CN107252093B - Guava leaf rich in soluble polyphenol and flavonoid aglycone, preparation method and application - Google Patents

Guava leaf rich in soluble polyphenol and flavonoid aglycone, preparation method and application Download PDF

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CN107252093B
CN107252093B CN201710383696.4A CN201710383696A CN107252093B CN 107252093 B CN107252093 B CN 107252093B CN 201710383696 A CN201710383696 A CN 201710383696A CN 107252093 B CN107252093 B CN 107252093B
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enzymolysis
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hemicellulase
glucosidase
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CN107252093A (en
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吴振强
王露
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South China University of Technology SCUT
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    • A—HUMAN NECESSITIES
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    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract

The invention discloses guava leaves rich in soluble polyphenol and flavonoid aglycone, a preparation method and application thereof. The method comprises the steps of draining, drying, crushing and sieving the cleaned guava leaves to obtain the guava leaf parts; mixing the mixture with water, adjusting the pH value, adding enzyme, and performing enzymolysis reaction; and (5) drying the enzymolysis reaction system to obtain the guava leaf product rich in soluble polyphenol and flavonoid aglycone. The guava leaf tea prepared by the preparation method has the advantages that the content of soluble polyphenol in the guava leaf is greatly improved, the flavonoid glycoside components in the guava leaves are degraded into flavonoid aglycone components with stronger functional activity, the content of aglycone such as quercetin and kaempferol is improved, and the antioxidation and DNA damage resistance effects of the guava leaves are improved. Therefore, the guava leaves rich in soluble polyphenol and flavonoid aglycone have great potential in application in the food field and/or the health-care product field.

Description

Guava leaf rich in soluble polyphenol and flavonoid aglycone, preparation method and application
Technical Field
The invention belongs to the field of food, and particularly relates to guava leaves rich in soluble polyphenol and flavonoid aglycone, a preparation method and application thereof.
Background
Guava leaves as a substance used as both medicine and food have been used for many years, and have multiple curative effects of resisting oxidation, inhibiting DNA damage, reducing blood sugar, resisting inflammation, inhibiting bacteria, reducing blood pressure, protecting heart and the like. Many studies have shown that the main bioactive functional components in guava leaves include polyphenols, which can eliminate the body damage caused by the damage of antioxidant defense system caused by excess oxygen or nitrogen free radicals in the body. The plant polyphenol substances mainly exist in the plant body in three forms (free state, conjugated state and bound state), and bound polyphenol is usually combined with polysaccharide and protein on the plant cell wall in a chemical bond form, so that the extraction is difficult, and the utilization rate of the polyphenol active substances of the guava leaves is low.
Therefore, it is necessary to promote the release of soluble polyphenol and flavonoid aglycone from guava leaves so as to fully utilize the polyphenol active substances in the guava leaves.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of guava leaves rich in soluble polyphenol and flavonoid aglycone.
The invention also aims to provide a guava leaf product obtained by the preparation method.
The invention also aims to provide application of the guava leaf product.
The purpose of the invention is realized by the following technical scheme: a method for preparing guava leaf rich in soluble polyphenol and flavonoid aglycone comprises the following steps:
(1) draining, drying and crushing cleaned guava leaves, sieving the crushed guava leaves, and removing stems of the guava leaves to obtain guava leaf parts with basically consistent sizes;
(2) mixing the guava leaf part finally obtained in the step (1) with water, adjusting the pH value, adding enzyme, and carrying out enzymolysis reaction;
(3) and (3) drying the system obtained after the enzymolysis reaction in the step (2) to obtain the guava leaf product rich in soluble polyphenol and flavonoid aglycone.
The drying condition in the step (1) is preferably drying at 50-80 ℃ to constant weight; more preferably dried at 60 c to constant weight.
The sieve in the step (1) is preferably a sieve with 4 meshes of pore size.
The amount of the water in the step (2) is suitable for dispersing the guava leaves finally obtained in the step (1) so as to be beneficial to carrying out enzymolysis reaction; preferably, the mass of the guava leaves is 4 times of that of the guava leaves finally obtained in the step (1).
The pH value in the step (2) is 4.5-6.0; preferably 5 to 5.5.
The temperature of the enzymolysis reaction in the step (2) is 45-55 ℃; preferably 50 deg.c.
The time of the enzymolysis reaction in the step (2) is preferably 5-8 hours of each enzyme reaction; more preferably 6 hours per enzyme reaction.
The enzyme in the step (2) is at least one of cellulase, hemicellulase, beta-glucosidase and xylanase; preferably, a combination of cellulase, hemicellulase and β -glucosidase is used.
The cellulase is preferably cellulase with the enzyme activity of 8000U/g.
The hemicellulase is preferably hemicellulase with enzyme activity of 8000U/g.
The beta-glucosidase is preferably beta-glucosidase with the enzyme activity of 8000U/g.
The xylanase is preferably xylanase with the enzyme activity of 8000U/g.
The mass dosage of the cellulase is preferably 0.5 percent of the mass of the guava leaf part.
The mass consumption of the hemicellulase is preferably 0.5% of the mass of the leaf part of the guava.
The mass dosage of the beta-glucosidase is preferably 0.5 percent of the mass of the leaf part of the guava.
The mass usage amount of the xylanase is preferably 0.5% of the mass of the guava leaf part.
The specific process of the enzymolysis reaction is preferably shown in steps 1), 2) or 3), and most preferably step 1):
1) firstly, adding cellulase for carrying out first enzymolysis to inactivate the cellulase; adding hemicellulase for second enzymolysis to inactivate the hemicellulase; finally, adding beta-glucosidase to carry out third enzymolysis, and inactivating the beta-glucosidase;
2) adding hemicellulase for first enzymolysis to inactivate the hemicellulase; adding cellulase for second enzymolysis to inactivate the cellulase; finally, adding beta-glucosidase to carry out third enzymolysis, and inactivating the beta-glucosidase;
3) firstly, adding beta-glucosidase to carry out first enzymolysis, and inactivating the beta-glucosidase; adding cellulase for second enzymolysis to inactivate the cellulase; and finally adding hemicellulase for carrying out third enzymolysis to inactivate the hemicellulase.
In the steps 1), 2) and 3),
the reaction conditions of the first enzymolysis, the second enzymolysis and the third enzymolysis are preferably respectively carried out for 6 hours at 50 ℃;
the inactivation condition is preferably 80 ℃ for 10 min;
the mass dosage of the cellulase is preferably 0.5 percent of the mass of the guava leaf part;
the mass usage amount of the hemicellulase is preferably 0.5% of the mass of the leaf part of the guava;
the mass dosage of the beta-glucosidase is preferably 0.5 percent of the mass of the leaf part of the guava.
The drying temperature in the step (3) is preferably 50-70 ℃; more preferably 60 deg.c.
The drying time in the step (3) is preferably at least 12 hours; more preferably 16 h.
A guava leaf product rich in soluble polyphenol and flavonoid aglycone is prepared by the above preparation method.
The guava leaf product rich in soluble polyphenol and flavonoid aglycone is applied to the food field and/or the health-care product field; it can be directly eaten; can also be further processed into various foods, such as guava leaf tea beverage rich in soluble polyphenol and flavonoid aglycone, guava leaf biscuit rich in soluble polyphenol and flavonoid aglycone, nutritious food bar, etc.
Compared with the prior art, the invention has the following advantages and effects:
the preparation method provided by the invention releases the difficult-to-extract and insoluble bound polyphenol in the guava leaves through multiple enzyme hydrolysis, and converts the difficult-to-extract and soluble polyphenol; degrading macromolecular functional components of guava leaves into aglycones with stronger absorption capacity and higher functional activity, such as micromolecule quercetin, kaempferol and the like; and the enzymolysis reaction time is short, the condition is mild, the efficiency is high, and the method can be used for processing and enhancing the effect of medicinal plants. The preparation method has effects of improving antioxidant ability of folium Psidii Guajavae product, inhibiting DNA injury ability, reducing blood sugar and cholesterol, and preventing cardiovascular disease and cerebrovascular disease.
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FIG. 1 is a graph showing the results of measuring the total soluble polyphenol content and the insoluble polyphenol content of guava leaves in different examples.
FIG. 2 is a graph showing the results of measuring the total soluble flavone content and the insoluble flavone content of guava leaves in different examples.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the embodiments of the present invention are not limited thereto.
Example 1
(1) Preparing a guava leaf matrix: drying cleaned guava leaves in a 60 ℃ oven for 16h, rubbing and sieving with a sieve with the aperture of 4 meshes, wherein the sieved guava leaves are the enzymatic hydrolysis matrix; adding water to the enzymatic reaction substrate, the amount of water (pH 5.5 with citric acid) being 80% of the total weight;
(2) and (3) performing enzymatic hydrolysis reaction: then, uniformly mixing cellulase (8000U/g, the same below) and the guava leaves finally obtained in the step (1), placing the mixture in a triangular flask, performing enzyme treatment for 6 hours in a water bath at 50 ℃, then placing the mixture in an oven at 80 ℃ for 10min (inactivating the cellulase), and cooling to room temperature; adding hemicellulase (8000U/g, the same below), mixing, performing enzymolysis in 50 deg.C water bath for 6 hr, placing in 80 deg.C oven for 10min (inactivating hemicellulase), and cooling to room temperature; continuing adding beta-glucosidase (8000U/g, the same below), mixing, performing enzymolysis for 6h in 50 deg.C water bath, and placing in 80 deg.C oven for 10min (to inactivate beta-glucosidase); wherein, the mass dosages of the cellulase, the hemicellulase and the beta-glucosidase are respectively equal to 0.5 percent of the mass of the sieved guava leaves (namely the enzymatic hydrolysis matrix) obtained in the step (1).
(3) Treating guava leaf products after enzyme hydrolysis: and (3) drying the guava leaves hydrolyzed by various enzymes in an oven at 60 ℃ for 16h to obtain the guava leaf product rich in soluble polyphenol and flavonoid aglycone.
Example 2
(1) Preparing a guava leaf matrix: essentially the same as in step (1) of example 1, except that the pH was adjusted to 5.5 with citric acid.
(2) Multi-enzyme hydrolysis reaction: then, uniformly mixing the hemicellulase and the guava leaves finally obtained in the step (1), placing the mixture in a triangular flask, performing enzyme treatment for 6 hours in a water bath at 50 ℃, placing the mixture in an oven at 80 ℃ for 10min (inactivating the hemicellulase), and cooling to room temperature; then adding cellulase, mixing well, performing enzymolysis for 6h in 50 deg.C water bath, placing in 80 deg.C oven for 10min (inactivating cellulase), and cooling to room temperature; continuing to add the beta-glucosidase, mixing uniformly, performing enzymolysis for 6h in a water bath at 50 ℃, and placing in an oven at 80 ℃ for 10min (inactivating the beta-glucosidase); wherein, the mass dosages of the hemicellulase, the cellulase and the beta-glucosidase are respectively equal to 0.5 percent of the mass of the sieved guava leaves (namely the enzymatic hydrolysis matrix) obtained in the step (1).
(3) Treating guava leaf products after enzyme hydrolysis: and (3) drying the guava leaves hydrolyzed by various enzymes in an oven at 60 ℃ for 16h to obtain the guava leaf product rich in soluble polyphenol and flavonoid aglycone.
Example 3
(1) Preparing a guava leaf matrix: same as in step (1) of example 2.
(2) Various enzymatic hydrolysis reactions: uniformly mixing xylanase (8000U/g) and the guava leaves finally obtained in the step (1), placing the mixture in a triangular flask, performing enzyme treatment for 6 hours in a water bath at 50 ℃, placing the mixture in an oven at 80 ℃ for 10min (inactivating xylanase), and cooling to room temperature; then adding cellulase, mixing well, performing enzymolysis for 6h in 50 deg.C water bath, placing in 80 deg.C oven for 10min (inactivating cellulase), and cooling to room temperature; adding hemicellulase, mixing, performing enzymolysis for 6 hr in 50 deg.C water bath, and placing in 80 deg.C oven for 10min (inactivating hemicellulase); wherein, the mass dosages of the xylanase, the cellulase and the hemicellulase are respectively equal to 0.5 percent of the mass of the sieved guava leaves (namely the enzymatic hydrolysis matrix) obtained in the step (1).
(3) Treating guava leaf products after enzyme hydrolysis: and (3) drying the guava leaves hydrolyzed by various enzymes in an oven at 60 ℃ for 16h to obtain the guava leaf product rich in soluble polyphenol and flavonoid aglycone.
Example 4
(1) Preparing a guava leaf matrix: same as in step (1) of example 2.
(2) Various enzymatic hydrolysis reactions: uniformly mixing beta-glucosidase and guava leaves finally obtained in the step (1), placing the mixture in a triangular flask, carrying out enzyme treatment for 6 hours in a water bath at 50 ℃, placing the mixture in an oven at 80 ℃ for 10min (inactivating the beta-glucosidase), and cooling to room temperature; then adding cellulase, mixing well, performing enzymolysis for 6h in 50 deg.C water bath, placing in 80 deg.C oven for 10min (inactivating cellulase), and cooling to room temperature; adding hemicellulase, mixing, performing enzymolysis for 6 hr in 50 deg.C water bath, and placing in 80 deg.C oven for 10min (inactivating hemicellulase); wherein the mass dosages of the beta-glucosidase, the cellulase and the hemicellulase are respectively equal to 0.5% of the mass of the sieved guava leaves (namely the enzymatic hydrolysis matrix) obtained in the step (1).
(3) Treating guava leaf products after enzyme hydrolysis: and (3) drying the guava leaves hydrolyzed by various enzymes in an oven at 60 ℃ for 16h to obtain the guava leaf product rich in soluble polyphenol and flavonoid aglycone.
Effects of the embodiment
Detection method
The guava leaf products prepared in the examples 1 to 4 and the untreated guava leaves were pulverized by a pulverizer, and passed through a 40-mesh sieve for extraction and detection of the following components:
extraction of soluble polyphenol: respectively taking 1.0g of the guava leaf products prepared in the examples 1 to 4, putting the 1.0g of the guava leaf products into a 50mL colorimetric tube, adding 25mL of 50% (v/v) methanol solution, leaching the guava leaf products in a water bath at 45 ℃ for 1h, filtering the guava leaf products by using 0.45 mu m filter paper, carrying out rotary evaporation on filtrate at 37 ℃ for 30min by using a vacuum rotary evaporator to remove the methanol to obtain concentrated solution, adding 40mL of distilled water into the concentrated solution, then adding 10mL of hexane for degreasing, extracting the mixture for 3 times by using 70mL of ethyl acetate, combining the extracts, and carrying out vacuum rotary drying at 35 ℃ to remove the ethyl acetate. Finally, 5mL of 50% (v/v) methanol is added for dissolution, thus obtaining the soluble polyphenol extracting solution. Storing at-20 deg.C for polyphenol content analysis and HPLC quantitative analysis.
② extraction of insoluble bound polyphenol: and (3) adding 40mL of distilled water into the residue of the guava leaves after the soluble polyphenol is extracted in the step (I) to remove the organic solvent, filtering, drying at 60 ℃ to constant weight, and recording the weight of the residue. Adding 40mL of 4M NaOH solution, extracting at room temperature for 4h, adjusting pH to about 2 with concentrated hydrochloric acid (concentration is 37%), adding 70mL of ethyl acetate, extracting for 3 times, combining extracts, performing vacuum spin-drying at 35 ℃, removing ethyl acetate, and finally adding 5mL of 50% methanol for dissolving to obtain the insoluble bound polyphenol extracting solution. Storing at-20 deg.C for polyphenol content analysis and HPLC quantitative analysis.
③ detecting the polyphenol content: respectively sucking 100 μ L of the above extractive solutions, and diluting to appropriate concentration. Taking 1mL of diluted sample solution or gallic acid standard solution (10-100 μ g/mL), sequentially adding 0.5mL of Folin phenol reagent, mixing, reacting for 3-8min, adding 1.5mL of 20% (w/v) Na2CO3Adding water to the solution until the volume is 10mL, fully oscillating, mixing uniformly, and standing for 30 min. A blank reagent is used as a control, and the absorbance at 760nm is measured.
Fourthly, measuring the flavone content: respectively sucking 100 μ L of the above extractive solutions, and diluting to appropriate concentration. Taking 1mL of diluted sample solution or rutin standard solution (10-100 μ g/mL), sequentially adding 0.3mL of 5% (w/v) NaNO2Mixing the solutions, and standing for 5 min. 0.3mL of 10% (w/v) AlCl was added3Mixing the solutions, and standing for 6 min. Then 2mL of 4% (w/v) NaOH solution is added and mixed evenly, 70% (v/v) ethanol solution is added to the mixture to reach the constant volume of 10mL, the mixture is fully shaken and kept stand for 10 min. The absorbance at 510nm was measured against a blank reagent.
Detecting flavonoid aglycone (quercetin and kaempferol): respectively sucking the above extractive solutions, filtering with 0.45 μm filter paper, collecting the clear liquid, filtering with 0.22 μm organic microporous filter membrane, and analyzing the filtrate by HPLC. The specific analysis conditions were: high performance liquid chromatography system (Waters 2695) with ultraviolet detector (Waters 2998), wavelength of 350nm, column temperature of 30 deg.C, and C18 chromatographic column. The mobile phases used were: a-0.1% (v/v) formic acid aqueous solution, B-acetonitrile solution, flow rate of 0.8mL/min, sample size of 10. mu.L. Detection conditions are as follows: gradient elution-0 min, 85% A + 15% B, 5min, 85% A + 15% B, 10min, 80% A + 20% B, 20min, 65% A + 35% B, 30min, 50% A + 50% B, 31min, 20% A + 80% B, 40min, 20% A + 80% B, 45min, 85% A + 15% B, 50min, 85% A + 15% B (all by volume). The analysis time was 50 min.
Detecting the oxidation resistance:
a: DPPH radical scavenging ability
Respectively sucking 100 μ L of the above extractive solutions, and diluting to appropriate concentration. 100 μ L of diluted sample solution or vitamin C standard solution (5-30 μ g/mL) was added with 400 μ L of DPPH-methanol reagent, and the mixture was left standing at 30 ℃ in the dark for 30 min. And (3) taking water as a negative control and VC as a positive control, and determining the light absorption value at 510 nm. The DPPH free radical scavenging ability of the sample is expressed by VC, namely the number of mmol/L of the sample per g of guava leaves is equivalent to VC.
b：ABTS+Radical scavenging ability
Respectively sucking 100 μ L of the above extractive solutions, and diluting to appropriate concentration. Adding 50 μ L diluted sample solution or vitamin C standard solution (5-30 μ g/mL) into 400 μ L ABTS+(7mM ABTS and 2.45mM K2S2O8Mixing in a volume ratio of 2:1, standing in the dark for 16h) the reagent, and standing in the dark for 30min at 30 ℃. And (3) taking water as a negative control and VC as a positive control, and determining the light absorption value at 510 nm. ABTS of samples+The free radical scavenging capacity is expressed by VC, namely the number of mmol/L of VC is equivalent to that of a guava leaf sample per g.
Sixthly, detecting the antioxidant capacity:
respectively taking 2 μ L of diluted soluble polyphenol and insoluble bound polyphenol extractive solution sample (2mg/mL) and quercetin standard solution (2mg/mL), adding 5 μ L of pMD 18-T plasmid DNA (200ng/μ L), and 10 μ L of Fenton reagent (50 m/μ L)MVC、80mM FeCl3And 30mM H2O2) Mixing with pipette, and standing in dark at 37 deg.C for 30 min. And (3) taking PBS buffer solution as a blank control and quercetin as a positive control, then loading the mixed solution into 1% agarose gel for electrophoresis, observing the DNA gel after electrophoresis under an ultraviolet condition, and calculating the proportion of the helical DNA in the total DNA. The calculation formula of the DNA damage inhibition rate is as follows:
Figure BDA0001305774570000071
second, the detection result
As shown in FIGS. 1 and 2, it was found that the soluble polyphenol content and the soluble flavone content of the guava leaf products were significantly increased by the enzyme treatment in the order of example 1 and 2. While the enzyme reaction sequence of example 1 is that after the guava leaves are treated by cellulase, hemicellulase and beta-glucosidase, the soluble polyphenol content of the guava leaves is the highest (the soluble polyphenol content of the guava leaves treated in example 1 is increased by 94.74 percent, the soluble flavone content is increased by 89.48 percent and the insoluble bound polyphenol content is obviously reduced compared with the untreated group), which shows that the soluble polyphenol release of the guava leaves can be promoted by various enzyme treatments. The polyphenol release efficiency is the worst after xylanase, cellulase and hemicellulase are sequentially added in the example 3; however, the soluble polyphenols obtained from the guava leaves treated with β -glucosidase and then treated as in example 4 were less efficient than those obtained from examples 1 and 2, but better than those obtained from example 3, compared to the untreated group.
Detecting flavone aglycone (quercetin and kaempferol) content by high performance liquid chromatography. After the sequential addition of multiple enzymes in example 1 for co-hydrolysis, the contents of quercetin and kaempferol were highest, 248.95mg/100g DM and 11.35mg/100g DM, respectively, and were increased by 1.97 times and 1.82 times, respectively, compared with those of the untreated group. While the total antioxidant activity and the DNA damage inhibition effect of the soluble polyphenol extract of the guava leaves treated by the enzymes in the examples 1 and 2 are obviously improved, wherein the guava leaves treated by the enzymes (0.5 percent of cellulase, 0.5 percent of hemicellulase and 0.5 percent of beta-glucosidase) in the example 1The biological activity is highest. DPPH, ABTS+The radical scavenging ability was equivalent to 74.29mmol VC/g DM, 77.41mmol VC/g DM. The inhibition rate of DNA damage reaches 81.23%. The guava leaves treated by the method of example 3 had the lowest activity. The test data are shown in Table 1.
TABLE 1
Figure BDA0001305774570000072
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (6)

1. A preparation method of guava leaves rich in soluble polyphenol and flavonoid aglycone is characterized by comprising the following steps:
(1) draining, drying, crushing and sieving the cleaned guava leaves, and removing the stems of the guava leaves to obtain the guava leaves with basically consistent sizes;
(2) mixing the guava leaf part finally obtained in the step (1) with water, adjusting the pH value, adding enzyme, and carrying out enzymolysis reaction;
(3) drying the system subjected to the enzymolysis reaction in the step (2) to obtain a guava leaf product rich in soluble polyphenol and flavonoid aglycone;
the specific process of the enzymolysis reaction is shown in steps 1), 2) or 3):
1) firstly, adding cellulase for carrying out first enzymolysis to inactivate the cellulase; adding hemicellulase for second enzymolysis to inactivate the hemicellulase; finally, adding beta-glucosidase to carry out third enzymolysis, and inactivating the beta-glucosidase;
2) adding hemicellulase for first enzymolysis to inactivate the hemicellulase; adding cellulase for second enzymolysis to inactivate the cellulase; finally, adding beta-glucosidase to carry out third enzymolysis, and inactivating the beta-glucosidase;
3) firstly, adding beta-glucosidase to carry out first enzymolysis, and inactivating the beta-glucosidase; adding cellulase for second enzymolysis to inactivate the cellulase; finally adding hemicellulase for carrying out third enzymolysis to inactivate the hemicellulase;
the cellulase has enzyme activity of 8000U/g;
the hemicellulase is the hemicellulase with the enzyme activity of 8000U/g;
the beta-glucosidase is beta-glucosidase with the enzyme activity of 8000U/g;
the mass consumption of the cellulase is equivalent to 0.5 percent of the mass of the guava leaf part;
the mass consumption of the hemicellulase is equivalent to 0.5 percent of the mass of the guava leaf part;
the mass usage amount of the beta-glucosidase is equivalent to 0.5 percent of the mass of the guava leaf part;
the pH value in the step (2) is 4.5-6.0;
the temperature of the enzymolysis reaction in the step (2) is 45-55 ℃;
and (3) the time of the enzymolysis reaction in the step (2) is counted by 5-8 hours of each enzyme reaction.
2. The method of claim 1, wherein the guava leaves are enriched in soluble polyphenols and flavonoid aglycones, and further comprising the steps of:
the drying condition in the step (1) is drying at 50-80 ℃ to constant weight;
the drying temperature in the step (3) is 50-70 ℃.
3. The method of claim 1, wherein the guava leaves are enriched in soluble polyphenols and flavonoid aglycones, and further comprising the steps of:
in the steps 1), 2) and 3),
the reaction conditions of the first enzymolysis, the second enzymolysis and the third enzymolysis are respectively 6 hours of reaction at 50 ℃;
the inactivation condition is that the treatment is carried out for 10min at 80 ℃.
4. A guava leaf product rich in soluble polyphenol and flavonoid aglycone is characterized in that: the preparation method of any one of claims 1 to 3.
5. Use of the guava leaf product enriched in soluble polyphenols and flavonoid aglycones as claimed in claim 4 for the preparation of a food product.
6. Use of the guava leaf product enriched in soluble polyphenols and flavonoid aglycones as claimed in claim 4 for the preparation of a health product.
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