CN107325071A - A kind of preparation method and purposes of the Hickory Leaves pinostrobin with hypoglycemic effect - Google Patents

Abstract

The invention relates to a preparation method and application of hickory coniferin with hypoglycemic effect, and the mass percentage of coniferin containing is more than 90%. The hickory yew coniferin can significantly reduce the blood sugar level of diabetic mice, has a significant curative effect on diabetes, especially type II diabetes, and can be used in medicines and health care products whose main function is to prevent and treat diabetes.

Description

A kind of preparation method and application of hickory carya phyllocinin with hypoglycemic effect

technical field

The invention relates to the technical field of medicines, in particular to a preparation method and application of hickory pecan yewetine with hypoglycemic effect.

Background technique

Diabetes is a metabolic disease caused by excess accumulation of glucose in the blood, which can compromise immune function. Also because of its longer course of disease, the troubles of the human body follow. Diabetics are easily infected with various infectious diseases caused by colds, pneumonia, and tuberculosis, and are difficult to cure. Severe cases may cause damage to multiple organs throughout the body, blindness, kidney disease and other complications, endangering human health. Diabetes is generally divided into type 1 diabetes, type 2 diabetes, gestational diabetes and other special types of diabetes. Type 2 diabetes accounts for approximately 95% of diabetic patients. With the improvement of residents' living standards this year, the number of diabetic patients has also increased greatly, which has also led to the advent of a large number of hypoglycemic drugs. However, most of the commonly used clinical diabetes treatment drugs are chemical synthetic drugs. There are many adverse reactions. Natural medicines are animal medicines, herbal medicines and mineral medicines with certain pharmacological activities proved by the modern medicine system.

The development of society has made people pay more and more attention to the negative effects of chemicals. In this context, Chinese herbal medicine has irreplaceable advantages. Therefore, the development of Chinese herbal medicines for the prevention and treatment of diabetes and its complications not only meets the market demand but also has good social benefits.

The ob/ob mouse is a congenitally obese mouse obtained by knocking out the leptin gene. Its pathogenesis is similar to that of human obesity. It is one of the more commonly used genetic obesity models and is also commonly used in animal experiments for type 2 diabetes.

Hickory is a deciduous tree of the Juglandaceae family. Most of its leaves are harvested in summer and autumn. It is used as medicine to clear away heat, detoxify, kill insects and relieve itching. It is mainly used for the treatment of toe wet itching and skin ringworm syndrome. Studies have found that hickory leaf extract contains a large number of antioxidant active substances, such as flavonoids, which have anti-aging, anti-tumor, and anti-angiogenesis effects. Flavonoids are one of the main active ingredients of medicinal plants, which have broad-spectrum pharmacological activity and low toxicity. Studies have shown that hickory carya pine can reduce blood sugar levels in model ob/ob mice, and can be used to prevent and treat hyperglycemia, especially It is a medicine and health food with type 2 diabetes as its main function.

Contents of the invention

The purpose of the present invention is to provide a preparation method of the hickory nut leaf extract bulbocenadin with hypoglycemic effect, and also provide its application.

The technical solution adopted by the present invention to solve its technical problems is:

The hickory nut leaf extract ballconiferin with hypoglycemic effect contains more than 90% of the ballconiferin by mass.

A kind of preparation method of the hickory nut yew coniferin with hypoglycemic effect, described preparation method comprises the following steps:

(1) Put hickory leaves in an oven to dry, crush and sieve, add 75-95% ethanol to reflux extraction, each time for 30-120 minutes, repeat 1-3 times, filter, and combine the filtrates;

(2) Concentrate the filtrate of step (1), add 0.7mL, 1mol/L sodium hydroxide solution to every 4mL filtrate, mix well, then add polyamide and mix thoroughly, concentrate under reduced pressure and put on the column, at 4-12BV/h The flow rate is successively eluted with water, 20% ethanol, 40% ethanol, and 60% ethanol, and each solvent is eluted for 6-10 times of column volume, and the eluate of 60% ethanol is collected, concentrated under reduced pressure to dryness to obtain hickory leaves crude extract;

(3) Add methanol to the pecan leaf crude extract in step (2) according to the ratio of 5-35mL methanol per 1g of pecan leaf crude extract, and extract with petroleum ether in a ratio of 1:1, repeat 3-10 times , and then concentrate the petroleum ether extract to dryness under reduced pressure at 30-50°C to obtain the yellow crude extract of hickory nut leaves;

(4) Add 60vol%-90% aqueous ethanol solution to the yellow crude extract of pecan leaves in step (3) in a ratio of 1g:8-12mL, and dissolve it in a water bath at 70-90°C. Place in a refrigerator at 4° C. for 12-24 hours for crystallization, and filter through filter paper to obtain hickory coniferin crystals, the content of which is more than 90%.

Preferably, the oven drying temperature in step (1) is 30-80°C.

Preferably, the sieve mesh in step (1) is 40-100 mesh.

As a preference, the extraction condition in step (1) is 60-120° C. under normal pressure.

As preferably, the consumption of 70-95% ethanol aqueous solution in step (1) is to add 10mL per 1g hickory leaf pulverization

As a preference, the amount of polyamide used in step (2) is to add 5 g of polyamide per 100 mL of alkalized filtrate.

A use of hickory jaggery with hypoglycemic effect, the hickory jaggery prepared above and pharmaceutical or food approved adjuvant, using modern preparation technology, made hickory jelly jelly as the The main capsules, tablets, powders, oral liquids, soft capsules, pills, gels, sustained-release preparations and other dosage forms and health food.

Beneficial effect of the present invention: the present invention uses the leaves of Juglandaceae hickory as raw material to prepare its primary extract, and further purifies to obtain the hickory leaf condylopin with hypoglycemic effect. This method is simple and easy, and further expands the The scope of application of walnut leaves has very huge market potential and economic value.

detailed description

The present invention will be described in further detail below through specific examples, but the present invention is not limited to these examples.

Example 1

Hickory leaves were dried at 40°C, crushed, passed through a 40-mesh sieve, weighed 100g of the powder, added 1L of 85% ethanol solution for reflux extraction, twice, the extraction conditions were normal pressure 80°C, 60min each time, and the filtrates were combined. Add 0.7mL, 1mol/L sodium hydroxide solution to each 4mL filtrate to alkalize for 12h. After alkalization, add 5g polyamide (100-200 mesh) to the sample solution per 100mL and mix evenly. Concentrate to dryness under reduced pressure and perform chromatography. The column (15mm×350mm i.d) was sequentially eluted with 10BV of water, 20% ethanol, 40% ethanol, and 60% ethanol, and the 60% ethanol eluate was collected and concentrated to dryness under reduced pressure to obtain the crude extract of hickory nut leaves.

The above-mentioned hickory leaf crude extract is dissolved according to the ratio of adding 5mL of methanol per 1g of hickory leaf crude extract, and then extracted 10 times with an equal volume of petroleum ether, and the petroleum ether product is evaporated to dryness under reduced pressure to obtain hickory leaf Yellow crude extract. Dissolve the yellow crude extract of pecan leaves in 80% ethanol at 90°C, filter while hot, and quickly put the filtrate in a refrigerator at 4°C for 12 hours to crystallize. The crystallization solution is filtered through a filter paper to obtain condyloconolin crystals of the hickory leaf extract, and the content of condyloconolin is more than 90%.

Example 2

Place hickory leaves in a 35°C oven to dry, crush, pass through an 80-mesh sieve, weigh 200g of hickory leaf powder, add 2L volume concentration of 95% ethanol for reflux extraction, and the extraction conditions are atmospheric pressure 60°C, 2 times, 30min each time , combine the filtrates, add 0.7mL, 1mol/L sodium hydroxide solution to each 4mL filtrate to alkalinize for 12h, after alkalization, add 5g polyamide (100-200 mesh) to the sample solution per 100mL and mix evenly, concentrate to dryness under reduced pressure , upper chromatographic column (15mm * 350mm i.d), with the water of 10BV, 20% ethanol, 40% ethanol, 60% ethanol elution successively, collect 60% ethanol eluate, concentrate under reduced pressure to dryness, obtain pecan leaf crude extract.

The above-mentioned hickory leaf crude extract is dissolved according to the ratio of adding 20mL of methanol per 1g of hickory leaf crude extract, and then extracted 10 times with an equal volume of petroleum ether, and the petroleum ether product is evaporated to dryness under reduced pressure to obtain hickory leaf Yellow crude extract. Dissolve the yellow crude extract of pecan leaves in 80% ethanol at 90°C, filter while hot, and quickly put the filtrate in a refrigerator at 4°C for 12 hours to crystallize. The crystallization solution is filtered through a filter paper to obtain condyloconolin crystals of the hickory leaf extract, and the content of condyloconolin is more than 90%.

Example 3

Hickory leaves are placed in an oven to dry at 80°C, crushed through a 100-mesh sieve, weigh 300g of hickory leaf powder, add 3L of ethanol with a volume concentration of 75% for reflux extraction, and the extraction conditions are 120°C at normal pressure, 120 minutes each time, Repeat once, filter, and combine the filtrates;

(2) Concentrate the filtrate of step (1), add 0.7mL, 1mol/L sodium hydroxide solution for every 4mL of filtrate, add polyamide after alkalization for 12h and mix thoroughly, add 5g of polyamide for every 100mL of alkalized filtrate Concentrate under reduced pressure and put on the column, with a flow rate of 12BV/h, sequentially elute with water, 20% ethanol, 40% ethanol, and 60% ethanol, each solvent is eluted for 6-10 times the column volume, and the elution of 60% ethanol is collected. liquid, concentrated under reduced pressure to dryness to obtain the crude extract of pecan leaves;

(3) Add methanol to the pecan leaf crude extract of step (2) according to the ratio of 35mL methanol for every 1g pecan leaf crude extract, and extract with sherwood oil 1:1 ratio, repeat 3 times, then add petroleum Concentrate the ether extract to dryness under reduced pressure at 30-50°C to obtain the yellow crude extract of pecan leaves;

(4) Add 90% ethanol aqueous solution to the yellow crude extract of pecan leaves in step (3) in a ratio of 1g:12mL, and dissolve it in a water bath at 70°C. After the dissolution, quickly put it in a refrigerator at 4°C and place After 24 hours, carry out crystallization, and filter with filter paper to obtain hickory coniferin crystals, the content of which is more than 90%.

See Shen Yong et al. Determination of 5 flavonoid aglycones in hickory leaves by high performance liquid chromatography [J]. Journal of Pharmaceutical Analysis, 2013-05.

Study on the effect of hickory carya phyllopine on the mouse model of hyperglycemia induced by alloxan

1. Experimental method:

50 male ICR mice were randomly divided into 6 groups, including 10 blank control groups, and 8 mice in each of the other groups. After fasting for 12 hours, 40 model mice were injected with alloxan solution 80mg/kg into the tail vein to make a model. (The normal control group was injected with the same amount of normal saline), blood glucose was measured 5 days later, and the model was successfully established when the blood glucose was >11.1mmol/L. Then the model mice were randomly divided into 5 groups, which were the hyperglycemia model group, the low-dose group of 50mg/kg hickory cypressin, the middle-dose group of 100mg/kg, the high-dose group of 200mg/kg, and the 50mg/kg metformin positive group. medicine group. During the experiment, the normal control group and the model group were given 0.5% CMC-Na orally, and the other groups were given intragastric administration according to the dose, once a day, for 14 consecutive days. After 14 days, the blood glucose, body weight and serum fructosamine of mice in each group were measured and recorded. The experimental results were statistically processed using SPSS 18.0 statistical software. The experimental data are expressed as mean ± standard deviation, using one-way analysis of variance, when P < 0.05, there is a significant difference.

2. Experimental results

The results are shown in Table 1 and Table 2. From the table, it can be known that hickory carya pachypinin can significantly reduce the blood sugar content of mice, and has a significant effect on the mouse model of hyperglycemia induced by alloxan.

Table 1 The effect of hickory phyllopine on hyperglycemia model mice

Note: Compared with the normal group, ^** P<0.01; compared with the model group, ^# P<0.05, ^## P<0.01.

Table 2 Effects of different dosages of total flavonoids from pecan leaves on the body weight of hyperglycemic mice

Note: Compared with the normal group, ^** P<0.01; compared with the model group, ^# P<0.05.

Study on the effect of hickory phyllocinin on ob/ob obese mouse model

1. Experimental method

Take 40 8-week-old ob/ob male mice, feed them adaptively for 1 week, and then divide the mice into a low-dose group of 50 mg/kg of hickory leaf pinecone, a medium-dose group of 100 mg/kg, and a high-dose group of 200 mg/kg. Dosage group, 50mg/kg berberine positive drug group, and C57 mice as normal control group, mice in each group were administered orally once a day. 28 days in a row. After 28 days, the blood glucose of mice in each group was measured and recorded. The experimental results were statistically processed using SPSS 18.0 statistical software. The experimental data are expressed as mean ± standard deviation, using one-way analysis of variance, when P < 0.05, there is a significant difference.

2. Experimental results

The experimental results are shown in Table 3. It can be seen from the table that hickory pine pinulin can significantly reduce the blood sugar content of mice, and has a good effect on obese mice, that is, type II diabetes model mice.

Table 3 The effect of hickory phyllopine on ob/ob obesity model mice

Note: Compared with the normal group, ^** P<0.01; compared with the model group, ^# P<0.05, ^## P<0.01.

The above-described embodiment is only a preferred solution of the present invention, and does not limit the present invention in any form. There are other variations and modifications on the premise of not exceeding the technical solution described in the claims.

Claims (7)

1. a kind of preparation method of the hickory carya coniferin with hypoglycemic effect, it is characterized in that, described preparation method comprises the following steps: (1) Put hickory leaves in an oven to dry, crush and sieve, add 75-95% ethanol to reflux extraction, each time for 30-120 minutes, repeat 1-3 times, filter, and combine the filtrates; (2) Concentrate the filtrate of step (1), add 0.7mL, 1mol/L sodium hydroxide solution to every 4mL filtrate, mix well, then add polyamide and mix thoroughly, concentrate under reduced pressure and put on the column, at 4-12BV/h The flow rate is successively eluted with water, 20% ethanol, 40% ethanol, and 60% ethanol, and each solvent is eluted for 6-10 times of column volume, and the eluate of 60% ethanol is collected, concentrated under reduced pressure to dryness to obtain hickory leaves crude extract; (3) Add methanol to the pecan leaf crude extract in step (2) according to the ratio of 5-35mL methanol per 1g of pecan leaf crude extract, and extract with petroleum ether in a ratio of 1:1, repeat 3-10 times , and then concentrate the petroleum ether extract to dryness under reduced pressure at 30-50°C to obtain the yellow crude extract of hickory nut leaves; (4) Add 60vol%-90% aqueous ethanol solution to the yellow crude extract of pecan leaves in step (3) in a ratio of 1g:8-12mL, and dissolve it in a water bath at 70-90°C. Place in a refrigerator at 4° C. for 12-24 hours for crystallization, and filter through filter paper to obtain hickory coniferin crystals, the content of which is more than 90%. 2. The preparation method of a kind of hickory carya coniferin with hypoglycemic effect according to claim 1, characterized in that, the oven drying temperature in step (1) is 30-80°C. 3. the preparation method of a kind of hickory pine pinulin with hypoglycemic effect according to claim 1, is characterized in that, step (1) sieve mesh is 40-100 mesh. 4. A kind of preparation method of hickory nut echinocin with hypoglycemic effect according to claim 1, it is characterized in that, the extraction condition in step (1) is normal pressure 60-120 ℃. 5. the preparation method of a kind of hickory nut leaf pinol with hypoglycemic effect according to claim 1, is characterized in that, in the step (1), 70-95% ethanol aqueous solution consumption is every 1g hickory nut leaf pulverization thing Add 10 mL. 6. the preparation method of a kind of hickory carya coniferin with hypoglycemic effect according to claim 1, is characterized in that, the consumption of polyamide is that the filtrate after every 100mL basification adds 5g polyamide amides. 7. the purposes of a kind of hickory yew coniferin with hypoglycemic effect, it is characterized in that, the hickory jaggery yew pinin prepared according to claim 1 and the adjuvant approved on the medicament or food adopt modern preparation technology to make Capsules, tablets, powders, oral liquids, soft capsules, pills, gels, sustained-release preparations and other dosage forms and health food mainly based on hickory phyllopine.