CN110585424A - Preparation method of leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases - Google Patents
Preparation method of leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases Download PDFInfo
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- CN110585424A CN110585424A CN201910880264.3A CN201910880264A CN110585424A CN 110585424 A CN110585424 A CN 110585424A CN 201910880264 A CN201910880264 A CN 201910880264A CN 110585424 A CN110585424 A CN 110585424A
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- leech
- polypeptide
- blood
- leech polypeptide
- liquid
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- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
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Abstract
The invention relates to a preparation method of leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases, which is prepared by crushing dried spina date seeds, angelica sinensis, ligusticum wallichii and dried longan, soaking the crushed dried spina date seeds, the soaked dried spina date seeds and the soaked dried longan in primary pulp liquor, and adding leech polypeptide refined extract to prepare the leech polypeptide medicinal liquor. The leech polypeptide medicated wine prepared by the method has the effects of improving sleep quality, relieving dizziness and headache symptoms, enhancing blood supply and nourishing capability of tissues and organs, accelerating blood flow, promoting blood circulation, reducing blood viscosity, regulating blood fat, reducing cholesterol and fat content in plasma, strongly removing garbage toxin in blood, dissolving thrombus, softening blood vessels, improving immunity, eliminating atherosclerotic plaque, preventing vascular rupture and the like, and can effectively prevent and treat cardiovascular and cerebrovascular diseases.
Description
Technical Field
The invention relates to medicinal liquor, in particular to a preparation method of leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases.
Background
Cardiovascular and cerebrovascular diseases are common diseases seriously harming human health, and are rapidly increased year by year, the death rate and the disability rate are high, and the number of people dying from the cardiovascular and cerebrovascular diseases in each year in the world is up to 1500 thousands of people, and the people live at the first cause of death. Cardiovascular and cerebrovascular diseases and complications thereof become the first killers endangering human life, and the diseases are mostly related to thrombus, blood coagulation and blood viscosity. The prevention and treatment of cardiovascular and cerebrovascular diseases are mainly to prevent and treat hypertension, hyperlipidemia, diabetes, coronary heart disease, cerebral arteriosclerosis and the like, which are risk factors of the cardiovascular and cerebrovascular diseases, and thrombus and blood fat are common diseases of middle-aged and elderly people over 50 years old and seriously threaten human health. Clinical treatment mostly depends on drugs strictly, the existing oral thrombolytic products have the defects of poor performance, large side effect of chemical thrombolytic drugs, incomplete thrombolysis, large side effect on human bodies and the like, and the side effect of various lipid-lowering and cholesterol-lowering drugs limits the treatment and application range.
The leech polypeptide is a natural substance with active ingredients extracted from leech saliva or fresh leech extract of bloodsucker golden-edged, and is a small molecular protein (polypeptide) consisting of 65-66 amino acids, which is also called hirudin. The molecular weight of the natural hirudin is only 7000 daltons, the hirudin is very soluble in water and normal saline, is very stable at normal temperature, and the activity of the hirudin is not influenced by simple temperature rise (100 ℃ water bath) or pH change (1-13). Hirudin is a thrombin natural specific inhibitor with the strongest effect discovered so far, is not damaged by heat or ethanol, and has extremely strong inhibition effect on thrombin, so that the hirudin has special effect on human cardiovascular and cerebrovascular diseases, especially thrombotic diseases such as cerebral thrombosis, and the prevention and treatment effect of the hirudin is incomparable with any substance. Clinical studies show that the main pharmacological actions of hirudin are: (1) anticoagulation: leeches have found substances which inhibit different links of the coagulation mechanism, such as the thrombin inhibitors hirudin family, the coagulation factor Xa inhibitors antistasin, ghilante; a factor VIII inhibitor; platelet glycoprotein inhibitors decorsin, ornatin; collagen-induced platelet aggregation inhibitors Calin, Lapp, etc.; (2) antithrombotic effect: the most fundamental process of intravascular thrombosis is coagulation by thrombin. The leech has the function of directly dissolving thrombus, not only can be combined with free thrombin in plasma, but also can neutralize the thrombin combined with fibrin, so the hirudin has the effect on various thrombotic diseases, and particularly has obvious effect on disseminated intravascular coagulation; (3) the anti-tumor effect is as follows: the high anticoagulation effect of Hirudo is beneficial to the invasion of anticancer pharmacologically active substances (such as manganese, magnesium, zinc, etc.) and immunocompetent cells into cancer tissues to kill cancer cells; (4) blood fat reducing effect: animal experiments show that the leech can obviously reduce serum cholesterol, triglyceride and low-density lipoprotein of high-fat animals and improve the level of the high-density lipoprotein; (5) has effects on cerebral hematoma and subcutaneous hematoma: experiments prove that the thrombin is one of neurotoxic mediums, low dose can protect neurons and glial cells from cell death caused by damage such as hypoglycemia and ischemia, and large dose can cause encephaledema, irreversible damage of brain cells and brain cell apoptosis.
The hirudin (Hirudo polypeptide) has good anticoagulant and antithrombotic effects, and can be used for relieving atherosclerosis and spasm, lowering blood pressure adhesion, and relieving hypertension and hyperlipidemia. Meanwhile, related researches show that the anticoagulation effect of the natural hirudin is very specific, the anticoagulation effect is stronger than that of heparin, and the natural hirudin does not depend on the participation of AT III and heparin cofactor II when inhibiting the thrombin activity. Compared with other anticoagulants such as heparin, the natural hirudin has more reliable anticoagulation effect. Because of these advantages, natural hirudins, which are direct thrombin inhibitors, have been proposed to replace conventional heparins in europe and north america for the conventional prevention and treatment of cardiovascular diseases.
The leech is utilized to prevent and treat diseases, and the traditional methods comprise three methods, wherein the first method is to directly decoct and eat leeches, the second method is to grind dried leeches into powder and eat the powder, and the third method is to soak leeches in white spirit or honey water and eat the powder; the edible method has the advantages of low cost, but the disadvantages of serious damage to the active ingredients of the leech, low extraction rate and utilization rate of the hirudin and partial loss of the hirudin.
Disclosure of Invention
The invention aims to provide a preparation method of leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases, which adopts leech polypeptide extracted from leech with golden edge and Chinese medicinal materials to prepare the medicinal liquor together.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
(1) according to the mass ratio of spina date seed, Chinese angelica, szechuan lovage rhizome, dried longan and virgin pulp wine of 1-2:3-4:3-4:2-3:150 and 180, crushing the dried spina date seed, Chinese angelica, szechuan lovage rhizome and dried longan, putting the crushed dried spina date seed, Chinese angelica, dried szechuan lovage rhizome and dried longan into the virgin pulp wine with the alcoholic strength of more than 40 degrees, sealing and soaking for 1-2 months, stirring in the soaking process, and filtering to obtain filtrate;
(2) and (2) extracting leech polypeptide refined extract, mixing the leech polypeptide refined extract with the filtrate obtained in the step (1), and uniformly stirring to prepare medicinal liquor with the antithrombin activity content of not less than 4.0U per milliliter of liquor, thus obtaining the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases.
The preparation method of the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
selecting healthy adult leech living bodies with blood sucking leeches and golden edges, purifying and culturing the leech living bodies for 30-40 days without feeding, keeping clean running water in the period, then transferring the leech living bodies to an extraction workshop, feeding the leech living bodies with the inducing liquid for 65-70 minutes until the leech living bodies are full, and removing the residual inducing liquid and feeding equipment;
secondly, zinc sulfate solution with the mass fraction of 0.3 to 0.5 percent is injected as emetic, the weight ratio of the emetic to the leech living body after the leech is fully absorbed by the induced liquid is 1:5 to 6, and the leech is kept stand for 30 to 50 minutes to ensure that the leech fully absorbed by the induced liquid reversely ejects saliva in the body; collecting the saliva as leech polypeptide stock solution; culturing the leeches with golden edges of the bloodsucking leeches after the original liquid is extracted again for the next extraction or preparing the leeches into dry products for sale;
thirdly, filtering, removing impurities, concentrating the stock solution, adding glacial acetic acid, uniformly stirring to ensure that the stock solution is acidic, controlling the pH to be 3-4, standing for 5 hours at the ambient temperature of 20 ℃, centrifuging at low speed, removing precipitates to obtain clear solution, heating the clear solution to 60 ℃, keeping the temperature for 30 minutes, standing for 30 minutes, naturally cooling to normal temperature, centrifuging by using a high-speed centrifuge, removing residues to obtain liquid, adding sodium hydroxide to neutralize the liquid until the pH value is 6.5-7, and thus obtaining crude leech polypeptide extract;
adding ethanol with the volume of 5 times of the crude leech polypeptide extracting solution into the crude leech polypeptide extracting solution, centrifuging at a high speed to remove precipitates, filtering clear liquid by using a molecular sieve, keeping filtrate with the molecular weight of 300-10000, freezing and concentrating, and performing cobalt radiation sterilization to obtain the leech polypeptide refined extracting solution.
The semen Ziziphi Spinosae used in the invention is dry mature seed of Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H.F.Chou belonging to Rhamnaceae. The spina date seed has the effects of nourishing heart and tonifying liver, calming heart and tranquilizing mind, arresting sweating and promoting fluid production, and is mainly used for treating dysphoria and insomnia, palpitation and dreaminess, body deficiency and hyperhidrosis, body fluid deficiency and thirst. The spina date seed total glycosides contained in the spina date seeds can effectively reduce cholesterol contained in serum after entering a body, and can improve the content of high-density lipoprotein. Experiments also find that the effect of the wild jujube not only can regulate blood pressure and blood fat, but also has good effects of preventing and treating arteriosclerosis. The semen Ziziphi Spinosae contains alkaloid, saponin and flavone. Has tranquilizing, hypnotic, analgesic, anticonvulsive, blood lipid reducing, platelet aggregation inhibiting, immunity enhancing, blood pressure lowering, anoxia resisting, myocardial ischemia resisting, arrhythmia resisting, antiaging, and radioprotective effects.
Angelica sinensis (Oliv.) Diels is dried root of Angelica sinensis (Oliv.) Diels, Umbelliferae. Sweet, pungent and warm in nature. Meridian tropism enters liver, heart and spleen meridians. The functional indications are as follows: tonify blood, activate blood, regulate menstruation, alleviate pain, moisten intestines to relieve constipation. Can be used for treating blood deficiency, sallow complexion, giddiness, palpitation, menoxenia, amenorrhea, dysmenorrhea, asthenia cold, abdominal pain, constipation due to intestinal dryness, rheumatic arthralgia, traumatic injury, superficial infection, pyocutaneous disease. The wine-soaked Chinese angelica can activate blood and dredge channels, and is used for treating amenorrhea and dysmenorrhea, rheumatic arthralgia and traumatic injury. The radix Angelicae sinensis has effects of inhibiting platelet aggregation, resisting thrombi, resisting myocardial ischemia and anoxia, and dilating peripheral blood vessel. Reducing blood pressure, reducing blood lipid, resisting inflammation and protecting liver. The sodium ferulate extract in angelica and the polysaccharide contained in angelica have very obvious stimulation effect on a phagocyte system in vivo after entering a body, and can well regulate and stimulate the recovery of the immunoregulatory function in vivo; the volatile oil contained in radix Angelicae sinensis can act on cardiovascular system well to reduce contraction frequency of cardiopalmus. The angelica can be taken daily or soaked in water for drinking, so that cardiovascular diseases can be well treated.
Ligusticum wallichii, dried rhizome of Ligusticum chuanxiong Hort (or Ligusticum wallichi Franch.) belonging to Umbelliferae. Has effects of promoting blood circulation, activating qi-flowing, dispelling pathogenic wind and relieving pain, and can be used for treating blood stasis, qi stagnation, pain, headache, and rheumatalgia. Rhizoma Ligustici Chuanxiong contains alkaloid (such as ligustrazine), volatile oil (mainly including ligustilide, cholestene, etc.), phenolic substance (such as ferulic acid), lactone, vitamin A, folic acid, sucrose, sterol, fatty oil, etc. Ligustrazine can dilate coronary artery, increase coronary blood flow, improve myocardial blood oxygen supply, and reduce myocardial oxygen consumption; ligustrazine can dilate cerebral vessels, reduce vascular resistance, remarkably increase blood flow of brain and limbs, and improve microcirculation; can reduce the surface activity of blood platelet, inhibit blood platelet aggregation, and prevent thrombosis; the water decoction has a sedative effect on the central nervous system of animals and has an obvious and lasting antihypertensive effect; can accelerate the absorption of local hematoma of fracture and promote the formation of callus; has effect in resisting vitamin E deficiency; can inhibit multiple bacilli; has antihistaminic and choleretic effects.
Dried longan, also known as fructus Alpinae Oxyphyllae and arillus longan, is the sun-dried mature fruit of longan, arillus longan of Dimocarpus longan, Lour, Sapindaceae, longan. Has mild nature and sweet taste, contains chemical components such as thiamine, riboflavin, nicotinic acid, ascorbic acid and the like, and has the effects of tonifying qi and enriching blood, soothing nerves and calming mind, and nourishing blood and preventing miscarriage. Is suitable for insomnia, amnesia, spleen deficiency, diarrhea, listlessness, etc. The longan contains rich glucose, sucrose, protein and the like, has higher iron content, and can promote the regeneration of hemoglobin to enrich the blood while improving heat energy and supplementing nutrition. Experimental research shows that the longan pulp has the functions of tonifying the whole body, is particularly beneficial to brain cells, and can enhance memory and eliminate fatigue. The longan contains a large amount of elements such as iron, potassium and the like, and can promote the regeneration of hemoglobin to treat palpitation, insomnia and amnesia caused by anemia. The arillus longan has effects of reducing blood lipid and increasing coronary blood flow. Has strong inhibiting effect on brain B type monoamine oxidase (MAO-B), which is a flavoprotein closely related to the aging process, and has the effects of reducing blood fat, protecting heart and delaying aging. Longan is fresh, sweet and tasty, has high nutritive value, and has the functions of protecting blood vessels and preventing angiosclerosis and fragility for middle-aged and elderly people.
The leech polypeptide refined extract used in the invention is a natural substance with active ingredients extracted from leech saliva of leech with golden edge of bloodsucking leech, and is a small molecular protein (polypeptide) consisting of 65-66 amino acids, which is also called hirudin. Animal experiments and clinical researches show that the leech polypeptide can effectively prevent blood coagulation and thrombosis, and prevent further blood stasis phenomena such as thrombin activation and platelet reaction catalyzed by thrombin. In addition, it can inhibit thrombin-induced fibroblast proliferation and thrombin stimulation of endothelial cells. Compared with heparin, it has less consumption, no bleeding and no dependence on endogenous auxiliary factor. At present, the natural hirudin is a specific natural medicine for preventing and treating human cardiovascular and cerebrovascular diseases, can prevent thrombosis, promote wound healing, and has the effects of reducing blood fat, reducing blood pressure, expanding peripheral blood vessels and accelerating blood circulation; the natural medicine has the advantages of special effect and natural medicine for preventing and treating human cardiovascular and cerebrovascular diseases, and wide application prospect in the fields of medicine, beauty and skin care products, health care products, biochemical reagents and the like.
The invention has the beneficial effects that:
1. the extraction method of the leech polypeptide refined extract adopts the inducing liquid and the emetic to extract the saliva of the leech (leech gilt edge), can avoid the problem of high death rate of the leech caused by a generally adopted manual extrusion method, and has the advantages of high activity, good quality and high antithrombin activity content of the leech polypeptide extracted by the extraction method limited by the invention.
2. The invention relates to a preparation method of leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases, which is prepared by the steps of scientifically mixing spina date seed, Chinese angelica, Szechuan lovage rhizome and dried longan, soaking in original pulp liquor, and adding leech polypeptide refined extract for preparation. The used spina date seed has the effects of nourishing heart, tonifying liver, calming heart, soothing nerves, reducing cholesterol, and regulating blood pressure and blood fat; angelica sinensis has the effects of activating blood and stimulating menstrual flow, inhibiting platelet aggregation and resisting thrombus; the rhizoma ligustici wallichii is capable of activating blood and promoting qi, dispelling wind and relieving pain, reducing blood pressure and benefiting gallbladder, and preventing thrombus; the dried longan has the effects of tonifying qi and enriching blood, tranquilizing the mind, reducing blood fat and increasing coronary blood flow; the compatibility of the traditional Chinese medicinal materials can play roles of activating blood circulation, dredging channels, tranquilizing mind, resisting thrombus, regulating blood fat and the like, the prepared medicinal liquor contains rich alkaloid, glycosides, trace elements and other substances, can tonify liver and kidney, resist myocardial ischemia, resist aging, enhance human immunity, accelerate blood flow, resist coagulation and prevent thrombus, prevent and treat cardiovascular and cerebrovascular diseases, and is added with leech polypeptide refined extract for preparation, so that the prepared leech polypeptide medicinal liquor has the antithrombin activity content of not less than 4.0U per milliliter of liquor, has the effects of improving sleep quality, relieving dizziness and headache symptoms, enhancing the blood supply and nourishing capacity of tissue organs, accelerating blood flow, promoting blood circulation, reducing blood viscosity, regulating blood fat, reducing the contents of cholesterol and fat in plasma, strongly clearing garbage toxin in blood, dissolving thrombus, softening blood vessels and improving immunity, eliminating atherosclerotic plaque, preventing blood vessel rupture, and effectively preventing and treating cardiovascular disease and cerebrovascular disease.
Detailed Description
Example 1
A method for preparing leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
(1) according to the mass ratio of spina date seed, Chinese angelica, ligusticum wallichii, dried longan and raw pulp wine of 1:3:3:2:150, crushing the dried spina date seed, Chinese angelica, ligusticum wallichii and dried longan, putting the crushed dried spina date seed, Chinese angelica, ligusticum wallichii and dried longan into the raw pulp wine with the alcoholic strength of 40 degrees, sealing and soaking for 2 months, stirring in the soaking process, and filtering to obtain filtrate;
(2) and (2) extracting leech polypeptide refined extract, mixing the leech polypeptide refined extract with the filtrate obtained in the step (1), and uniformly stirring to prepare medicinal liquor with the antithrombin activity content of not less than 4.0U per milliliter of liquor, thus obtaining the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases.
The preparation method of the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
selecting healthy adult leech living bodies with blood sucking leeches and golden edges, purifying and culturing for 30 days without feeding, keeping clean running water in the period, then transferring the leeches to an extraction workshop, feeding an inducing liquid for 65 minutes until the leech is full, and removing the residual inducing liquid and feeding equipment; the inducing liquid is prepared by 0.5 part of glucose powder, 1.0 part of sodium chloride, 1 part of citric acid and 100 parts of water in parts by weight;
secondly, zinc sulfate solution with the mass fraction of 0.3 percent is injected as an emetic, the weight ratio of the emetic to the leech living body after the leech is fully absorbed with the induced liquid is 1:5, and the leech is kept stand for 50 minutes to ensure that the leech fully absorbed with the induced liquid reversely spits out saliva in the body; collecting the saliva as leech polypeptide stock solution; culturing the leeches with golden edges of the bloodsucking leeches after the original liquid is extracted again for the next extraction or preparing the leeches into dry products for sale;
thirdly, filtering, removing impurities, concentrating the stock solution, adding glacial acetic acid, uniformly stirring to ensure that the stock solution is acidic, controlling the pH to be 3.8, standing for 5 hours at the ambient temperature of 20 ℃, centrifuging at low speed, removing precipitates to obtain clear solution, heating the clear solution to 60 ℃, keeping the temperature for 30 minutes, standing for 30 minutes, naturally cooling to normal temperature, centrifuging by using a high-speed centrifuge, removing residues to obtain liquid, adding sodium hydroxide to neutralize the liquid until the pH value is 6.5, and obtaining the leech polypeptide extract;
adding ethanol with the volume of 5 times of the crude leech polypeptide extracting solution into the crude leech polypeptide extracting solution, centrifuging at a high speed to remove precipitates, filtering clear liquid by using a molecular sieve, keeping filtrate with the molecular weight of 300-10000, freezing and concentrating, and performing cobalt radiation sterilization to obtain the leech polypeptide refined extracting solution.
Example 2
A method for preparing leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
(1) according to the mass ratio of spina date seed, Chinese angelica, ligusticum wallichii, dried longan and virgin pulp wine of 1.5:3.5:3.5:2.5:170, crushing the dried spina date seed, Chinese angelica, ligusticum wallichii and dried longan, putting the crushed dried spina date seed, Chinese angelica, ligusticum wallichii and dried longan into the virgin pulp wine with the alcoholic strength of 52 degrees, sealing and soaking for 1.5 months, stirring in the soaking process, and filtering after soaking to obtain filtrate;
(2) and (2) extracting leech polypeptide refined extract, mixing the leech polypeptide refined extract with the filtrate obtained in the step (1), and uniformly stirring to prepare medicinal liquor with the antithrombin activity content of not less than 4.0U per milliliter of liquor, thus obtaining the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases.
The preparation method of the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
selecting healthy adult leech living bodies with blood sucking leeches and golden edges, purifying and culturing for 35 days without feeding, keeping clean running water in the period, then transferring the leeches to an extraction workshop, feeding the inducing liquid for 68 minutes until the leech is full, and removing the residual inducing liquid and feeding equipment; the inducing liquid is prepared from 0.6 part of glucose powder, 0.8 part of sodium chloride, 1.2 parts of citric acid and 110 parts of water in parts by weight;
secondly, zinc sulfate solution with the mass fraction of 0.4 percent is injected as an emetic, the weight ratio of the amount of the emetic to the leech living body after the leech is fully absorbed with the induced liquid is 1:5.5, and the leech is kept stand for 40 minutes to ensure that the leech fully absorbed with the induced liquid reversely spits out the saliva in the body; collecting the saliva as leech polypeptide stock solution; culturing the leeches with golden edges of the bloodsucking leeches after the original liquid is extracted again for the next extraction or preparing the leeches into dry products for sale;
thirdly, filtering, removing impurities, concentrating the stock solution, adding glacial acetic acid, uniformly stirring to ensure that the stock solution is acidic and has a pH value of 4, standing for 5 hours at the ambient temperature of 20 ℃, centrifuging at a low speed, removing precipitates to obtain a clear solution, heating the clear solution to 60 ℃, keeping the temperature for 30 minutes, standing for 30 minutes, naturally cooling to the normal temperature, centrifuging by using a high-speed centrifuge, removing residues to obtain a liquid, adding sodium hydroxide to neutralize the liquid to a pH value of 7, and thus obtaining the golden-edged leech polypeptide extracting solution;
adding ethanol with the volume of 5 times of the crude leech polypeptide extracting solution into the crude leech polypeptide extracting solution, centrifuging at a high speed to remove precipitates, filtering clear liquid by using a molecular sieve, keeping filtrate with the molecular weight of 300-10000, freezing and concentrating, and performing cobalt radiation sterilization to obtain the leech polypeptide refined extracting solution.
Example 3
A method for preparing leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
(1) according to the mass ratio of spina date seed, Chinese angelica, ligusticum wallichii, dried longan and raw pulp wine of 2:4:4:3:180, crushing the dried spina date seed, Chinese angelica, ligusticum wallichii and dried longan, putting the crushed dried spina date seed, Chinese angelica, ligusticum wallichii and dried longan into the raw pulp wine with the alcoholic strength of 52 degrees, sealing and soaking for 1 month, stirring in the soaking process, and filtering to obtain filtrate;
(2) and (2) extracting leech polypeptide refined extract, mixing the leech polypeptide refined extract with the filtrate obtained in the step (1), and uniformly stirring to prepare medicinal liquor with the antithrombin activity content of not less than 4.0U per milliliter of liquor, thus obtaining the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases.
The preparation method of the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
selecting healthy adult leech living bodies with blood sucking leeches and golden edges, purifying and culturing for 40 days without feeding, keeping clean running water in the period, then transferring the leeches to an extraction workshop, feeding the inducing liquid for 70 minutes until the leech is full, and removing the residual inducing liquid and feeding equipment; the inducing liquid is prepared by 1 part of glucose powder, 0.5 part of sodium chloride, 1.5 parts of citric acid and 120 parts of water in parts by weight;
secondly, zinc sulfate solution with the mass fraction of 0.5 percent is injected as an emetic, the weight ratio of the emetic to the leech living body after the leech is fully absorbed with the induced liquid is 1:6, and the leech is kept stand for 30 minutes to ensure that the leech fully absorbed with the induced liquid reversely spits out saliva in the body; collecting the saliva as leech polypeptide stock solution; culturing the leeches with golden edges of the bloodsucking leeches after the original liquid is extracted again for the next extraction or preparing the leeches into dry products for sale;
thirdly, filtering, removing impurities, concentrating the stock solution, adding glacial acetic acid, uniformly stirring to ensure that the stock solution is acidic, controlling the pH to be 3.0, standing for 5 hours at the ambient temperature of 20 ℃, centrifuging at low speed, removing precipitates to obtain clear solution, heating the clear solution to 60 ℃, keeping the temperature for 30 minutes, standing for 30 minutes, naturally cooling to normal temperature, centrifuging by using a high-speed centrifuge, removing residues to obtain liquid, adding sodium hydroxide to neutralize the liquid until the pH value is 6.5-7, and obtaining the leech polypeptide extract;
adding ethanol with the volume of 5 times of the crude leech polypeptide extracting solution into the crude leech polypeptide extracting solution, centrifuging at a high speed to remove precipitates, filtering clear liquid by using a molecular sieve, keeping filtrate with the molecular weight of 300-10000, freezing and concentrating, and performing cobalt radiation sterilization to obtain the leech polypeptide refined extracting solution.
Example 4
A method for preparing leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
(1) according to the mass ratio of the spina date seed, the Chinese angelica, the ligusticum wallichii, the dried longan and the original pulp wine of 2:3:4:3:170, the dried spina date seed, the Chinese angelica, the ligusticum wallichii and the dried longan are crushed and then put into the original pulp wine with the alcoholic strength of 52 degrees, the mixture is sealed and soaked for 2 months, stirring is carried out during the soaking process, and the filtrate is obtained after the soaking;
(2) and (2) extracting leech polypeptide refined extract, mixing the leech polypeptide refined extract with the filtrate obtained in the step (1), and uniformly stirring to prepare medicinal liquor with the antithrombin activity content of not less than 4.0U per milliliter of liquor, thus obtaining the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases.
The preparation method of the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases comprises the following steps:
selecting healthy adult leech living bodies with blood sucking leeches and golden edges, purifying and culturing for 40 days without feeding, keeping clean running water in the period, then transferring the leeches to an extraction workshop, feeding the inducing liquid for 70 minutes until the leech is full, and removing the residual inducing liquid and feeding equipment; the inducing liquid is prepared from 0.7 part of glucose powder, 0.8 part of sodium chloride, 1 part of citric acid and 110 parts of water in parts by weight;
secondly, zinc sulfate solution with the mass fraction of 0.3 percent is injected as an emetic, the weight ratio of the amount of the emetic to the leech living body after the leech is fully absorbed by the induced liquid is 1:6, and the leech is kept stand for 40 minutes to ensure that the leech fully absorbed by the induced liquid reversely spits out saliva in the body; collecting the saliva as leech polypeptide stock solution; culturing the leeches with golden edges of the bloodsucking leeches after the original liquid is extracted again for the next extraction or preparing the leeches into dry products for sale;
thirdly, filtering, removing impurities, concentrating the stock solution, adding glacial acetic acid, uniformly stirring to ensure that the stock solution is acidic, controlling the pH to be 3-4, standing for 5 hours at the ambient temperature of 20 ℃, centrifuging at low speed, removing precipitates to obtain clear solution, heating the clear solution to 60 ℃, keeping the temperature for 30 minutes, standing for 30 minutes, naturally cooling to normal temperature, centrifuging by using a high-speed centrifuge, removing residues to obtain liquid, adding sodium hydroxide to neutralize the liquid until the pH value is 6.5-7, and obtaining the leech polypeptide extract;
adding ethanol with the volume 5 times that of the crude leech polypeptide extracting solution into the crude leech polypeptide extracting solution, wherein the volume concentration of the ethanol is more than 40%, centrifuging at a high speed to remove precipitates, filtering clear liquid by using a molecular sieve, keeping filtrate with the molecular weight of 300-10000, freezing and concentrating, and performing cobalt radiation sterilization to obtain the leech polypeptide refined extracting solution.
The antithrombin activity content of each ml of the medicinal liquor prepared by the invention is not lower than 4.0U, and the physicochemical index and the microbial index meet the regulations in the following tables 1 and 2.
Claims (3)
1. The preparation method of the leech polypeptide medicinal liquor for preventing and treating the cardiovascular and cerebrovascular diseases is characterized by comprising the following steps:
(1) according to the mass ratio of spina date seed, Chinese angelica, szechuan lovage rhizome, dried longan and virgin pulp wine of 1-2:3-4:3-4:2-3:150 and 180, crushing the dried spina date seed, Chinese angelica, szechuan lovage rhizome and dried longan, putting the crushed dried spina date seed, Chinese angelica, dried szechuan lovage rhizome and dried longan into the virgin pulp wine with the alcoholic strength of more than 40 degrees, sealing and soaking for 1-2 months, stirring in the soaking process, and filtering to obtain filtrate;
(2) and (2) extracting leech polypeptide refined extract, mixing the leech polypeptide refined extract with the filtrate obtained in the step (1), and uniformly stirring to prepare medicinal liquor with the antithrombin activity content of not less than 4.0U per milliliter of liquor, thus obtaining the leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases.
2. The method for preparing leech polypeptide medicinal liquor for preventing and treating cardiovascular and cerebrovascular diseases according to claim 1, wherein the extraction method of the leech polypeptide refined extract comprises the following steps:
selecting healthy adult leech living bodies with blood sucking leeches and golden edges, purifying and culturing the leech living bodies for 30-40 days without feeding, keeping clean running water in the period, then transferring the leech living bodies to an extraction workshop, feeding the leech living bodies with the inducing liquid for 65-70 minutes until the leech living bodies are full, and removing the residual inducing liquid and feeding equipment;
secondly, zinc sulfate solution with the mass fraction of 0.3 to 0.5 percent is injected as emetic, the weight ratio of the emetic to the leech living body after the leech is fully absorbed by the induced liquid is 1:5 to 6, and the leech is kept stand for 30 to 50 minutes to ensure that the leech fully absorbed by the induced liquid reversely ejects saliva in the body; collecting the saliva as leech polypeptide stock solution; culturing the leeches with golden edges of the bloodsucking leeches after the original liquid is extracted again for the next extraction or preparing the leeches into dry products for sale;
thirdly, filtering, removing impurities, concentrating the stock solution, adding glacial acetic acid, uniformly stirring to ensure that the stock solution is acidic, controlling the pH to be 3-4, standing for 5 hours at the ambient temperature of 20 ℃, centrifuging at low speed, removing precipitates to obtain clear solution, heating the clear solution to 60 ℃, keeping the temperature for 30 minutes, standing for 30 minutes, naturally cooling to normal temperature, centrifuging by using a high-speed centrifuge, removing residues to obtain liquid, adding sodium hydroxide to neutralize the liquid until the pH value is 6.5-7, and thus obtaining crude leech polypeptide extract;
adding ethanol with the volume of 5 times of the crude leech polypeptide extracting solution into the crude leech polypeptide extracting solution, centrifuging at a high speed to remove precipitates, filtering clear liquid by using a molecular sieve, keeping filtrate with the molecular weight of 300-10000, freezing and concentrating, and performing cobalt radiation sterilization to obtain the leech polypeptide refined extracting solution.
3. The preparation method of the leech polypeptide medicinal liquor for preventing and treating the cardiovascular and cerebrovascular diseases according to claim 2, wherein the extraction method of the leech polypeptide refined extract comprises the step of preparing the inducing liquid from 0.5 to 1 part by weight of glucose powder, 0.5 to 1.0 part by weight of sodium chloride, 1 to 1.5 parts by weight of citric acid and 100 to 120 parts by weight of water.
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Application publication date: 20191220 |
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