阅读说明：本技术 一种调节糖脂代谢中药提取物的制备方法 (Preparation method of traditional Chinese medicine extract for regulating glycolipid metabolism ) 是由 刘湘 李伟 邢琪昌 彭科 于 2021-01-13 设计创作，主要内容包括：本发明属于中药复方有效成分提取分离技术领域,具体是涉及到一种调节糖脂代谢中药提取物的制备方法,步骤为,将桑叶和葛根粉末混合,加入乙醇溶液,调节pH值至2以下,加热浸提,过滤,滤液回收,过柱,洗脱除杂,收集洗脱液,浓缩,干燥,得到调节糖脂代谢中药提取物,所述调节糖脂代谢中药提取物中,1-脱氧野尻霉素的含量为2-4mg/g,槲皮素的含量为40-80mg/g,葛根素的含量为150-200mg/g,本发明具有降低血糖、调节血脂、保护胰岛等作用。(The invention belongs to the technical field of extraction and separation of active ingredients of a traditional Chinese medicine compound, and particularly relates to a preparation method of a traditional Chinese medicine extract for regulating glycolipid metabolism, which comprises the steps of mixing mulberry leaf powder and kudzuvine root powder, adding an ethanol solution, regulating the pH value to be below 2, heating, leaching, filtering, recovering filtrate, passing through a column, eluting to remove impurities, collecting eluent, concentrating and drying to obtain the traditional Chinese medicine extract for regulating glycolipid metabolism, wherein in the traditional Chinese medicine extract for regulating glycolipid metabolism, the content of 1-deoxynojirimycin is 2-4mg/g, the content of quercetin is 40-80mg/g, and the content of puerarin is 150-200 mg/g.)

1. A preparation method of a traditional Chinese medicine extract for regulating glycolipid metabolism is characterized by mixing mulberry leaf and kudzu root powder, adding an ethanol solution, regulating the pH value to be below 2, heating and leaching, filtering, recovering filtrate, passing through a column, eluting to remove impurities, collecting eluent, concentrating and drying to obtain the traditional Chinese medicine extract for regulating glycolipid metabolism, wherein the content of 1-deoxynojirimycin in the traditional Chinese medicine extract for regulating glycolipid metabolism is 2-4mg/g, the content of quercetin is 40-80mg/g, and the content of puerarin is 150-200 mg/g.

2. The method for preparing the extract of a Chinese medicinal material for regulating glycolipid metabolism according to claim 1, characterized in that the substance for regulating pH value is concentrated hydrochloric acid.

3. The method for preparing the extract of Chinese traditional medicine for regulating glycolipid metabolism as claimed in claim 1 or 2, wherein the weight ratio of mulberry leaf to kudzu root is 2: 1.

4. The method for preparing the extract of a Chinese medicine for regulating glycolipid metabolism according to claim 1 or 2, characterized in that the concentration of ethanol is 60-80%.

5. The method for preparing a Chinese medicinal extract for regulating glycolipid metabolism according to claim 4, wherein the weight of said ethanol is 20 times of the total weight of mulberry leaf and kudzu root.

6. The method for preparing the extract of Chinese herbal medicine for regulating glycolipid metabolism as claimed in claim 1, characterized by regulating pH value to 2.

7. The method for preparing the extract of Chinese herbal medicine for regulating glycolipid metabolism as claimed in claim 1, characterized in that the temperature of the heating extraction is 80 ℃.

8. The method for preparing the extract of Chinese traditional medicine for regulating glycolipid metabolism as claimed in claim 1, wherein the column is AB-8 macroporous resin column.

9. The method for preparing the extract of Chinese herbal medicine for regulating glycolipid metabolism as claimed in claim 1, wherein the elution and impurity removal are carried out by eluting with 5BV of pure water and then with 50% by volume of ethanol at 5 BV.

Technical Field

The invention belongs to the technical field of extraction and separation of active ingredients of a traditional Chinese medicine compound, and particularly relates to a preparation method of a traditional Chinese medicine extract for regulating glycolipid metabolism.

Background

The folium Mori is dry leaf of Morus alba L of Moraceae, and has effects of dispelling pathogenic wind and heat, clearing lung-heat, moistening dryness, removing liver fire, and improving eyesight. Pharmacological experiments show that the active ingredients such as flavone, flavonoid glycoside, polyphenol, polysaccharide, alkaloid and the like in the mulberry leaves have the effects of reducing blood sugar, improving insulin resistance and the like. The radix Puerariae is dried root of Pueraria lobata Ohwi of Leguminosae, and has effects of relieving muscles and fever, promoting fluid production to quench thirst, etc., and pharmacological experiments show that active ingredients such as flavone and flavonoid glycoside in radix Puerariae have effects of resisting diabetes, protecting nerve, etc. They are usually combined for treating diabetes.

The mulberry leaves-kudzuvine roots are common medicinal and edible medicaments, the dosage of the mulberry leaves-kudzuvine roots for treating type 2 diabetes is large, the conventional extraction and purification process mostly takes mulberry leaf alkaloid as a main investigation target, a large amount of active ingredients such as flavone, flavonoid glycoside and polyphenol are abandoned, the blood sugar reducing effect is good, but the curative effects such as lipid regulation, antioxidation and the like cannot be guaranteed.

Disclosure of Invention

The invention aims to solve the technical problem of providing a preparation method of a traditional Chinese medicine extract for regulating glycolipid metabolism, and the traditional Chinese medicine extract has the effects of reducing blood sugar, regulating blood fat, protecting pancreatic islets and the like.

The invention relates to a preparation method of a traditional Chinese medicine extract for regulating glycolipid metabolism, which comprises the steps of mixing mulberry leaf powder and kudzuvine root powder, adding an ethanol solution, regulating the pH value to be below 2, heating and leaching, filtering, recovering filtrate, passing through a column, eluting to remove impurities, collecting eluent, concentrating and drying to obtain the traditional Chinese medicine extract for regulating glycolipid metabolism, wherein the content of 1-deoxynojirimycin in the traditional Chinese medicine extract for regulating glycolipid metabolism is 2-4mg/g, the content of quercetin is 40-80mg/g, and the content of puerarin is 150-200 mg/g.

Preferably, the substance for adjusting the pH value is concentrated hydrochloric acid.

Preferably, the weight ratio of the mulberry leaves to the kudzuvine roots is 2: 1.

Preferably, the ethanol concentration is 60-80%.

Preferably, the weight of the ethanol is 20 times of the total weight of the mulberry leaves and the kudzuvine roots.

Preferably, the pH is adjusted to 2.

Preferably, the temperature of the heated leaching is 80 ℃.

Preferably, the column is an AB-8 macroporous resin column.

Preferably, the elution and impurity removal method comprises the steps of eluting and removing impurities by 5BV pure water and then eluting by using 5BV 50% ethanol by volume concentration.

The traditional Chinese medicine extract for regulating glycolipid metabolism is rich in active ingredients such as mulberry leaf-pueraria flavone, flavonoid glycoside and alkaloid, and animal experiments show that the extract can obviously reduce blood sugar and blood fat of a T2DM rat and has a protection effect on damaged pancreatic islets, and is expected to be developed to treat type 2 diabetes.

The invention adopts acidic ethanol for heating extraction, compared with simple ethanol extraction, the invention enriches the alkaloid of mulberry leaves and simultaneously reserves the effective components of flavone of mulberry leaves, polyphenol, pueraria flavone and the like as much as possible, and animal experiments prove that the extract can reduce blood sugar, regulate blood fat and protect pancreas islet and has anti-diabetic activity.

The invention obtains a compound extract capable of regulating the T2DM rat glycolipid metabolism function by leaching the mulberry leaves-kudzu roots with acidic ethanol solution, and has the advantage that the antidiabetic activity of the compound extract is stronger than that of a single medicinal material extract or the traditional mulberry leaves-kudzu roots extract, probably because active ingredients such as mulberry leaf alkaloid, flavone, polyphenol, kudzu root flavone and the like have synergistic effect, and the compound extract has the action advantage of multi-component multi-target multi-action approach.

Drawings

FIG. 1 is a graph of HE staining of mouse islets (200-fold).

Detailed Description

Example 1

A preparation method of a traditional Chinese medicine extract for regulating glycolipid metabolism comprises the steps of grinding 2 equal parts of mulberry leaves and 1 equal part of kudzu roots into coarse powder, uniformly mixing, adding 20 times of ethanol solution with the weight volume concentration of 60% -80%, regulating the pH value of the solution to 2.0 by using concentrated hydrochloric acid, heating to 80 ℃, leaching for 2 times, 1 hour each time, filtering, combining filtrates, recovering ethanol from the filtrates until no alcohol smell exists, passing through an AB-8 macroporous resin column, eluting by 5BV pure water to remove impurities, eluting by 5BV 50% ethanol (volume concentration), collecting eluent, concentrating under reduced pressure, and drying.

The product is characterized in that: the brown powder contains 2-4mg/g, 40-80mg/g, and 150-200mg/g of 1-deoxynojirimycin, quercetin, and puerarin respectively, as determined by high performance liquid chromatography.

Comparative example 1

Compared with example 1, the pH is not adjusted by concentrated hydrochloric acid, extracted by ethanol solution, and extracted 2 times by heating to 80 ℃, and the rest is the same as example 1.

Experimental example 1

The experimental scheme is as follows: 24 male SD rats were housed in a room for acclimatization for 7 days. Rats were randomly divided into 6 normal groups and 18 model groups. Normal group was given normal diet and molding group was given high fat diet. Fasting is carried out for 12h in the fourth week, 1% streptozotocin is injected into the abdominal cavity of the molding group, the dosage is 40mg/kg, and the normal group is given equal volume of 0.1mol/L citric acid buffer solution. After 3 days, determining the fasting blood glucose of the rat, wherein the blood glucose value is between 11.1 mmol/L and 33.3mmol/L, and the molding is successful. Successfully modeled rats are randomly divided into 3 groups, 6 rats are respectively given with 0.25g/kg of mulberry leaf and kudzuvine root compound extract (administration group) and distilled water, the normal group is given with equal amount of distilled water for 1 time per day, the administration is performed by gastric perfusion for 42 days, the fasting blood sugar and blood fat are measured on the 43 th day, and pancreas tissues are taken for HE staining to observe the islet morphology.

Test results

Blood glucose was significantly elevated in the model group compared to the normal group, while blood glucose was significantly reduced in the administered group compared to the model group, and the results are shown in table 1.

Table 1 effect of herbal extracts on fasting plasma glucose in T2DM rats (mmol/L, x ± s, n ═ 6)

Compared with the normal group, the blood lipid indexes of the model group are obviously changed, and the indexes of the administration group (the example 1 group) are obviously improved compared with the indexes of the model group, and the results are shown in table 2.

Table 2 effect of herbal extracts on T2DM rat plasma lipids (mmol/L, x ± s, n ═ 6)

Morphological changes of pancreatic islets

In T2DM rats induced by streptozotocin in combination with high fat diet, the number of islets decreased or were atrophied, and the islet cells within the islets were partially denatured, which was improved in the group of example 1, as shown in fig. 1.

Those of ordinary skill in the art will understand that: the discussion of any embodiment above is meant to be exemplary only, and is not intended to intimate that the scope of the disclosure, including the claims, is limited to these examples; within the spirit of the present disclosure, features from the above embodiments or from different embodiments may also be combined, steps may be implemented in any order, and there are many other variations of different aspects of one or more embodiments in this application as described above, which are not provided in detail for the sake of brevity.

It is intended that the one or more embodiments of the present application embrace all such alternatives, modifications and variations as fall within the broad scope of the appended claims. Therefore, any omissions, modifications, substitutions, improvements, and the like that may be made without departing from the spirit and principles of one or more embodiments of the present disclosure are intended to be included within the scope of the present disclosure.