CN110384721B - Method for extracting anti-inflammatory effective part of striga asiatica - Google Patents
Method for extracting anti-inflammatory effective part of striga asiatica Download PDFInfo
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- 230000003110 anti-inflammatory effect Effects 0.000 title claims abstract description 11
- 240000001705 Striga asiatica Species 0.000 title claims description 24
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- 239000000243 solution Substances 0.000 claims abstract description 17
- 239000003814 drug Substances 0.000 claims abstract description 16
- 239000000843 powder Substances 0.000 claims abstract description 13
- 239000007788 liquid Substances 0.000 claims abstract description 11
- 238000000605 extraction Methods 0.000 claims abstract description 10
- 238000001914 filtration Methods 0.000 claims abstract description 7
- 239000011259 mixed solution Substances 0.000 claims abstract description 7
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 5
- 108010059892 Cellulase Proteins 0.000 claims abstract description 4
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- 238000001035 drying Methods 0.000 claims abstract description 4
- 239000000706 filtrate Substances 0.000 claims abstract description 4
- 238000010298 pulverizing process Methods 0.000 claims abstract 2
- 238000004108 freeze drying Methods 0.000 claims description 4
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- 238000002156 mixing Methods 0.000 claims description 4
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- 238000002474 experimental method Methods 0.000 description 10
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- 241000700159 Rattus Species 0.000 description 6
- 229940113118 carrageenan Drugs 0.000 description 6
- 235000010418 carrageenan Nutrition 0.000 description 6
- 229920001525 carrageenan Polymers 0.000 description 6
- 239000000679 carrageenan Substances 0.000 description 6
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
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- 206010061218 Inflammation Diseases 0.000 description 4
- 240000003705 Senecio vulgaris Species 0.000 description 4
- 210000000683 abdominal cavity Anatomy 0.000 description 4
- 230000004856 capillary permeability Effects 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 241000801118 Lepidium Species 0.000 description 3
- 244000211187 Lepidium sativum Species 0.000 description 3
- 235000007849 Lepidium sativum Nutrition 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 241000722721 Capparis Species 0.000 description 2
- 240000001307 Myosotis scorpioides Species 0.000 description 2
- 208000002193 Pain Diseases 0.000 description 2
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- 241000416918 Polyphylla Species 0.000 description 2
- 230000001760 anti-analgesic effect Effects 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
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- 241000873224 Capparaceae Species 0.000 description 1
- 244000140995 Capparis spinosa Species 0.000 description 1
- 235000017336 Capparis spinosa Nutrition 0.000 description 1
- 208000032912 Local swelling Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 240000006694 Stellaria media Species 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
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- 238000005520 cutting process Methods 0.000 description 1
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- 210000005069 ears Anatomy 0.000 description 1
- 229960003699 evans blue Drugs 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 210000000548 hind-foot Anatomy 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- CGIGDMFJXJATDK-UHFFFAOYSA-N indomethacin Chemical group CC1=C(CC(O)=O)C2=CC(OC)=CC=C2N1C(=O)C1=CC=C(Cl)C=C1 CGIGDMFJXJATDK-UHFFFAOYSA-N 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000001543 one-way ANOVA Methods 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- VOKSWYLNZZRQPF-GDIGMMSISA-N pentazocine Chemical compound C1C2=CC=C(O)C=C2[C@@]2(C)[C@@H](C)[C@@H]1N(CC=C(C)C)CC2 VOKSWYLNZZRQPF-GDIGMMSISA-N 0.000 description 1
- 229960005301 pentazocine Drugs 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000004080 punching Methods 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- Natural Medicines & Medicinal Plants (AREA)
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- Alternative & Traditional Medicine (AREA)
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- Pain & Pain Management (AREA)
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- General Chemical & Material Sciences (AREA)
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Abstract
本发明申请属于药物提取技术领域，具体公开了一种独行千里抗炎有效部位的提取方法，包括以下步骤；(1)取独行千里根茎干燥并粉碎得粉末；(2)将步骤(1)中的粉末过筛，向粉末中加入硫酸溶液，加入纤维素酶混匀，置于恒温箱内酶解；(3)将步骤(2)中的混合液过滤收集滤液，将酶解后的独行千里药渣加入8倍量0.25％的硫酸溶液，超声波提取多次，收集、合并多次提取得到的提取液；(4)将步骤(3)中的滤渣过滤得到药液，将药液浓缩成稠膏；(5)将步骤(4)中的膏状物冷冻干燥并粉碎成干粉。本发明主要用于提取独行千里内的提取物，解决了现有技术中直接给患者使用独行千里的根或叶入药会对患者的康复造成影响的问题。The application of the present invention belongs to the technical field of drug extraction, and specifically discloses a method for extracting the effective anti-inflammatory parts of Radix vulgaris, comprising the following steps: (1) drying and pulverizing the rhizome of Radix Radix et Rhizoma to obtain powder; (2) extracting the rhizome from step (1) sieve the powder, add sulfuric acid solution to the powder, add cellulase to mix well, and place it in an incubator for enzymatic hydrolysis; (3) filter the mixed solution in step (2) to collect the filtrate, and separate the enzymatic hydrolysis for thousands of miles. Adding 8 times the amount of 0.25% sulfuric acid solution to the medicinal residue, ultrasonic extraction for many times, collecting and merging the extract obtained by multiple extractions; (4) filtering the filter residue in step (3) to obtain medicinal liquid, and concentrating the medicinal liquid into a thick (5) freeze-dry the paste in step (4) and pulverize it into dry powder. The present invention is mainly used for extracting the extract within the loneliness, and solves the problem in the prior art that directly using the roots or leaves of the loneliness as medicine will affect the recovery of the patient.
Description
Technical Field
The invention belongs to the technical field of medicine extraction, and particularly discloses a method for extracting an anti-inflammatory effective part of striga asiatica.
Background
Qianli, the name of Chinese medicine. Is root and leaf of Capparis Spinosa Makino of Capparidaceae. Distributed in southeast to south China. Has the effects of promoting blood circulation, removing blood stasis, dispelling pathogenic wind and relieving pain. It is commonly used for activating blood and dissipating blood stasis, dispelling wind and alleviating pain.
In the prior art, when the single-row groundsel is used, the roots and the leaves of the single-row groundsel are usually cut into slices or sections to be filled with medicine, the effective ingredients in the single-row groundsel cannot be accurately judged during use, when a patient takes the medicine, different medicine effects can be brought by too much or too little effective ingredients of the medicine, and the different medicine effects easily cause influence on the rehabilitation of the patient.
Disclosure of Invention
The invention aims to provide an extraction method of an anti-inflammatory effective part of striga asiatica to solve the problem that the recovery of a patient is affected by directly taking roots or leaves of the striga asiatica into a medicine in the prior art.
In order to achieve the purpose, the technical scheme of the invention is as follows: a method for extracting antiinflammatory effective component from herba Senecionis Scandentis comprises the following steps;
(1) cleaning and drying roots and stems of the medicinal material striga asiatica and crushing the dried roots and stems to obtain powder;
(2) sieving the powder in the step (1), adding a sulfuric acid solution with the mass being 8 times that of the powder, adding cellulase accounting for 1.2 percent of the mass of the strigosae asiatica, uniformly mixing to obtain a mixed solution, and placing the mixed solution in a constant temperature box for enzymolysis;
(3) filtering the mixed solution in the step (2), collecting filtrate, adding 8 times of 0.25% sulfuric acid solution into the enzymolysis-processed herba strigae Asiaticae dregs, performing ultrasonic extraction for multiple times, collecting and combining the extracting solutions obtained by multiple times of extraction;
(4) filtering the filter residue obtained in the step (3) by using three layers of gauze, performing suction filtration by using three layers of filter paper to obtain a liquid medicine, and evaporating and concentrating the liquid medicine in a rotary evaporator to obtain thick paste;
(5) and (4) freeze-drying and crushing the paste in the step (4) into dry powder.
Further, the mass fraction of the sulfuric acid solution in the step (2) is 0.25%, and the pH is 4.
Further, the temperature in the constant temperature box in the step (2) is 35 ℃, and the enzymolysis time is 3 hours.
Further, ultrasonic extraction is performed for 2 times in the step (3), and the extraction time is 30min each time.
Further, in the step (5), the mixture was freeze-dried for 12 hours and then pulverized.
The working principle and the beneficial effects of the technical scheme are as follows:
(1) multiple tests of the inventor show that the extract of the striga asiatica has better anti-inflammatory and analgesic effects;
(2) the extract of the striga asiatica linn extracted by the method has high yield and improves the utilization rate of the striga asiatica linn.
Detailed Description
The following is further detailed by way of specific embodiments:
the method for extracting the anti-inflammatory effective part of the striga asiatica in the embodiment comprises the following steps: the method comprises the steps of cleaning and drying 2kg of roots and stems of the medicinal material of the striga asiatica, crushing the roots and stems of the striga asiatica, sieving the dried roots and stems of the striga asiatica, sieving the ground roots and stems of the striga asiatica by a 24-mesh sieve, adding 8 times of 0.25% sulfuric acid solution with the pH value of 4.0, adding 1.2% (mass percentage of the medicinal material) of cellulase, uniformly mixing the mixture, placing the mixture in a constant temperature box at 35 ℃ for enzymolysis for 3 hours, filtering the mixture, collecting filtrate, adding 8 times of 0.25% sulfuric acid solution into the dregs of the striga asiatica after the enzymolysis, carrying out ultrasonic extraction for 2 times, wherein the extraction time is 30min each time, collecting and combining the extracted liquid obtained after 3 times of extraction, filtering the liquid by three layers of gauze, carrying out suction filtration by using three layers of filter paper to obtain liquid medicine, evaporating and concentrating the liquid medicine into thick paste by a rotary evaporator, and carrying out freeze drying in a freeze dryer for 12 hours to form dry powder for experiments.
In this example, the striga asiatica extract obtained by the ultrasonic method was dissolved in distilled water, ground, and prepared into a solution with a desired concentration to perform the following experiments, and the mouse was used as an experimental subject, and the doses of the experimental extract of the mouse were 80mg/kg and 160mg/kg, specifically as follows:
first, experiment method
1.1 xylene experiments 60 mice were taken and randomly divided into 6 groups: model control group, indomethacin group, high and low dose group of extract of herba Senecionis Scandentis, each group contains 10 animals, and male and female are half each. The administration was performed by gavage at the dose shown in Table 1, and the model control group was administered with distilled water of equal volume for 1 time/day for 5 days continuously. 30min after the last administration, uniformly coating 30 mu L of dimethylbenzene on the front and back surfaces of the right ear of the mouse, dislocation of the cervical vertebra of the mouse after 30min, cutting off the left and right auricles, punching a round ear piece on the same part of the left and right ears by using a hole puncher with the diameter of 7mm, weighing, and calculating the inhibition rate of the swelling degree by taking the mass difference of the left and right ear pieces as the swelling degree.
1.2 grouping and administration method of capillary permeability test animals are the same as 1. The final administration is carried out for 30min, then the mouse is intravenously injected with 0.6% Evans blue solution 10 mL/kg-1, and then the intraperitoneal injection is carried out with 0.6% glacial acetic acid 10 mL/kg-130min later, dislocation and sacrifice, open the abdominal cavity, wash the abdominal cavity with 5mL of normal saline, suck the washing liquid out, combine, 3000 r.min-1Centrifuging for 15min, collecting supernatant, measuring absorbance A at 595nm wavelength, and calculating inhibition rate.
1.3 grouping administration method of acetic acid writhing experiment animals is the same as 1, 30min after the last administration, and 10 mL/kg of 0.6% glacial acetic acid solution is injected into abdominal cavity of mouse-1The latency period for the mice to develop writhing response and the number of writhing responses within 20min were recorded.
1.4 carrageenan-induced paw swelling experiment of rats 40 rats were selected, 8 rats were selected in each group, the grouping and administration methods were the same as 1, 30min after the last administration, 0.1mL of 1% carrageenan physiological saline was subcutaneously injected into the right hind paw, and paw volumes were measured with a paw swelling meter at 1, 4, and 6h before and after the inflammation, respectively, so that the difference between the paw volumes before and after the inflammation represents the swelling degree.
1.5 Hot plate experiment female mice are placed in a preheated (55 +/-0.5) DEG C hot plate instrument, the foot latent period after the mice lick is used as a pain threshold index, 50 mice with pain threshold values of 5-30 s before the experiment are selected as qualified mice, the animal grouping administration method is the same as 1, the stomach is filled according to the dosage of table 5 for administration, the blank control group is administered with equal volume distilled water solution, the pain threshold value is measured for 2 times before the first administration and is separated for 5min, the average value is taken as the basic pain threshold value, in order to prevent feet from being scalded, 60s is set as the cut-off time, the pain threshold value is recorded as 60s when the time exceeds 60s, then the administration is carried out according to table 5, and the hot plate pain threshold values of the animals 30min and 60min after the last administration are recorded.
1.6 statistical analysis of data to
Mean comparisons were analyzed by One-way ANOVA. Statistical processing is done by SPSS 19.0 software. The difference is statistically significant when P is less than 0.05.
Second, experimental results
2.1 effects of paraxylene on ear swelling in mice; compared with the model control group, the high and low doses of the extract of the single-element groundsel herb can obviously inhibit the mouse ear swelling caused by dimethylbenzene, and the table 1 shows.
TABLE 1 Effect of Lepidium striifolia extract on ear swelling in mice: (
n＝10)
P <0.05, P <0.01 compared to model control.
2.2 effects on acetic acid-induced increase in capillary permeability in mice; compared with the model control group, the extract of the striga asiatica can obviously inhibit the increase of the capillary permeability of mice, and the table 2 shows.
Table 2 independent rowInfluence of Stellaria extract on acetic acid-induced increase in permeability of capillary vessels in abdominal cavity of mouse (A)
n＝10)
P <0.05, P <0.01 compared to model control.
2.3 Effect on carrageenan-induced foot swelling in rats; after carrageenan-induced inflammation, the degree of swelling of the foot of the model control group is continuously increased, and the degree of swelling of the foot of the striga asiatica maxim extract group reaches a peak at 4 hours and then begins to decline. Compared with a model control group, the swelling degree of each experimental group has no significant difference 1h after inflammation; the extract has obvious inhibiting effect at high dose within 4 h; at 6h, the higher and lower doses of the extract of the Capparis polyphylla can inhibit the swelling of feet, as shown in Table 3.
TABLE 3 Effect of Lepidium Sativum extract on carrageenan-induced swelling of rat feet: (
n＝8)
P <0.05, P <0.01 compared to model control.
2.4 influence on acetic acid induced writhing experiment of mice; compared with a model control group, the high dose of the extract can obviously prolong the incubation time of the writhing of the mice caused by acetic acid; the high and low dose of the extract reduced the number of writhing in mice, as shown in Table 4.
TABLE 4 Effect of Lepidium Sativum extract on acetic acid-induced writhing in mice (
n＝10)
P <0.05, P <0.01 compared to model control.
2.5 impact on hotplate experiments; the comparison of the basic pain threshold values of all experimental groups has no statistical difference; compared with the blank control group, pain thresholds of the pentazocine and striga asiatica high-dose groups are obviously prolonged at 60min (P is less than 0.01), and pain thresholds of the extract low-dose groups are obviously prolonged at 60min, which is shown in Table 5.
TABLE 5 Effect of Lepidium strife extract on pain threshold in Hot plate mice (
n＝8)
P <0.01 compared to the blank control group.
The following conclusions can be drawn from the above experimental data:
1: the striga asiatica extract can obviously inhibit the increase of the capillary permeability of mice caused by acetic acid.
2: the Lepidium japonicum extract can inhibit mouse ear swelling caused by xylene.
3: the extract of the striga asiatica can obviously prolong the latency time of writhing reaction of the mice caused by acetic acid and reduce the times of writhing of the mice.
4: the herba Senecionis Scandentis extract can significantly improve pain threshold of hot plate test of mice 30min after administration.
5: the extract of the Capparis polyphylla can inhibit the swelling of the feet of the rat caused by the carrageenan.
The above results show that: the Lepidium Sativum extract has certain anti-inflammatory and analgesic effects.
The foregoing is merely an example of the present invention and common general knowledge of known specific structures and features of the embodiments is not described herein in any greater detail. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent.
Claims (2)
1. A method for extracting anti-inflammatory effective parts of striga asiatica, which is characterized by comprising the following steps;
(1) cleaning and drying roots and stems of the medicinal material striga asiatica and crushing the dried roots and stems to obtain powder;
(2) sieving the powder in the step (1), adding 0.25% sulfuric acid solution in an amount which is 8 times the mass of the powder, adding cellulase accounting for 1.2% of the mass of the mile, uniformly mixing to obtain a mixed solution, and placing the mixed solution in a constant temperature box for enzymolysis; wherein the mass fraction of the sulfuric acid solution is 0.25 percent, and the pH value is 4; the temperature in the constant temperature box is 35 ℃, and the enzymolysis time is 3 hours;
(3) filtering the mixed solution in the step (2), collecting filtrate, adding 8 times of 0.25% sulfuric acid solution into the residue of the herba Senecionis Scandentis after enzymolysis, performing ultrasonic extraction for 2 times, wherein the extraction time is 30min each time, collecting and mixing the extractive solutions obtained by multiple times of extraction;
(4) filtering the extracting solution in the step (3) by using three layers of gauze, performing suction filtration by using three layers of filter paper to obtain a liquid medicine, and evaporating and concentrating the liquid medicine in a rotary evaporator to obtain thick paste;
(5) and (4) freeze-drying and crushing the paste in the step (4) into dry powder.
2. The method for extracting the effective anti-inflammatory component of the striga asiatica according to claim 1, wherein the pulverizing step (5) is performed after freeze-drying for 12 hours.
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| CN101780124A (en) * | 2010-03-12 | 2010-07-21 | 华南农业大学 | Capparis acutifolia sweet exact and preparation method and application thereof |
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| CN101780124A (en) * | 2010-03-12 | 2010-07-21 | 华南农业大学 | Capparis acutifolia sweet exact and preparation method and application thereof |
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| 独行千里化学成分研究;陈熔等;《广东医药学院学报》;19901231;第6卷(第1期);第12-13页 * |
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