TWI610681B - A manufacture method of a black garlic extraction for anti hemoglobin glycation and reducing blood sugar - Google Patents

Abstract

The invention relates to a method for preparing anti-hemoglobin saccharification and hypoglycemic extract of black garlic, which comprises: (a) an extraction step of extracting black garlic head in a different polar alcohol solvent to obtain a black garlic filtrate; (b) a concentration step The excess solvent contained in the black garlic is removed to obtain a black garlic extract; the extract has anti-hemoglobin saccharification and blood sugar lowering effect.

Description

Anti-hemoglobin saccharification and blood sugar lowering extract of black garlic

The invention discloses a method for preparing anti-hemoglobin saccharification and blood sugar lowering extract, in particular to a method for preparing anti-hemoglobin saccharification and blood sugar lowering extract with black garlic as source.

Diabetes (DM) is a worldwide multi-pathogenic metabolic disease. It is estimated that there are about 200 million people with diabetes worldwide, and the number of patients has increased year by year. The cause of diabetes is complicated, mainly due to insufficient insulin secretion or insulin resistance, which makes the glucose in the blood unable to be effectively utilized, resulting in the rise of blood sugar in patients, which can be divided into the following. Type: Type 1 diabetes, caused by insufficient insulin secretion, occurs in children, the mechanism is unknown, may be caused by autologous immunity caused by pancreatic β-cell defects; type 2 diabetes, also known as non-insulin dependent Diabetes, the more common type of diabetes, occurs mostly in adults. The sensitivity to insulin in the human body decreases, resulting in metabolic imbalance and relative insulin deficiency. The dystrophic diabetes occurs in undeveloped countries. Protein deficiency causes pancreatic fibrosis, affecting its cellular function and causing disease; gestational diabetes, which is diabetes in women during pregnancy; and recurrent diabetes, complicated by pancreatic injury or pathology such as acute pancreatitis Diabetes and so on.

The type of diabetes in Taiwan is mainly type 2 diabetes, which accounts for about 95% of the diabetic population. The main cause of the disease is insulin resistance caused by factors such as obesity, excessive food intake or family history, which affects insulin-dominated body. Normal metabolic effects, such as regulation of glucose concentration, synthesis of triglycerides, and release of free fatty acids, cause elevated blood sugar and hyperlipidemia. Therefore, patients with type 2 diabetes are prone to cardiovascular disease and other diseases. Lesions (such as kidney disease, neurological diseases and visual diseases) are one of the top ten causes of death among Chinese people.

At present, the diagnosis and treatment of diabetes can not achieve the effect of radical cure. It can only control body weight with regular exercise and balanced diet, delay the deterioration of the disease, and then supplement the drug to lower blood sugar and improve insulin resistance. It is used clinically to treat diabetes. Drugs such as thiazolidinediones (the mechanism of action is to increase the sensitivity of insulin receptors); drugs such as biguanides (the mechanism of action is inhibition of glucose absorption) or α-glucosidase inhibitors (the mechanism of action is to reduce hepatic saccharide decomposition); However, this clinical drug can only cure the symptoms and can cause serious side effects. For example, taking biguanides may cause fatal lactic acidosis and liver disease problems; therefore, if you can lock up the high risk group of diabetes - Obese people, taking preventive measures to correct insulin resistance as soon as possible, can avoid diabetes.

Introduction of α-Glucosidase (AG): Sugar is the main source of energy for the human body, accounting for 50-60% of the total calories. Among them, starch and sucrose are the most important sugars in food. After ingestion, the sugar is first decomposed into disaccharide by saliva and gastric juice or amylase in insulin. After entering the small intestine, the α-glucosidase secreted by the intestinal epithelial cells hydrolyzes the disaccharide into a monosaccharide such as glucose. Fructose thus increases the concentration of glucose in the blood. When the α-glucosidase activity in the small intestine wrinkles is too strong, the glucose concentration in the blood will increase rapidly after meals. Therefore, if the decomposition and absorption of sugars can be slowed down, the rate of blood sugar rise can be stabilized, and the resistance can be achieved. color The effect of saccharification and blood sugar lowering.

α-Glucosidase inhibitor (AGI) introduces: AGI belongs to competitive inhibition and reversible competition with α-glucosidase on intestinal villi, which inhibits the decomposition process of starch and sucrose, thereby prolonging glucose in The absorption rate of the digestive tract makes the postprandial blood glucose level not rise rapidly and causes hyperglycemia. Clinically applied AGIs include Acarbose and Voglibose. AGI in plants is mostly derived from polysaccharides, cellulose, pectin, oligosaccharides, isoflavones, saponins, alkaloids, polyphenols, glycopeptides, and sugar alcohols.

Black garlic is related to the research of this application

According to Japanese literature, the antioxidant activity of black garlic in bioconversion is 10 times higher than that of common garlic; blood flow velocity is also increased. Black garlic is a bio-converted product of fresh garlic. The bio-converted garlic has higher total phenolics, flavonoids and SOD-like content than raw garlic, and the main ingredient is Allicin, and for the sulfur in garlic, including DAS , DADS, DATS content have a lifting effect (Xiao, 2008). Studies have shown that the anti-oxidant activity test of raw garlic and black garlic in vivo and in vitro experiments found that the effect of black garlic was significantly higher than that of raw garlic; and in the feeding experiment of type 2 diabetic mice, it was found that the liver of the rats fed the black garlic group was related. Antioxidant enzyme activity and active substances are significantly higher than the raw garlic group, so it is speculated that black garlic has higher antioxidant capacity than raw garlic and can avoid complications of diabetes (Lee et al., 2009).

Polyphenols related research

Polyphenols (Lin, 2002) refer to a number of phenolic hydroxyl groups in the molecule, which are related to antioxidants in the body and contain a general term for such plants. Polyphenols are widely distributed in plants, and their types are more than 8,000, ranging from phenolic acid (C6H5COOH), flavonoids (C6-C3-C6) to tannins, depending on the degree of polymerization of phenol. Various combinations Most of them are in the form of glycoforms. The structure of flavonoids is a polyhydric phenol, and thus has a hydrogen-donating ability to sequester active oxygen radicals such as a hydroxyl group and a superoxide anion. The most significant physiological action of polyphenols is anti-oxidation. Excessive free radicals such as active oxygen in the body cause oxidation of lipids in the cell membrane and blood lipids, causing arteriosclerosis, debilitating heart disease and stroke. The strongest antioxidants in plant polyphenols are flavonoids, which inhibit the peroxidation of lipids in the body by capturing free radicals and chelation of metal ion catalysts for peroxidation.

According to the rationale of the present invention, the polyphenolic substance of black garlic is an effective antioxidant, and the present invention discloses that it has the effects of inhibiting the inhibition of α-glucosidase activity, and can achieve anti-hemoglobin saccharification and blood sugar lowering effect, thereby completing the present invention. .

The main object of the present invention is to provide a method for preparing a black garlic hypoglycemic extract, which can improve the effect of the black garlic extract on inhibiting α-glucosidase activity. In order to achieve the foregoing object, the technical means used in the present invention comprises: a method for preparing a black garlic hypoglycemic extract, comprising: an extraction step of extracting black garlic in an alcohol solvent to obtain a black garlic head; The filtrate; the concentration step removes excess solvent contained in the black garlic filtrate to obtain a black garlic extract.

Another object of the present invention is to provide a health food for improving the anti-hemoglobin saccharification and blood sugar lowering effect of the black garlic head for enhancing the activity of inhibiting α-glucosidase activity, characterized in that it comprises black garlic and a pharmaceutically acceptable carrier, Excipient or diluent.

The above and other objects, features and advantages of the present invention will become more <RTIgt; The agent extracts black garlic. The alcohol solvent has lower polarity and higher volatility than water, and has better compatibility with the α-glucosidase component contained in black garlic, so it is extracted by alcohol. The obtained black garlic extract has a better hypoglycemic effect. In this embodiment, the sample of the black garlic is placed in a solvent (such as water, 50% ethanol, 95% ethanol, ethyl acetate, acetone, n-hexane, etc.), and extracted in a high temperature oscillating manner, and extracted for a suitable time ( Preferably, after 1 to 24 hours), excess impurities or precipitates are removed by centrifugation to obtain a black garlic filtrate. Wherein, the weight ratio of the sample of the black garlic to the solvent is preferably 1:10, so that the black garlic sample can be completely dissolved in the solvent, and the invention is extracted in a water bath thermostat, and the preferred extraction temperature is 50-85. °C, after a suitable time, the impurities contained in the centrifuge were precipitated, and the residual matter was removed by filtration through a filter paper to collect the black garlic filtrate of the present invention.

This concentration step removes the excess solvent obtained in the extraction step by heating or depressurization. The filtrate of the black garlic is concentrated by a decompression method, and the decompression method can be operated by a vacuum concentrator at a certain temperature, so that the alcohol solvent contained in the black garlic filtrate can be completely volatilized, this embodiment The action temperature of the vacuum concentrator is 50 ° C, concentrated and then dissolved in a commercial solvent to a suitable concentration, such as 0.1 g / ml (g / ml), the black garlic extract of the present invention, the black The garlic extract can be further diluted to a suitable concentration with a buffer solution for subsequent testing. The commercial solvent used in this embodiment is Dimethysulfoxide (DMSO), and the buffer solution is a monophosphate buffer solution (concentration of 0.1 M, pH 6). .8).

The composition analysis of the polyphenols and flavonoids of the black garlic of the present invention is determined by the Folin-Ciocalteau reagent as total phenolics, and the result is 215-358 mg/g; the flavonoid is analyzed by the (2-aminoetheyl) diphenylborate analysis method. (favonoids) analysis, the result is 15 ~ 69mg / g.

The black garlic extract of the invention is obtained by extracting different polar solvents at a moderate temperature, and the different polar solvents have different extraction effects on the hypoglycemic components contained in the black garlic, and the black obtained by the alcohol solvent extraction The garlic extract has a better inhibitory effect on α-glucosidase than the extract obtained by the prior art. In order to confirm that the black garlic extract of the present invention has an inhibitory activity against α-glucosidase, the extract obtained by using the black garlic of the present invention in different polar solvents and temperatures is subjected to analysis of extract composition and activity. The main method is to compare the extraction effect of black garlic on the extraction solvent or the extraction temperature.

Take appropriate amount of black garlic, which is divided into arrays using a variety of extraction solvents, including water, 50% ethanol, 95% ethanol, ethyl acetate, acetone and n-hexane or different extraction temperatures; in order to avoid test errors, Except for solvent selection and extraction temperature difference in the extraction process of each group of black garlic, the other steps were carried out by the same treatment method, and the activity of the extracts obtained from each group was calculated.

In this example, the inhibition activity of each group of extracts for α-glucosidase was analyzed by means of an absorbance test, which was carried out by an analytical method proposed by Oki et al. in 1999 and by Shim et al. Modified, sampling the extract from each group, adding α-glucosidase solution and PNP-G respectively for reaction. In this example, the reaction was carried out at 37 ° C for 5-20 minutes and then a stop reagent was added to measure the extracts of each group. The absorbance of 400 nm; wherein the extracts of each group are first diluted to 50-100 μg/ml with the phosphate buffer solution, and then added with a ratio of about 1/2 times the α-glucosidase solution and 4 times the PNP- G (10 mM) is a reaction, and the α-glucosidase is the main action enzyme of the sugar digestion reaction, and if the activity of the α-glucosidase can be inhibited, the increase of blood sugar can be retarded or lowered; this embodiment is in the reaction After 20 minutes, a sodium carbonate solution (Na ₂ CO ₃ ; concentration: 0.1 M, pH 6.8) was added to deactivate the α-glucosidase to terminate the reaction, and the absorbance of each group of extracts was measured by a spectrophotometer. . The measured absorbance was brought into the following formula, and the inhibition rate of each group of extracts for the α-glucosidase was calculated, and the inhibition rate was calculated as (100%)=B-(CA)/B x 100%. Wherein, A is the background absorbance, that is, the absorbance measured in the state in which the extracts of each group are not reacted with α-glucosidase; B is the absorbance measured by the reaction of the phosphate buffer solution in place of each group of extracts. Quality; and C is the absorbance of each group of extracts.

The results in Table 1 show that the extract obtained by extracting black garlic with water, 50% ethanol and 95% ethanol increases the inhibition rate of α-glucosidase with the increase of extraction temperature, and the extraction temperature is 50-80 °C. The inhibition rate is above 60%; and the extract obtained by extraction with ethyl acetate, acetone and n-hexane has an inhibition rate of less than 60%. It can be seen that the black garlic extract obtained by extracting and extracting the temperature with water or alcohol solvent at 50-80 ° C has a better blood sugar lowering effect.

The black garlic hypoglycemic extract of the present invention is subjected to an in vitro test for the inhibition activity of α-glucosidase, and the in vitro test of the present embodiment is referred to the “Evaluation Method for Promoting Iron Absorption Function of Healthy Foods” by the Department of Health of the Executive Yuan. Simulating the reaction of gastrointestinal digestion In the test, the quantitative simulated gastric juice and the simulated pancreatic juice were added to each group of formulas for reaction, and the inhibition rate of each group of formulas for the α-glucosidase was calculated after a certain reaction time. More specifically, the simulated gastric juice is prepared by adding 1 gram of pepsin to a 0.1 N (equivalent concentration) hydrochloric acid solution in the same manner as the pepsin solution in the living body; and the simulated pancreatic juice is modeled after the living body. A pancreatin-bile suspension is prepared by adding 0.2 g of pancreatic enzyme and 1.2 g of bile extract to a 1 M (mole concentration) sodium carbonate solution; the simulated gastric juice and the simulated pancreatic juice are respectively Each group of formulas reacted for 2 to 6 hours [at 37 ° C (body temperature of simulated organisms)], and then the reaction was terminated by heating at high temperature for several minutes, and the inhibitory activities of each group of formulas for α-glucosidase were measured.

Extracting the extract of black garlic with water, 50% ethanol and 95% ethanol to simulate the intestinal environment. The inhibition rate of α-glucosidase is shown by the results in Table 2, except that the extraction temperature is less than 50% at low temperature (25 ° C). The inhibition rate of the remaining groups increased with the increase of extraction temperature, and the inhibition rate of the extraction temperature at high temperature (80 °C) was maintained above 75%.

It can be confirmed from Tables 1 and 2 that the black garlic is extracted with different polar solvents at different temperatures. The obtained extract can effectively increase the inhibitory activity against α-glucosidase and achieve anti-hemoglobin saccharification and blood sugar lowering effects. Accordingly, the present invention discloses a pharmaceutical composition in which black garlic can be utilized for producing an inhibitory activity of α-glucosidase, and a novel use for producing a health food for anti-hemoglobin saccharification and blood sugar lowering.

The invention discloses a method for preparing a black garlic head blood sugar lowering extract, which is characterized in that the extract obtained by extracting the solvent with different polarities at different temperatures can effectively improve the action activity of the black garlic extract, and the extract has the function of promoting inhibition of α-glucoside. Enzymes, to prevent blood sugar from rising to achieve anti-hemoglobin saccharification and blood sugar lowering effect.

The black garlic of the present invention is prepared into a pharmaceutical composition by the addition of an excipient used in the conventional art, wherein the excipient includes the use of the black garlic itself, the black garlic extract, and the like, and a conventional oral dosage form. Agents include: binders, disintegrators and lubricants. Adhesives suitable for pharmaceutical compositions and dosage forms: corn starch, potato starch or other starches, sodium alginate, natural and synthetic gums, powdered tragacanth, guar gum, cellulose and its derivatives, ethyl cellulose , cellulose acetate, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, microcrystalline cellulose, and a mixture of their excipients, the components used in the coating layer include: methyl cellulose (Methyl cellulose), Hydroxymethyl propyl cellulose, Hydroxymethyl propyl cellulose, Polyvinyl alcohol, Polyvinyl pyrrolidone, Cellulose acetate Acetate phthalate) or other material suitable for use in the present invention, methylcellulose, to prepare a pharmaceutical composition. The pharmaceutical composition of the black garlic or the extract thereof according to the present invention may be added with a pharmaceutically acceptable additive.

The invention has the beneficial effects that the health product can be obtained by mixing the black garlic head with the base ingredients which are generally consumed in daily consumption, so that the extract can be obtained and fed in a more appropriate concentration and a more convenient packaging type, thereby enabling Make up for the civilized diseases caused by the imbalance of modern people's diet, and provide health foods that are used as daily maintenance to improve blood sugar. The present invention enhances the effect of inhibiting α-glucosidase activity to achieve anti-hemoglobin saccharification and blood sugar lowering effect. The preferred embodiment comprises black garlic itself, or black garlic extract and a base ingredient. Wherein, the binder component may be a material selected from the group consisting of starch, beans, and grains. And the extract and The base component can be further mixed and processed to form a tablet, a block and a powder. In addition, the base component may also be a liquid, and the extract is formulated with a liquid carrier to form a health care product in the form of a liquid containing or not containing alcohol.

The invention relates to a black garlic health care product which is resistant to hemoglobin saccharification and blood sugar lowering. It should be understood by those skilled in the art that any technical solution of the present invention can be used for concentrated extraction of black garlic, additional additives, and products of different types (eg, gelatinous). Modifications by means of alcohol or non-alcoholic beverages may result in the protection of the present invention.

While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims.

Claims (5)

The invention relates to a method for preparing black garlic anti-hemoglobin saccharification and blood sugar lowering extract, comprising the steps of: (a) extracting black garlic head in water and/or alcohol solvent, and temperature system of water and/or alcohol solvent in extraction step; 50-80 ° C to obtain a black garlic filtrate; (b) concentration step, the excess solvent contained in the black garlic filtrate is removed to obtain an anti-hemoglobin saccharification and blood sugar lowering extract. The method for producing a black garlic anti-hemoglobin saccharification and blood sugar lowering extract according to the first aspect of the invention, wherein the alcohol solvent is ethanol. A use of the black garlic anti-hemoglobin saccharification and hypoglycemic extract obtained in the first aspect of the patent application for the preparation of a pharmaceutical composition for inhibiting α-glucosidase activity. A health food for inhibiting α-glucosidase activity, comprising the black garlic anti-hemoglobin saccharification and hypoglycemic extract obtained according to claim 1 of the patent application, and a food-acceptable carrier, excipient, and diluent Or base material. The health food according to claim 4, wherein the base component is a liquid, and the extract is combined with a liquid carrier to prepare a health care product containing a liquid form containing alcohol or not containing alcohol.