8.8%.
The spleen index of rats in the flavone extract group, the comparative example 1 group, the comparative example 2 group, the comparative example 4 group and the example 1 group was significantly reduced compared to the model group, and the comparative example 1 group and the example 1 group were very significantly different from the model group.
Table 6 results of fat coefficients for each experimental group
| Group of | Perirenal fat coefficient/%) | Epididymal fat coefficient/%) | Total fat coefficient/% |
| Blank group | 2.16±0.45## | 1.58±0.16## | 3.83±0.64# |
| Model set | 2.91±0.41** | 1.86±0.23** | 4.79±0.45* |
| Flavone extract group | 2.43±0.46# | 1.55±0.14## | 4.19±0.75 |
| Comparative example 1 group | 2.29±0.39## | 1.46±0.16## | 3.83±0.50# |
| Comparative example 2 group | 2.47±0.36# | 1.60±0.14# | 4.45±0.74 |
| Comparative example 3 group | 2.93±0.36** | 1.84±0.24** | 4.81±0.42* |
| Comparative example 4 group | 2.42±0.32# | 1.51±0.12## | 3.93±0.69 |
| EXAMPLE 1 group | 2.18±0.26## | 1.49±0.15## | 3.78±0.68# |
Notes comparison of P with blank group<0.05，**P<0.01; comparison with model group#P<0.05，##P<0.01。
As can be seen from the above table, the epididymal fat index, perirenal fat index and total fat index of the model group were all increased and statistically significant (P <0.01 or P <0.05) compared to the blank group. Compared with the model group, the comparative example 1 group, the comparative example 2 group, the comparative example 4 group and the example 1 group all have a reduction effect on the perirenal fat factor, and the comparative example 1 group and the example 1 have a very significant difference (P <0.01), which is reduced by 21.3% and 25.1%, respectively, and the comparative example 4 group and the flavone extract group have a significant difference (P < 0.05). And there was a significant difference between the comparative example 4 group and the flavone extract group.
Compared with the model group, the epididymis fat coefficient reduction of each group has very obvious difference (P is less than 0.01), and the total fat coefficient of each group has a reduction effect, but the reduction effect of the group in example 1 is more obvious and is reduced by 21.1%, so that the hawthorn flavone microcapsules prepared in the group in example 1 can have a weight-losing effect.
The tests prove that the addition of the casein phosphopeptide not only improves the embedding rate of the core material and the stability of the microcapsule structure, but also has the synergistic effect with hawthorn flavone. Meanwhile, the synergistic interaction effect between the plant selenium polypeptide and the hawthorn flavone is also proved, in the embodiment 1, the addition amount of the hawthorn flavone is reduced, and the plant selenium polypeptide is added, so that the microcapsule has more excellent effects of resisting oxidation and reducing blood fat than the single hawthorn flavone. Therefore, the hawthorn flavone microcapsule prepared by the invention has better application prospects in the aspects of oxidation resistance and blood fat reduction.
The effects of oxidation resistance and blood fat reduction which are basically the same as those of the embodiment 1 can be achieved within the mass ratio range of other wall material and core material raw materials and within other parameter ranges of process parameters of the preparation method.
The plant selenium polypeptide can be prepared by adopting a commercial product or a conventional extraction method, and has no obvious influence on the efficacy of the prepared hawthorn flavone microcapsule.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents or improvements made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (9)
1. A hawthorn flavone microcapsule comprises a core material and a wall material, and is characterized in that the core material comprises hawthorn flavone extract and plant selenium polypeptide, and the wall material comprises casein phosphopeptide, calcium chloride, gelatin and sodium carboxymethylcellulose; wherein the mass ratio of the casein phosphopeptide to the calcium chloride to the gelatin to the sodium carboxymethylcellulose is 4-6:4-6:75-85: 8-12; the mass ratio of the hawthorn flavone extract to the plant selenium polypeptide is 200-400: 1; the mass ratio of the core material to the wall material is 1: 6-8.
2. The hawthorn flavone microcapsule of claim 1, wherein the preparation method of the hawthorn flavone extract comprises: freezing fructus crataegi with water content of 5-15%, pulverizing, and sieving to obtain fructus crataegi powder; adding 30-40 times of anhydrous ethanol, ultrasonic extracting at 50-60 deg.C for 30-40min, and filtering to obtain residue and filtrate; adding 15-20 times of anhydrous ethanol into the residue, ultrasonic extracting at 50-60 deg.C for 30-40min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone extract.
3. The method for preparing hawthorn flavone microcapsules of any one of claims 1 to 2, which is characterized by comprising the following steps:
weighing the components according to a designed ratio, and respectively dissolving the weighed casein phosphopeptide, calcium chloride, gelatin and sodium carboxymethylcellulose in water to respectively obtain a casein phosphopeptide solution, a calcium chloride solution, a gelatin solution and a sodium carboxymethylcellulose solution;
secondly, dissolving the weighed hawthorn flavone extract into an ethanol solution with the mass concentration of 70-90% to obtain a hawthorn flavone solution;
dissolving the weighed plant selenium polypeptide in water to obtain a plant selenium polypeptide aqueous solution;
step four, uniformly mixing the hawthorn flavone solution and the plant selenium polypeptide aqueous solution, adjusting the pH value to 4.8-5.2, and stirring at 50-70 ℃ for 2-4h to obtain a core material;
and step five, adding an emulsifier into the gelatin solution, uniformly mixing to obtain a gelatin emulsion, adding the core material into the gelatin emulsion, uniformly mixing, adding the sodium carboxymethylcellulose solution, uniformly mixing, adjusting the pH to 4.3-4.7, adding the casein phosphopeptide solution and the calcium chloride solution at the temperature of 0-5 ℃, adjusting the pH to 8-9, curing for 50-70min, centrifuging, and freeze-drying to obtain the hawthorn flavone microcapsule.
4. The method for preparing hawthorn flavone microcapsules of claim 3, wherein in the first step, the concentration of the casein phosphopeptide solution is 15-25mg/mL, the concentration of the calcium chloride solution is 15-25mg/mL, the concentration of the gelatin solution is 12-18mg/mL, and the concentration of the sodium carboxymethylcellulose solution is 1.2-1.8 mg/mL.
5. The preparation method of the hawthorn flavone microcapsule as claimed in claim 3, wherein the concentration of the hawthorn flavone solution is 35-45mg/mL, and the concentration of the plant selenium polypeptide aqueous solution is 180-220 μ g/mL.
6. The method for preparing hawthorn flavone microcapsules of claim 3, wherein the emulsifier is Tween 80.
7. The method for preparing hawthorn flavone microcapsules of claim 6, wherein the addition amount of the emulsifier is 0.8-1.2% of the mass of the gelatin solution.
8. Use of the hawthorn flavone microcapsules of any one of claims 1-2 in the preparation of anti-oxidant and hypolipidemic drugs.
9. The use of the hawthorn flavone microcapsules of any one of claims 1-2 in the preparation of an antioxidant, antilipemic-assisted health food.
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