CN108969580B - Preparation method and application of blue cloth total tannin - Google Patents
Preparation method and application of blue cloth total tannin Download PDFInfo
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- CN108969580B CN108969580B CN201811079304.6A CN201811079304A CN108969580B CN 108969580 B CN108969580 B CN 108969580B CN 201811079304 A CN201811079304 A CN 201811079304A CN 108969580 B CN108969580 B CN 108969580B
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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)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P7/00—Drugs for disorders of the blood or the extracellular fluid
- A61P7/06—Antianaemics
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- 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
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
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- 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
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
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- 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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- 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
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
-
- 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
- A61K2236/55—Liquid-liquid separation; Phase separation
Abstract
A preparation method and application of total tannin of blue cloth belongs to the technical field of traditional Chinese medicines, and the preparation method comprises the following steps: 1) taking the raw medicinal materials of the blue linen, crushing the medicinal materials into powder, soaking the medicinal materials in acetone, 2) carrying out ultrasonic extraction, 3) extracting the medicinal materials by using water saturated ethyl acetate with 2 times of volume, 4) adding a gelatin solution for precipitation, 5) carrying out ultrasonic extraction by using acetone, 6) carrying out vacuum drying to obtain the purified blue linen positive tannin. The preparation method of the total tannin of the blue-cloth Zhengtan comprises the steps of ultrasonic extraction, solvent extraction, gelatin precipitation, vacuum drying and the like, and the prepared purified blue-cloth Zhengtan has obvious effect on the medicine for tonifying qi and blood.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to a preparation method and application of total tannin of blue-cloth euonymus.
Background
The blue cloth is the dried whole plant of the rosaceous plant wayside or velveteen wayside, is perennial herb, has wide resource distribution, is usually collected in summer and autumn, is subjected to impurity removal, is cleaned, and is dried or dried to obtain the blue cloth. Has the functions of replenishing qi, invigorating spleen, replenishing blood, nourishing yin, moistening lung and eliminating phlegm. It is commonly indicated for deficiency of qi and blood, cough due to consumptive disease, spleen deficiency and leukorrhagia. Modern researches show that the blue cloth contains chemical components such as tannins, flavonoids, phenylpropanoids, triterpenoids, volatile oils, amino acids, inorganic elements and the like, wherein the tannins are important active ingredients.
Modern researches find that the blue-cloth normal water extract has good blood replenishing effect, but due to the complexity of the components of the traditional Chinese medicine, the effective components are not extracted and separated, the traditional Chinese medicine is not easy to research, and the effect of the finished medicine is not outstanding.
Disclosure of Invention
The invention aims to overcome the difficulties of the background technology and provides an extraction and preparation method of total tannin of aleppo avens and application of the total tannin to qi and blood supplementation.
In order to achieve the purpose, the technical scheme is as follows:
a preparation method of blue cloth positive total tannin comprises the following steps:
1) taking 100g of the blue cloth crude drug, crushing the blue cloth crude drug into particles, and soaking the particles in 50 percent acetone solution which is 10 times of the weight of the drug for 12 hours;
2) ultrasonically extracting the blue cloth and the soak solution for 10-30 min, filtering, collecting the filtrate, removing acetone in the filtrate, concentrating, adding water to 500ml, standing, filtering, and collecting the filtrate;
3) extracting the blue-cloth normal ultrasound extracting solution by using water saturated ethyl acetate with the volume 2 times that of the blue-cloth normal ultrasound extracting solution until an ethyl acetate layer is colorless, collecting the extracting solution, removing the ethyl acetate, adding water to 300mL after concentration, standing, filtering, and collecting filtrate;
4) slowly adding 90mL of gelatin solution into the blue cloth positive extract, standing, filtering, and collecting precipitate;
5) adding 250mL of 90% acetone solution into the blue cloth normal gelatin precipitate, performing ultrasonic extraction, and removing acetone to obtain a concentrated solution;
6) and vacuum drying the concentrated solution obtained in the step 5) to obtain the purified ceratin.
And removing the solvent in the steps 2), 3) and 5) by adopting a 45 ℃ reduced pressure concentration method.
The concentration of the gelatin solution in the step 4) is 3.3%.
The temperature of vacuum drying in the step 6) is 45 ℃, and the time is 10-20 h.
An application of the blue-cloth healthy total tannin prepared by the method in the aspect of medicaments for replenishing qi and blood.
The beneficial effect who adopts above-mentioned scheme does: the preparation method of the total tannin of the blue-cloth Zhengtan comprises the steps of crushing, ultrasonic extraction, solvent extraction, gelatin precipitation, vacuum drying and the like, and the prepared purified blue-cloth Zhengtan has obvious effect on the medicine for replenishing qi and blood.
Detailed Description
The present invention is further described with reference to the following examples, but the present invention is not limited to the following examples, and it is anticipated that one skilled in the art may make various modifications in combination with the prior art.
Example 1
A preparation method of blue cloth positive total tannin comprises the following steps:
1) taking 100g of the blue cloth crude drug, crushing the blue cloth crude drug into particles, and soaking the particles in 50 percent acetone solution which is 10 times of the weight of the drug for 12 hours;
2) ultrasonically extracting the blue cloth and the soak solution for 10-30 min, filtering, collecting the filtrate, removing acetone in the filtrate, concentrating, adding water to 500ml, standing, filtering, and collecting the filtrate;
3) extracting the blue-cloth normal ultrasound extracting solution by using water saturated ethyl acetate with the volume 2 times that of the blue-cloth normal ultrasound extracting solution until an ethyl acetate layer is colorless, collecting the extracting solution, removing the ethyl acetate, adding water to 300mL after concentration, standing, filtering, and collecting filtrate;
4) slowly adding 90mL of gelatin solution into the blue cloth positive extract, stirring, standing, filtering, and collecting precipitate;
5) adding 250mL of 90% acetone solution into the blue cloth normal gelatin precipitate, performing ultrasonic extraction, and removing acetone to obtain a concentrated solution;
6) and vacuum drying the concentrated solution obtained in the step 5) to obtain the purified ceratin.
And removing the solvent in the steps 2), 3) and 5) by adopting a 45 ℃ reduced pressure concentration method.
The concentration of the gelatin solution in the step 4) is 3.3%.
The temperature of vacuum drying in the step 6) is 45 ℃, and the time is 10-20 h.
An application of the blue-cloth healthy total tannin prepared by the method in the aspect of medicaments for replenishing qi and blood.
Example 2
1. Material
1.1. Animal female Kunming breed mice, weight (20 + -2) g, supplied by Tianqin biotechnology, Inc., Changsha, under the certification number SCXK (Xiang) 2014-0011.
1.2. The blue-cloth positive medicinal material is collected from liquidambar formosana town of Zunyi county in Guizhou province, and is identified as the whole grass of Geum japonicum Thunb var chinensis F.bollle by doctor of Feichun university of medical academy of Zunyi Feichun, Lianzhi Yaowuqiang, Chinense Memo, Geum japonicum Thunb; cyclophosphamide for injection (CTX, shendy pharmaceutical ltd, jiang su); pharmaceutical gelatin (melena); RE-2000A rotary evaporator (Shanghai Yangrong Biochemical Instrument factory); a vacuum pump of SHB-III type (Zhengzhou great wall science, Ltd.); 2500Y type Chinese medicine grinder (platinum Europe hardware products Co., Yongkang); full-automatic blood cell analyzer.
2. Method of producing a composite material
2.1. Preparation of the total tannin of the herba Gei, 100g of herba Gei, is taken and crushed into coarse particles, and the ratio of 1: adding 50% acetone solution into 10 material-liquid ratio, soaking overnight, ultrasonic extracting for 10min, filtering the liquid medicine, recovering acetone from the filtrate at 45 deg.C under reduced pressure until no acetone smell, pouring out the concentrated solution, adding water to 500mL, standing for 1h, filtering to remove precipitate, collecting the filtrate, extracting the filtrate with 2 times volume of water saturated ethyl acetate until the ethyl acetate layer is colorless, collecting the extract, recovering ethyl acetate at 45 deg.C under reduced pressure until the ethyl acetate layer is complete, pouring out the concentrated solution, adding water to 300mL, standing for 1h, filtering to obtain precipitate, slowly adding 90mL gelatin solution (5 g gelatin dissolved in 150mL water) into the filtrate, standing for 1h, collecting the precipitate, ultrasonic extracting the precipitate with 250mL 90% acetone solution for 2h at 250mL, maintaining the water temperature below 20 deg.C, recovering acetone at 45 deg.C under reduced pressure until the acetone is complete to obtain the concentrated solution, vacuum drying the concentrated solution at 45 deg.C for 12h to obtain purified tannin (determining tannin content according to 2015, its purity is greater than 98%).
2.2. Experimental group administration and animal model preparation mice were fed for 1 week, and then randomly divided into a normal control group, a cyclophosphamide model group and a blue-cloth positive tannin group by weight, each group containing 10 mice. From the day of the experiment, the blue cloth positive tannin group was gavaged with 20mg/kg of aqueous blue cloth positive tannin solution for 13 days continuously, and the other groups of mice were gavaged with physiological saline of the same volume. And 8d, beginning the gavage, i.e. performing intraperitoneal injection of cyclophosphamide on the mice of the other groups except the normal control group at the dose of 100mg/kg to establish a blood deficiency model, and performing intraperitoneal injection of equal volume of normal saline for 3d continuously on the mice of the normal control group.
2.3. Peripheral hemogram was measured 2h after the last gavage, and the eyeballs were picked up and blood was taken into a blood collection tube containing EDTA anticoagulant, and peripheral blood leukocytes (WBC), Red Blood Cells (RBC), Hemoglobin (HGB), Platelets (PLT) were measured with a fully automatic blood cell analyzer.
3. Results
3.1. Influence of total tannin of aleppon avens on peripheral hemogram of cyclophosphamide-induced blood deficiency mouse
As shown in table 1, peripheral blood leukocytes, erythrocytes, hemoglobin, and platelets were significantly decreased in the model group mice as compared with those in the normal group (P < 0.01); compared with the model group, the blue-cloth positive tannin can obviously increase the reduced WBC, RBC and HGB (P < 0.01) and has the effect of increasing PLT (P < 0.05).
TABLE 1 comparison of peripheral hemograms (. + -. s, n = 8) for groups of mice
| Group of | Blood platelets (10)9/L） | Hemoglobin (g/L) | Red blood cells (10)12/L) | White blood cells (10)9/L) |
| Normal control group | 925.5±101.8** | 174.5±3.4** | 11.2±0.2** | 3.3±0.9** |
| Model set | 544.0±33.3 | 154.4±2.8 | 9.8±0.3 | 0.8±0.1 |
| Blue cloth total tannin group | 704.1±142.9* | 167.8±7.6** | 10.8±0.6** | 1.3±0.3** |
Note: compared with the model groupP<0.05, **P<0.01。
Claims (1)
1. The application of the total tannin of the blue linen in the aspect of preparing the medicine for replenishing qi and blood is characterized in that: the method comprises the following steps:
1) taking 100g of the blue cloth crude drug, crushing the blue cloth crude drug into powder, and soaking the powder in 50% acetone solution which is 10 times of the weight of the drug for 12 hours;
2) ultrasonically extracting the blue cloth and the soak solution for 10-30 min, filtering, collecting the filtrate, removing acetone in the filtrate, concentrating, adding water to 500ml, standing, filtering, and collecting the filtrate;
3) extracting the final filtrate obtained in the step 2 by using 2 times volume of water saturated ethyl acetate until an ethyl acetate layer is colorless, collecting extract liquor, removing ethyl acetate, adding water to 300mL after concentration, standing, filtering, and collecting filtrate;
4) slowly adding 90mL of gelatin solution with the concentration of 3.3% into the filtrate obtained in the step 3, stirring, standing, filtering, and collecting precipitate;
5) adding 250mL of 90% acetone solution into the blue cloth normal gelatin precipitate, performing ultrasonic extraction, and removing acetone to obtain a concentrated solution;
6) vacuum drying the concentrated solution obtained in the step 5) for 10-20h at the temperature of 45 ℃ to obtain purified blue cloth positive tannin;
wherein the solvent is removed in the steps 2), 3) and 5) by adopting a method of decompression concentration at 45 ℃.
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Non-Patent Citations (3)
| Title |
|---|
| Role of nitric oxide in the vasorelaxant and hypotensive effects of;Yi-Wu Xie等;《Journal of Ethnopharmacology》;20071231(第109期);第128-133页 * |
| 基于地榆鞣质与皂苷治疗白细胞减少症相恶关系的欣生胶囊成药性研究;苏柘僮;《中国博士学位论文全文数据库 医药卫生科技辑》;20130315(第03期);第14-15、25-26页，尤其是第14页最后1段，第15页第1行，第25页"2"，第26页最后1段 * |
| 蓝布正补血作用的实验研究;彭泽萍等;《黔南民族医专学报》;20110930;第24卷(第3期);第161-163页，尤其是第161页左栏第1段、右栏"1.2.1实验药物的提取"，第163页左栏"3讨论" * |
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Address after: 563000 No. 201, Dalian Road, Zunyi, Guizhou Patentee after: ZUNYI MEDICAL University Address before: 563000 No. 201, Dalian Road, Huichuan District, Zunyi City, Guizhou Province Patentee before: ZUNYI MEDICAL University |
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Effective date of registration: 20220129 Address after: 557500 Industrial Park, Jinzhong Economic Development Zone, Yangwu Town, Danzhai County, Qiandongnan Miao and Dong Autonomous Prefecture, Guizhou Province Patentee after: Guizhou kebeian biopharmaceutical Co.,Ltd. Address before: 563000 No. 201, Dalian Road, Zunyi, Guizhou Patentee before: ZUNYI MEDICAL University |