TW201924709A - A manufacture method of a sprout black garlic extraction for anti hemoglobin glycation and reducing blood sugar - Google Patents

Abstract

Polysaccharides are broken down into disaccharides by amylase in saliva, gastric juice or insulin after intake. Subsequently, in the small intestine, disaccharides are further broken down into monosaccharides by intestinal epithelial cells secreted [alpha]-glucosidase and lead the rising of blood glucose concentration. However, over activity of [alpha]-glucosidase in the small intestine folds will cause the blood glucose concentration rising sharply after meal. Slow down the decomposition and absorption of carbohydrates thus can stabilize the increasing rate of blood sugar concentration that avoid hemoglobin glycosylation and reducing the blood sugar. In this study, Sprout black garlic extracts prepared from a variety of solvents at different temperatures were subjected to analyze the effects in inhibition of [alpha]-glucosidase activity in vitro and in simulated gastrointestinal environment. Our results demonstrated that 60-100 DEG C water or alcohol extracts show the effects of suppressing [alpha]-glucosidase activity in vitro. In simulated gastrointestinal environment, the [alpha]-glucosidase activity was also suppressed by the sprout black garlic extracts. The inhibition effects of the extracts were increased with the increasing of their corresponding extraction temperature. Extract from temperature at 100 DEG C, showed an inhibition rate over 98%. The present study reveals the strength of the composition of the sprout black garlic in developing of the [alpha]-glucosidase activity inhibition pharmaceutics, and the anti-hemoglobin glycosylation and hypoglycemic health care foods.

Description

Anti-hemoglobin saccharification and blood sugar lowering extract of germinated 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 germinated 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.

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. A variety of different compounds, mostly in the form of glycosides. 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.

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. Raw garlic, and the main ingredient is Allicin, and has a lifting effect on the content of sulphide in garlic, including DAS, DADS, DATS (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).

At present, the methods, products and physiological activities of making germinated black garlic with germinated garlic are not disclosed in domestic and foreign literatures and patent reports. The research team has been researching black garlic for many years. The technical field related to the present invention is the Republic of China invention patent No. 106116027 "a kind of germinated black garlic food", and the application for the Republic of China invention patent No. 104105785 "a black garlic anti-hemoglobin The method for producing saccharification and hypoglycemic extracts (patent approved), the present invention is a request for deepening revision of the research team's many years of research experience.

The phenolic substance of the germinated black garlic is an effective antioxidant, and the invention discloses that the invention 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. invention.

The invention discloses the anti-oxidation/free radical scavenging ability of germinated black garlic, and the prior literature indicates that black garlic has better antioxidant power, polyphenols and flavonoid content than white garlic (Choi et al., 2014); During germination, plants are susceptible to bacterial, viral and even insect feeding, so plants during germination produce a variety of phytoalexin-like chemicals to protect themselves (Sritongtae et al., 2017).

The invention discloses that the white garlic is germinated under appropriate temperature and humidity, and then fermented (biotransformation) The germinated black garlic was prepared, and the blood sugar test was carried out with the germinated black garlic.

The main object of the present invention is to provide a method for preparing a germinated black garlic hypoglycemic extract, which can improve the effect of the germinated 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 germinated black garlic hypoglycemic extract, comprising: an extraction step of extracting germinated black garlic in a solvent to obtain a germinated black The garlic filtrate; the concentration step removes the excess solvent contained in the germinated black garlic filtrate to obtain a germinated black garlic extract.

Another object of the present invention is to provide a health food for germinated black garlic for improving the effect of inhibiting α-glucosidase activity to achieve anti-hemoglobin saccharification and blood sugar lowering, characterized in that it comprises germinated black garlic and medicinally acceptable Carrier, excipient or diluent.

The above and other objects, features and advantages of the present invention will become more apparent and understood. In the environment, the garlic is germinated 2-5 cm, and then fermented in a temperature of 40-90 ° C, humidity 40-90%, 20-45 days to make germinated black garlic.

The germinated black garlic extract of the present 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 germinated black garlic. In order to confirm that the germinated black garlic extract of the present invention has an inhibitory activity against α-glucosidase, the extract obtained by using the germinated black garlic of the present invention in different polar solvents and temperatures is subjected to analysis of extract composition and activity. This embodiment mainly compares the extraction effects of germinated black garlic on the extraction solvent or the extraction temperature.

The germinated black garlic is treated with an appropriate weight of solvent (eg water, 50% ethanol, 95% ethanol, B) Ethyl acetate, acetone, n-hexane, etc. are extracted by means of constant temperature shaking in a water bath. After extraction for a suitable period of time (preferably 1 to 12 hours), the excess impurities or precipitates are removed by a centrifugal filtration method to obtain a germination. Black garlic filtrate. Wherein, the weight ratio of the sample of the germinated black garlic to the solvent is preferably 1:10, so that the black garlic sample can be completely dissolved in the solvent, and the extraction temperature of the invention is 25 to 100 ° C, and then centrifuged after appropriate time. The impurities contained in the machine were precipitated, and the residual substance was removed by filtration through a filter paper to collect the germinated black garlic filtrate of the present invention, and the extract was freeze-dried, wherein the extraction ratio ranged from 55 to 66%. The ability of each group to remove DPPH free radicals and chelate ferrous ions ranged from 35-55% to 45-65%. The extracts of each group were determined by Folin-Ciocalteau reagent to have a total phenol content range of 55±10 mg gallic acid/g. Dry weight; flavonoid content in the range of 3-7 mg rutin/g dry weight by (2-aminoetheyl) diphenylborate analysis. Since the present invention has been subjected to a number of tests and found to have a dose effect, the sampling used for each group was 0.1 g. The following is only the performance of the examples.

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. (1999) and Shim et al. (2003). modify. The extracts from each group were sampled and added with α-glucosidase solution and PNP-G respectively. 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 at 400 nm. Absorbance value; 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 ( The reaction is carried out at 10 mM, 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 for 20 minutes in the reaction. Thereafter, a sodium carbonate solution (Na ₂ CO ₃ ; concentration: 0.1 M, pH 6.8) was added to terminate the reaction by inactivating the α-glucosidase, 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 germinated black garlic with water, 50% ethanol and 95% ethanol increased the inhibition rate of α-glucosidase with the increase of extraction temperature, and the extraction temperature was inhibited at 60 °C. The rate is above 60%, and it is known that the germinated black garlic extract obtained by water or alcohol solvent extraction and extraction temperature control above 60 ° C has a better blood sugar lowering effect.

The germinated 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 the reference patent application. In the method of No. 99115592, the simulated gastric juice is prepared by formulating a pepsin solution in a living body, and adding 1 gram of pepsin to a 0.1 N (equivalent concentration) hydrochloric acid solution; and the simulated pancreatic juice is modeled after the gallbladder in 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 The samples were co-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 activity of the samples for α-glucosidase was measured.

Extracting the extract of germinated 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 other groups increased with the increase of extraction temperature, and the extraction temperature was higher than 85% at 80 °C and 100 °C (with thermal destructive properties).

As can be confirmed from Tables 1 and 2, the present invention discloses a pharmaceutical composition which 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 germinated black garlic hypoglycemic extract, which is utilized in The extract obtained by extracting the solvent with different polarities at the same temperature has the function of inhibiting α-glucosidase, preventing blood sugar from rising, and achieving anti-hemoglobin saccharification and blood sugar lowering effect.

The germinated 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 germinated black garlic itself, the germinated black garlic extract and the like and a conventional oral dosage form. The excipients include: binders, disintegrating agents 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 germinated 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 extract product can be obtained and fed in a more suitable concentration and a more convenient packaging form by combining the germinated black garlic head with the health care product prepared from the base ingredients of the daily daily consumption. It can 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 improves the effect of inhibiting the activity of α-glucosidase to achieve the anti-hemoglobin saccharification and blood sugar lowering effect. The preferred embodiment comprises the germinated black garlic itself, or the germinated 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 form and a powder form. 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 germinated black garlic health product which is resistant to hemoglobin saccharification and blood sugar lowering, and has the ability Those skilled in the art should understand that any technical solution of the present invention is to produce germinated black garlic, germinated black garlic concentrated extract, additional additives, and different types of products (eg, gelatinous, alcoholic or not). Alcoholic beverages may be modified to the extent that they fall within the scope 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 germinated black garlic anti-hemoglobin saccharification and blood sugar lowering extract, the steps comprising the steps of: (a) extracting germinated black garlic in water and/or an alcohol solvent, and performing water and/or alcohol solvent in the extraction step; The temperature system is 50-80 ° C to obtain a germinated black garlic filtrate; (b) the concentration step is to remove excess solvent contained in the germinated black garlic filtrate to obtain an anti-hemoglobin saccharification and blood sugar lowering extract. A method for producing a germinated 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 germinated 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, which comprises the germinated black garlic anti-hemoglobin saccharification and hypoglycemic extract obtained according to claim 1 of the patent application, and a food acceptable carrier, excipient, and dilution Agent or base. 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.