CN110384721A - A kind of extracting method of the anti-inflammatory active component of acuteleaf caper root - Google Patents

Abstract

The application of the present invention belongs to the technical field of medicine extraction, and specifically discloses a method for extracting the effective anti-inflammatory part of Dixing Qianli. sieve the powder, add sulfuric acid solution to the powder, add cellulase to mix, and place in an incubator for enzymolysis; (3) filter the mixed solution in step (2) to collect the filtrate, and enzymatically hydrolyze the Add 8 times the amount of 0.25% sulfuric acid solution to the dregs, ultrasonically extract multiple times, collect and combine the extracts obtained by multiple extractions; (4) filter the filter residue in step (3) to obtain the medicinal solution, and concentrate the medicinal solution to thicken paste; (5) the paste in step (4) is freeze-dried and crushed into dry powder. The present invention is mainly used for extracting the extract of Lonely Thousand Miles, and solves the problem in the prior art that directly using the roots or leaves of Lonely Thousand Miles as medicine will affect the rehabilitation of patients.

Description

A kind of extraction method of anti-inflammation effective part

technical field

The invention belongs to the technical field of medicine extraction, and specifically discloses a method for extracting anti-inflammation effective parts.

Background technique

Traveling thousands of miles alone, the name of traditional Chinese medicine. It is the root and leaves of the vine of the cauliflower plant. Distributed in southeast to south of my country. It has the effects of promoting blood circulation and removing blood stasis, expelling wind and relieving pain. Commonly used in promoting blood circulation and dispelling blood stasis, expelling wind and relieving pain.

In the prior art, when using Lonely Thousand Miles, the roots and leaves of Lonely Thousand Miles are usually cut into slices or segments to be used as medicine, and the active ingredients in it cannot be accurately judged during use. Rarely will bring different drug effects, and different drug effects are likely to affect the recovery of patients.

Contents of the invention

The purpose of the present invention is to provide a method for extracting the effective anti-inflammatory parts of Lonely Thousand Miles, so as to solve the problem in the prior art that directly using the roots or leaves of Lonely Thousand Miles as medicine will affect the recovery of patients.

In order to achieve the above object, the technical solution of the present invention is: a method for extracting effective anti-inflammatory parts of a single journey, comprising the following steps;

(1) Take the rhizome of the medicinal material "Lonely Thousand Miles", wash, dry and pulverize to obtain powder;

(2) Sieve the powder in step (1), add 8 times the mass of sulfuric acid solution to the powder, add 1.2% cellulase by mass and mix to obtain a mixed solution, and place it in an incubator for enzymatic hydrolysis;

(3) Filter the mixed solution in step (2), collect the filtrate, add 8 times the amount of 0.25% sulfuric acid solution to the enzymatically hydrolyzed medicinal residue of Du Xingqianli, perform ultrasonic extraction multiple times, collect and combine the extraction obtained by multiple extractions liquid;

(4) Filter the filter residue in step (3) with three layers of gauze, then use three layers of filter paper to obtain the medicinal solution, evaporate and concentrate it into a thick paste in a rotary evaporator;

(5) The paste in step (4) is freeze-dried and pulverized into dry powder.

Further, the mass fraction of the sulfuric acid solution in step (2) is 0.25%, and the pH is 4.

Further, in step (2), the temperature in the incubator is 35° C., and the enzymatic hydrolysis time is 3 hours.

Further, in step (3), ultrasonic extraction is performed twice, and the time of each extraction is 30 min.

Further, in the step (5), crushing is carried out after freeze-drying for 12 hours.

The working principle and beneficial effects of this technical solution are:

(1) Through multiple tests by the inventor, it is concluded that the extract of Lonely Thousand Miles has a good anti-inflammatory and analgesic effect;

(2) The yield of the extract obtained by adopting this method to extract the extract of Lone Walker is higher, which improves the utilization rate of Lone Walker.

Detailed ways

The following is further described in detail through specific implementation methods:

A method for extracting the anti-inflammatory effective part of the present embodiment, comprising the following steps: take 2 kg of the rhizome of the medicinal material, the root of the plant, washed and dried, then pulverized, passed through a 24-mesh sieve, and added 8 times the amount of 0.25% sulfuric acid solution with a pH of 4.0 , add 1.2% (accounting for the percentage of medicinal material mass) cellulase and mix well, place it in a 35°C incubator for enzymolysis for 3 hours, filter, collect the filtrate, add 8 times the amount of 0.25% sulfuric acid to the enzymatically hydrolyzed medicinal residue Solution, ultrasonic extraction 2 times, the time of each extraction is 30min, collect and merge the extract obtained from 3 times of extraction, filter with three layers of gauze, and then filter with three layers of filter paper to obtain the medicinal solution, evaporate and concentrate in a rotary evaporator to form a thick The paste was freeze-dried in a freeze dryer for 12 hours to form a dry powder for experimentation.

In this example, the extract obtained by the ultrasonic method was dissolved in distilled water and ground, and prepared into a solution of the required concentration to carry out the following tests. The mice were used as the experimental subjects, and the dose of the mouse experimental extract was 80mg/kg, 160mg /kg, as follows:

1. Experimental method

1.1 Xylene experiment 60 mice were randomly divided into 6 groups: model control group, indomethacin group, high-dose and low-dose groups of Duxingqianli extract, 10 mice in each group, half male and half male. The doses in Table 1 were given by intragastric administration, and the model control group was given an equal volume of distilled water solution, once a day, for 5 consecutive days. 30 minutes after the last administration, take 30 μL of xylene and apply it evenly on the front and back sides of the right ear of the mice. After 30 minutes, the mice were killed by cervical dislocation, and the left and right auricles were cut off, and round ear pieces were punched on the same part of the left and right ears with a puncher with a diameter of 7 mm. , weighed, and the difference between the mass of the left and right ear pieces was regarded as the degree of swelling, and the inhibition rate of the degree of swelling was calculated.

1.2 The animal grouping and administration method of the capillary permeability experiment are the same as in 1. 30 minutes after the last administration, the mice were given tail vein injection of 0.6% Evans blue solution 10mL·kg-1, followed by intraperitoneal injection of 0.6% glacial acetic acid 10mL·kg ^-1 . After 30 minutes, the mice were killed by dislocation, the abdominal cavity was opened, and the abdominal cavity was washed with 5mL normal saline , suck out the washing solution, combine, centrifuge at 3000r·min ^-1 for 15min, take the supernatant and measure the absorbance A value at a wavelength of 595nm, and calculate the inhibition rate.

1.3 The administration method of animals in groups of acetic acid writhing experiment is the same as that in 1. 30 minutes after the last administration, the mice were intraperitoneally injected with 0.6% glacial acetic acid solution 10mL·kg ^-1 , and the incubation period of mice writhing reaction and the mice writhing within 20 minutes were recorded. number of responses.

1.4 Carrageenan-induced swelling of rat paws 40 rats were taken, 8 in each group, grouped and administered in the same way as in 1. 30 minutes after the last administration, 1% carrageenin physiological salt was subcutaneously injected into the right hind paw Aqueous solution 0.1mL, respectively before inflammation and 1, 4, 6 hours after inflammation, paw swelling meter was used to measure the volume of paw, so that the difference of paw volume before and after inflammation indicated the degree of swelling.

1.5 Hot plate experiment Place the female mice on a preheated (55±0.5)°C hot plate apparatus, take the incubation period of the mouse licking the hind paw as the pain threshold index, and select 50 mice with a pain threshold of 5-30s before the experiment as qualified The administration method of mice and animals is the same as 1, and the doses in Table 5 are given by intragastric administration. The blank control group is given an equal volume of distilled water solution. Before the first administration, the pain threshold is measured twice with an interval of 5 minutes, and the average value is taken as the basic pain threshold. , in order to prevent foot scald, set 60s as the cut-off time, exceed 60s and record as the pain threshold of 60s, then administer according to Table 5, record the animal hot plate pain threshold of 30 and 60min after the last administration.

1.6 Statistical analysis data to Means were compared using One-way ANOVA analysis. Statistical processing was completed by SPSS 19.0 software. P<0.05 was regarded as a statistically significant difference.

2. Experimental results

2.1 The effect on ear swelling of mice induced by xylene; compared with the model control group, high and low doses of the extract of Duxingqianli can significantly inhibit ear swelling of mice induced by xylene, see Table 1.

Table 1 The effect of the extract of a thousand miles alone on ear swelling in mice ( n=10)

Compared with the model control group, *P<0.05, **P<0.01.

2.2 The effect on the increase of capillary permeability of mice induced by acetic acid; compared with the model control group, the extract of Duxingqianli can significantly inhibit the increase of capillary permeability of mice, see Table 2.

Table 2 The effect of the extract of Duxingqianli on the increase of acetic acid-induced peritoneal capillary permeability in mice ( n=10)

Compared with the model control group, *P<0.05, **P<0.01.

2.3 Effects on carrageenan-induced paw swelling in rats; after carrageenan-induced inflammation, the paw swelling of the model control group continued to increase, and in the Duxingqianli extract group, the paw swelling reached a peak at 4 hours and then began to decline. Compared with the model control group, after 1 hour of inflammation, there was no significant difference in the swelling degree of each experimental group; at 4 hours, the high dose of the extract had a significant inhibitory effect; at 6 hours, the high and low doses of Duxingqianli extract could inhibit the paw swelling, see table 3.

Table 3 The effect of the extract of Lone Walking Thousand Miles on paw swelling of rats induced by carrageenan ( n=8)

Compared with the model control group, *P<0.05, **P<0.01.

2.4 Effects on acetic acid-induced writhing experiments in mice; compared with the model control group, high doses of extracts can significantly prolong the incubation time of mice writhing caused by acetic acid; high and low doses of extracts can reduce the number of writhing times in mice, see Table 4.

Table 4 The effect of the extract of Duxingqianli on the writhing test of mice induced by acetic acid ( n=10)

Compared with the model control group, *P<0.05, **P<0.01.

2.5 The effect on the hot plate test; there was no statistical difference in the basic pain threshold of each experimental group; compared with the blank control group, the pain threshold of the pentazocine and Duxingqianli high-dose groups were significantly prolonged at 30 and 60 minutes (P<0.01) , the pain threshold of the low-dose extract group was significantly prolonged at 60 minutes, as shown in Table 5.

Table 5 The effect of the extract of Lone Walking Thousand Miles on the pain threshold of mice in the hot plate experiment ( n=8)

Compared with blank control group, **P<0.01.

The following conclusions can be drawn from the above experimental data:

1: The extract of Lone Walking Thousand Miles can significantly inhibit the increase of capillary permeability in mice induced by acetic acid.

2: The extract of Alone Thousand Miles can inhibit mouse ear swelling caused by xylene.

3: The extract of Lonely Thousand Miles can significantly prolong the latency time of acetic acid-induced writhing reaction in mice and reduce the number of writhing in mice.

4: The extract of Alone Thousand Miles can significantly increase the pain threshold of the mouse hot plate test after administration for 30 and 60 minutes.

5: The extract of Lonely Thousand Miles can inhibit the paw swelling of rats induced by carrageenan.

The above results show that the extract of Duxingqianli has certain anti-inflammatory and analgesic effects.

What is described above is only an embodiment of the present invention, and common knowledge such as specific structures and characteristics known in the scheme are not described here too much. It should be pointed out that for those skilled in the art, under the premise of not departing from the structure of the present invention, several modifications and improvements can also be made, and these should also be regarded as the protection scope of the present invention, and these will not affect the implementation of the present invention. Effects and utility of patents.

Claims (5)

1. a kind of extracting method of the anti-inflammatory active component of acuteleaf caper root, which is characterized in that include the following steps；

(1) it takes medicinal material acuteleaf caper root rhizome clean dry and crushes to obtain powder；

(2) powder in step (1) is sieved, 0.25% sulfuric acid solution of 8 times of quality is added into powder, addition, which accounts for, walks alone The cellulase of a thousand li quality 1.2% mixes to obtain mixed liquor, is placed in enzymatic hydrolysis in insulating box；

(3) mixed liquor in step (2) is filtered, collects filtrate, 8 times of amounts 0.25% are added in the acuteleaf caper root dregs of a decoction after enzymatic hydrolysis Sulfuric acid solution, ultrasonic wave extraction is multiple, collect, merge repeatedly extract obtained extracting solution；

(4) filter residue in step (3) is used into three layers of filtered through gauze, then filters to obtain medical fluid with three layers of filter paper, in rotary evaporation Instrument is concentrated by evaporation into thick paste；

(5) paste in step (4) is freeze-dried and is ground into dry powder.

2. a kind of extracting method of the anti-inflammatory active component of acuteleaf caper root according to claim 1, which is characterized in that step (2) mass fraction of sulfuric acid solution is 0.25%, pH 4 in.

3. a kind of extracting method of the anti-inflammatory active component of acuteleaf caper root according to claim 1, which is characterized in that step (2) temperature in insulating box is 35 DEG C, enzymolysis time 3h.

4. a kind of extracting method of the anti-inflammatory active component of acuteleaf caper root according to claim 1, which is characterized in that step (3) ultrasonic wave extraction 2 times in, the time extracted every time are 30min.

5. a kind of extracting method of the anti-inflammatory active component of acuteleaf caper root according to claim 1, which is characterized in that step (5) it is crushed after having suffered freeze-drying 12h.