CN1450077A - Method for extracting polygonum cuspidatum tannic acid - Google Patents

Abstract

The present invention relates to an extraction method of Chinese medicinal material polygonum cuspidatum extract-polygonum cuspidatum tannin. The polygonum cuspidatum tannin as main component of polygoum cuspidatum extract has alpha-glycosidase inhibiting activity, can be used as main active component for preparing medicine for reducing blood sugar, also can be used as raw material for preparing medicine for curing diabetes and obesity. Its extraction method in cludes the following steps: pulverizing raw material, using water and methyl alcohol or water-containing alcohol to make extraction under the condition of heating, filtering, using petroleum ether to degrease the filtrate, using active carbon to decolour, the concentrating at normal pressure or reduced pressure, making the concentrated filtrate pass through ion exchange resin.

Description

Extraction method of Polygonum cuspidatum extract Polygonum cuspidatum tannin

technical field

The invention relates to a method for extracting Polygonum cuspidatum tannic acid from Polygonum cuspidatum extract, which belongs to the technical field of chemical extraction and preparation of Chinese herbal medicines.

Background technique

The medicinal Polygonum cuspidatum is a plant of the genus Polygonaceae, and the medicinal part is the rhizome. The main chemical components are very wide and many, including: emodin, chrysophanol, glucoside, resveratrol glycoside, glycosides, amino acids, trace elements, etc., and are rich in condensed tannins and polysaccharides.

The tannin or tannic acid in the extract of Polygonum cuspidatum has the inhibitory activity of diabetic α-glucosidase, so it can be used as a hypoglycemic drug.

Contents of the invention

The object of the present invention is to provide a Polygonum cuspidatum extract with α-glucosidase inhibitory activity, the effective part of which is water-soluble Polygonum cuspidatum tannic acid with good hypoglycemic activity.

Another object of the present invention is to provide a preparation process of Polygonum cuspidatum extract.

The above object of the present invention is achieved through the following technical solutions.

A method for extracting Polygonum cuspidatum extract Polygonum cuspidatum tannin of the present invention is characterized in that the method comprises the following process steps:

a. The raw medicinal material Polygonum cuspidatum is crushed and extracted with water, methanol or water-containing alcohol under heating conditions.

The time is 2-30 hours, and the amount of solvent is 3-10 times that of the original medicinal material;

b. filter or centrifuge, discard the filter residue, get the filtrate or supernatant, and put the filtrate or supernatant at temperature

Keep warm at 50-100°C for 5-30 minutes, and place in the refrigerator overnight;

c. Put it into a centrifuge, centrifuge at 2000-5000rmp for 5-20 minutes, and take the supernatant;

d. Degrease the supernatant with petroleum ether for 2-5 times in a separating funnel;

e. decolorize the degreased extract with activated carbon, and then remove low molecular weight impurities by ultrafiltration;

f. Strongly acidic cation exchange resin is applied to the resulting filtrate, and then eluted with distilled water, and the eluent adjusts its

After the pH reaches neutral, heat to 60-100°C under normal pressure or reduced pressure to concentrate, and then use an ultrafiltration membrane

Carry out ultrafiltration, and after drying, a brown solid powder can be obtained, and its main component is Polygonum cuspidatum tannic acid.

The extraction method of another kind of Polygonum cuspidatum extract Polygonum cuspidatum tannin of the present invention is characterized in that, the method comprises the following process steps:

a. Soak the raw medicinal material Polygonum cuspidatum in water, ethanol or water-containing alcohol, macerate it after soaking, crush it with a masher, and put it in a heating bar

Under certain conditions, extract for 2-30 hours, and the amount of solvent used is 3-10 times that of the original medicinal material;

b. Filtrate, discard the filter residue, take the filtrate and heat it to 50-100°C under normal pressure or reduced pressure, and concentrate to obtain the extract;

c. Add water to dissolve, heat to reflux with chloroform, discard the chloroform layer, and extract the resulting aqueous layer with ethyl acetate for 2-3 times,

Take the ethyl acetate layer, and then extract it with NaOH aqueous solution for 2-3 times, take the NaOH aqueous solution, and adjust the pH with HCl

When the value reaches 2-6, add 4-10% gelatin aqueous solution to obtain a precipitate;

d. The precipitate is dissolved with 30-70% acetone aqueous solution, then centrifuged with a centrifuge, the precipitate is discarded, and the acetone solution is taken.

Liquid, it is concentrated by distillation under reduced pressure, and reclaims acetone, obtains colloidal solid;

e. Dissolve the obtained colloidal solid in water, then extract with ethyl acetate, and concentrate the obtained ethyl acetate layer under reduced pressure.

Shrink, and reclaim ethyl acetate, obtain concentrated jelly;

f. The jelly is dissolved by adding appropriate amount of water, filtered, and the precipitated insoluble matter is filtered off to obtain a clear liquid, which is freeze-dried

Obtain orange-yellow powder, the main component of which is Polygonum cuspidatum tannic acid.

Polygonum cuspidatum tannic acid prepared by the method of the present invention has α-glucosidase inhibitory activity, and its action mechanism is that polygonum cuspidatum tannic acid interacts with many enzymes in the digestive tract, especially the inhibitory activity on α-glucosidase, thereby regulating the body The absorption of nutrients such as glucose makes it anti-nutritive. For obese diabetic patients, knotweed tannic acid can not only effectively lower blood sugar, but also reduce its absorption of nutrients. Therefore, knotweed tannic acid can be used as Glycosidase inhibitor drugs can also be used as the main component of slimming tea.

The extraction process of the method of the invention has the advantages of simple equipment, easy operation of separation and extraction, and low cost.

Polygonum cuspidatum tannic acid, the preparation of the present invention, can reduce plasma blood sugar concentration as an inhibitor.

The following test is the hypoglycemic effect of Polygonum cuspidatum tannic acid inhibitor on animal mice.

Experiment 1, the effect of one administration of Polygonum cuspidatum tannic acid inhibitor on the blood sugar content of normal mice: 45 mice were randomly divided into 5 groups, and they were fed with normal saline, Baitangping (100mg/kg) and tannic acid inhibitor respectively. (50mg/kg, 100mg/kg, 150mg/kg). After fasting for 3 hours, the blood was collected by docking the tail to measure the plasma glucose concentration. The results showed that both doses of 100 and 150 mg/kg could significantly reduce blood sugar in normal mice (see Table 1).

Table 1 Inhibitors once administered to the impact of blood sugar levels in normal mice group no Dose (mg/kg) Blood glucose level (mmol/L) normal saline 10 - 7.80±0.48 sugar apple 10 100 6.31±0.50 ² Inhibitor 889 50100150 7.22±1.11 ¹ 5.50±0.49 ² 4.54±0.69 ²

Compared with normal saline group: ¹ p<0.005, ² p<0.001

The above-mentioned sugar apple contrast reagent is a product of German Bayer company.

Test 2, the effect of continuous multiple administration of Polygonum cuspidatum tannic acid inhibitors on the blood sugar content of normal mice: 30 mice were randomly divided into 3 groups, which were administered with normal saline, Baitangping (100mg/kg), Polygonum cuspidatum tanned Acid inhibitor (100mg/kg) for eight consecutive days. After fasting for 3 hours after the last administration, the blood glucose concentration was measured. It can be seen that multiple administrations to mice also caused the reduction of blood sugar in mice (see Table 2).

The influence of table 2 inhibitor continuous multiple administration on the blood sugar content of normal mice group no Blood glucose level (mmol/L) normal saline 10 6.30±0.89 sugar apple 10 5.41±0.31 ¹ Inhibitor 10 4.98±0.89 ²

Compared with normal saline group: ¹ p<0.005, ² p<0.001

The above-mentioned sugar apple contrast reagent is a product of German Bayer company.

Test 3, the influence of inhibitor on the blood sugar content of alloxan diabetic mice: 30 alloxan diabetic mice were divided into 3 groups (the difference in blood sugar value between the groups was not more than 0.5mmol/L), and they were fed with normal saline, Sugar apple (100mg/kg), inhibitor (100mg/kg), for eight consecutive days. After fasting for 3 hours after the last administration, blood was taken to measure blood sugar. The results showed that the inhibitor could significantly reduce the blood sugar of alloxan diabetic mice (see Table 3). The influence of table 3 inhibitor on the blood sugar content of alloxan diabetic mice group no Blood glucose level (mmol/L) normal saline 10 11.42±1.31 sugar apple 10 8.32±1.23 ¹ Inhibitor 10 6.85±0.98 ¹

Compared with normal saline group: ¹ p<0.001

The above-mentioned sugar apple contrast reagent is a product of German Bayer company.

Detailed ways

Now the embodiment of the extraction method of Polygonum cuspidatum tannic acid of the present invention is described in the following.

Embodiment 1: get 300g Chinese medicine Rhizoma Polygonum cuspidatum and add 1200ml distilled water, macerate and pulverize with a masher, keep warm at 60°C for 24 hours, then separate with a centrifuge, the centrifuge speed is 5000rpm, centrifugation time 10 minutes, discard the filter residue, Take the supernatant. The supernatant was heated to 80° C. for 10 minutes, and then placed in the refrigerator overnight. Then put it into a centrifuge, centrifuge at 5000rpm for 10 minutes, and take the supernatant. The supernatant was defatted 3-4 times with petroleum ether (30-60bp) in a separatory funnel. The degreased extract was decolorized through an activated carbon column. Then ultrafiltration removes low-molecular-weight impurities, and the resulting filtrate is placed on 732 strong acidic cation exchange resin, and then eluted with distilled water. Adjust the pH of the eluate to neutral, then heat it to 60-100°C under normal pressure or reduced pressure for concentration, and then use an ultrafiltration membrane (molecular weight cut-off 5000dalton) for ultrafiltration concentration to remove low-molecular impurities. After drying Brown solid powder can be obtained, the main component of which is Polygonum cuspidatum tannic acid.

Example 2: 300g Polygonum cuspidatum coarse powder is cold-soaked with 1500ml95% ethanol for 24hr, filtered, the filter residue is discarded, the filtrate is taken, the filtrate is heated to 80°C under reduced pressure, concentrated to obtain an extract, the extract is dissolved in water, refluxed with chloroform, discarded Chloroform layer, the obtained water layer was extracted 2-3 times with ethyl acetate, the ethyl acetate layer was taken, and then extracted 2-3 times with NaOH, the NaOH aqueous solution was taken, the pH value was adjusted to 4 with HCl, and 5% gelatin aqueous solution was added, get sediment. The precipitate was dissolved with 50% acetone aqueous solution, then centrifuged with a centrifuge at 5000rmp for 10 minutes, discarded the precipitate, took the acetone solution, concentrated it by distillation under reduced pressure, and recovered the acetone to obtain a colloidal solid. The obtained jelly solid was dissolved in water, and then extracted with ethyl acetate, the obtained ethyl acetate layer was concentrated under reduced pressure, and the ethyl acetate was recovered to obtain a concentrated gum. Add appropriate amount of water to dissolve the obtained jelly, filter, and filter out the precipitated insoluble matter to obtain a clear solution, freeze-dry the clear solution to obtain an orange-yellow powder, the main component of which is Polygonum cuspidatum tannic acid.

Claims (3)

1. the extracting method of a Rhizoma Polygoni Cuspidati extract giant knotweed tannic acid is characterized in that, this method comprises following processing step:

A. the crude drug giant knotweed is through after pulverizing, and water, methyl alcohol or aqueous alcohol are extracted under heating condition, during extraction

Between 2-30 hour, solvent load be crude drug 3-10 doubly;

B. filter or centrifugation, abandon filter residue, get filtrate or supernatant liquid, with filtrate or supernatant liquid in temperature

50-100 ℃ is incubated 5-30 minute, and refrigerator is placed and spent the night;

C. put into separating centrifuge,, get supernatant liquid 2000-5000rmp centrifugation 5-20 minute;

D. supernatant liquid is used petroleum ether degreasing 2-5 time in separatory funnel;

E. with the extracting solution activated carbon decolorizing after the degreasing, low molecular weight impurities is removed in ultrafiltration then;

F. gained filtrate is gone up storng-acid cation exchange resin, use the distilled water wash-out then, elutriant is regulated it

PH is heated to 60-100 ℃ and concentrates to neutrality under normal pressure or decompression, use ultra-filtration membrane then

Carry out ultrafiltration, can obtain the brown solid powder after drying, its main component is the giant knotweed tannic acid.

2. the extracting method of a Rhizoma Polygoni Cuspidati extract giant knotweed tannic acid is characterized in that, this method comprises following processing step:

A. with crude drug giant knotweed elder generation water, ethanol or aqueous alcohol are soaked, and macerate the back and pulverize with stamp mill, in heating

Under the condition, extracted 2-30 hour, solvent load is 3-10 a times of crude drug;

B. filter, abandon filter residue, get filtrate and under normal pressure or decompression, be heated to 50-100 ℃, concentrate medicinal extract;

C. be dissolved in water, use the chloroform reflux, abandon chloroform layer, gained water layer ethyl acetate extraction 2-3

Inferior, get ethyl acetate layer, use NaOH aqueous solution extraction 2-3 time again, get the NaOH aqueous solution, use

HCl transfers pH value to 2-6, adds the 4-10% aqueous gelatin solution, gets throw out;

D. precipitation is used the whizzer centrifugation then with the dissolving of 30-70% aqueous acetone solution, abandons precipitation, gets third

Ketone solution concentrates its underpressure distillation, and reclaims acetone, obtains the colloidal solid thing;

E. with gained colloidal solid thing water dissolution, use ethyl acetate extraction again, the gained ethyl acetate layer is reduced pressure

Concentrate, and reclaim ethyl acetate, obtain spissated jelly;

F. jelly adds the suitable quantity of water dissolving, filters, and elimination precipitation insolubles obtains clear liquid, and clear liquid is freezing dried

Dry orange-yellow powder, its main component is the giant knotweed tannic acid.

3. the tannic acid product that extracting method extracted of a kind of Rhizoma Polygoni Cuspidati extract giant knotweed tannic acid according to claim 1 and 2 owing to have alpha-glucoside inhibiting activity, can be used for making the raw material of the medicine for the treatment of diabetes, obesity etc.