CN115025142A - Mangosteen shell polyphenol capable of reducing blood sugar and preparation method thereof - Google Patents

Mangosteen shell polyphenol capable of reducing blood sugar and preparation method thereof Download PDF

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CN115025142A
CN115025142A CN202210427094.5A CN202210427094A CN115025142A CN 115025142 A CN115025142 A CN 115025142A CN 202210427094 A CN202210427094 A CN 202210427094A CN 115025142 A CN115025142 A CN 115025142A
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mangosteen shell
polyphenol
mangosteen
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刘晓珍
陈宏著
李福香
陈家杰
项君涵
李琳
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Dongguan University of Technology
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/38—Clusiaceae, Hypericaceae or Guttiferae (Hypericum or Mangosteen family), e.g. common St. Johnswort
    • 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
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
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    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/55—Liquid-liquid separation; Phase separation
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention discloses a preparation method of mangosteen shell polyphenol for reducing blood sugar and a composition for reducing blood sugar; the preparation method comprises the following steps: a. pulverizing the mangosteen shell to obtain mangosteen shell powder; b. extracting mangosteen shell polyphenol with acetone; c. filtering and collecting filtrate; d. repeating the steps b and c, and concentrating to obtain a concentrated solution; e. separating and purifying the concentrated solution by a column; f. and (5) drying and packaging. The mangosteen shell polyphenol obtained by the invention has good activity of inhibiting alpha-amylase, can effectively reduce the hydrolysis speed of starch, prevents the appearance of hyperglycemia caused by eating, and has good blood sugar reducing effect.

Description

Mangosteen shell polyphenol capable of reducing blood sugar and preparation method thereof
Technical Field
The invention relates to the field of extraction of plant polyphenol; in particular to mangosteen shell polyphenol for reducing blood sugar and a preparation method thereof.
Background
Mangosteen (Garciniamangostana L.) also called mangosteen, mangosteen and phoenix fruit belongs to the gamboge genus of the gambogaceae family, is an allopolyploid fruit hybridized among Malaysia and Indonesia, is mainly distributed in southeast Asia countries, and is planted in Guangdong, Hainan, Fujian, Taiwan and other provinces in China. The mangosteen serving as a medicine-food homologous fruit is cold in nature and has the effects of clearing heat, reducing internal heat, expelling toxin, beautifying, reducing phlegm, stopping vomiting and the like.
Mangosteen is mainly eaten fresh, a large amount of shells generated after eating are discarded, and excessive byproducts are accumulated due to the utilization of mangosteen for producing fruit wine, preserves, jam, mashed fruits and other products in the food processing industry, so that high resource waste and environmental pollution are generated. The waste such as pomace, pericarp and seeds is used as a continuous source of organic pollution, which causes serious environmental problems. The Food and Agriculture Organization (FAO) estimates that agricultural waste and loss production across the food supply chain may reach 25% to 30%. These by-products contain a large amount of vitamins, minerals, fiber and a large amount of biologically active compounds. In response to environmental issues, there is an urgent need to develop a sustainable way of processing organic waste products and high value-added inputs. The mangosteen shell accounts for about 60% of the fresh mangosteen fruit, and is rich in xanthone, flavonoid and other active substances. The mangosteen skin is used as a folk medicine in southeast Asia to treat abdominal pain, diarrhea, dysentery, infectious wound, suppuration and chronic ulcer.
Methods for extracting strawberry polyphenol from strawberry and loquat polyphenol from loquat have been disclosed; however, since chemical components of polyphenols contained in various plants and fruits are not completely the same, a method for extracting polyphenols from different plants cannot be generally known, and a method for extracting mangosteen husk polyphenols from mangosteen husks is not disclosed.
The polyphenols are important functional active ingredients in mangosteen shell, especially xanthenone such as alpha-mangostin and gamma-mangostin, and have anticancer, antibacterial and blood sugar lowering effects. Researches find that xanthenone, flavonoid, saponin and the like in the mangosteen shell can reduce blood sugar by inhibiting alpha-amylase and alpha-glucosidase. Manaharan et al studied the inhibition of α -amylase and α -glucosidase by crude ethanol extract of mangosteen shell and determined that the acting substances were non-tannin substances with EC50 of 41.7 and 200.0mg/mL, respectively. Adnyana et al compared the inhibitory effect of acarbose, alpha-mangostin, total oxa-mangostin and mangosteen shell ethanol crude extract on alpha-amylase, the IC50 of the acarbose, alpha-mangostin, total oxa-mangostin and mangosteen shell ethanol crude extract are 14.33, 29.67, 517.85 and 105.36 mu g/mL respectively, and the inhibitory activity of the mangosteen shell ethanol crude extract on alpha-amylase is significantly lower than that of standard acarbose and alpha-mangostin and higher than that of oxa-anthracenone. Ibrahim and the like find that aliphatic substitution on C-2, C-4 and C-8 structures of xanthenone, the length of side chains on the C-4 and C-8 structures is longer, and hydroxyl substitution on the C-5 structure and His-299 form hydrogen bonds, so that the inhibition effect of the xanthenone on alpha-amylase can be enhanced. Chen et al verified by enzymatic hydrolysis that gamma-mangostin has higher inhibitory activity on alpha-glucosidase and alpha-amylase than acarbose. It was found that after administering alpha-mangostin for 40 weeks to mice with type 2 diabetes induced by high fat diet and Streptozotocin (STZ), fasting blood glucose level, glycosylation end product and glycosylated hemoglobin A1C can be improved, serum insulin level and glucose tolerance can be increased, and the blood glucose lowering mechanism is to inhibit C-C chemokine receptor 2 signal related to macrophage recruitment induced by obesity, lower ketohexokinase level, inhibit secreted acid sphingomyelinase playing a role in regulating the activity of hepatic phosphoadenosine activated protein kinase and threonine protein kinase, and enhance the expression of leptin and glucose transporter 4 in adipocytes. The gamma-mangostin can lower the blood sugar of diabetic mice in both short time (2h) and long time (28d), and the long-term intake of 4 mg/kg and 8mg/kg of the gamma-mangostin can lower the fasting blood sugar and has no liver toxicity and kidney toxicity of the diabetic mice, and the action mechanism of the gamma-mangostin can be the translocation mediated by adenosine phosphate activated protein kinase and peroxisome proliferator activated receptor gamma, which is different from insulin signal transduction.
Disclosure of Invention
The invention aims to solve the technical problem of providing a preparation method of mangosteen shell polyphenol which is beneficial to large-scale production operation, has high product purity and has the function of reducing blood sugar.
The purpose of the invention can be achieved by adopting the following technical scheme:
a preparation method of mangosteen shell polyphenol for reducing blood sugar comprises the following steps: the method comprises the following steps:
a. crushing the mangosteen shell by using a multifunctional crusher to obtain mangosteen shell powder;
b. adding acetone into the mangosteen shell powder, and homogenizing by using a high-speed disperser while performing ice bath for 1-5 minutes;
c. centrifuging for 5-15 min, filtering and collecting filtrate;
d. repeating the steps b and c for 2-4 times, combining the filtrates, and concentrating to obtain a concentrated solution;
e. dissolving the concentrated solution in an acidified acetone solution; then loading the sample to a macroporous resin chromatographic column; eluting with distilled water after sample loading, and finishing water elution when the color of the elution effluent is the same as that of the blank group when the elution is detected by a phenol-sulfuric acid method; then adopting ethanol water solution for elution;
f. collecting eluate, concentrating by rotary evaporation under vacuum condition, and vacuum freeze drying the concentrate; subpackaging the collected freeze-dried powder of the eluent, and storing in a light-resistant dryer; obtaining mangosteen shell polyphenol for reducing blood sugar.
Preferably, the amount of acetone added in the step b is 3-7 times of the weight of the mangosteen shell powder added in the step a.
Preferably, the acetone in step b is 30-90% by volume acetone solution in water.
Preferably, the filtrates in step d are combined and concentrated to obtain a concentrated solution; the resulting concentrate volume was 20% of the combined filtrate volume.
Preferably, the acidified acetone solution in step e is a mixed solution of acetone and acetic acid in a volume ratio of (99-99.9): 1.
Preferably, the loading in step e is from 1 to 2g per 100g of macroporous resin.
Preferably, said macroporous resin in step e is at least one of HPD300, AB-8 or NKA-9.
Preferably, the ethanol aqueous solution in the step e is 20-80% ethanol aqueous solution by volume ratio.
Preferably, the ethanol aqueous solution in the step e is 60% ethanol aqueous solution by volume.
The invention also provides a hypoglycemic composition; the composition specifically comprises the mangosteen shell polyphenol capable of reducing blood sugar prepared by the preparation method.
Compared with the prior art, the invention has the beneficial effects that:
the mangosteen shell polyphenol is obtained by adopting the technologies of acetone extraction and chromatographic column liquid-phase separation, has the speed of inhibiting amylase from hydrolyzing starch, and can effectively prevent the rapid rise of blood sugar after meals. In the preparation method provided by the invention, the mangosteen shell polyphenol with high activity is obtained by selecting the type of the special resin filler and the concentration of the specific eluent.
Drawings
In order to clearly illustrate the specific embodiments of the present invention and certain detection techniques employed in the experiments, the following description of the embodiments and techniques employed will be made primarily by way of the accompanying drawings.
FIG. 1 shows a gallic acid standard curve;
FIG. 2 shows the elution effect of different resins on mangosteen shell polyphenols;
FIG. 3 is a graph of the fit of the inhibitory effect of mangosteen shell polyphenols eluted with 60% ethanol concentration as a function of concentration;
detailed description of the preferred embodiment
The specific implementation of the present invention is explained by way of examples, and except that the technology used for detection does not limit the present invention in any way, some solutions in the described examples belong to some examples of the present invention, and those skilled in the art can obtain examples without inventive efforts, and all of them belong to the protection scope of the present invention.
Example 1
A method for preparing mangosteen shell polyphenol for reducing blood sugar; comprises the following steps:
a. crushing the mangosteen shell by using a multifunctional crusher to obtain mangosteen shell powder;
b. adding 30-90% acetone 3-7 times the amount of the mixture, and homogenizing with a high-speed disperser in ice bath for 1-5 min;
c, centrifuging at 3500r/min for 5-15 min, and filtering;
d. repeating the steps b and c for 2-4 times, combining the filtrates, and concentrating by 20% volume to obtain a concentrated solution;
e. dissolving the concentrated solution in an acidified acetone solution (the ratio of acetone to acetic acid is 99-99.9: 1, v/v), and loading according to the proportion of 1-2g of sample per 100g of macroporous resin; eluting with distilled water at flow rate of 1.0mL/min to remove water soluble protein, saccharide, organic acid, etc. in the crude extract, and finishing water elution when the color of the eluate is the same as or equivalent to that of the blank group in phenol-sulfuric acid method (at least 3 times of column volume). Then, elution was carried out with 20% -80% (v/v) ethanol aqueous solution of three column volumes, respectively.
f. The eluate was collected, concentrated by rotary evaporation under vacuum at 45 ℃ and the concentrate was freeze-dried under vacuum. And (4) subpackaging the collected elution freeze-dried powder and storing in a light-resistant dryer.
Example 2
Purity and efficacy determinations
10.0mg of mangosteen shell polyphenol purified product was weighed and dissolved in 10mL of deionized water. The purity calculation method of the purified polyphenol is shown as the following formula:
Figure BDA0003608851000000061
in the formula:
c-polyphenol concentration, mg/mL;
v-volume of polyphenols, mL;
m-powder mass after lyophilization, mg.
1. Determination of mangosteen shell polyphenol content
The Folin phenol method is used for determining the content of total phenols in the bamboo shells of Doiscorea polystachya, namely, gallic acid is used as a standard reference substance, the absorbance value of each sample is determined by determining a standard curve of the gallic acid, the concentration of the sample is calculated by a regression equation, and the determination is repeated for 3 times. And then converted to mg GAE/g by the formula, and the results are expressed as mean. + -. standard deviation.
(1) Determination of gallic acid Standard Curve
Gallic acid solutions were prepared at 1mg/mL and diluted to concentration gradients of 0, 20, 40, 60, 80, 100, 150, 200, 300, 400, 500, and 600 μ g/mL. 200. mu.L of each dilution and 800. mu.L of distilled water were transferred to a 10mL test tube, and 200. mu.L of Folin's phenol reagent was added thereto and shaken. Standing in dark for 6min, adding 7% Na 2 CO 3 Shaking the solution 2mL and 1.6mL of deionized water uniformly, standing for 90min in the dark, and measuring the absorbance value by using an ultraviolet spectrophotometer at 760 nm. The above operation was repeated 2 times. Recording the absorbance corresponding to the concentration of gallic acid, and drawing a gallic acid standard curve by using the mass concentration x of gallic acid as an abscissa and the corresponding absorbance value y as an ordinate, as shown in fig. 1.
(2) Determination of polyphenol content
Diluting the free phenol extract and the combined phenol extract of the mangosteen shell by 125 times and 5 times respectively, then transferring 200 mu L of the diluted solution into a 10mL test tube respectively, adding 800 mu L of distilled water, adding 200 mu L of forlin phenol reagent, and shaking up. Standing in dark for 6min, adding 7% Na 2 CO 3 Shaking the solution 2mL and 1.6mL of deionized water uniformly, standing for 90min in a dark place, and measuring the absorbance value by using an ultraviolet spectrophotometer under 760 nm. Repeating the above operation for 3 times, obtaining the average value of the absorbance values, substituting the average value of the absorbance values of the sample to be detected into the gallic acid standard curve, and calculating the polyphenol concentration according to the equation. And then calculating the polyphenol content according to the following formula:
Figure BDA0003608851000000071
total phenol content ═ free phenol content + bound phenol content
In the formula:
Figure BDA0003608851000000081
the average of the absorbance measurements of the sample at 760 nm;
A 0 : absorbance of deionized water at 760 nm;
m: the weight of the weighed mangosteen skin crushed sample, g;
v: the volume of the extracting solution is constant;
n: dilution times of the extracting solution;
2. determination of alpha-amylase activity inhibition by mangosteen shell polyphenol
2.1 materials and instruments
Potato starch, mangosteen shell polyphenol, alpha-amylase (sigma), maltose, sodium carbonate solution, phosphate buffer solution, 4-hydroxybenzoyl hydrazine (Michalin), distilled water and ultraviolet spectrophotometer
2.2 determination of Activity of mangosteen Shell Polyphenol in inhibiting alpha-Amylase
150mg (0.15g) of potato starch (5mg/mL) is weighed firstly, added into 30mL of PBS, added into a rotor to be gelatinized for 30min at 100 ℃ in a water-proof way, and then stirred at 37 ℃ continuously to prepare the mangosteen shell polyphenol concentrations of 1000 mug/mL, 500 mug/mL, 250 mug/mL, 125 mug/mL, 62.5 mug/mL, 31.25 mug/mL and 0 mug/mL. The alpha-amylase was prepared at a concentration of 0.2u/mL, 7mg (0.007g) of alpha-amylase was added to 7mL of PBS and subjected to vortex centrifugation at 4000r/min for 10min, and 1mL of the supernatant was dissolved in 24mL of PBS. Adding 0.4 mL of mangosteen shell polyphenol solution with corresponding concentration into 7 centrifuge tubes with 5mL, respectively adding a magnetic rotor, oscillating and mixing uniformly, adding 1mL of potato starch with the concentration of 5mg/mL, placing the centrifuge tubes into a glass dish, adding about 300mL of distilled water, and placing the centrifuge tubes in a water bath at 37 ℃ under 250r for heat preservation. A2 mL centrifuge tube of 30 pieces was prepared, and 0.6mL sodium carbonate solution was added to inactivate the enzyme for use. Using a stopwatch to time, 1mL of amylase solution with the concentration of 0.2u/mL is added every 34s, then the interval is 4min, 8min and 12min, 0.3mL of enzymolysis solution is added into a 2mL centrifuge tube to inactivate enzyme (the solution is rinsed for three times and is evenly shaken). Opening the water bath, preheating at 97 ℃, transferring 3mL of PAHBAH color developing agent to a 5mL centrifuge tube, adding 0.1mL of enzyme killing solution, rinsing for three times, oscillating uniformly, and making two groups in parallel. And (3) placing the 5mL centrifuge tube in an iron frame for fixation, placing the centrifuge tube in a water bath at 97 ℃ for 7min, taking out the centrifuge tube after the water bath, and cooling the centrifuge tube to room temperature in a dark place. The uv spectrophotometer was turned on, the wavelength was set at 410nm, the sample was poured into a cuvette, and the data was measured and recorded.
3 results and analysis
3.1 elution Effect of different resins on free phenols from mangosteen Shell
The mangosteen shell free phenol was purified using 3 different polarity macroporous resins. Wherein, the adsorption effect of HPD300 and AB-8 is obviously higher than that of NKA-9. (NKA-9 is source leaf S25839; AB-8 is source leaf S30931; HPD300 is source leaf S25183; the source leaf is Shanghai source leaf Biotech Co., Ltd.; the above resin is produced by this Co., Ltd.)
Purification Effect of 3.2 HPD300 type resin on free Polyphenol of mangosteen Shell
As can be seen from table 1, in this example, the method of example 1 is adopted, and the concentration of ethanol elution is changed, so as to obtain different polyphenol contents in the final product under different concentrations, and the purity of the mangosteen shell free polyphenol is increased from 31.42% (the concentration is the concentration before column purification) to 62.31%, which is improved by 1.98 times; thus, the polyphenol obtained when the concentration of ethanol is 60% has the highest purity; meanwhile, the method also shows that the content of impurities (such as polysaccharide impurities and the like) in the sample after purification and freeze-drying is reduced, and the alpha-amylase activity inhibition capability of mangosteen shell polyphenol is improved.
TABLE 1 elution Effect of ethanol of different concentrations on mangosteen shell polyphenols
Figure BDA0003608851000000091
3.3 determination of alpha-Amylase Activity inhibition by mangosteen Shell free phenols
TABLE 2 median inhibitory concentration of ethanol-eluted polyphenols on alpha-amylase
Figure BDA0003608851000000101
The method can be obtained by Graphpadprism nonlinear fitting, wherein the alpha-amylase is subjected to the mangosteen shell polyphenol with different concentrationsIC 50 The value (i.e. the concentration of polyphenol required for inhibiting half of the activity of alpha-amylase) is 1755, 178, 170 and 504 mu g/mL, wherein the mangosteen shell polyphenol eluted by 60% ethanol has better inhibiting effect which is lower than the IC of acarbose on the alpha-amylase 50 The value was 22. mu.g/mL. In the present invention, the mangosteen shell polyphenol obtained is not a pure product but a mixture, and when different polyphenols are eluted with ethanol of different concentrations due to different adsorption capacities on the resin, the obtained mangosteen shell polyphenol is different not only in purity but also in type, and as can be seen from table 2, when combined with HPD300 type resin, when eluted with ethanol of 60% concentration, the obtained resin has the most efficient α -amylase inhibitor effect in type and concentration. The detection data show that the polyphenol prepared by the method can better inhibit the activity of alpha-amylase, is a better natural alpha-amylase inhibitor, and can relieve the degradation of starch when being added into food, so that the blood sugar is prevented from rising too fast after eating. Therefore, the method has good application prospect in the fields of food and medicine.
TABLE 3.60% ethanol concentration eluted mangosteen Shell Polyphenol inhibition of alpha-amylase
Figure BDA0003608851000000102
As shown in fig. 3, the specific values are shown in table 3; the macroporous resin is HPD300, the eluent is 60% ethanol, the polyphenol prepared by elution has the change of the alpha-amylase inhibition rate which is increased along with the increase of the polyphenol concentration, and the fitted curve shows that the polyphenol is increased in an S-shaped curve; the experiment shows that the mangosteen shell polyphenol eluted with the concentration of 0 percent ethanol has the inhibition rate on alpha-amylase, namely IC 50 149.3 μ g/mL; that is, it was shown that when the food contains mangosteen shell polyphenol at a concentration of 149.3 μ g/mL, the hydrolysis rate of α -amylase is suppressed by half, and the pressure of blood sugar elevation is effectively relieved.
All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims (10)

1. A preparation method of mangosteen shell polyphenol for reducing blood sugar comprises the following steps: the method is characterized by comprising the following steps:
a. crushing the mangosteen shell by using a multifunctional crusher to obtain mangosteen shell powder;
b. adding acetone into the mangosteen shell powder, and homogenizing by using a high-speed disperser while performing ice bath, wherein the homogenizing time is 1-5 minutes;
c. centrifuging for 5-15 min, filtering and collecting filtrate;
d. repeating the steps b and c for 2-4 times, combining the filtrates, and concentrating to obtain a concentrated solution;
e. dissolving the concentrated solution in an acidified acetone solution; then loading the sample to a macroporous resin chromatographic column; eluting with distilled water after sample loading, and ending water elution when the color of the elution effluent is the same as that of the blank group when the elution is detected by a phenol-sulfuric acid method; then adopting ethanol water solution for elution;
f. collecting eluate, concentrating by rotary evaporation under vacuum condition, and vacuum freeze drying the concentrate; subpackaging the collected freeze-dried powder of the eluent, and storing in a light-resistant dryer; obtaining mangosteen shell polyphenol for reducing blood sugar.
2. The method for preparing mangosteen shell polyphenol as claimed in claim 1, which comprises the steps of: the method is characterized in that: and (c) adding acetone in the step (b) in an amount which is 3-7 times of the weight of the mangosteen shell powder in the step (a).
3. The method for preparing mangosteen shell polyphenol as claimed in claim 1, which comprises the steps of: the method is characterized in that: the acetone in the step b is acetone aqueous solution with the volume concentration of 30-90%.
4. The method for preparing mangosteen shell polyphenol as claimed in claim 1, which comprises the steps of: the method is characterized in that: combining the filtrates in the step d, and concentrating to obtain a concentrated solution; the resulting concentrate volume was 20% of the combined filtrate volume.
5. The method for preparing mangosteen shell polyphenol for lowering blood glucose according to claim 1: the method is characterized in that: the acidified acetone solution in the step e is a mixed solution of acetone and acetic acid in a volume ratio of (99-99.9): 1.
6. The method for preparing mangosteen shell polyphenol for lowering blood glucose according to claim 1: the method is characterized in that: the sample loading amount in the step e is 1-2g per 100g of macroporous resin.
7. The method for preparing mangosteen shell polyphenol as claimed in claim 1, which comprises the steps of: the method is characterized in that: the macroporous resin in the step e is at least one of HPD300, AB-8 or NKA-9.
8. The method for preparing mangosteen shell polyphenol as claimed in claim 1, which comprises the steps of: the method is characterized in that: the ethanol water solution in the step e is 20-80% of ethanol water solution by volume ratio.
9. The method for preparing mangosteen shell polyphenol for lowering blood glucose according to claim 1: the method is characterized in that: the ethanol water solution in the step e is 60% ethanol water solution by volume ratio.
10. A hypoglycemic composition: characterized by comprising the mangosteen husk polyphenol prepared by the preparation method of any one of claims 1 to 9.
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CN111357909A (en) * 2020-04-02 2020-07-03 吉林农业科技学院 Purification process and determination experimental method of mangosteen polysaccharide hypoglycemic solid beverage

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