CN110643655A - Hawthorn flavone chelated sugar and preparation method and application thereof - Google Patents
Hawthorn flavone chelated sugar and preparation method and application thereof Download PDFInfo
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- CN110643655A CN110643655A CN201911013910.2A CN201911013910A CN110643655A CN 110643655 A CN110643655 A CN 110643655A CN 201911013910 A CN201911013910 A CN 201911013910A CN 110643655 A CN110643655 A CN 110643655A
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- polysaccharide
- hawthorn
- hawthorn flavone
- sugar
- flavone
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/18—Preparation of compounds containing saccharide radicals produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention provides a hawthorn flavone chelated sugar and a preparation method and application thereof, wherein the preparation method of the hawthorn flavone chelated sugar is to hydrolyze agaric polysaccharide and tremella polysaccharide to obtain polysaccharide hydrolysate; uniformly mixing the hawthorn flavone and the polysaccharide hydrolysate, and adding UDP-glycosyltransferase to carry out saccharification reaction to obtain the hawthorn flavone chelated sugar. According to the invention, hydrolysis monosaccharide of agaric polysaccharide and tremella polysaccharide is selected to chelate and modify hawthorn flavone, and glycosyl is added on a matrix of hawthorn flavonoid compound, so that the solubility of the hawthorn flavonoid compound is greatly improved, the problem of poor water solubility of the hawthorn flavonoid compound is solved, the bioavailability of the hawthorn flavone is improved, the tremella polysaccharide, agaric polysaccharide and hawthorn flavone are synergistic, and the prepared hawthorn flavone chelate has excellent effects of resisting oxidation and reducing blood fat.
Description
Technical Field
The invention relates to the technical field of flavone compound modification preparation, in particular to hawthorn flavone chelated sugar and a preparation method and application thereof.
Background
Hawthorn (Hawthorn), also known as fructus Rhododendri Mucronulati, Rhus, Rosaceae, Maloideae, Crataegus, in different areas with different names, such as red jujube, annatto, Hawthorn etc., larch trees, up to 6 meters. The hawthorn fruit can be eaten raw or used as products such as preserved fruit cakes and the like, has high medicinal value and is a plant which is special in China and can be used for medicinal and edible purposes. The hawthorn has very high treatment efficacy in the aspects of preventing and treating cardiovascular and cerebrovascular diseases, improving heart activity, reducing blood pressure and blood fat, promoting urination and the like; the flavonoids and vitamin C contained in the hawthorn can strengthen the immune system of the organism and has the effects of delaying senility and resisting tumors; the hawthorn can accelerate the fluidity of the blood circulation system of the human body and promote blood circulation to remove blood stasis; meanwhile, the hawthorn also has good curative effects on asthma, cough with excessive phlegm, abdominal pain and diarrhea.
As a functional factor, the flavonoid substance is an active ingredient with higher content in the hawthorn, exists in the form of flavonoid glycoside substance, has good effects in various aspects such as blood pressure reduction, blood fat reduction, cancer resistance, oxidation resistance and the like, and has important significance in resisting aging, treating cardiovascular diseases and the like. However, most hawthorn flavonoids are alcohol-soluble substances, and the water solubility is poor, so that the hawthorn flavonoids are poorly absorbed and utilized by human bodies, the effect is not obvious, and the application of the hawthorn flavonoids in actual production is greatly limited.
Disclosure of Invention
The invention provides a hawthorn flavone chelating sugar and a preparation method and application thereof, aiming at the problem that the existing hawthorn flavone compound is low in water solubility, so that the hawthorn flavone bioavailability is low.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
a preparation method of hawthorn flavone chelated sugar comprises the following steps: hydrolyzing the agaric polysaccharide and the tremella polysaccharide to obtain polysaccharide hydrolysate; uniformly mixing the hawthorn flavone and the polysaccharide hydrolysate, and adding UDP-glycosyltransferase to carry out glycosylation reaction to obtain the hawthorn flavone chelated sugar.
In the prior art, hydrolysis monosaccharide of agaric polysaccharide and tremella polysaccharide is selected to chelate and modify hawthorn flavone, and glycosyl is added to a matrix of hawthorn flavonoid, so that the solubility of the hawthorn flavonoid is greatly improved, the problem of poor water solubility of the hawthorn flavonoid is solved, the bioavailability of the hawthorn flavone is improved, the tremella polysaccharide, the agaric polysaccharide and the hawthorn flavone are synergistic, and the prepared hawthorn flavone chelate has excellent effects of resisting oxidation and reducing blood fat.
The hawthorn flavonoid compound has the effects of reducing blood pressure and blood fat, increasing coronary flow, improving myocardial blood and oxygen supply and the like for cardiovascular systems, and has good curative effects on cardiovascular diseases such as hypertension, hyperlipidemia, coronary heart disease, angina pectoris and the like. The agaric polysaccharide and the tremella polysaccharide have the effects of reducing blood fat, reducing blood sugar, resisting oxidation, enhancing immunity and the like, and the tremella polysaccharide can directly reach the large intestine and be used as a nutrient substance of bifidobacterium and lactobacillus, so that the formation of ideal intestinal microflora is promoted, the functions of intestines and stomach are improved, and the functions of balancing gastric acid secretion and protecting and repairing gastric mucosa are realized; the agaric polysaccharide can promote gastrointestinal movement and enhance gastrointestinal functions, and the agaric polysaccharide can promote the absorption and utilization of hawthorn flavone by intestines and stomach in a synergistic manner, so that the bioavailability of the hawthorn flavone is further improved.
Preferably, the preparation method of the hawthorn flavone chelated sugar comprises the following steps:
step one, dissolving agaric polysaccharide and tremella polysaccharide in water, and uniformly mixing to obtain polysaccharide solution; dissolving hawthorn flavone in 70-90% ethanol solution to obtain hawthorn flavone solution;
adding mixed enzyme into the polysaccharide solution, and performing enzymolysis for 2-4h at 40-55 ℃ to obtain polysaccharide hydrolysate;
and step three, adding a hawthorn flavone solution into the polysaccharide enzymatic hydrolysate, uniformly mixing to obtain a mixed solution, adjusting the pH of the mixed solution to 3.5-5.5, adding UDP-glycosyltransferase, reacting at 40-65 ℃ for 3-6h, and performing spray drying to obtain the hawthorn flavone chelating sugar.
The preferable preparation method of the hawthorn flavone chelate sugar has the advantages of mild conditions, high glycosylation conversion rate, high solubility of the prepared hawthorn flavone chelate and wide application prospect.
Preferably, the mass ratio of the hawthorn flavone to the agaric polysaccharide to the tremella polysaccharide is 1:1-1.5: 1-1.5.
The optimized mass ratio of the hawthorn flavone to the agaric polysaccharide to the tremella polysaccharide can improve the glycosylation conversion rate of the hawthorn flavone and the solubility of the hawthorn flavone, and can also enable the hawthorn flavone, the agaric polysaccharide and the tremella polysaccharide to synergistically exert the maximum effects of resisting oxidation and reducing blood fat.
Preferably, the preparation method of the hawthorn flavone comprises the following steps: the preparation method of the hawthorn flavone comprises the following steps: freezing fructus crataegi with water content of 2-5%, pulverizing, and sieving to obtain fructus crataegi powder; adding 65-75% ethanol solution 10-50 times the weight of the hawthorn powder, performing ultrasonic extraction at 40-80 deg.C for 2-3 times for 5-15min, centrifuging, mixing supernatants, and concentrating to obtain hawthorn flavone.
Optionally, the power of the ultrasonic extraction is 100-.
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.
Preferably, in the step one, the mass concentration of the polysaccharide solution is 3-5 g/mL; the concentration of the hawthorn flavone solution is 35-45 mg/mL.
The preferable concentration of the polysaccharide solution is beneficial to hydrolyzing the polysaccharide into monosaccharide, and the preferable concentration of the hawthorn flavone solution is beneficial to improving the chelation of the monosaccharide and the hawthorn flavone and improving the glycosylation efficiency.
Preferably, in the second step, the mixed enzyme is endoglucanase I, beta-glucosidase, pectinase and cellulase in a mass ratio of 22-27:20-25:19-23: 25-29.
The preferred mixed enzyme can hydrolyze the agaric polysaccharide and the tremella polysaccharide into monosaccharides with smaller molecular weights, and after the agaric polysaccharide and the tremella polysaccharide are chelated with hawthorn flavone, the water solubility of the hawthorn flavone is remarkably improved, and the bioavailability of the hawthorn flavone is improved; the high glycosylation of hawthorn flavonoids can be realized by matching UPD-glycosyltransferase, and the conversion rate of glycosylated flavonoids reaches more than 95%.
Preferably, in the second step, the addition amount of the mixed enzyme is 5-10 wt% of the mass of the polysaccharide solution.
Preferably, in step three, the mass-to-volume ratio of the UPD-glycosyltransferase to the mixed solution is 0.5 to 3:1, wherein the unit of mass is gram and the unit of volume is liter.
The preferable adding amount of UPD-glycosyltransferase can improve the glycosyl conversion rate of hawthorn flavone, and is beneficial to realizing the complete glycosylation of hawthorn flavonoid compounds.
The invention also provides the hawthorn flavone chelated sugar which is prepared by the preparation method of the hawthorn flavone chelated sugar.
The invention also provides the application of the hawthorn flavone chelated sugar in the health-care food for resisting oxidation and reducing blood fat.
According to the hawthorn flavone chelated sugar prepared by the invention, the agaric polysaccharide and the hydrolyzed monosaccharide of the tremella polysaccharide are chelated with hawthorn flavone compounds, so that the activity and the solubility of hawthorn flavone are obviously improved, the hawthorn flavone chelated sugar is easy to absorb by a human body, the agaric polysaccharide and the tremella polysaccharide also have the functions of regulating intestines and stomach and promoting hawthorn flavone absorption, the bioavailability of the hawthorn flavone is further improved, and the hawthorn flavone chelated sugar has a wide application prospect in the fields of anti-oxidation and blood fat reduction health-care foods.
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 chelated sugar comprises the following steps:
step one, weighing hawthorn flavone, agaric polysaccharide and tremella polysaccharide according to the mass ratio of 1:1.2: 1.2;
step two, dissolving the weighed agaric polysaccharide and tremella polysaccharide in water, and uniformly mixing to obtain a polysaccharide solution with the mass concentration of 4 g/mL; dissolving the weighed hawthorn flavone in 90% ethanol solution to obtain hawthorn flavone solution with mass concentration of 40 mg/mL;
adding mixed enzyme of endoglucanase I, beta-glucosidase, pectinase and cellulase with the mass ratio of 22:23:19:29 into the polysaccharide solution, and performing enzymolysis at 50 ℃ for 3 hours to obtain polysaccharide enzymolysis liquid; the adding amount of the mixed enzyme is 8 wt% of the mass of the polysaccharide solution;
step four, adding a hawthorn flavone solution into the polysaccharide enzymatic hydrolysate, uniformly mixing to obtain a mixed solution, adjusting the pH of the mixed solution to 4.5, adding 1.5g of UPD-glycosyltransferase into each 1L of the mixed solution, and reacting at 55 ℃ for 5 hours to obtain a hawthorn flavone chelated sugar solution;
step five, drying the hawthorn flavone chelated sugar solution by adopting a spray drying method to obtain hawthorn flavone chelated sugar, wherein the drying conditions are as follows: the sample injection concentration is 10g/100mL, the sample injection speed is 5.5mL/min, and the air inlet temperature is 150 ℃.
The preparation method of the hawthorn flavone extract comprises the following steps: freezing and pulverizing fructus crataegi with water content of 2-5%, and sieving with 80 mesh sieve to obtain fructus crataegi powder; adding 70% ethanol solution 35 times the weight of the fructus crataegi powder, ultrasonic extracting at 55 deg.C and 200W for 10min, and filtering to obtain primary filtrate and primary residue; adding 20 times of 70% ethanol solution into the primary filter residue, performing ultrasonic extraction at 55 deg.C and 200W for 10min, and filtering to obtain secondary filtrate and secondary filter residue; adding 10 times of 70% ethanol solution into the secondary filter residue, ultrasonic extracting at 55 deg.C and 200W for 10min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone.
Example 2
A preparation method of hawthorn flavone chelated sugar comprises the following steps:
step one, weighing hawthorn flavone, agaric polysaccharide and tremella polysaccharide according to the mass ratio of 1:1: 1.5;
step two, dissolving the weighed agaric polysaccharide and tremella polysaccharide in water, and uniformly mixing to obtain a polysaccharide solution with the mass concentration of 3 g/mL; dissolving the weighed hawthorn flavone into an ethanol solution with the mass concentration of 70% to obtain a hawthorn flavone solution with the mass concentration of 35 mg/mL;
adding mixed enzyme of endoglucanase I, beta-glucosidase, pectinase and cellulase with the mass ratio of 25:20:21:27 into the polysaccharide solution, and performing enzymolysis at 50 ℃ for 3 hours to obtain polysaccharide enzymolysis liquid; the adding amount of the mixed enzyme is 10 wt% of the mass of the polysaccharide solution;
step four, adding a hawthorn flavone solution into the polysaccharide enzymatic hydrolysate, uniformly mixing to obtain a mixed solution, adjusting the pH of the mixed solution to 5.5, adding 3g of UPD-glycosyltransferase into each 1L of the mixed solution, and reacting at 40 ℃ for 6 hours to obtain a hawthorn flavone chelated sugar solution;
step five, drying the hawthorn flavone chelated sugar solution by adopting a spray drying method to obtain hawthorn flavone chelated sugar, wherein the drying conditions are as follows: the sample injection concentration is 10g/100mL, the sample injection speed is 5.5mL/min, and the air inlet temperature is 150 ℃.
The preparation method of the hawthorn flavone extract comprises the following steps: freezing and pulverizing fructus crataegi with water content of 2-5%, and sieving with 80 mesh sieve to obtain fructus crataegi powder; adding 65% ethanol solution 50 times the weight of the fructus crataegi powder, performing ultrasonic extraction at 80 deg.C and 100W for 5min, and filtering to obtain primary filtrate and primary residue; adding 30 times of 65% ethanol solution into the primary filter residue, performing ultrasonic extraction at 80 deg.C and 100W for 5min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone.
Example 3
A preparation method of hawthorn flavone chelated sugar comprises the following steps:
step one, weighing hawthorn flavone, agaric polysaccharide and tremella polysaccharide according to the mass ratio of 1:1.5: 1;
step two, dissolving the weighed agaric polysaccharide and tremella polysaccharide in water, and uniformly mixing to obtain a polysaccharide solution with the mass concentration of 5 g/mL; dissolving the weighed hawthorn flavone in an ethanol solution with the mass concentration of 80% to obtain a hawthorn flavone solution with the mass concentration of 45 mg/mL;
adding mixed enzyme of endoglucanase I, beta-glucosidase, pectinase and cellulase with the mass ratio of 27:25:23:25 into the polysaccharide solution, and performing enzymolysis at 50 ℃ for 3 hours to obtain polysaccharide enzymolysis liquid; the adding amount of the mixed enzyme is 5 wt% of the mass of the polysaccharide solution;
step four, adding a hawthorn flavone solution into the polysaccharide enzymatic hydrolysate, uniformly mixing to obtain a mixed solution, adjusting the pH of the mixed solution to 3.5, adding 0.5g of UPD-glycosyltransferase into each 1L of the mixed solution, and reacting at 65 ℃ for 3 hours to obtain a hawthorn flavone chelated sugar solution;
step five, drying the hawthorn flavone chelated sugar solution by adopting a spray drying method to obtain hawthorn flavone chelated sugar, wherein the drying conditions are as follows: the sample injection concentration is 10g/100mL, the sample injection speed is 5.5mL/min, and the air inlet temperature is 150 ℃.
The preparation method of the hawthorn flavone extract comprises the following steps: freezing and pulverizing fructus crataegi with water content of 2-5%, and sieving with 80 mesh sieve to obtain fructus crataegi powder; adding 75% ethanol solution 40 times the weight of the fructus crataegi powder, ultrasonic extracting at 40 deg.C and 300W for 15min, and filtering to obtain primary filtrate and primary residue; adding 15 times of 75% ethanol solution into the primary filter residue, ultrasonic extracting at 40 deg.C under 300W for 15min, filtering, mixing filtrates, and concentrating to obtain fructus crataegi flavone.
In examples 1-3, the freeze-pulverization of Hawthorn fruit comprises charging liquid nitrogen into a pulverizer, and pulverizing, wherein the charging amount of liquid nitrogen is 5-20% (v/m), the pulverization conditions are 20000-30000rpm/min, and the pulverization time is 1-4 min.
Comparative example 1
The comparative example provides a preparation method of hawthorn flavone chelated sugar, which is the same as the preparation method of example 1, except that the tremella polysaccharide in example 1 is replaced by ganoderma lucidum polysaccharide.
Comparative example 2
The comparative example provides a preparation method of hawthorn flavone chelated sugar, which is the same as the preparation method of example 1, except that hawthorn flavone in example 1 is replaced by cortex mori radicis flavone.
Research on blood fat reducing function
54 Wistar rats were selected and randomly divided into 6 groups of 9 rats each, which were a blank group, a model group, a hawthorn flavone group, a comparative example 1 group, a comparative example 2 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.
The blank group had free access to basal diet, and the remaining 4 groups had free access to 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 group is used for gastric lavage by using the hawthorn flavone extracted in the example 1, and the single administration dosage is 100mg/Kg of body weight; group of comparative example 1, the hawthorn flavone chelating sugar prepared in comparative example 1 is used for gastric perfusion, and the single administration dosage is 50mg/Kg body weight; group of comparative example 2, the hawthorn flavone chelating sugar prepared in comparative example 2 is used for gastric perfusion, and the single administration dosage is 50mg/Kg of body weight; example 1 group, the administration of single dose of 50mg/Kg body weight by intragastric administration of the hawthorn flavone chelate sugar prepared in example 1; 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
| Group of | Quantity (only) | Feed and gavage amount |
| Blank group | 9 | Common feed plus 1mL/100g distilled water |
| Model set | 9 | High fat feed +1mL/100g distilled water |
| Hawthorn flavone group | 9 | High fat feed +100mg/kg |
| Comparative example 1 group | 9 | High fat feed + comparative example 1 chelated sugar 50mg/kg |
| Comparative example 2 group | 9 | High fat feed + comparative example 2 chelated sugar 50mg/kg |
| EXAMPLE 1 group | 9 | High fat diet + example 1 chelated sugar 50mg/kg |
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 lipid 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
| Group of | TC(mmol/l) | TG(mmol/l) | HDL-C(mmol/l) | LDL-C(mmol/l) | AI |
| Blank group | 2.13±0.21## | 0.99±0.07## | 0.77±0.13## | 0.26±0.07## | 2.03±0.72## |
| Model set | 2.7±0.15** | 1.41±0.17** | 0.45±0.10** | 0.42±0.08** | 4.38±1.51** |
| HawFlavone group | 2.36±0.22# | 1.06±0.27## | 0.50±0.14* | 0.41±0.09** | 4.02±1.72** |
| Comparative example 1 group | 2.21±0.21# | 1.03±0.23## | 0.55±0.11* | 0.37±0.08** | 3.98±1.64** |
| Comparative example 2 group | 2.25±0.19# | 1.05±0.22## | 0.57±0.13* | 0.35±0.07** | 3.94±1.59** |
| EXAMPLE 1 group | 2.10±0.16## | 0.91±0.14## | 0.78±0.12*# | 0.26±0.03## | 2.33±0.29## |
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 in the example 1 group have extremely obviously reduced TC and TG (P <0.01), obviously increased HDL-C content (P <0.05), obviously reduced LDL-C content, increased HDL-C content of hawthorn flavone group, comparative example 1 group and comparative example 2 group, and reduced LDL-C content, but the difference has no statistical significance (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, which has higher atherosclerosis risk, and the AI value of the rats of the group of example 1 is reduced by 46.8% after the treatment by the drug 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 hawthorn flavonoids, the group of comparative example 1 and the group of comparative example 2.
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 |
| Hawthorn flavone group | 55.42±10.30 | 226.50±42.32 | 1.74±0.35 | 131.06±18.37 |
| Comparative example 1 group | 57.69±7.51 | 221.59±24.11 | 1.61±0.38 | 134.78±16.58 |
| Comparative example 2 group | 59.69±5.37 | 217.89±25.21 | 1.73±0.32 | 136.47±17.56 |
| EXAMPLE 1 group | 69.23±12.36# | 212.89±28.05 | 1.42±0.19# | 141.25±31.39 |
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, only the CAT activity of the example 1 group is obviously higher than that of the model group, and the MDA content is obviously lower than that of the model group. This shows that the example 1 group has a more excellent effect of improving the oxidation resistance of the body than the comparative example 1 group and the comparative example 2 group.
TABLE 4 serum antioxidant index of each experimental group
| Group of | CAT(U/ml) | GPX(U/ml) | MDA(nmol/ml) | SOD(U/ml) |
| Blank group | 7.85±2.80## | 2243.30±177.36 | 11.81±1.34 | 486.79±57.82# |
| Model set | 3.73±0.34** | 2065.97±195.45 | 12.68±0.89 | 401.18±24.95* |
| Hawthorn flavone group | 4.34±1.49** | 2086.01±210.40 | 11.88±1.58 | 445.08±76.00 |
| Comparative example 1 group | 5.17±0.96* | 2068.52±122.61 | 11.81±1.28 | 458.02±56.58 |
| Comparative example 2 group | 4.54±1.23** | 2075.46±213.67 | 11.73±1.72 | 453.53±68.75 |
| EXAMPLE 1 group | 7.25±2.49## | 2376.57±75.84## | 9.76±0.82# | 449.69±58.47 |
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 and the GPX activity of the example 1 group are obviously improved, the MDA content is obviously reduced, and the comparative example 1 group and the comparative example 2 group are not obviously changed. This shows that example 1 has a better effect of improving the oxidation resistance of the body than the comparative example 1 and comparative example 2.
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 |
| Hawthorn flavone group | 2.53±0.14 | 0.21±0.01# | 0.32±0.04 | 0.51±0.02 |
| Comparative example 1 group | 2.51±0.19 | 0.20±0.01# | 0.33±0.02 | 0.52±0.03 |
| Comparative example 2 group | 2.50±0.17 | 0.21±0.02# | 0.32±0.03 | 0.54±0.02 |
| EXAMPLE 1 group | 2.42±0.08# | 0.19±0.01## | 0.34±0.03 | 0.52±0.02 |
Notes comparison of P with blank group<0.05，**P<0.01; comparison with model group#P<0.05，##P<0.01。
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 coefficient of the group in example 1 has significant difference (P <0.05), and is reduced by 10.37%.
Spleen indices of rats in the hawthorn flavone group, the comparative example 1 group, the comparative example 2 group and the example 1 group were significantly reduced compared to the model group, and the spleen index of the example 1 group was more significantly reduced by 20.8% compared to the comparative example 1 group, the comparative example 2 group and the hawthorn flavone extract group.
Table 6 results of fat coefficients for 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, 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 a model group, the group in the embodiment 1 has a reducing effect on the perirenal fat coefficient, has a very significant difference (P <0.01), and is reduced by 23.7%; the hawthorn flavone group, the comparative example 1 group and the comparative example 2 group also have a reducing effect on perirenal fat coefficient, and have significant difference (P <0.05), but have no very significant difference.
Compared with the model group, the fat coefficient of epididymis is reduced in each group, but compared with the group of the comparative example 1, the group of the comparative example 2 and the hawthorn flavone group, the reduction effect of the group of the example 1 is more obvious, the reduction effect is reduced by 19.4%, and the difference is very significant (P is less than 0.01). Compared with the model group, although the total fat coefficient of each group is reduced, the difference of only the group in the example 1 has statistical significance (P <0.05), and is only 3.93%, thereby indicating that the hawthorn flavone microcapsules prepared in the group in the example 1 can have the weight-reducing effect.
The tests prove that the tremella polysaccharide, the agaric polysaccharide and the hawthorn flavone have a synergistic effect, and the tremella polysaccharide and the agaric polysaccharide hydrolyzed monosaccharide are adopted to glycosylate the hawthorn flavone, so that the bioavailability of the hawthorn flavone is remarkably improved, and the effects of resisting oxidation and reducing blood fat of the hawthorn flavone are further improved. The hawthorn flavone chelated sugar prepared by the invention is proved to have better application prospects in the aspects of oxidation resistance and blood fat reduction.
Within other parameter ranges of the preparation method of the hawthorn flavone chelating sugar, which is defined by the invention, the effects of resisting oxidation and reducing blood fat which are basically the same as those of the embodiment 1 can be achieved.
The tremella polysaccharide and the agaric polysaccharide can be prepared from commercial products or conventional extraction methods in the field, and have no obvious influence on the efficacy of the prepared hawthorn flavone chelating sugar.
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 (10)
1. The preparation method of the hawthorn flavone chelated sugar is characterized by comprising the following steps: hydrolyzing the agaric polysaccharide and the tremella polysaccharide to obtain polysaccharide hydrolysate; uniformly mixing the hawthorn flavone and the polysaccharide hydrolysate, and adding UDP-glycosyltransferase to carry out glycosylation reaction to obtain the hawthorn flavone chelated sugar.
2. The method for preparing hawthorn flavone chelated sugar as claimed in claim 1, comprising the following steps:
step one, dissolving agaric polysaccharide and tremella polysaccharide in water, and uniformly mixing to obtain polysaccharide solution; dissolving hawthorn flavone in 70-90% ethanol solution to obtain hawthorn flavone solution;
adding mixed enzyme into the polysaccharide solution, and performing enzymolysis for 2-4h at 40-55 ℃ to obtain polysaccharide hydrolysate;
and step three, adding a hawthorn flavone solution into the polysaccharide enzymatic hydrolysate, uniformly mixing to obtain a mixed solution, adjusting the pH of the mixed solution to 3.5-5.5, adding UDP-glycosyltransferase, reacting at 40-65 ℃ for 3-6h, and performing spray drying to obtain the hawthorn flavone chelating sugar.
3. The method for preparing hawthorn flavone chelated sugar as claimed in claim 1 or 2, wherein the mass ratio of hawthorn flavone, agaric polysaccharide and tremella polysaccharide is 1:1-1.5: 1-1.5.
4. The method for preparing hawthorn flavone chelated sugar as claimed in claim 1 or 2, wherein the preparation method of hawthorn flavone is as follows: freezing fructus crataegi with water content of 2-5%, pulverizing, and sieving to obtain fructus crataegi powder; adding 65-75% ethanol solution 10-50 times the weight of the hawthorn powder, performing ultrasonic extraction at 40-80 deg.C for 2-3 times for 5-15min, filtering, mixing filtrates, and concentrating to obtain hawthorn flavone.
5. The method for preparing hawthorn flavone chelated sugar as claimed in claim 2, wherein in step one, the mass concentration of the polysaccharide solution is 3-5 g/mL; the concentration of the hawthorn flavone solution is 35-45 mg/mL.
6. The method for preparing hawthorn flavone microcapsules of claim 2, wherein in the second step, the mixed enzyme is endoglucanase I, beta-glucosidase, pectinase and cellulase in a mass ratio of 22-27:20-25:19-23: 25-29.
7. The method for preparing hawthorn flavone chelated sugar as claimed in claim 6, wherein the adding amount of the mixed enzyme is 5-10 wt% of the mass of the polysaccharide solution.
8. The method for preparing hawthorn flavone chelated sugar as claimed in claim 2, wherein in step three, the mass-to-volume ratio of UPD-glycosyltransferase to the mixed solution is 0.5-3:1, wherein the unit of mass is gram and the unit of volume is liter.
9. A hawthorn flavone chelated sugar, which is characterized by being prepared by the preparation method of hawthorn flavone chelated sugar as described in any one of claims 1 to 8.
10. The use of the hawthorn flavone chelate sugar of claim 9 in an antioxidant, hypolipidemic health food.
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| CN114712346B (en) * | 2022-04-20 | 2023-08-08 | 深圳海创生物科技有限公司 | Application of hawthorn polyphenol, hawthorn polysaccharide or composition in preparation of medicines or foods with blood fat reducing and/or weight losing effects |
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