CN101199581A - Effective parts of chickpea germ and bean sprouts and their preparation method and application - Google Patents

Abstract

The present invention relates to an effective part of chickpea germ and bean sprouts and its preparation method and application. The effective parts of total saponins and isoflavones are obtained from chickpea bean germ and bean sprouts by alcohol extraction or water extraction. The effective part has been proved by the preliminary activity screening test: it has the function of protein tyrosine phosphatase (PTP1B) inhibitor. The effective part is used in the application of medicine or health product for treating diabetes, and the effective part or its derivative is used as a pharmaceutical composition.

Description

Effective parts of chickpea germ and bean sprouts and their preparation method and application

technical field

The invention relates to an effective part extracted from chickpea germ and bean sprouts or dried chickpea germ powder and bean sprout powder, and the effective part is used as preparation for treating hypoglycemia, especially for treating type 2 diabetes.

Background technique

Plant extract is a new industry, it has only been ten years since its appearance, but there has been a frenzy of "returning to nature" all over the world. Chinese herbal medicine produces a sense of security, which makes the development of plant extracts rapid and unstoppable. The plant extract market is closely related to the herbal drug market. In 2005, the global herbal drug market was 26 billion US dollars, of which Europe: 34.5% (Germany and France accounted for 65%), North America: 21%, Asia: 26%, Japan: 11.3%. The growth rate of the global herbal medicine market is 10% to 20%, and the growth rate of the global plant extract market is 15% to 20%. The main active countries of plant extracts are the United States, Europe (of which Germany and France are the most active), China, India, South Korea, Japan, Brazil and other countries. The United States is the last country in the world to recognize herbal medicines, but at present, the consumption of herbal extracts in the United States accounts for more than 60% of the world's total. In 2005, the consumption of plant extracts in the United States was about 2.8 billion US dollars. The American "Food Encyclopedia" conducted a survey of 2,000 health food stores, and the results showed that 7.4% of them used extracts as the type of use. In 1992, the sales of herbal medicines in the United States was 1.2 billion US dollars, and reached 7 billion US dollars in 2005, with an average annual growth rate of more than 10%. In 2005, the Asian herbal extract market was US$1.5 billion, China was about US$400 million, accounting for about 26.7%; Japan was US$320 million, accounting for 21%. It can be seen that Chinese herbal medicines have yet to be further developed. At present, there are more than 80 kinds of plant extracts. The natural plant extracts in the North American market mainly include Ginkgo biloba extract - Ginkgo brass, green tea extract - Tea polyphenols, drunken pepper extract - Kava Kava, etc. There are many kinds of domestic natural plant extracts, and the existing varieties can be divided into 3 categories: single herbal extracts, such as Citrus aurantium, Angelica, Astragalus, Schisandra, Ganoderma lucidum, Tribulus terrestris, Magnolia officinalis, Acanthopanax, Hypericum perforatum, Red axle Grass, Ginkgo biloba and other extracts; compound Chinese medicine extracts, such as Buzhong Yiqi Fang extract, etc.; purified extracts, including active parts and monomeric compounds, such as soybean isoflavones, ginsenosides, tea catechins, and quinoa Retrol, huperzine A and so on.

Chickpea (Cicer arietinum L.) originated in West Asia and the Mediterranean coast, and the global planting area has reached 150 million mu. It is currently the second largest edible bean crop in the world. With a growth history of 10 years, it is a commonly used medicinal material in Uyghur medicine. It has been recorded in the Uyghur medicine sub-volume of "Drug Standards" and "Uyghur Medicine Chronicles" of the Ministry of Health of the People's Republic of China. For thin body, low libido, loss of appetite, skin itching and diabetes and other diseases.

Chickpeas are rich in nutrients: every 100 grams of dried beans contains about 10 grams of water, 23 grams of crude protein, 47.3 grams of starch, 5.8 grams of soluble sugar, 5.3 grams of fat, 6.3 grams of crude fiber, and 3.2 grams of ash. Chickpeas are rich in trace elements, among which the vitamin content is 42 mg, which is higher than most beans; the iron content is 47 mg, which is 90% higher than other beans, which makes eating chickpea products can effectively lower the body weight. Serum lipid levels, reduce cholesterol accumulation, strengthen the regulation of the central nervous system of middle-aged and elderly people, and have obvious preventive effects on high blood pressure and heart disease. A comparative study shows that chickpea is the highest in terms of protein efficacy ratio, biological value and digestibility indicators among chickpea, soybean, and mung bean. In 1956, Kleinster, a famous American medical expert, confirmed that chickpea has obvious curative effect on more than 70 severe malnutrition diseases through a large amount of clinical experience.

The medicinal value of chickpeas has also been scientifically proven: it has the functions of strengthening the body, enhancing memory, treating and preventing diabetes, etc. In addition, chickpea seeds can also prevent and treat gallbladder disease, can be used as diuretics, lactogens, treat insomnia, and prevent skin diseases. According to the analysis of existing data, chickpea has a good hypoglycemic and blood lipid-lowering effect on hyperglycemic mice, can reduce the concentration of blood sugar, insulin and triglyceride in serum, and enhance the activity of glucokinase, suggesting that chickpea as a functional Food plays an important role in diabetes prevention and treatment. Studies have also shown that chickpea has good anticancer activity. Giron-Calle J et al. found that the ethanol and acetone extracts of chickpea can almost completely inhibit the growth of tumor cells, showing good antitumor activity. Studies have found that components such as isoflavones and saponins in chickpea have good blood lipid-lowering effects, and these components are mainly concentrated in chickpea germ. Therefore, the inventors have conducted research on reducing blood sugar on chickpea sprouts. The study adopted the method of intragastric administration, and found that the chickpea bean sprouts extract had a significant hypoglycemic effect on alloxan hyperglycemia model mice. Although the model group animals gradually decreased their blood sugar over time, the blood sugar levels of the administration group There is a very significant difference between the percentage decrease and its comparison.

Since the occurrence and development of type 2 diabetes can be caused by the interaction of genetic and environmental factors, among them, the disorder of insulin signal transduction plays an important role in the pathogenesis. Protein tyrosine phosphatase 1B (Protein tyrosine phosphatase PTP1B) is a key negative regulator of insulin signal transduction. PTP1B is the representative of intracellular PTP. It is the first mammalian protein tyrosine phosphatase identified and purified. It consists of 435 amino acid residues and has a molecular weight of about 50KD. Protein tyrosine phosphatase 1B is active in various human tissues. It is ubiquitously present and expressed in cells, and exerts physiological functions by dephosphorylating and inactivating activated insulin receptors and insulin receptor substrates (IRs-1). It mediates the balance between phosphorylation and dephosphorylation of tyrosine residues, preventing insulin signaling. The content and activity of specific protein tyrosine phosphatase in patients with type 2 diabetes and insulin resistance both increased. When protein tyrosine phosphatase was deleted, not only insulin sensitivity increased, but also the symptoms of diabetes improved. Mice deficient in protein tyrosine phosphatase 1B are more insulin sensitive and have better glycemic control than their wild-type control mice , and have better resistance to diet-induced obesity. Therefore, inhibition of the action of protein tyrosine phosphatase 1B has become a novel approach for the treatment of insulin resistance (IR), type II diabetes and obesity.

Contents of the invention

The purpose of the present invention is to provide an effective part of total saponins and isoflavones obtained from fresh chickpea bean germ, bean sprouts or dried chickpea germ powder and bean sprout powder by solvent extraction method. The activity screening test proves that: it has the function of protein tyrosine phosphatase (PTP1B) inhibitor, and the effective part is used for the application of medicine or health care product for treating diabetes, and at the same time, the effective part or its derivatives can be used as the drug composition application.

According to the present invention, the solvent extraction method (alcohol extraction method or water extraction method) is used to prepare the effective parts of chickpea germ and bean sprouts and the preparation method and application thereof. The effective parts contain isoflavones and saponins, and the specific operations are as follows: conduct:

a, germinate the chickpeas by a conventional method;

b. Add 2-10 times the volume of methanol or ethanol or isopropanol or butanol to fresh chickpea germ and bean sprouts or dry chickpea germ powder and bean sprout powder, the concentration of which is 10-100%.

c. Extract 1-15 times with ultrasonic wave or cold dipping or heating and reflux, each time for 1-3 hours, wherein the ultrasonic extraction temperature is 20-40°C, and the reflux extraction temperature is 60-80°C, filter with filter paper, combine the filtrate, reduce Concentrate under pressure until there is no alcohol smell, and obtain the effective part of the alcohol extract;

d, or add 2-10 times the volume of purified water to fresh chickpea germ and bean sprouts or dry chickpea germ powder and bean sprout powder;

e. Use ultrasound or cold soaking or heating and reflux extraction for 1-15 times, each time for 2-4 hours, wherein the ultrasonic extraction temperature is 20-60 ° C, and the reflux extraction temperature is 60-100 ° C. Filter with gauze and filter paper respectively, and combine the filtrates , concentrated under reduced pressure to obtain the effective fraction of the water extract.

The effective parts are total saponins and isoflavones.

Prepare the effective part as functional food according to conventional methods, or make capsules or tablets or oral liquids or granules or injections or sustained-release preparations or controlled-release preparations according to conventional pharmaceutical methods; or make capsules by combining the effective parts with Chinese and Western medicines Or tablets or oral liquids or granules or injections or sustained-release preparations or controlled-release preparations.

A method for preparing the effective parts of chickpea germ and bean sprouts, the effective parts are prepared by alcohol extraction or water extraction, and the specific operation is carried out according to the following steps:

a, germinate the chickpeas by a conventional method;

b. Add 2-10 times the volume of methanol or ethanol or isopropanol or butanol to fresh chickpea germ and bean sprouts or dry chickpea germ powder and bean sprout powder, the concentration of which is 10-100%.

c. Extract 1-15 times with ultrasonic wave or cold dipping or heating and reflux, each time for 1-3 hours, wherein the ultrasonic extraction temperature is 20-40°C, and the reflux extraction temperature is 60-80°C, filter with filter paper, combine the filtrate, reduce Concentrate under pressure until there is no alcohol smell, and obtain the effective part of the alcohol extract;

d, or add 2-10 times the volume of purified water to fresh chickpea germ and bean sprouts or dry chickpea germ powder and bean sprout powder;

e. Use ultrasound or cold soaking or heating and reflux extraction for 1-15 times, each time for 2-4 hours, wherein the ultrasonic extraction temperature is 20-60°C, and the reflux extraction temperature is 60-100°C, filter with gauze and filter paper respectively, and combine The filtrate was concentrated under reduced pressure to obtain the effective fraction of the water extract.

In the preparation method, the optimal concentration of alcohol in step b is 60-70%

In the preparation method, in step c, the temperature of ultrasonic extraction of alcohol is preferably 30-35°C, and the temperature of reflux extraction is preferably 70-80°C.

In the preparation method, the ultrasonic extraction temperature in step e is preferably 30-40°C, and the reflux extraction temperature is preferably 80-96°C.

The invention relates to the use of the chickpea germ and the effective parts of the bean sprouts as a medicine for treating diabetes and hypoglycemic function or as a hypoglycemic functional food.

The preparation method for extracting the hypoglycemic active ingredient from fresh chickpea bean germ, bean sprout segment or dried chickpea germ powder and bean sprout powder and its application in functional food and medicine include the following process steps:

Preparation of chickpea germ, bean sprouts

Wash the selected bean raw materials with water and put them into a container. A washing device is required on the top of the container, and a water hole with a diameter of 5cm-6cm should be reserved on the bottom edge and a vent hole. Then add an appropriate amount of water for soaking to facilitate its germination. Usually soaking for 2h-3h can ensure that the raw beans can absorb enough water. The raw beans that have been soaked and absorbed enough water are poured with water once every 5h-6h at room temperature. The bean sprouts can grow to 6cm-8cm long, and the longest should not exceed 10cm;

Preparation of Chickpea Germ and Alcoholic Extract of Bean Sprout

Place the fresh germs and bean sprouts of chickpea (about 2 mm in length each) or dry germs and bean sprouts (25 mesh) in a round bottom flask, add alcohol with a concentration of 10-100%, and use ultrasound or cold soak Or heat and reflux extraction for 1-15 times, each time for 1 hour, filter through filter paper, combine the filtrate, concentrate under reduced pressure until there is no alcohol smell, and obtain the alcohol extract;

Preparation of Chickpea Germ and Bean Sprout Water Extract

Put fresh germs, bean sprouts (each segment about 2mm long) or dried chickpea germs, bean sprouts (25 mesh) in a round bottom flask, add pure water, and extract 1- 15 times, each time for 2 hours, filter with gauze and filter paper respectively, combine the filtrate, concentrate under reduced pressure to obtain the water extract;

The effective parts of chickpea germ and bean sprouts include: total saponins, isoflavones and other compounds.

The effective parts of chickpea germ and bean sprouts are used as preparations for lowering blood sugar. Chickpea germ and water extract of bean sprouts can be used as raw materials for pharmaceutical preparations and food, which will play a positive role in reducing processing costs, improving the quality of finished products, and stabilizing the curative effect. Basic research on effective substances to expand the safety of the drug.

Detailed ways

Embodiment 1 (alcoholic extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a hole with a diameter of 5 cm and a vent hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 2 hours to ensure that the raw beans can absorb enough water. After soaking, the raw beans that have absorbed enough water are poured with water every 5 hours at room temperature. When the bean sprouts grow to 6cm long, the longest does not exceed 10cm;

Preparation of chickpea germ and bean sprout alcohol extract:

Sieve the dried chickpea germ and bean sprout powder to 25 meshes, place 5 grams each in a round bottom flask, add 2 times volume concentration of 10% methanol, take 15 times with ultrasonic waves, each time for 1 hour, filter with filter paper, Combine the filtrates, control the temperature at 20°C by ultrasonic waves, concentrate under reduced pressure until there is no alcohol smell, and obtain the effective fraction of the alcohol extract.

Embodiment 2 (water extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a water hole with a diameter of 6 cm and a ventilation hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 3 hours to ensure that the raw beans can absorb enough water. After soaking the raw beans that have absorbed enough water, pour water every 6 hours at room temperature. When the bean sprouts grow to 7cm long, the longest does not exceed 10cm;

Preparation of aqueous extracts of chickpea germ and bean sprouts:

Sieve the dried chickpea germ powder and bean sprout powder to 25 meshes, place 5 grams each in a round-bottomed flask, add 2 times the volume of pure water, extract 15 times with ultrasonic waves, each time for 2 hours, and use gauze and filter paper respectively Filtrate, combine the filtrates, control the temperature at 20° C. with ultrasonic waves, and concentrate under reduced pressure to obtain the effective fraction of the water extract.

The effective parts of Examples 1 and 2 include: compounds such as total saponins, isoflavones, etc., which are used as the preparation of drugs for treating diabetes and lowering blood sugar or as the preparation of functional health food for lowering blood sugar.

The filter method is:

Use p-nitrophenyl phosphate disodium (pNPP) as a substrate, and use the positive drug sodium vanadate as a control, use a microplate reader to perform high-throughput screening of PTP1B enzyme inhibitors, and hydrolyze the phosphate group of pNPP according to PTP1B Instead, a color reaction was produced to measure the activity of PTP1B. The composition of the enzyme reaction system is as follows: buffer (50mM HEPES, pH7.3, 100mM sodium chloride, 0.1% bovine serum albumin, and 1mM dithiothreitol), PTP1B fusion protein, pNPP, PTP1B specific inhibitor vanadic acid Sodium (100 μg/mL). After mixing the reaction system, place it at 37°C for 30 minutes, add 1M sodium hydroxide to terminate the reaction, set the colorimeter to measure the absorption value (A) at 405 wavelength, and calculate the enzyme activity after subtracting the background value from the measurement result.

result:

Chickpea germ powder water extract group, alcohol extract group, bean sprout powder water extract group, alcohol extract group can significantly inhibit the activity of protein tyrosine phosphatase 1B and regulate blood sugar, the results are as follows:

blank   Bean sprouts powder water extract   Bean sprouts powder alcohol extract   Germ Powder Water Extract   Alcoholic Extract of Germ Powder Concentration (μg/mL) A _(0min) A _(30min) inhibition rate (%) 0.0610.64   1000.080.09297.93 750.0550.0699.13   1000.0540.0796.50   1000.0520.0698.62

The effective parts of chickpea germ and bean sprouts also have hypoglycemic effect on alloxan hyperglycemia model mice.

Embodiment 3 (alcoholic extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a hole with a diameter of 5.5 cm and a vent hole, and then add water for soaking, so as to facilitate Its germination, usually soaked for 2.5 hours, can ensure that the raw beans can absorb enough water. The raw beans that have absorbed enough water after soaking are poured with water every 5.5 hours at room temperature. When the bean sprouts grow to 8cm long, it is enough. No more than 10cm in length;

Preparation of Alcohol Extract from Chickpea Germ and Bean Sprout

Cut the chickpea germ and bean sprouts into sections, each section is about 2 mm long, put 5 grams each in a round bottom flask, add 5 times the volume concentration of 50% ethanol, and extract 5 times with heating and reflux, 2 hours each time , filter paper, combine the filtrates, control the reflux temperature at 80°C, concentrate under reduced pressure until there is no alcohol smell, and obtain the effective fraction of the alcohol extract.

Embodiment 4 (water extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a hole with a diameter of 5.5 cm and a vent hole, and then add water for soaking, so as to facilitate Its germination, usually soaked for 2.5 hours, can ensure that the raw beans can absorb enough water. The raw beans that have absorbed enough water after soaking are poured with water every 5.5 hours at room temperature. When the bean sprouts grow to 8cm long, it is enough. No more than 10cm in length;

Preparation of Chickpea Germ and Bean Sprout Water Extract

Cut the chickpea germ and bean sprouts into sections, each section is about 2 mm long, put 5 grams each in a round bottom flask, add 5 times the volume of pure water, extract 5 times with heating and reflux, each time for 3 hours, use Filter through gauze and filter paper, combine the filtrates, control the reflux temperature at 60°C, and concentrate under reduced pressure to obtain the effective fraction of the water extract.

The active ingredients in Examples 3 and 4 include: total saponins and isoflavones used as a drug for treating diabetes and lowering blood sugar or used as a functional health food for lowering blood sugar.

Screening method is identical with the screening method in embodiment 1, and its result:

Chickpea germ water extract group, alcohol extract group, bean sprout water extract group, alcohol extract group can significantly inhibit the activity of protein tyrosine phosphatase 1B and regulate blood sugar, the results are as follows:

blank Bean sprouts water extract   Alcoholic Extract of Bean Sprouts   Aqueous Germ Extract   Germ Alcohol Extract Concentration (μg/mL) A _(0min) A _(30min) inhibition rate (%) 0.0610.64 1000.0780.08698.62   1000.0650.07298.79 500.0750.09896.03   1000.060.07397.75

The active ingredients of chickpea germ and bean sprouts also have hypoglycemic effect on alloxan hyperglycemia model mice.

Embodiment 5 (alcoholic extract)

Germinate chickpeas in the usual way;

The selected chickpea raw materials are washed with water and put into a container. The top of the container is required to have a washing device, and a water hole with a diameter of 6 cm and a ventilation hole should be reserved on the bottom edge. Then add water for soaking to facilitate its germination. Usually soak for 2 hours to ensure that the raw bean grains can absorb enough water. After soaking the raw bean grains that have absorbed enough water, pour water once every 5.5 hours at room temperature, and the bean sprouts grow to 7cm long is enough, the longest is not more than 10cm;

Preparation of Chickpea Germ and Alcoholic Extract of Bean Sprout

Each 5 grams of dried chickpea germ powder and bean sprout powder (25 orders) is placed in a round-bottomed flask, adding 8 times the volume concentration is 70% isopropanol, extracting 8 times with cold soaking, each 3 hours, Filter through filter paper, combine the filtrates, concentrate under reduced pressure until there is no alcohol smell, and obtain the effective part of the alcohol extract.

Embodiment 6 (water extract)

Germinate chickpeas in the usual way;

The selected chickpea raw materials are washed with water and put into a container. The top of the container is required to have a washing device, and a water hole with a diameter of 6 cm and a ventilation hole should be reserved on the bottom edge. Then add water for soaking to facilitate its germination. Usually soak for 2 hours to ensure that the raw bean grains can absorb enough water. After soaking the raw bean grains that have absorbed enough water, pour water once every 5.5 hours at room temperature, and the bean sprouts grow to 7cm long is enough, the longest is not more than 10cm;

Preparation of Chickpea Germ and Bean Sprout Water Extract

Put 5 grams each of dried chickpea germ powder and bean sprouts powder (25 mesh) into a round bottom flask, add 8 times the volume of pure water, extract 8 times by cold soaking, each time for 4 hours, and use gauze and filter paper respectively Filter, combine the filtrates, and concentrate under reduced pressure to obtain the effective fraction of the water extract.

The active ingredients in Examples 5 and 6 include: total saponins and isoflavones for use in the preparation of drugs for treating diabetes to lower blood sugar or in the preparation of functional health food for lowering blood sugar.

Screening method is identical with the screening method in embodiment 1, and its result:

Chickpea germ water extract group, alcohol extract group, bean sprout water extract group, alcohol extract group can significantly inhibit the activity of protein tyrosine phosphatase 1B and regulate blood sugar, the results are as follows:

blank Bean sprouts powder water extract Alcoholic extract of bean sprouts powder Germ powder water extract Alcoholic extract of germ powder Concentration (μg/mL) A _(0min) A _(30min) 0.0610.64 1000.0720.079 1000.0540.063 500.0730.088 2500.0590.071

Inhibition rate(%) 98.79 98.4 97.41 97.92

The active ingredients of chickpea germ and bean sprouts also have hypoglycemic effect on alloxan hyperglycemia model mice.

Embodiment 7 (alcoholic extract)

To germinate chickpeas the conventional way:

The selected chickpea raw materials are washed with water and put into a container. The top of the container is required to have a washing device, and a water hole with a diameter of 5 cm and a ventilation hole should be reserved on the bottom edge. Then add water for soaking to facilitate its germination. Usually soak for 3 hours to ensure that the raw bean grains can absorb enough water. After soaking, the raw bean grains that have absorbed enough water are poured with water every 6 hours at room temperature, and the bean sprouts grow to 8cm. Long time is enough, the longest is no more than 10cm;

Preparation of chickpea germ and bean sprout alcohol extract:

Cut the chickpea germ and bean sprouts into sections, each section is about 2 mm long, put 5 grams each in a round bottom flask, add 10 times the volume concentration of 100% butanol, and use ultrasonic extraction 10 times, each time for 1.5 hours , filtered through filter paper, combined the filtrates, ultrasonically controlled the temperature at 35°C, and concentrated under reduced pressure until there was no alcohol smell, and the effective fraction of the alcohol extract was obtained.

Embodiment 8 (water extract)

To germinate chickpeas the conventional way:

The selected chickpea raw materials are washed with water and put into a container. The top of the container is required to have a washing device, and a water hole with a diameter of 5 cm and a ventilation hole should be reserved on the bottom edge. Then add water for soaking to facilitate its germination. Usually soak for 3 hours to ensure that the raw bean grains can absorb enough water. After soaking, the raw bean grains that have absorbed enough water are poured with water every 6 hours at room temperature, and the bean sprouts grow to 8cm. Long time is enough, the longest is no more than 10cm;

Preparation of Chickpea Germ and Bean Sprout Water Extract:

Cut the chickpea germ and bean sprouts into sections, each section is about 2 mm long, put 5 grams each into a round bottom flask, add 8 times the volume of pure water, and use ultrasonic extraction for 10 times, each time for 2 hours, and use gauze , filter paper, combine the filtrates, control the temperature at 35° C. with ultrasonic waves, and concentrate under reduced pressure to obtain the effective fraction of the water extract.

The active ingredients of Examples 7 and 8 include: compounds such as total saponins and isoflavones, used as a drug for treating diabetes and lowering blood sugar or as a functional health food for lowering blood sugar.

Screening method is identical with the screening method in embodiment 1, and its result:

Chickpea germ water extract group, alcohol extract group, bean sprout water extract group, alcohol extract group can significantly inhibit the activity of protein tyrosine phosphatase 1B and regulate blood sugar, the results are as follows:

blank   Bean sprouts water extract   Alcoholic Extract of Bean Sprouts   Aqueous Germ Extract   Germ Alcohol Extract Concentration (μg/mL) A _(0min) A _(30min) inhibition rate (%) 0.0610.64   1000.0780.09197.90 750.0570.06299.23   1000.0550.07196.51   1000.0530.06098.72

The active ingredients of chickpea germ and bean sprouts also have hypoglycemic effect on alloxan hyperglycemia model mice.

Embodiment 9 (alcohol extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a water hole with a diameter of 6 cm and a ventilation hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 3 hours to ensure that the raw bean grains can absorb enough water. After soaking, the raw bean grains that have absorbed enough water are poured with water every 5 hours at room temperature. When the bean sprouts grow to 6cm long, the longest does not exceed 10cm;

Preparation of Chickpea Germ and Alcoholic Extract of Bean Sprout

Place 5 grams each of dried chickpea germ powder and bean sprout powder (25 mesh) in a round-bottomed flask, add 4 times the volume concentration of 40% ethanol, extract 3 times with heating and reflux, each time for 3 hours, filter through filter paper , combine the filtrates, control the reflux temperature at 75°C, concentrate under reduced pressure until there is no alcohol smell, and obtain the effective fraction of the alcohol extract.

Embodiment 10 (water extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a water hole with a diameter of 6 cm and a ventilation hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 3 hours to ensure that the raw bean grains can absorb enough water. After soaking, the raw bean grains that have absorbed enough water are poured with water every 5 hours at room temperature. When the bean sprouts grow to 6cm long, the longest does not exceed 10cm;

Preparation of aqueous extracts of chickpea germ and bean sprouts

Cut the chickpea germ and bean sprouts into sections, each section is about 2 mm long, put 5 grams each in a round bottom flask, add 4 times the volume of pure water, extract 3 times with heating and reflux, each time for 4 hours, use Filter through gauze and filter paper, combine the filtrates, control the reflux temperature at 96°C, and concentrate under reduced pressure to obtain the effective fraction of the water extract.

The active ingredients of Examples 9 and 10 include: compounds such as total saponins and isoflavones, which are used as a drug for treating diabetes and lowering blood sugar or as a functional health food for lowering blood sugar.

Screening method is identical with the screening method in embodiment 1, and its result:

Chickpea germ water extract group, alcohol extract group, bean sprout water extract group, alcohol extract group can significantly inhibit the activity of protein tyrosine phosphatase 1B and regulate blood sugar, the results are as follows:

blank Bean sprouts water extract   Alcoholic Extract of Bean Sprouts   Aqueous Germ Extract Germ Alcohol Extract Concentration (μg/mL) A _(0min) A _(30min) inhibition rate (%) 0.0610.64 1000.0760.08498.53   1000.0640.07398.59 500.0720.09596.13 1000.0620.07197.55

The active ingredients of chickpea germ and bean sprouts also have hypoglycemic effect on alloxan hyperglycemia model mice.

Embodiment 11 (alcoholic extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a hole with a diameter of 5 cm and a vent hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 2.5 hours to ensure that the raw beans can absorb enough water. After soaking, the raw beans that have absorbed enough water are poured with water every 5.5 hours at room temperature. When the bean sprouts grow to 8cm long, the longest No more than 10cm;

Preparation of Chickpea Germ and Alcoholic Extract of Bean Sprout

Cut the fresh chickpea germ section and bean sprouts into sections, each section is about 2 mm long, put 5 grams each in a round bottom flask, add 5 times the volume concentration of 60% isopropanol, and use ultrasonic extraction for 3 times, Filter with filter paper for 1.5 hours each time, combine the filtrates, control the temperature at 30°C with ultrasonic waves, concentrate under reduced pressure until there is no alcohol smell, and obtain the effective fraction of the alcohol extract.

Embodiment 12 (water extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a hole with a diameter of 5 cm and a vent hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 2.5 hours to ensure that the raw beans can absorb enough water. After soaking, the raw beans that have absorbed enough water are poured with water every 5.5 hours at room temperature. When the bean sprouts grow to 8cm long, the longest No more than 10cm;

Preparation of Chickpea Germ and Bean Sprout Water Extract

Cut fresh chickpea germ segments and bean sprouts into segments, each segment is about 2 mm long, put 5 g each in a round bottom flask, add 5 times the volume of pure water, and use ultrasonic extraction for 15 times, each time for 2 hours, Filter with gauze and filter paper respectively, combine the filtrates, control the temperature at 40°C with ultrasonic waves, and concentrate under reduced pressure to obtain the effective fraction of the water extract.

The active ingredients of Examples 11 and 12 include: total saponins, isoflavones and other compounds used for the preparation of medicines for treating diabetes and hypoglycemia or for the preparation of functional health food for hypoglycemia.

Screening method is identical with the screening method in embodiment 1, and its result:

Chickpea germ water extract group, alcohol extract group, bean sprout water extract group, alcohol extract group can significantly inhibit the activity of protein tyrosine phosphatase 1B and regulate blood sugar, the results are as follows:

blank Bean sprouts water extract Alcoholic Extract of Bean Sprouts   Aqueous Germ Extract   Germ Alcohol Extract Concentration (μg/mL) A _(0min) A _(30min) inhibition rate (%) 0.0610.64 1000.0650.07798.41 1000.0610.07298.51 500.0710.07998.65   1000.0530.06698.54

The active ingredients of chickpea germ and bean sprouts also have hypoglycemic effect on alloxan hyperglycemia model mice.

Embodiment 13 (alcoholic extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a hole with a diameter of 5 cm and a vent hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 3 hours to ensure that the raw beans can absorb enough water. After soaking the raw beans that have absorbed enough water, pour water once every 6 hours at room temperature. When the bean sprouts grow to 8cm long, the longest does not exceed 10cm;

Preparation of Chickpea Germ and Alcoholic Extract of Bean Sprout

Cut fresh chickpea germ segments and bean sprouts into segments, each segment is about 2 mm long, put 5 g each in a round bottom flask, add 7 times volume concentration of 90% methanol, extract 10 times with ultrasonic waves, each time After 1 hour, filter with filter paper, combine the filtrates, control the ultrasonic temperature at 25°C, concentrate under reduced pressure until there is no alcohol smell, and obtain the effective fraction of the alcohol extract.

Embodiment 14 (water extract)

Germinate chickpeas in the usual way;

Wash the selected chickpea raw materials and put them into the container. The top of the container is required to have a washing device, and the bottom edge should leave a hole with a diameter of 5 cm and a vent hole, and then add water for soaking, so as to facilitate it. For germination, usually soak for 3 hours to ensure that the raw beans can absorb enough water. After soaking the raw beans that have absorbed enough water, pour water once every 6 hours at room temperature. When the bean sprouts grow to 8cm long, the longest does not exceed 10cm;

Preparation of Chickpea Germ and Bean Sprout Water Extract

Cut fresh chickpea germ segments and bean sprouts into segments, each segment is about 2 mm long, put 5 g each in a round bottom flask, add 10 times the volume of pure water, and use ultrasonic extraction for 5 times, each time for 2 hours, Filter with gauze and filter paper respectively, combine the filtrates, control the ultrasonic temperature at 33°C, and concentrate under reduced pressure to obtain the effective fraction of the water extract.

The effective parts of Examples 13 and 14 include: total saponins and isoflavones, which are used as a medicine for treating diabetes and lowering blood sugar or as a functional health food for lowering blood sugar.

Screening method is identical with the screening method in embodiment 1, and its result:

Chickpea germ water extract group, alcohol extract group, bean sprout water extract group, alcohol extract group can significantly inhibit the activity of protein tyrosine phosphatase 1B and regulate blood sugar, the results are as follows:

blank   Bean sprouts water extract   Alcoholic Extract of Bean Sprouts Germ water extract   Germ Alcohol Extract Concentration (μg/mL) A _(0min) A _(30min) inhibition rate (%) 0.0610.64   1000.0570.06698.04   1000.0610.06998.21 500.0670.07898.59   1000.0500.06198.24

The active ingredients of chickpea germ and bean sprouts also have hypoglycemic effect on alloxan hyperglycemia model mice.

Claims (8)

1. a kind of employing solvent extraction method to prepare chickpea germ and bean sprouts effective part, it is characterized in that carry out according to the following steps: a, the chickpeas are germinated by a conventional method; b. Add 2-10 times the volume of methanol or ethanol or isopropanol or butanol with a concentration of 10-100% to fresh chickpea germ and bean sprouts or dried chickpea germ powder and bean sprouts powder; c. Use ultrasound or cold soaking or heating and reflux extraction for 1-15 times, each time for 1-3 hours, wherein the ultrasonic extraction temperature is 20-40 ° C, the reflux extraction temperature is 60-80 ° C, filter, combine the filtrates, and concentrate under reduced pressure Until there is no alcohol smell, the effective part of the alcohol extract is obtained; d, or add 2-10 times the volume of purified water to fresh chickpea germ and bean sprouts or dry chickpea germ powder and bean sprout powder; e. Use ultrasound or cold soaking or heating and reflux extraction for 1-15 times, each time for 2-4 hours, wherein the ultrasonic extraction temperature is 20-60 ° C, the reflux extraction temperature is 60-100 ° C, filter, combine the filtrates, and concentrate under reduced pressure Obtain the effective part of the water extract. 2. The effective parts of chickpea germ and bean sprouts according to claim 1, characterized in that the effective parts are total saponins and isoflavones. 3. The effective parts of chickpea germ and bean sprouts according to claim 1, characterized in that the effective parts are prepared as functional food according to conventional methods or made into capsules or tablets or oral liquids or granules or injections according to conventional pharmaceutical methods Or slow-release preparation or controlled-release preparation; or make capsules or tablets or oral liquids or granules or injections or slow-release preparations or controlled-release preparations by combining the effective parts with Chinese and Western medicines. 4. A preparation method for chickpea germ and bean sprouts effective parts, characterized in that the effective parts are prepared by solvent extraction, that is, alcohol extraction or water extraction, and the specific operations are carried out according to the following steps: a, the chickpeas are germinated by a conventional method; b. Add 2-10 times the volume of methanol or ethanol or isopropanol or butanol to fresh chickpea germ and bean sprouts or dried chickpea germ powder and bean sprout powder, the concentration of which is 10-100%; c. Use ultrasound or cold soaking or heating and reflux extraction for 1-15 times, each time for 1-3 hours, wherein the ultrasonic extraction temperature is 20-40 ° C, the reflux extraction temperature is 60-80 ° C, filter, combine the filtrates, and concentrate under reduced pressure Until there is no alcohol smell, the total saponins and isoflavones of the effective parts of the alcohol extract are obtained; d, or add 2-10 times the volume of purified water to fresh chickpea germ and bean sprouts or dry chickpea germ powder and bean sprout powder; e. Use ultrasound or cold soaking or heating and reflux extraction for 1-15 times, each time for 2-4 hours, wherein the ultrasonic extraction temperature is 20-60 ° C, the reflux extraction temperature is 60-100 ° C, filter, combine the filtrates, and concentrate under reduced pressure The total saponins and isoflavones in the effective parts of the water extract are obtained. 5. The preparation method according to claim 4, characterized in that the optimal concentration of alcohol in step b is 60-70%. 6. The preparation method according to claim 4, characterized in that in step c, the ultrasonic extraction temperature of alcohol is preferably 30-35°C, and the reflux extraction temperature is preferably 70-80°C. 7. The preparation method according to claim 4, characterized in that the ultrasonic extraction temperature in step e is preferably 30-40°C, and the reflux extraction temperature is preferably 80-96°C. 8. The use of the chickpea germ and the effective parts of the bean sprouts as a hypoglycemic drug for treating diabetes or as a hypoglycemic functional food.