CN110787197A - Quinoa bran extract and extraction method and application thereof - Google Patents

Quinoa bran extract and extraction method and application thereof Download PDF

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CN110787197A
CN110787197A CN201911095870.0A CN201911095870A CN110787197A CN 110787197 A CN110787197 A CN 110787197A CN 201911095870 A CN201911095870 A CN 201911095870A CN 110787197 A CN110787197 A CN 110787197A
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quinoa bran
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董琦
王洪伦
胡娜
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Northwest Institute of Plateau Biology of CAS
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Abstract

Experiments prove that the quinoa bran extract can effectively inhibit the activity of α -glucosidase, and the inhibition capability of the quinoa bran 75% ethanol extract and the ethyl acetate extract is superior to that of the conventional α -glucosidase inhibitor, so that the quinoa bran extract can be used for preparing products for regulating carbohydrate metabolism disorder, reducing postprandial blood sugar, preventing and/or treating impaired glucose tolerance and/or diabetes and complications thereof, can be used for preparing medicaments for preventing and/or treating diseases taking α -glucosidase as a target point, such as obesity, hypertension, chronic hepatitis B, AIDS, tumors and the like, and can be further used for preparing α -glucosidase inhibitor with safety and high efficiency.

Description

Quinoa bran extract and extraction method and application thereof
Technical Field
The invention relates to the field of plant extraction, in particular to a quinoa bran extract and an extraction method and application thereof.
Background
Chenopodium quinoa Willd (the name of Chenopodium quinoa Willd) is a plant of Chenopodiaceae, is native to Andes mountain area in south America, is a main traditional food for indigenous residents in printed and native areas, and has an edible and planting history of more than 5000-7000 years. In the early 60 s of the 20 th century, chenopodium quinoa resources are introduced in China, and large-scale chenopodium quinoa planting is not carried out until 2008. In recent years, the planting area of quinoa in China is rapidly enlarged, and the planting area in 2015 is close to 3333hm2The Qinghai area begins to be planted in a large range, the Chenopodium quinoa planting area in China is large, and the Chenopodium quinoa planting area is mainly distributed in Shanxi, Gansu, Qinghai, Jilin and other areas.
Quinoa has unique and comprehensive nutritional value, contains rich total phenols and flavonols, contains the highest content of protein and carbohydrate in the nutritional ingredients, and contains water-soluble substances with higher content of sucrose and oxalic acid. On the other hand, the fat contains unsaturated fatty acids 83%, and has antioxidant, immunoregulatory, and anticancer activities.
Quinoa bran (namely testa sojae atricolor of quinoa) contains a large amount of quinoa saponin, and the saponin is bitter in taste, and is usually removed by washing or grinding before eating, so that a large amount of quinoa bran can be produced in the actual processing process of quinoa. At present, the quinoa bran is mainly used for extracting quinoa saponin, and the utilization mode is very limited. If the quinoa bran can be developed to be applied to other fields, the comprehensive utilization value of the quinoa can be improved better.
Disclosure of Invention
The invention mainly solves the technical problem of providing the quinoa bran extract, the extraction method and the application thereof, and can effectively inhibit the activity of α -glucosidase.
In order to solve the technical problems, the invention adopts a technical scheme that:
provides the application of the quinoa bran extract in preparing products for inhibiting the activity of glucosidase.
Including but not limited to pharmaceuticals, nutraceuticals, foods, and the like.
Experimental data show that the quinoa bran extract has a good inhibition effect on the activity of α -glucosidase, so that the quinoa bran extract can be used for preparing products with related effects, and is safe and effective.
Further, the product is a product for regulating disturbance of carbohydrate metabolism.
The glycochemical metabolic disorder refers to glycometabolism disorder, and glycometabolism disorder (glucose metabolism disorders) refers to abnormal structure, function and concentration of hormones or enzymes regulating metabolism of glucose, fructose, galactose and the like, or pathophysiological changes of tissues and organs, and blood sugar is monitored to increase blood sugar. It is usually caused by some diseases, obesity, high fat diet, etc., or by structural abnormality, function and concentration abnormality of hormone or enzyme for regulating sugar metabolism caused by congenital factors or pathological changes of tissues and organs.
β -glucosidase can participate in cellulose metabolism and various physiological and biochemical pathways, α -glucosidase directly participates in starch and glycogen metabolism pathways, and abnormal functions of the enzyme can cause metabolic diseases, and the enzyme is also an action target of various medicines and inhibitors and is used for regulating the glycochemistry metabolism in a human body.
Further, the product is a product with the function of reducing blood sugar.
Further, the product is a product for reducing postprandial blood glucose.
The quinoa bran extract can slow down the speed of decomposing starch into glucose and reduce and delay the absorption of glucose by small intestine by inhibiting α -glucosidase on intestinal mucosa so as to reduce blood sugar, has obvious effect on postprandial hyperglycemia, and can be used for preparing products for reducing postprandial blood sugar.
Further, the product is a product for preventing and/or treating impaired glucose tolerance and/or diabetes and complications thereof.
The quinoa bran extract has good inhibition effect on the activity of α -glucosidase, can effectively improve impaired glucose regulation, prevent and/or treat impaired glucose tolerance and/or diabetes, can also prevent or improve diabetic complications at the same time, and can be used for preparing related products.
Further, the product is a product for preventing and/or treating one or more symptoms of obesity, hypertension, chronic hepatitis B, AIDS and tumors; further, the product is at least one of a medicine, a health product and a food.
α -glucosidase plays an important role in the process of food absorption, and food can be digested and absorbed after the glucosidase is combined with the food. α -glucosidase inhibitor has a glucose-reducing mechanism that the speed of decomposing starch into glucose is slowed down and the glucose absorption of small intestines is reduced and delayed by inhibiting α -glucosidase on intestinal mucosa so as to reduce blood sugar and have obvious effect on postprandial hyperglycemia.
Further, the product is a glucosidase inhibitor.
In a specific embodiment of the invention, the glucosidase inhibitor is an α -glucosidase inhibitor.
Experiments show that different extracts of the quinoa bran have inhibitory activity on α -glucosidase, the inhibitory capacity of the 75% ethanol extract is better than that of the existing α -glucosidase inhibitor acarbose, and the quinoa bran can be used for preparing α -glucosidase inhibitors.
The invention also provides a composition, which comprises more than 50% of quinoa bran extract by weight; further comprises more than 70 percent of quinoa bran extract by weight.
The invention also provides a product which is characterized by consisting of the quinoa bran extract and pharmaceutically acceptable auxiliary materials or carriers;
the quinoa bran extract can be mixed with pharmaceutically acceptable auxiliary additive components to prepare α -glucosidase inhibitor medicines according to corresponding conventional pharmaceutical preparation methods, for example, the quinoa bran extract can be mixed with commonly used auxiliary additive components such as disintegrants, excipients, lubricants, binders, fillers and the like which can be accepted in oral preparations to prepare solid oral preparations such as tablets, pills, capsules or various corresponding sustained-release agents, controlled-release agents and the like according to conventional operation methods and processes, and can be mixed with conventional surfactants such as solubilizers, emulsifiers, wetting agents, foaming or defoaming agents and the like, diluents, preservatives, stabilizers, flavoring agents, thickening agents and the like to prepare oral medicines in liquid preparations such as water aqua, syrup and the like according to corresponding conventional methods.
Further, the product comprises 50-80% of quinoa bran extract, 10-25% of microcrystalline cellulose, 9-22% of medicinal starch and 1-3% of magnesium stearate.
In the above-mentioned use or composition or product, the quinoa bran extract is one or more of a quinoa bran water extract, a quinoa bran ethyl acetate extract and a quinoa bran ethanol extract.
Further, the quinoa bran ethanol extract is an extract extracted by 65-95% ethanol.
Further, the quinoa bran ethanol extract is an extract extracted by 75% ethanol.
The invention also provides an extraction method of the quinoa bran extract, which comprises the following steps: ultrasonically extracting quinoa bran with a solvent;
further, when the ultrasonic extraction is carried out: the feed-liquid ratio of the quinoa bran to the solvent is 1 (10-20) g/mL, the time is 0.5-1.5 h, the temperature is 20-40 ℃, and the ultrasonic power is 20-50 KHZ; the optimal time is 1.0h, the temperature is 30 ℃, and the ultrasonic power is 40 KHZ;
further, the solvent is one or more of water, ethyl acetate and ethanol; further, the ethanol is 65-95% ethanol water solution; further, the ethanol is a 75% ethanol aqueous solution.
The invention has the beneficial effects that:
(1) the invention successfully extracts the corresponding quinoa bran extracts by water, ethyl acetate and ethanol respectively, and has simple preparation method and convenient operation.
(2) The quinoa bran extract can effectively inhibit the activity of α -glucosidase, and the inhibition capability of the quinoa bran 75% ethanol extract and the ethyl acetate extract is superior to that of the conventional α -glucosidase inhibitor, so that the quinoa bran extract can be used for preparing α -glucosidase inhibitor products, and can also be used for preparing medicaments for preventing and/or treating diseases taking α -glucosidase as a target, such as medicaments for treating diseases of impaired glucose tolerance, diabetes, hypertension, obesity, chronic hepatitis B, AIDS, tumors and the like.
Detailed Description
The technical solutions of the present invention are described clearly and completely below, and it is obvious that the described embodiments are some, not all embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Instruments and reagents
Yeast α -glucosidase (sigma, 1kN, USA);
4-Nitrobenzene- α -D-glucopyranoside (pNPG) (Aladdin reagent, Inc.);
acarbose (Shanghai-derived leaf science and technology Co., Ltd.);
other reagents are domestic analytical pure reagents and are obtained by commercial purchase;
enzyme-labeling instrument (BioTek company, model: EPOCH 2);
constant temperature oscillator (THOMO SHAKER, model: BE-9010).
Example 1 quinoa bran extract
Precisely weighing 10g of quinoa bran, respectively extracting with water, ethyl acetate and 75% ethanol under the conditions of a feed-liquid ratio of 1:20g/mL, a time of 1h, a temperature of 30 ℃ and an ultrasonic power of 40KHZ to obtain samples, carrying out reduced pressure concentration on an extracting solution at 40 ℃, and carrying out freeze drying to obtain a quinoa bran water extract, an ethyl acetate extract and a 75% ethanol extract.
Example 2 quinoa bran extract
Precisely weighing 10g of quinoa bran, respectively extracting with water, ethyl acetate and 75% ethanol at a feed-liquid ratio of 1:10g/mL for 0.5h, at 20 ℃ and an ultrasonic power of 30KHZ to obtain samples, concentrating the extract at 30 ℃ under reduced pressure, and freeze-drying to obtain a quinoa bran water extract, an ethyl acetate extract and a 75% ethanol extract.
Example 3 Tribulus terrestris bran extract
Precisely weighing 10g of quinoa bran, respectively extracting with water, ethyl acetate and 75% ethanol under the conditions of a feed-liquid ratio of 1:15g/mL, a time of 1.5h, a temperature of 40 ℃ and an ultrasonic power of 50KHZ to obtain samples, carrying out reduced pressure concentration on an extracting solution at 60 ℃, and carrying out freeze drying to obtain a quinoa bran water extract, an ethyl acetate extract and a 75% ethanol extract.
Example 4 quinoa bran extract
Precisely weighing 10g of quinoa bran, respectively extracting with water, ethyl acetate and 75% ethanol under the conditions of a feed-liquid ratio of 1:15g/mL, a time of 1h, a temperature of 30 ℃ and an ultrasonic power of 40KHZ to obtain samples, carrying out reduced pressure concentration on an extracting solution at 40 ℃, and carrying out freeze drying to obtain a quinoa bran water extract, an ethyl acetate extract and a 75% ethanol extract.
Example 5 quinoa bran extract
Precisely weighing 10g of quinoa bran, respectively extracting with water, ethyl acetate and 75% ethanol under the conditions of a feed-liquid ratio of 1:10g/mL, a time of 1.5h, a temperature of 40 ℃ and an ultrasonic power of 50KHZ to obtain samples, carrying out reduced pressure concentration on an extracting solution at 40 ℃, and carrying out freeze drying to obtain a quinoa bran water extract, an ethyl acetate extract and a 75% ethanol extract.
Example 6
A hypoglycemic drug comprises the following components in percentage by weight: 80% of quinoa bran water extract, 10% of microcrystalline cellulose, 9% of medicinal starch and 1% of magnesium stearate.
Example 7
A hypoglycemic drug comprises the following components in percentage by weight: 50% of quinoa bran ethyl acetate extract, 25% of microcrystalline cellulose, 22% of medicinal starch and 3% of magnesium stearate.
Example 8
A hypoglycemic drug comprises the following components in percentage by weight: 70% of quinoa bran, 75% of ethanol extract, 15% of microcrystalline cellulose, 13% of medicinal starch and 2% of magnesium stearate.
Example 9
The water extract or ethyl acetate extract or 75% ethanol extract of quinoa bran used in this example was the extract prepared in example 1.
1. Preparation of reagents
(1) Preparation of 0.1M phosphate buffer: a nail polish solution 51mL (NaH) is measured2PO4·2H23.1206g of O, constant volume of 100mL), 49mL of B solution (Na)2HPO4·12H2Weighing 7.1644g, diluting to 100mL), adding 100mL of water, and mixing to obtain a phosphate buffer solution with the pH value of 6.8.
(2) Yeast α -glucosidase 100U/mL enzyme stock solution was diluted to 20U/mL enzyme solution with phosphate buffer (pH 6.8), frozen for use, and diluted to 1U/mL with phosphate buffer (pH 6.8) before use.
(3) Substrate pNPG preparation: accurately weighing 15.02mg of pNPG, adding a proper amount of sodium phosphate buffer solution for dissolving, and then fixing the volume to 100mL to prepare 0.5mmol/L mother liquor.
(4) Preparing an acarbose inhibitor: 253mg of acarbose was precisely weighed, and the volume was adjusted to 10mL with DMSO to prepare 25.3 mg/mL.
(5)0.1mol/L of Na2CO3Preparation: weighing 1.06g of Na2CO3Adding a proper amount of distilled water to dissolve in a beaker, and fixing the volume to 100 mL.
2. Preparation of inhibitors
Precisely weighing 2.000g of quinoa bran water extract or ethyl acetate extract or 75% ethanol extract into a 10mL volumetric flask, dissolving with a small amount of DMSO, and fixing the volume to scale, wherein the water extract is not diluted, and the ethyl acetate extract or 75% ethanol extract is diluted by 10 times with DMSO, and the concentrations are respectively 200, 20 and 20mg/mL as inhibitors.
3. Activity inhibition effect experiment of quinoa bran extract on α -glucosidase
The principle is that hydrolysis of 4-nitrobenzene- α -D-glucoside (pNPG) by α -glucosidase can produce p-nitrophenol, which is specifically absorbed at 405nm, so that the activity of α -glucosidase can be detected by detecting the production amount of p-nitrophenol.
The experiment is divided into a blank group, a control group, a sample blank group and a sample group, and the preparation method comprises the following steps: adding samples of each reactant in a 96-well plate according to the dosage in the table 1, wherein each group comprises 3 parallel samples, uniformly mixing an inhibitor, DMSO, a buffer solution and an enzyme solution, preserving the temperature in a constant temperature oscillator at 37 ℃ for 10min, taking out after the completion, adding 50 mu L of 0.5mmol/L pNPG solution, fully and uniformly mixing, carrying out water bath reaction at 37 ℃ for 20min, adding 50 mu L of 0.1mol/L Na after the completion2CO3And stopping the reaction of the solution to obtain each test group (blank group, control group, sample blank group and sample group).
Because PNPG can be hydrolyzed under the action of α -glucosidase to generate glucose and PNP, PNP has maximum absorption at 405nm, and absorbance of PNP is measured by using an enzyme-labeling instrument, and the inhibition rate and IC50 value of α -glucosidase of each sample can be calculated according to a formula.
The formula:
Figure BDA0002268319450000071
wherein Ac is blank absorbance, ABAs is the absorbance of the control group, As is the absorbance of the sample group, ASBThe absorbance values are for sample blanks.
TABLE 1 metering and sequence of addition of the reactants (. mu.L)
Figure BDA0002268319450000072
Figure BDA0002268319450000081
The results are shown in table 2, the water, 75% ethanol and ethyl acetate extracts of the quinoa bran all have inhibitory activity to α -glucosidase, and the inhibitory capacity is that 75% ethanol extract > ethyl acetate extract > water extract in sequence, wherein the inhibitory activity of 75% ethanol extract to α -glucosidase is obviously stronger than that of the commercial α -glucosidase inhibitor acarbose.
TABLE 2 IC50 value of quinoa bran extract to α -glucosidase
Figure BDA0002268319450000082
Acarbose is an α -glucosidase inhibitor, which can inhibit the activity of α -glucosidase such as maltase, isomaltase, glucoamylase and sucrase, so that the rate of starch decomposition into oligosaccharides such as maltose (disaccharide), maltotriose and dextrin (oligosaccharide) and further glucose decomposition is reduced, and the rate of sucrose decomposition into glucose and fructose is reduced, thereby causing the absorption of glucose in intestinal tract to be slowed down, relieving postprandial hyperglycemia and achieving the effect of reducing blood sugar.
From the experimental results, the water, 75% ethanol and ethyl acetate extracts of the quinoa bran can effectively inhibit the activity of α -glucosidase, can be used for preparing α -glucosidase inhibitors, and can also be used for preparing medicaments for treating diseases taking α -glucosidase as a target point, such as medicaments for treating diseases of impaired glucose tolerance, diabetes, hypertension, obesity, chronic hepatitis B, AIDS, tumors and the like, and in the extracts obtained by extracting the three solvents, the 75% ethanol and ethyl acetate extracts have better inhibition capability on the activity of α -glucosidase than water extracts and are better than acarbose.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.

Claims (10)

1. Use of quinoa bran extract in preparing products for inhibiting glucosidase activity is provided.
2. Use according to claim 1, characterized in that the product is a product for regulating disorders of carbohydrate metabolism.
3. Use according to claim 1, characterized in that the product is a product having a hypoglycemic effect; further, the product is a product for reducing postprandial blood glucose.
4. Use according to claim 1, characterized in that the product is a product for the prevention and/or treatment of impaired glucose tolerance and/or diabetes and its complications.
5. The use according to claim 1, wherein the product is a product for the prevention and/or treatment of one or more of obesity, hypertension, chronic hepatitis B, AIDS, tumors; further, the product is at least one of a medicine, a health product and a food.
6. The use according to claim 1, wherein the product is a glucosidase inhibitor, and further wherein the glucosidase inhibitor is α -glucosidase inhibitor.
7. A composition comprising more than 50% by weight of a quinoa bran extract; further comprises more than 70 percent of quinoa bran extract by weight.
8. A product is characterized by consisting of a quinoa bran extract and a pharmaceutically acceptable auxiliary material or carrier; further comprises 50 to 80 percent of quinoa bran extract, 10 to 25 percent of microcrystalline cellulose, 9 to 22 percent of medicinal starch and 1 to 3 percent of magnesium stearate.
9. The use or composition or product as claimed in any one of claims 1 to 8, wherein the quinoa bran extract is one or more of quinoa bran water extract, quinoa bran ethyl acetate extract and quinoa bran ethanol extract; further, the quinoa bran ethanol extract is an extract extracted by 65-95% ethanol; further, the quinoa bran ethanol extract is an extract extracted by 75% ethanol.
10. The extraction method of the quinoa bran extract is characterized by comprising the following steps: ultrasonically extracting quinoa bran with a solvent; further, when the ultrasonic extraction is carried out: the feed-liquid ratio of the quinoa bran to the solvent is 1 (10-20) g/mL, the time is 0.5-1.5 h, the temperature is 20-40 ℃, and the ultrasonic power is 20-50 KHZ; the optimal time is 1.0h, the temperature is 30 ℃, and the ultrasonic power is 40 KHZ;
further, the solvent is one or more of water, ethyl acetate and ethanol; further, the ethanol is 65-95% ethanol water solution; further, the ethanol is a 75% ethanol aqueous solution.
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Cited By (3)

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CN110591410A (en) * 2019-10-18 2019-12-20 忻州师范学院 Quinoa pigment and extraction method and application thereof
CN112716937A (en) * 2021-02-19 2021-04-30 中国农业大学 Active ingredient composition with synergistic blood sugar reducing function and preparation method thereof
CN113244280A (en) * 2021-06-08 2021-08-13 天津现代创新中药科技有限公司 Chenopodium quinoa willd extract composition and preparation method and application thereof

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