A hyperglycemia regulating composition and preparation method thereof

Abstract

The invention provides a hyperglycemia regulating composition, comprising 5-15 parts by weight of a mulberry leaf total alkaloid extract, 5-10 parts by weight of a cinnamon extract, 0.5-1.5 parts by weight of a 6kDa-6.5kDa loquat leaf extract, 0.5-1.5 parts by weight of a 7.5kDa-8.5kDa loquat leaf extract, 1-2 parts by weight of a 10kDa-11kDa loquat leaf extract and 0.5-2 parts by weight of a 1800-2000kDa loquat leaf extract. The hyperglycemia regulating composition of the present invention improves the inhibitory effect on α-glucosidase and α-amylase by reasonably combining the components of mulberry leaf total alkaloid extract, cinnamon extract and loquat leaf extract of different molecular weights in a weight ratio, and loquat leaf extract can directly lower blood sugar and assist weight loss, cinnamon extract mainly lowers fasting blood sugar, and mulberry leaf total alkaloid extract significantly lowers postprandial blood sugar, with the incidental effect of lowering fasting blood sugar. After combination, the absorption of food sugar is prevented from the source, and postprandial blood sugar is directly reduced, while obesity is prevented at the same time.

Description

Technical Field

[0001] The invention relates to the application field of plant extracts, and in particular to a hyperglycemia regulating composition and a preparation method thereof. Background Art

[0002] Mulberry leaf extract has multiple biological activities such as anti-oxidation, anti-tumor, anti-virus, and immune regulation. Mulberry leaves are often used to extract alkaloids and flavonoids. Mulberry leaf extract is mostly used in the preparation of health foods. However, due to the difference in the combination of mulberry leaf extract and other components in health foods, the efficacy of each component cannot be better exerted. Summary of the invention

[0003] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a hyperglycemia regulating composition and a preparation method thereof.

[0004] To achieve the above-mentioned purpose, the technical scheme adopted by the present invention is: a hyperglycemia regulating composition, which comprises 5 to 15 parts by weight of mulberry leaf total alkaloid extract, 5 to 10 parts by weight of cinnamon extract, 0.5 to 1.5 parts by weight of 6kDa to 6.5kDa loquat leaf extract, 0.5 to 1.5 parts by weight of 7.5kDa to 8.5kDa loquat leaf extract, 1 to 2 parts by weight of 10kDa to 11kDa loquat leaf extract and 0.5 to 2 parts by weight of 1800 to 2000kDa loquat leaf extract.

[0005] The above-mentioned hyperglycemia regulating composition reasonably combines mulberry leaf total alkaloid extract, cinnamon extract and loquat leaf extract, and improves the inhibitory effect on α-glucosidase and α-amylase by selecting components with molecular weights of 6kDa~6.5kDa, 7.5kDa~8.5kDa, 10kDa~11kDa, and 1800~2000kDa of loquat leaf extract, and reasonably combines the weight proportions of components of loquat leaf extract with different molecular weights. In addition, loquat leaf extract can directly lower blood sugar and assist in weight loss, cinnamon extract mainly lowers fasting blood sugar, and mulberry leaf total alkaloid extract significantly lowers postprandial blood sugar, and has the incidental effect of lowering fasting blood sugar. After combination, it prevents the absorption of food sugar from the source, directly reduces postprandial blood sugar, and also has the effect of preventing obesity.

[0006] Preferably, the preparation method of the mulberry leaf total alkaloid extract comprises: mixing mulberry leaf powder with 50% to 70% ethanol aqueous solution at a solid-liquid ratio of 1 g: (10 to 20) mL, and performing ultrasonic-assisted extraction for 25 to 40 minutes at an extraction temperature of 65°C to 75°C.

[0007] Preferably, the preparation method of loquat leaf extract comprises the following steps:

[0008] (1) drying loquat leaf powder below 60°C and mixing it with 30% to 45% ethanol aqueous solution at a solid-liquid ratio of 1 g: (15 to 25) mL at 75 to 85°C for 1 to 2.5 hours to separate the solid and liquid, and collecting the solid phase to obtain the pre-treated residue;

[0009] (2) The pre-treated residue dried below 60°C was mixed with deionized water at a solid-liquid ratio of 1 g: (15-25) mL at 75-85°C for 1-2.5 hours for solid-liquid separation, and the supernatant was collected to obtain loquat leaf extract A;

[0010] (3) removing the protein and pigment from the extract by dynamic adsorption using polyamide resin and S-8 macroporous resin in sequence to obtain loquat leaf extract B;

[0011] (4) After the loquat leaf extract B was dialyzed with distilled water using a 3500Da dialysis bag, it was passed through a balanced DEAE-52 cellulose chromatography column, and 0.1-0.2 column volumes of distilled water were added to elute and collect fraction A, 0.1-0.2 column volumes of 0.095-0.105 mol / L sodium chloride solution were added to elute and collect fraction B, 0.1-0.2 column volumes of 0.29-0.31 mol / L sodium chloride solution were added to elute and collect fraction C, and 0.1-0.2 column volumes of 0.48-0.52 mol / L sodium chloride solution were added to elute and collect fraction D; the flow rate of the eluent was 2.5-3.5 mL / min;

[0012] (5) Removing the solvent from fraction A yields 6 kDa to 6.5 kDa loquat leaf extract, removing the solvent from fraction B yields 7.5 kDa to 8.5 kDa, removing the solvent from fraction C yields 10 kDa to 11 kDa loquat leaf extract, and removing the solvent from fraction C yields 1800 to 2000 kDa.

[0013] Preferably, in step (2), the solid residue obtained after solid-liquid separation can be recycled in step (2) and then combined with the supernatant.

[0014] Preferably, the hyperglycemia regulating composition comprises 8 to 12 parts by weight of mulberry leaf total alkaloid extract, 6 to 8 parts by weight of cinnamon extract, 0.6 to 1.0 parts by weight of 6kDa to 6.5kDa loquat leaf extract, 0.8 to 1.2 parts by weight of 7.5kDa to 8.5kDa loquat leaf extract, 1.2 to 1.8 parts by weight of 10kDa to 11kDa loquat leaf extract and 0.5 to 1.2 parts by weight of 1800 to 2000kDa loquat leaf extract.

[0015] When the weight of the components of the above-mentioned hyperglycemia regulating composition meets the above-mentioned conditions, the inhibitory effect on α-glucosidase and α-amylase is further improved, and the postprandial blood sugar can be more effectively reduced, and there is also a certain effect of reducing fasting blood sugar.

[0016] Preferably, in the hyperglycemia regulating composition, the weight ratio of 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract and 1800~2000kDa loquat leaf extract is (0.6~1):(0.8~1.2):(1.2~1.8):1.

[0017] Preferably, in the hyperglycemia regulating composition, the weight ratio of 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract and 1800~2000kDa loquat leaf extract is (0.6~0.8):(1~1.2):(1.5~1.8):1.

[0018] In the above-mentioned hyperglycemia regulating composition, by adjusting and optimizing the weight ratio of loquat leaf extracts with different molecular weights, the inhibitory effect on α-glucosidase and α-amylase is further improved, which can more effectively reduce postprandial blood sugar and also has a certain effect of reducing fasting blood sugar.

[0019] Preferably, in the hyperglycemia regulating composition, the weight ratio of 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract and 1800~2000kDa loquat leaf extract is 0.6:1.2:1.8:1.

[0020] In the above hyperglycemia regulating composition, when the weight ratio of 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract, and 1800~2000kDa loquat leaf extract is 0.6:1.2:1.8:1, the inhibitory effect on α-glucosidase and α-amylase is better. It can more effectively reduce postprandial blood sugar and also has a certain effect of reducing fasting blood sugar.

[0021] The present invention also provides a method for preparing any of the above-mentioned hyperglycemia regulating compositions, the method comprising the following steps:

[0022] (I) drying loquat leaf powder below 60° C. and mixing with 30% to 45% ethanol aqueous solution at a solid-liquid ratio of 1 g: (15 to 25) mL at 75 to 85° C. for 1 to 2.5 hours, and then separating the solid and the liquid, and collecting the solid phase to obtain a pre-treated residue;

[0023] (II) refluxing the pre-treated residue after drying at a temperature below 60°C with deionized water at a solid-liquid ratio of 1 g: (15-25) mL at 75-85°C for 1-2.5 hours to separate the solid and liquid, and collecting the supernatant to obtain loquat leaf extract A;

[0024] (III) removing protein and pigment from the extract by dynamic adsorption using polyamide resin and S-8 macroporous resin in sequence to obtain loquat leaf extract B;

[0025] (IV) After the loquat leaf extract B was dialyzed with distilled water using a 3500Da dialysis bag, it was passed through a balanced DEAE-52 cellulose chromatography column, and 0.1 to 0.2 column volumes of distilled water were added to elute and collect fraction A, 0.1 to 0.2 column volumes of 0.095 to 0.105 mol / L sodium chloride solution were added to elute and collect fraction B, 0.1 to 0.2 column volumes of 0.29 to 0.31 mol / L sodium chloride solution were added to elute and collect fraction C, and 0.1 to 0.2 column volumes of 0.48 to 0.52 mol / L sodium chloride solution were added to elute and collect fraction D;

[0026] (V) Removing the solvent from fraction A yields 6 kDa to 6.5 kDa loquat leaf extract, removing the solvent from fraction B yields 7.5 kDa to 8.5 kDa, removing the solvent from fraction C yields 10 kDa to 11 kDa loquat leaf extract, and removing the solvent from fraction C yields 1800 to 2000 kDa;

[0027] (VI) mixing mulberry leaf powder with 50-70% ethanol aqueous solution at a solid-liquid ratio of 1 g: (10-20) mL, performing ultrasonic-assisted extraction at an extraction temperature of 65°C-75°C for 25-40 min, and then removing the solvent to obtain a mulberry leaf total alkaloid extract;

[0028] (VII) Mixing mulberry leaf total alkaloid extract, cinnamon extract, 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract and 1800~2000kDa loquat leaf extract according to weight ratio.

[0029] The beneficial effects of the present invention are as follows: the present invention provides a hyperglycemia regulating composition and a preparation method thereof. The hyperglycemia regulating composition of the present invention reasonably combines mulberry leaf total alkaloid extract, cinnamon extract and loquat leaf extract, selects components with molecular weights of 6kDa-6.5kDa, 7.5kDa-8.5kDa, 10kDa-11kDa and 1800-2000kDa of loquat leaf extract, and reasonably combines the weight proportions of components of loquat leaf extract with different molecular weights, thereby improving the inhibitory effect on alpha-glucosidase and alpha-amylase, and loquat leaf extract can directly lower blood sugar and assist in weight loss, cinnamon extract mainly lowers fasting blood sugar, mulberry leaf total alkaloid extract significantly lowers postprandial blood sugar, and has an incidental effect of lowering fasting blood sugar. After combination, the absorption of food sugar is blocked from the source, and postprandial blood sugar is directly lowered, while obesity is also prevented. DETAILED DESCRIPTION

[0030] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0031] Example 1

[0032] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition includes 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 1 part by weight of 1800~2000kDa loquat leaf extract.

[0033] The preparation method of the hyperglycemia regulating composition of this embodiment comprises the following steps:

[0034] (I) drying loquat leaf powder below 60°C and mixing it with 35% ethanol aqueous solution at a solid-liquid ratio of 1 g:20 mL at 80°C for 2 hours, centrifuging to separate the solid and liquid, and collecting the solid phase to obtain a pre-treated residue;

[0035] (II) the pre-treated residue dried below 60°C was mixed with deionized water at a solid-liquid ratio of 1 g:20 mL at 80°C for 2 hours for solid-liquid separation, and the supernatant was collected to obtain loquat leaf extract A;

[0036] (III) The protein and pigment in the extract are removed by dynamic adsorption of polyamide resin and S-8 macroporous resin to obtain loquat leaf extract B; a certain amount of polyamide resin and S-8 macroporous resin are respectively taken, soaked in anhydrous ethanol for 24 hours to remove the residual organic matter in the resin, filtered and washed with deionized water until there is no ethanol taste, and then soaked in 5% by weight NaOH and 5% by weight HCl for 3 hours respectively, washed with deionized water until neutral, and dried; the polyamide resin and S-8 macroporous resin are respectively loaded into columns by wet method, and the loquat leaf extract A is passed through the polyamide resin and S-8 macroporous resin columns and eluted with deionized water respectively;

[0037] (IV) After the loquat leaf extract B was dialyzed with distilled water using a 3500Da dialysis bag, it was passed through a balanced DEAE-52 cellulose chromatography column, and 0.15 column volumes of distilled water were added to elute and collect fraction A, 0.15 column volumes of 0.1 mol / L sodium chloride solution were added to elute and collect fraction B, 0.15 column volumes of 0.30 mol / L sodium chloride solution were added to elute and collect fraction C, and 0.15 column volumes of 0.5 mol / L sodium chloride solution were added to elute and collect fraction D;

[0038] (V) Removing the solvent from fraction A yields 6 kDa to 6.5 kDa loquat leaf extract, removing the solvent from fraction B yields 7.5 kDa to 8.5 kDa, removing the solvent from fraction C yields 10 kDa to 11 kDa loquat leaf extract, and removing the solvent from fraction C yields 1800 to 2000 kDa;

[0039] (VI) mixing mulberry leaf powder with 65% ethanol aqueous solution at a solid-liquid ratio of 1 g:15 mL, performing ultrasonic-assisted extraction at an extraction temperature of 70°C for 35 min, and then removing the solvent to obtain a mulberry leaf total alkaloid extract;

[0040] (VII) Mixing mulberry leaf total alkaloid extract, cinnamon extract, 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract and 1800~2000kDa loquat leaf extract according to weight ratio.

[0041] Example 2

[0042] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.8 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1 part by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 1 part by weight of 1800~2000kDa loquat leaf extract. The mulberry leaf total alkaloid extract of this embodiment is the same as that of Example 1, and the loquat leaf extract of this embodiment is the same as that of Example 1.

[0043] Example 3

[0044] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.5 parts by weight of 10kDa~11kDa loquat leaf extract and 1 part by weight of 1800~2000kDa loquat leaf extract. The mulberry leaf total alkaloid extract of this embodiment is the same as that of Example 1, and the loquat leaf extract of this embodiment is the same as that of Example 1.

[0045] Example 4

[0046] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 0.5 parts by weight of 1800~2000kDa loquat leaf extract. The mulberry leaf total alkaloid extract of this embodiment is the same as that of Example 1, and the loquat leaf extract of this embodiment is the same as that of Example 1.

[0047] Example 5

[0048] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 1.2 parts by weight of 1800~2000kDa loquat leaf extract. The mulberry leaf total alkaloid extract of this embodiment is the same as that of Example 1, and the loquat leaf extract of this embodiment is the same as that of Example 1.

[0049] Example 6

[0050] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition comprises 5 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 0.5 parts by weight of 1800~2000kDa loquat leaf extract. The mulberry leaf total alkaloid extract of this embodiment is the same as that of Example 1, and the loquat leaf extract of this embodiment is the same as that of Example 1.

[0051] Example 7

[0052] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition comprises 15 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 0.5 parts by weight of 1800~2000kDa loquat leaf extract. The mulberry leaf total alkaloid extract of this embodiment is the same as that of Example 1, and the loquat leaf extract of this embodiment is the same as that of Example 1.

[0053] Example 8

[0054] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition comprises 5 parts by weight of mulberry leaf total alkaloid extract, 5 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 0.5 parts by weight of 1800~2000kDa loquat leaf extract. The mulberry leaf total alkaloid extract of this embodiment is the same as that of Example 1, and the loquat leaf extract of this embodiment is the same as that of Example 1.

[0055] Comparative Example 1

[0056] As a comparative example of the present invention, a hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 1 part by weight of 1800~2000kDa loquat leaf extract. That is, the only difference from Example 1 is that the mulberry leaf total alkaloid extract of Example 1 is replaced by the mulberry leaf extract.

[0057] Comparative Example 2

[0058] As a comparative example of the present invention, a hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 1 part by weight of 1800~2000kDa loquat leaf extract. That is, it does not contain 6kDa~6.5kDa loquat leaf extract.

[0059] Comparative Example 3

[0060] As a comparative example of the present invention, a hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.8 parts by weight of 10kDa~11kDa loquat leaf extract and 1 part by weight of 1800~2000kDa loquat leaf extract. That is, it does not contain 7.5kDa~8.5kDa loquat leaf extract.

[0061] Comparative Example 4

[0062] As a comparative example of the present invention, a hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract and 1 part by weight of 1800~2000kDa loquat leaf extract. That is, it does not contain 10kDa~11kDa loquat leaf extract.

[0063] Comparative Example 5

[0064] As a comparative example of the present invention, a hyperglycemia regulating composition comprises 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, 0.6 parts by weight of 6kDa~6.5kDa loquat leaf extract, 1.2 parts by weight of 7.5kDa~8.5kDa loquat leaf extract, and 1.8 parts by weight of 10kDa~11kDa loquat leaf extract. That is, it does not contain 1 part by weight of 1800~2000kDa loquat leaf extract.

[0065] Comparative Example 6

[0066] As a hyperglycemia regulating composition of an embodiment of the present invention, the hyperglycemia regulating composition includes 10 parts by weight of mulberry leaf total alkaloid extract, 8 parts by weight of cinnamon extract, and 4.6 parts by weight of loquat leaf extract.

[0067] Experimental methods

[0068] 1. Experimental materials

[0069] Food-grade mulberry leaf extract was purchased from Jiangsu Caiwei Biotechnology Co., Ltd., food-grade cinnamon extract was purchased from Guangzhou Huayu Biotechnology Co., Ltd., and loquat leaf extract was purchased from Xi'an Anao Biotechnology Co., Ltd.

[0070] The mulberry leaf total alkaloid extract is prepared in-house, and the preparation method is as shown in Example 1.

[0071] The loquat leaf extract was prepared in-house, and the preparation method was the same as in Example 1.

[0072] 2. Detection of Molecular Weight of Loquat Leaf Extract

[0073] Gel chromatography (GPC) was used to determine the molecular weight of each component of loquat leaf polysaccharide (i.e., fraction A, fraction B, fraction C, and fraction D). Chromatographic conditions: Agilent technologies 1200 series high performance liquid chromatograph, Shodex OhpakSB-804HQ (8.0×300mm) column, differential refractometer (RID), column temperature 25°C, mobile phase 0.1 mol / L NaCl solution, flow rate 0.6 mL / min, injection 20 mL.

[0074] A certain amount of dextran series standard products (molecular weight 3, 10, 40, 100 and 200 kDa) were weighed and prepared into 1 mg / mL solution with 0.1 mol / L NaCl solution. The solution was filtered through a 0.45 μm filter membrane and 20 μL of the solution was injected in order from small to large molecular weight. The retention time was used as the abscissa and the logarithm of the molecular weight (lg Mw) was used as the ordinate to obtain the standard curve of polysaccharide molecular weight Y=0.4302X+9.9086 (R 2 =0.999). At the same time, take an appropriate amount of sample and dissolve it in 0.1 mol / L NaCl solution to prepare a 1 mg / mL solution, filter it through a 0.45 μm filter membrane, and inject it in the same way. Calculate the molecular weight of the polysaccharide based on the peak area, number of peaks, and peak time on the gel chromatogram.

[0075] After testing, the molecular weight of fraction A was 6.18 kDa, the molecular weight of fraction B was 8.12 kDa, the molecular weight of fraction C was 10.13 kDa, and the molecular weight of fraction D was 2015.34 kDa.

[0076] 3. Determination of α-glucosidase inhibitory activity

[0077] 200 μL of the sample solution was thoroughly mixed with an equal volume of α-glucosidase (7000 U mL-1), reacted at 37℃ for 15 min, and then an equal volume of 2.5 mmol·L-1 p-nitrophenyl-α-D-pyranose solution was added, reacted at 37℃ for 20 min, and 5 mL of 0.10 mol·L-1 Na 2 CO 3 The solution was used to terminate the reaction, and the absorbance was measured at 400 nm. The α-glucosidase inhibition rate was calculated as follows:

[0078] α-Glucosidase inhibition rate (%) = (1-(A0-A1) / A2) × 100%

[0079] Wherein: A0 is the OD400 after the reaction of the mixture of sample solution, enzyme and nitrophenyl-α-D-pyranose glucose solution; A1 is the OD400 after the reaction of the buffer solution replacing the α-glucosidase solution; A2 is the OD400 after the reaction of the buffer solution replacing the sample solution.

[0080] 4. Determination of α-amylase inhibitory activity

[0081] 250 μL of the sample solution to be tested was thoroughly mixed with an equal volume of α-amylase solution (1.4 U mL-1), reacted at 37°C for 10 min, and then an equal volume of 1% starch solution was added thereto, reacted at 37°C for 10 min, 500 μL of DNS was added to the reaction system to terminate the reaction, and the mixture was quickly cooled to room temperature after boiling in a water bath for 5 min, and 5 mL of distilled water was added to dilute the mixture, and the absorbance was measured at 540 nm. The α-amylase inhibition rate was calculated by the formula shown in the following formula:

[0082] α-Amylase inhibition rate (%) = (1-(A0-A1) / A2) × 100%

[0083] Wherein: A0 is the OD540 after the reaction of the sample solution, enzyme and starch mixture; A1 is the OD540 after the reaction of the buffer solution replacing the α-amylase solution; A2 is the OD540 after the reaction of the buffer solution replacing the sample solution.

[0084] The hyperglycemia regulating compositions of Examples 1-8 and Comparative Examples 1-6 were prepared into 70 g / L solutions with deionized water, and used as test samples to test the inhibition rates of α-glucosidase and α-amylase.

[0085] The experimental results are shown in Table 1.

[0086] Table 1 α-glucosidase and α-amylase inhibition rates of the hyperglycemia regulating composition.

[0087]

[0088] As can be seen from Table 1, the hyperglycemia regulating composition of the present invention reasonably combines mulberry leaf total alkaloid extract, cinnamon extract and loquat leaf extract, and improves the inhibitory effect on α-glucosidase and α-amylase by selecting components with molecular weights of 6kDa~6.5kDa, 7.5kDa~8.5kDa, 10kDa~11kDa, and 1800~2000kDa of loquat leaf extract, and reasonably combines the weight proportions of components of loquat leaf extract with different molecular weights. In addition, loquat leaf extract can directly lower blood sugar and assist in weight loss, cinnamon extract mainly lowers fasting blood sugar, and mulberry leaf total alkaloid extract significantly lowers postprandial blood sugar, with the incidental effect of lowering fasting blood sugar. After combination, it prevents the absorption of food sugar from the source, directly reduces postprandial blood sugar, and also has the effect of preventing obesity.

[0089] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. A hyperglycemia regulating composition, characterized in that: The hyperglycemia regulating composition comprises 5 to 15 parts by weight of mulberry leaf total alkaloid extract, 5 to 10 parts by weight of cinnamon extract, 0.5 to 1.5 parts by weight of 6 kDa to 6.5 kDa loquat leaf extract, 0.5 to 1.5 parts by weight of 7.5 kDa to 8.5 kDa loquat leaf extract, 1 to 2 parts by weight of 10 kDa to 11 kDa loquat leaf extract, and 0.5 to 2 parts by weight of 1800 to 2000 kDa loquat leaf extract; The preparation method of loquat leaf extract comprises the following steps: (1) drying loquat leaf powder below 60°C and mixing it with 30% to 45% ethanol aqueous solution at a solid-liquid ratio of 1 g: (15 to 25) mL at 75 to 85°C for 1 to 2.5 hours to separate the solid and liquid, and collecting the solid phase to obtain the pre-treated residue; (2) The pre-treated residue dried below 60°C was mixed with deionized water at a solid-liquid ratio of 1 g: (15-25) mL at 75-85°C for 1-2.5 hours for solid-liquid separation, and the supernatant was collected to obtain loquat leaf extract A; (3) removing the protein and pigment from the extract by dynamic adsorption using polyamide resin and S-8 macroporous resin in sequence to obtain loquat leaf extract B; (4) After the loquat leaf extract B was dialyzed with distilled water using a 3500Da dialysis bag, it was passed through a balanced DEAE-52 cellulose chromatography column, and 0.1-0.2 column volumes of distilled water were added to elute and collect fraction A, 0.1-0.2 column volumes of 0.095-0.105 mol / L sodium chloride solution were added to elute and collect fraction B, 0.1-0.2 column volumes of 0.29-0.31 mol / L sodium chloride solution were added to elute and collect fraction C, and 0.1-0.2 column volumes of 0.48-0.52 mol / L sodium chloride solution were added to elute and collect fraction D; the flow rate of the eluent was 2.5-3.5 mL / min; (5) Removing the solvent from fraction A yields 6 kDa to 6.5 kDa loquat leaf extract, removing the solvent from fraction B yields 7.5 kDa to 8.5 kDa loquat leaf extract, removing the solvent from fraction C yields 10 kDa to 11 kDa loquat leaf extract, and removing the solvent from fraction C yields 1800 to 2000 kDa loquat leaf extract.

2. The hyperglycemia regulating composition according to claim 1, characterized in that: The preparation method of the mulberry leaf total alkaloid extract comprises: mixing mulberry leaf powder with an ethanol aqueous solution with a volume fraction of 50% to 70% at a solid-liquid ratio of 1 g: (10 to 20) mL, and performing ultrasonic-assisted extraction for 25 to 40 minutes at an extraction temperature of 65° C. to 75° C.

3. The hyperglycemia regulating composition according to claim 1, characterized in that: In step (2), the solid residue obtained after solid-liquid separation can be recycled through step (2) and then combined with the supernatant.

4. The hyperglycemia regulating composition according to claim 1, characterized in that: The hyperglycemia regulating composition comprises 8 to 12 parts by weight of mulberry leaf total alkaloid extract, 6 to 8 parts by weight of cinnamon extract, 0.6 to 1.0 parts by weight of 6kDa to 6.5kDa loquat leaf extract, 0.8 to 1.2 parts by weight of 7.5kDa to 8.5kDa loquat leaf extract, 1.2 to 1.8 parts by weight of 10kDa to 11kDa loquat leaf extract and 0.5 to 1.2 parts by weight of 1800 to 2000kDa loquat leaf extract.

5. The hyperglycemia regulating composition according to claim 1, characterized in that: In the hyperglycemia regulating composition, the weight ratio of 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract, and 1800~2000kDa loquat leaf extract is (0.6~1):(0.8~1.2):(1.2~1.8):

1.

6. The hyperglycemia regulating composition according to claim 1, characterized in that: In the hyperglycemia regulating composition, the weight ratio of 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract, and 1800~2000kDa loquat leaf extract is (0.6~0.8):(1~1.2):(1.5~1.8):

1.

7. The hyperglycemia regulating composition according to claim 1, characterized in that: In the hyperglycemia regulating composition, the weight ratio of 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract, and 1800~2000kDa loquat leaf extract is 0.6:1.2:1.8:

1.

8. The method for preparing the hyperglycemia regulating composition according to any one of claims 1 to 7, characterized in that: The preparation method comprises the following steps: (I) drying loquat leaf powder below 60° C. and mixing with 30% to 45% ethanol aqueous solution at a solid-liquid ratio of 1 g: (15 to 25) mL at 75 to 85° C. for 1 to 2.5 hours, and then separating the solid and the liquid, and collecting the solid phase to obtain a pre-treated residue; (II) refluxing the pre-treated residue after drying at a temperature below 60°C with deionized water at a solid-liquid ratio of 1 g: (15-25) mL at 75-85°C for 1-2.5 hours to separate the solid and liquid, and collecting the supernatant to obtain loquat leaf extract A; (III) removing protein and pigment from the extract by dynamic adsorption using polyamide resin and S-8 macroporous resin in sequence to obtain loquat leaf extract B; (IV) After the loquat leaf extract B was dialyzed with distilled water using a 3500Da dialysis bag, it was passed through a balanced DEAE-52 cellulose chromatography column, and 0.1 to 0.2 column volumes of distilled water were added to elute and collect fraction A, 0.1 to 0.2 column volumes of 0.095 to 0.105 mol / L sodium chloride solution were added to elute and collect fraction B, 0.1 to 0.2 column volumes of 0.29 to 0.31 mol / L sodium chloride solution were added to elute and collect fraction C, and 0.1 to 0.2 column volumes of 0.48 to 0.52 mol / L sodium chloride solution were added to elute and collect fraction D; (V) Removing the solvent from fraction A yields a 6 kDa to 6.5 kDa loquat leaf extract, removing the solvent from fraction B yields a 7.5 kDa to 8.5 kDa loquat leaf extract, removing the solvent from fraction C yields a 10 kDa to 11 kDa loquat leaf extract, and removing the solvent from fraction C yields a 1800 to 2000 kDa loquat leaf extract; (VI) mixing mulberry leaf powder with 50-70% ethanol aqueous solution at a solid-liquid ratio of 1 g: (10-20) mL, performing ultrasonic-assisted extraction at an extraction temperature of 65°C-75°C for 25-40 min, and then removing the solvent to obtain a mulberry leaf total alkaloid extract; (VII) Mixing mulberry leaf total alkaloid extract, cinnamon extract, 6kDa~6.5kDa loquat leaf extract, 7.5kDa~8.5kDa loquat leaf extract, 10kDa~11kDa loquat leaf extract and 1800~2000kDa loquat leaf extract according to weight ratio.

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