CN109758480B - Preparation method of Agkistrodon acutus lyophilized powder for rheumatoid arthritis - Google Patents
Preparation method of Agkistrodon acutus lyophilized powder for rheumatoid arthritis Download PDFInfo
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Abstract
The invention discloses a preparation method of a agkistrodon acutus freeze-dried powder for rheumatoid arthritis, which comprises the following steps: freezing fresh Agkistrodon acutus to make it in hibernation state; preprocessing the five-step snakes in a hibernation state; cutting off the head, laparotomizing to remove internal organs and scales, cleaning, and crushing the tissues in the Agkistrodon acutus body; spraying alcohol on the body of the Agkistrodon acutus with crushed body tissues; freeze-drying Agkistrodon acutus sprayed with alcohol; pulverizing lyophilized Agkistrodon acutus to obtain lyophilized powder for treating rheumatoid arthritis. The invention has the beneficial effects that: the preparation method of the Agkistrodon acutus freeze-dried powder for rheumatoid arthritis adopts a freeze-drying technology to completely dry Agkistrodon acutus, and then adopts a crushing technology to process the Agkistrodon acutus freeze-dried powder into the Agkistrodon acutus freeze-dried powder, so that the active ingredients of the Agkistrodon acutus freeze-dried powder are preserved to the maximum extent, and the preparation method has a higher rheumatoid treatment effect.
Description
Technical Field
The invention relates to the technical field of traditional Chinese medicines, in particular to a preparation method of a snake venom lyophilized powder for rheumatoid arthritis.
Background
The five-step snake is also called long-noded pit viper and agkistrodon acutus, belongs to the class of reptile, the family of agkistrodon acutus has a triangular head, the snout of the head is extended to the upper front, the top of the head is provided with symmetrical large phosphor, the head is dark brown, the back of the body is grey brown, two sides of the head are provided with a series of dark brown inverted V-shaped stripes, the abdomen is yellow-white, two sides of the head are provided with black round spots, the five-step snake is usually living in mountainous forests and fed by small mammals and birds, the five-step snake kills after being bitten by the five-step snake, the five-step snake can kill five steps of people obviously, even the most poisonous snake in the world is unlikely to poison in that short time, but the five-step snake belongs to blood circulation poison, and the bitten person is hard to feel painful, and.
The research of the five-step snake in the aspect of medicinal value really brings huge welfare to the people, the Shennong's herbal channel' two thousand years ago in China is recorded with the medicine use of the snake, the Lishizhen points out that the snake can treat serious diseases such as half-body withering, skin obstinate arthralgia and the like in the compendium of the herbal, and the research of the medicinal value of the snake brings convenience to the life of the people from ancient to present.
Rheumatoid arthritis is a chronic systemic disease mainly comprising inflammatory synovitis with unknown etiology and is characterized by polyarticular, symmetrical and invasive joint inflammation of hands and small joints, and joint deformity and function loss can be caused by frequent involvement of external organs of joints and positive serum rheumatoid factors. The Chinese rheumatoid arthritis patients are huge in population, the patients are frequently suffered from repeated and continuous diseases, some patients even lose mobility, the existing technology cannot achieve the expected treatment effect on the rheumatoid arthritis, radical treatment cannot be achieved, and long-term administration treatment causes many adverse effects on human bodies.
The five-step snake has remarkable curative effect on rheumatoid arthritis which is a worldwide problem, researches show that the five-step snake medicinal materials are dried before being powdered, the conventional drying method is drying and sun-drying, but the activity of the five-step snake can be preserved only by strictly controlling the drying temperature of the five-step snake in the drying process, even if the drying temperature of the five-step snake is controlled, the loss of active ingredients is large in the drying process, and the curative effect of the five-step snake in the treatment of the rheumatoid arthritis is weakened; when the drying temperature exceeds 70 ℃, active enzyme components contained in the agkistrodon acutus are damaged, and the drug effect is greatly reduced; the drying method is very easy to be polluted by flies and insects, is limited by weather, is easy to cause rancidity when not dried in time, and cannot ensure the quality of medicinal materials.
Disclosure of Invention
The invention aims to provide a method for preparing the freeze-dried powder of the long-noded pit viper, which can furthest preserve the active ingredients of the long-noded pit viper and enhance the medicinal effect of the long-noded pit viper in treating rheumatoid arthritis.
A preparation method of Agkistrodon acutus lyophilized powder for rheumatoid arthritis comprises the following steps:
(1) freezing fresh Agkistrodon acutus to make it in hibernation state; because the wild-living agkistrodon acutus contains parasites, the agkistrodon acutus selected by the application is an agkistrodon acutus monitored in the whole breeding process, no parasite exists on the agkistrodon acutus, and the agkistrodon acutus is healthy; because the five-step snake is a poisonous snake, the five-step snake is in a hibernation state by adopting a freezing mode, and is convenient for pretreatment;
(2) preprocessing the five-step snakes in a hibernation state; cutting off the head, laparotomizing to remove internal organs and scales, cleaning, and crushing the tissues in the Agkistrodon acutus body; the tissue in the Agkistrodon acutus is crushed: the bone in the snake body can be chopped by a knife, but the skin of the snake body is kept intact;
(3) spraying alcohol on the body of the Agkistrodon acutus with crushed body tissues; the spraying of alcohol is convenient to remove the fishy smell on the Agkistrodon acutus, destroy the tissues in the Agkistrodon acutus, and is convenient to fully contact with the broken tissues, so as to be beneficial to removing the fishy smell on the Agkistrodon acutus;
(4) freeze-drying Agkistrodon acutus sprayed with alcohol; drying the five-step snake by adopting a freeze-drying technology, and then placing the dried five-step snake in a freeze-drying instrument to room temperature, so that active ingredients contained in the five-step snake are reserved to the maximum extent;
(5) pulverizing lyophilized Agkistrodon acutus to obtain lyophilized powder for treating rheumatoid arthritis.
Further, in the step (1), the fresh agkistrodon acutus is frozen for 2-4 h at the temperature of 18-10 ℃ below zero.
Further, in the step (2), before the five-step snake is pretreated, the five-step snake in the hibernation state needs to be soaked in water until the five-step snake is softened. The five-step snake in the hibernation state is soaked in water to be slightly soft, so that the five-step snake is still in the hibernation state and has a slightly soft body, and the five-step snake is convenient to pretreat.
Further, in the step (3), the concentration of the alcohol sprayed is 70%. The snake is sprayed with high concentration alcohol, so that the consumption of alcohol can be reduced.
Further, in the step (4), the freeze-drying process is as follows: pre-lyophilizing Agkistrodon acutus sprayed with alcohol, and performing cold hydrazine treatment after the pre-lyophilizing treatment is finished. Pre-freezing the material to freeze water into ice, and dewatering the material with the ice being sublimated into vapor under relatively low pressure and relatively high vacuum degree.
Further, the pre-freeze drying treatment comprises the following steps: freezing for 2-4 h at-30 deg.C to-18 deg.C.
Further, the cold hydrazine treatment is as follows: cold hydrazine is carried out for 8 to 15 hours under the conditions that the temperature is 60 ℃ below zero to 30 ℃ below zero and the vacuum degree is 0.05 to 0.5 mbr.
Further, in the step (5), after the five-step snake is freeze-dried, taking out the five-step snake after the temperature of the environment where the five-step snake is located is restored to 15-35 ℃. After the five-step snakes are frozen and dried, the temperature of the five-step snakes is low, the five-step snakes are taken out, moisture in air can be absorbed due to temperature difference, the drug effect of the five-step snakes can be affected, and the five-step snakes are taken out when the temperature of the environment where the five-step snakes are located is recovered to 15-35 ℃.
Further, the pulverization is as follows: pulverizing with jet mill, ball mill or colloid mill. The crushing mode is adopted for crushing, high temperature is avoided in the crushing process, and the active ingredients in the agkistrodon acutus are kept to the maximum extent.
Furthermore, the fineness of the obtained Agkistrodon acutus freeze-dried powder for treating rheumatoid arthritis is 80-500 meshes.
The invention has the beneficial effects that: according to the preparation method of the five-step snake freeze-dried powder for rheumatoid arthritis, the five-step snake is hibernated firstly in the process of preparing the five-step snake freeze-dried powder, so that the safety of operators is guaranteed when the five-step snake is treated; before the five-step snake is frozen and dried, the fishy smell of the five-step snake is removed by alcohol spraying; the freeze-drying technology is adopted to completely dry the agkistrodon acutus, and then the agkistrodon acutus freeze-dried powder is processed by the normal-temperature crushing technology, so that the active ingredients of the agkistrodon acutus freeze-dried powder are preserved to the maximum extent, and the agkistrodon acutus freeze-dried powder has a higher rheumatoid treatment effect.
Detailed Description
Example one
A preparation method of Agkistrodon acutus lyophilized powder for rheumatoid arthritis comprises the following steps:
(1) freezing fresh Agkistrodon acutus to make it in hibernation state;
(2) preprocessing the five-step snakes in a hibernation state; cutting off the head, laparotomizing to remove internal organs and scales, cleaning, and crushing the tissues in the Agkistrodon acutus body;
(3) spraying alcohol on the body of the Agkistrodon acutus with crushed body tissues;
(4) freeze-drying Agkistrodon acutus sprayed with alcohol;
(5) pulverizing lyophilized Agkistrodon acutus to obtain lyophilized powder for treating rheumatoid arthritis.
Example two
A preparation method of Agkistrodon acutus lyophilized powder for rheumatoid arthritis comprises the following steps:
(1) freezing fresh Agkistrodon acutus at-18 deg.C for 2 hr to make it in hibernation state for pretreating Agkistrodon acutus;
(2) preprocessing the five-step snakes in a hibernation state; soaking Agkistrodon acutus in water to slight softness, cutting head, removing viscera and scale, cleaning, and crushing tissue of Agkistrodon acutus; when in crushing, the five-step snake can be chopped by a knife to destroy the tissues in the body, and because the skin toughness of the five-step snake is relatively high, the bone in the body is chopped, and the skin is kept complete;
(3) spraying 70% ethanol onto the body of Agkistrodon acutus with broken body tissue to remove fishy smell;
(4) freeze-drying Agkistrodon acutus sprayed with alcohol; pre-lyophilizing Agkistrodon acutus sprayed with alcohol at-30 deg.C for 2h, and cold-hydrazine treating at-60 deg.C under vacuum degree of 0.05mbr for 8 h;
(5) after the five-step snake is freeze-dried, taking out the five-step snake until the temperature of the environment of the five-step snake is restored to 15 ℃, and crushing the five-step snake by adopting air flow crushing to obtain the five-step snake freeze-dried powder with the fineness of 80 meshes for treating rheumatoid arthritis.
EXAMPLE III
A preparation method of Agkistrodon acutus lyophilized powder for rheumatoid arthritis comprises the following steps:
(1) freezing fresh Agkistrodon acutus at-15 deg.C for 3 hr to make it in hibernation state for pretreating Agkistrodon acutus;
(2) preprocessing the five-step snakes in a hibernation state; soaking Agkistrodon acutus in water to slight softness, cutting head, removing viscera and scale, cleaning, and crushing tissue of Agkistrodon acutus; when in crushing, the five-step snake can be chopped by a knife to destroy the tissues in the body, and because the skin toughness of the five-step snake is relatively high, the bone in the body is chopped, and the skin is kept complete;
(3) spraying 70% ethanol onto the body of Agkistrodon acutus with broken body tissue to remove fishy smell;
(4) freeze-drying Agkistrodon acutus sprayed with alcohol; pre-lyophilizing Agkistrodon acutus sprayed with alcohol at-25 deg.C for 3h, and cold-hydrazine treating at-45 deg.C under vacuum degree of 0.2mbr for 11 h;
(5) after the five-step snakes are freeze-dried, taking out the five-step snakes when the temperature of the environment of the five-step snakes is restored to 20 ℃, and crushing the five-step snakes in a ball milling crushing mode to obtain the five-step snake freeze-dried powder with the fineness of 200 meshes for rheumatoid arthritis.
Example four
A preparation method of Agkistrodon acutus lyophilized powder for rheumatoid arthritis comprises the following steps:
(1) freezing fresh Agkistrodon acutus at-10 deg.C for 4 hr to make it in hibernation state for pretreating Agkistrodon acutus;
(2) preprocessing the five-step snakes in a hibernation state; soaking Agkistrodon acutus in water to slight softness, cutting head, removing viscera and scale, cleaning, and crushing tissue of Agkistrodon acutus; when in crushing, the five-step snake can be chopped by a knife to destroy the tissues in the body, and because the skin toughness of the five-step snake is relatively high, the bone in the body is chopped, and the skin is kept complete;
(3) spraying 70% ethanol onto the body of Agkistrodon acutus with broken body tissue to remove fishy smell;
(4) freeze-drying Agkistrodon acutus sprayed with alcohol; pre-lyophilizing Agkistrodon acutus sprayed with alcohol at-18 deg.C for 4h, and cold-hydrazine treating at-30 deg.C under vacuum degree of 0.5mbr for 15 h;
(5) after the five-step snake is freeze-dried, taking out the five-step snake until the temperature of the environment of the five-step snake is restored to 35 ℃, and crushing the five-step snake by adopting a colloid mill to obtain the five-step snake freeze-dried powder with the fineness of 500 meshes for treating rheumatoid arthritis.
The invention relates to a specific implementation case of the five-step snake freeze-dried powder:
the treatment subjects were: 30 out-patient patients of the traditional Chinese medicine institute in Jingdezhen city are selected and all meet the latest diagnosis standard of rheumatoid arthritis of American society for radiology (ACR 2009). And randomly dividing the mixture into 3 groups according to a random number table method, wherein each group comprises 10 cases; 10 cases of blank group, 10 cases of commercial agkistrodon acutus powder group, and 10 cases of agkistrodon acutus freeze-dried powder group. Each group had 1 male and 9 female; age 29-54 years, mean 39 ± 16 years; the course of the disease is 60-120 months, and the average is 94 +/-22 months. Compared with the age, the sex and the course of disease among the groups, the traditional Chinese medicine composition has no statistical difference (P is more than 0.05) and is comparable.
The treatment method comprises the following steps: the blank group is administered with 5g of Amylum Nelumbinis Rhizomatis each time, 1 time per day, and is taken with warm water before bedtime, and 30 days is used as 1 treatment course. The commercially available Agkistrodon acutus powder is administered with pulverized dry Agkistrodon acutus powder 5g each time 1 time per day, and is taken with warm water before sleep for 30 days as 1 treatment course. The Agkistrodon (Agkistrodon) lyophilized powder is administered 5g each time 1 time per day with warm water before sleep for 30 days as 1 treatment course.
The observation method comprises the following steps: before and after 1 course of treatment, the symptom indexes of each group, including morning stiffness time, number of tenderness joints and number of swelling joints, were recorded.
The results of the experiment are shown in table 1:
TABLE 1
Note: p < 0.05, P < 0.01, compared to the group before treatment; compared with the commercial agkistrodon acutus powder after treatment,#P＜0.05，##P＜0.01。
as can be seen from Table 1, compared with the commercially available agkistrodon acutus, the agkistrodon acutus freeze-dried powder provided by the invention has the advantages that the content of active ingredients is increased, and the rheumatoid treatment effect is higher.
The present invention is not limited to the above-described preferred embodiments, and various other forms of the product can be obtained by anyone who has come within the teachings of the present invention, but any variation in the details thereof, which is the same as or similar to the present application, falls within the scope of the present invention.
Claims (5)
1. A preparation method of Agkistrodon acutus freeze-dried powder for rheumatoid arthritis is characterized by comprising the following steps:
(1) freezing fresh Agkistrodon acutus to make it in hibernation state; the selected agkistrodon acutus is monitored in the whole breeding process, no parasite exists on the agkistrodon acutus, and the agkistrodon acutus is healthy; the freezing is to freeze fresh agkistrodon acutus for 2-4 h at the temperature of 18-10 ℃ below zero;
(2) preprocessing the five-step snakes in a hibernation state; cutting off the head, laparotomizing to remove internal organs and scales, cleaning, and crushing the tissues in the Agkistrodon acutus body;
before the five-step snake is pretreated, the five-step snake in a hibernation state needs to be soaked in water until the five-step snake is softened; soaking the five-step snake in the hibernation state in water until the five-step snake is slightly soft, so that the five-step snake is still in the hibernation state and has a slightly soft body, and the five-step snake is convenient to pretreat;
the tissue in the Agkistrodon acutus is crushed: the bone in the snake body can be chopped by a knife, but the skin of the snake body is kept intact;
(3) spraying alcohol to the body of the Agkistrodon acutus with crushed body tissue, wherein the concentration of the sprayed alcohol is 70%;
(4) freeze-drying Agkistrodon acutus sprayed with alcohol; the freeze-drying treatment comprises the following steps: pre-lyophilizing Agkistrodon acutus sprayed with alcohol, and performing cold hydrazine treatment after the pre-lyophilizing treatment is finished;
(5) crushing the freeze-dried agkistrodon acutus to obtain the agkistrodon acutus freeze-dried powder for treating rheumatoid arthritis; the obtained lyophilized powder of Agkistrodon acutus for treating rheumatoid arthritis has fineness of 80-500 mesh.
2. The method for preparing the lyophilized powder of agkistrodon acutus for rheumatoid arthritis according to claim 1, wherein the pre-lyophilization treatment comprises: freezing for 2-4 h at-30 deg.C to-18 deg.C.
3. The method for preparing the lyophilized powder of agkistrodon acutus for rheumatoid arthritis according to claim 1, wherein the cold hydrazine treatment comprises: cold hydrazine is carried out for 8 to 15 hours under the conditions that the temperature is 60 ℃ below zero to 30 ℃ below zero and the vacuum degree is 0.05 to 0.5 mbr.
4. The method for preparing the lyophilized powder of Agkistrodon acutus for rheumatoid arthritis according to claim 1, wherein in the step (5), the Agkistrodon acutus is taken out after the lyophilization of the Agkistrodon acutus is finished and the temperature of the environment of the Agkistrodon acutus is restored to 15-35 ℃.
5. The method for preparing the lyophilized powder of agkistrodon acutus for rheumatoid arthritis according to claim 1, wherein the pulverization is as follows: pulverizing with jet mill, ball mill or colloid mill.
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| KR102340531B1 (en) * | 2019-07-02 | 2021-12-21 | 독립바이오제약(주) | Composition for preventing or treating rheumatoid arthritis comprising venom derived from Agkistrodon piscivorous piscivorous |
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| CN103251655A (en) * | 2013-05-23 | 2013-08-21 | 浙江中医药大学 | Preparation method of Chinese medicinal composition for treating rheumatoid arthritis |
| CN103919810A (en) * | 2014-03-27 | 2014-07-16 | 浙江中医药大学 | Method for preparing long-noded pit viper water-soluble total protein lyophilized powder |
| CN104622820A (en) * | 2015-03-05 | 2015-05-20 | 济南康众医药科技开发有限公司 | Freeze-drying method for animal medicines |
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