阅读说明：本技术 基于y分子筛载体的炉甘石止血复合物及其制备 (Calamine hemostatic compound based on Y molecular sieve carrier and preparation thereof ) 是由 王颖莉 张丽丽 杨冬花 张馨予 王家信 刘凯丽 王艳 于 2021-09-03 设计创作，主要内容包括：本发明公开了一种基于Y分子筛载体的炉甘石止血复合物,是将用于制备Y分子筛的含有硅源、钠源和铝源的碱性水溶液在从常温至60℃的至少3个温度阶段下持续搅拌一定时间,得到形成晶核的分子筛前驱体溶液,再加入棉纤维和炉甘石,于密闭反应釜中加热至90～100℃晶化反应得到的在棉纤维上合成Y分子筛,原位负载炉甘石药物的止血复合物。本发明通过Y分子筛-炉甘石-棉纤维多重止血材料的复合,得到了一种快速止血、凝血性能更好,凝血活酶时间明显缩短的止血材料,可以作为外用止血材料应用于各种创面的止血处理中。(The invention discloses a Y molecular sieve carrier-based calamine hemostatic compound, which is a hemostatic compound prepared by continuously stirring alkaline aqueous solution containing silicon source, sodium source and aluminum source for preparing a Y molecular sieve at least 3 temperature stages from normal temperature to 60 ℃ for a certain time to obtain a molecular sieve precursor solution for forming crystal nuclei, adding cotton fibers and calamine, heating to 90-100 ℃ in a closed reaction kettle, performing crystallization reaction to obtain a Y molecular sieve synthesized on the cotton fibers, and loading calamine medicine in situ. The invention obtains the hemostatic material with fast hemostasis, better blood coagulation performance and obviously shortened thromboplastin time by compounding the Y molecular sieve-calamine-cotton fiber multiple hemostatic materials, and can be used as an external hemostatic material to be applied to the hemostasis treatment of various wounds.)

1. A preparation method of a calamine hemostatic compound based on a Y molecular sieve carrier is prepared by the following steps:

1) adding a silicon source for preparing the Y molecular sieve into an alkaline aqueous solution containing a sodium source and an aluminum source, and mixing to obtain a molecular sieve precursor solution;

2) continuously stirring the molecular sieve precursor solution for a certain time at least 3 temperature stages from normal temperature to 60 ℃ to obtain a molecular sieve precursor solution for forming crystal nuclei;

3) adding cotton fiber and calamine into the molecular sieve precursor solution for forming the crystal nucleus, heating the solution to 90-100 ℃ in a closed reaction kettle for crystallization reaction, and preparing the calamine hemostatic compound based on the Y molecular sieve carrier.

2. The preparation method of claim 1, wherein the molecular sieve precursor solution is stirred at constant temperature of 25 ℃, 45 ℃ and 60 ℃ for 12-24 hours.

3. The process according to claim 1 or 2, wherein the molar ratio of Na is₂O∶Al₂O₃∶SiO₂∶H₂And mixing O = (0.5-0.9): (0.1-0.4): 1: (40-80) to obtain the molecular sieve precursor solution.

4. The process according to claim 1 or 2, wherein the molar ratio of m (calamine) to m (cotton fiber) to m (SiO) is in the range of m (calamine)₂) And (5) adding cotton fibers and calamine into the molecular sieve precursor solution, wherein the ratio of the cotton fibers to the calamine is (0.06-0.18) to (0.5-0.9) to 1.

5. The method according to claim 1 or 2, wherein the cotton fiber and the calamine are added to the molecular sieve precursor solution for forming crystal nuclei by adding a layer of cotton fiber and scattering a layer of calamine.

6. The method according to claim 1 or 2, wherein the silicon source is any one of silica sol, silica white, ethyl orthosilicate and sodium silicate.

7. The preparation method according to claim 1 or 2, characterized in that the aluminum source is any one of sodium metaaluminate and silica-alumina microspheres.

8. The method according to claim 1 or 2, wherein the crystallization reaction time is 24-36 hours.

9. A Y molecular sieve carrier-based calamine hemostatic composite prepared by the preparation method of any one of claims 1 to 8.

10. Use of a Y molecular sieve support based calamine haemostatic complex as defined in claim 9 for the preparation of a haemostatic material for external use.

Technical Field

The invention belongs to the technical field of biomedical materials, relates to a medical hemostatic material, and particularly relates to a hemostatic composite obtained by compounding multiple hemostatic materials and a preparation method thereof.

Background

Bleeding from trauma is a very common condition in everyday accidents, medical treatments and military wars. Researchers at home and abroad have conducted many studies on rapid hemostatic materials, and at an earlier stage, wounds were mainly treated with non-absorbable materials such as natural or artificial gauze and bandages.

In recent years, researchers have started with substances with low cost and high hemostatic speed, and modified to obtain a hemostatic dressing with improved performance, or combined with other substances to obtain a composite hemostatic material with high biosecurity or antibacterial performance.

Many scholars rapidly absorb water in blood by using substances with porous structures or large specific surface areas to achieve the effect of rapid hemostasis. For example, Sunweiqing and the like (preparation of MPH hemostatic microspheres and structural representation thereof [ J ] in China sanitary industry, 2015, 12(02): 144-. The oxidized nano-cellulose/sodium alginate composite hemostatic material researched by Liuchang yoga (preparation of the oxidized nano-cellulose/sodium alginate composite hemostatic material and performance research [ D ]. Haerbin Industrial university Polymer chemistry and Physics, 2016.) integrates the advantages of high crystallinity, high specific surface area, biocompatibility, biodegradability of sodium alginate, mild gel conditions and the like of the oxidized nano-cellulose material.

The molecular sieve as a porous material is aluminosilicate with a microporous structure, has a large specific surface area and a regular and ordered pore structure, is widely applied to the fields of catalysis of industrial production, cracking of petrochemical industry and the like due to excellent adsorption ion exchange performance and pore selection capacity, and is also concerned by researchers of medical materials due to strong adsorption capacity, a microporous structure with a molecular scale, an internal electric field effect and good biocompatibility.

The Y-type molecular sieve belongs to the faujasite molecular sieve, and has a pore channel system connected by double six-membered rings, larger pore volume and specific surface area, strong acidity and good thermal and hydrothermal stability. The cage formed by the multi-element rings and the cage penetrated by the pore canal provide a large number of cavities and spaces, so that the specific surface area of the cage is far larger than that of common material particles. When the wound bleeds, the liquid components in the blood can be quickly absorbed in a short time, and the concentrations of red blood cells, platelets, blood coagulation factors and the like in the blood are relatively increased; the molecular sieve produces exothermic phenomenon when absorbing liquid components, and the heat can enhance the aggregation speed of blood platelets and promote the formation of hemostatic scab on the surface of a wound. Binghongda et al (Zeolite has hemostatic effect on porcine fatal vascular injury model [ J ]. J. Western national defense medicine journal 2006(03): 164-.

At present, QuikClot containing zeolite as the active ingredient is produced by Z-medical company in the United states, and quick-acting hemostatic powder containing A-type calcium zeolite molecular sieve as the active ingredient is developed in China.

However, the biggest disadvantage of molecular sieves is the heat they give off during hemostasis, which can raise the temperature locally up to 100 ℃ and cause tissue or nerve burns. Liu Yu and the like (clinical follow-up of quick-acting hemostatic powder for treating acute traumatic hemorrhage [ J ]. Chinese general medicine 2010, 13(29): 3346 and 3347.) indicate that the clinical follow-up of the quick-acting hemostatic powder for treating acute traumatic hemorrhage also needs to pay attention to the problem that the product permeates into blood vessels to form thrombus when contacting with a wound surface, and the product needs to be thoroughly debrided and sutured after hemostasis.

CN 107661536A discloses a hierarchical pore composite hemostatic agent containing a molecular sieve, a preparation method and an application thereof, wherein the hemostatic agent has larger surface area and pore volume by utilizing hierarchical pore composite microspheres of Ca-MCM-48 mesoporous molecular sieve-chitosan and through the pore-forming effect of sesbania powder, so as to achieve the effect of rapid hemostasis; the introduction of chitosan solves the problem of large heat release of the molecular sieve in hemostasis; the microspheres are prepared by adopting a spray granulation method, and are beneficial to wound surface cleaning. However, there is still a possibility that the hemostatic particles in the form of microspheres will break up under pressure and enter the blood stream during use.

The scholars further propose a composite method of firmly growing the molecular sieve on the supporting carrier so as to solve the problem that the molecular sieve is washed away by blood. Van Jie et al (A very-bound and flexible mesoporous zeolite-zeolite hybrid magnetic [ J ]. Nature Communications, 2019, 10 (1)), prepared a kind of closely bound mesoporous Chabazite molecular sieve (mCHA) -cotton fiber hemostatic material, this hemostatic material regard cotton fiber as the structural support, through breaking the original growth skeleton, can increase the absorption and diffusion of multiple adsorbates, create the mesopore on the basis of microporous structure; the adopted binderless synthesis method can avoid the phenomenon that the active ingredients of the molecular sieve are diluted by the binder and block the pore channels; different from the method of directly dripping zeolite turbid liquid on cotton fibers, the adopted synthesis method realizes the tight combination of the active hydroxyl on the cotton fibers and the zeolite by utilizing the chemical bonding of the active hydroxyl on the cotton fibers and the zeolite, solves the problems of reduced hemostatic effect and thrombus formation caused by easy falling of the molecular sieve, effectively relieves the heat release problem of the zeolite, and improves the hemostatic performance and safety.

Furthermore, exposure of the wound surface to the air involves the risk of bacterial infection, which may cause inflammation and local vascular dilatation, which is not conducive to hemostatic healing of the wound, and may also cause complications during the wound healing period. Therefore, the ideal hemostatic material should have the efficacy of antisepsis and anti-inflammation on the basis of good blood coagulation performance.

Zn²⁺、Ag⁺The metal ions can combine with the bacterial membrane and membrane protein to destroy the structure of bacteria and the enzyme responsible for electron transfer, thus achieving the antibacterial effect. Hou et al (antibiotics of cellulose silicate on water adsorption, degradability, antibiotic effectiveness, hemostati)c performances and cell viability to microporous starch based hemostat[J]. Materials science &engineering, C. Materials for biological applications, 2017, 76: 340-349.) mesoporous calcium zinc silicate (m-ZCS) is synthesized by a template method and is doped into Microporous Starch (MS) to prepare the m-ZCS/MS composite material, and the doping of the m-ZCS not only obviously enhances the water absorption performance of the product, but also is beneficial to Zn²⁺The antibacterial effect of the hemostatic is improved, and the hemostatic agent which has high hemostatic effect, is degradable and has antibacterial activity is obtained.

The compound calamine lotion has the effects of bacteriostasis, itching relieving, mild antisepsis and convergence as a conventional anti-inflammatory medicament. The calamine mainly contains natural zinc carbonate, and is decomposed into zinc oxide after being calcined at high temperature. Galamina is sweet, warm and nontoxic in nature and taste, and recorded in Ben Cao gang mu, has the functions of stopping bleeding, eliminating swelling and toxicity, promoting granulation and improving eyesight, removing nebula and red, astringing and removing rotten, and is a common medicine for ophthalmology and skin surgery in traditional Chinese medicine. The calamine has the same function of inhibiting bacteria and is also a zinc element.

Disclosure of Invention

The invention aims to provide a Y molecular sieve carrier-based calamine hemostatic composite, which is a hemostatic material with rapid hemostasis, better blood coagulation performance and obviously shortened thromboplastin time through the compounding of a Y molecular sieve-calamine-cotton fiber multiple hemostatic material.

The calamine hemostatic compound based on the Y molecular sieve carrier is prepared by the following method:

1) adding a silicon source for preparing the Y molecular sieve into an alkaline aqueous solution containing a sodium source and an aluminum source, and mixing to obtain a molecular sieve precursor solution;

2) continuously stirring the molecular sieve precursor solution for a certain time at least 3 temperature stages from normal temperature to 60 ℃ to obtain a molecular sieve precursor solution for forming crystal nuclei;

3) adding cotton fiber and calamine into the molecular sieve precursor solution for forming the crystal nucleus, heating the solution to 90-100 ℃ in a closed reaction kettle for crystallization reaction, and preparing the calamine hemostatic compound based on the Y molecular sieve carrier.

Preferably, the molecular sieve precursor solution is stirred at constant temperature of 25 ℃, 45 ℃ and 60 ℃ for 12-24 hours.

Specifically, in the preparation method of the present invention, Na is preferably used in a molar ratio₂O∶Al₂O₃∶SiO₂∶H₂O = (0.5-0.9): (0.1-0.4): 1: (40-80) to obtain the molecular sieve precursor solution.

Furthermore, in the preparation method, m (calamine) to m (cotton fiber) to m (SiO) are adopted₂) And (5) adding cotton fibers and calamine into the molecular sieve precursor solution, wherein the ratio of the cotton fibers to the calamine is (0.06-0.18) to (0.5-0.9) to 1.

The silicon source may include, but is not limited to, silica sol, silica white, ethyl orthosilicate, sodium silicate and various conventional silicon source materials, and silica sol or silica white is preferably used in the present invention.

The aluminum source can include, but is not limited to, sodium metaaluminate, silica-alumina microspheres and other conventional aluminum source materials, and sodium metaaluminate is preferably used in the present invention.

Preferably, the method adopts a mode of adding a layer of cotton fiber and scattering a layer of calamine, and the cotton fiber and the calamine are added into a molecular sieve precursor solution for forming crystal nuclei.

Furthermore, the crystallization reaction time is preferably 24-36 h.

Washing the reaction product obtained by the preparation method with water, and drying to obtain the calamine hemostatic compound based on the Y molecular sieve carrier.

The calamine hemostatic composite based on the Y molecular sieve carrier prepared by the invention can be used as an external hemostatic material and applied to the hemostatic treatment of various wound surfaces.

The rapid hemostasis of wounds is a key treatment link in daily life and before wound hospital battle, and the efficient and safe hemostatic material is a main way for realizing rapid hemostasis. The invention is based on a 'molecular sieve/cotton fiber/calamine' triple hemostatic material, and prepares an antibacterial external hemostatic compound with high hemostatic performance and good biological safety by utilizing the strong adsorption capacity of a Y molecular sieve, the heat dissipation capacity of cotton fiber and the anti-inflammatory and antibacterial performance of calamine.

According to the method, a large number of crystal nuclei are formed in advance in a molecular sieve precursor solution before crystallization reaction, cotton fibers and the calamine are added for in-situ synthesis, the crystal nuclei grow gradually into nano-microcrystals, and finally the Y molecular sieve which grows on the cotton fibers and supports the calamine medicine and is smaller in average particle size and better in crystallinity is obtained.

Compared with the conventional hemostatic medicaments, the activated partial thromboplastin time (APPT) of the Y molecular sieve carrier-based calamine hemostatic composite prepared by the invention is obviously shortened, the Prothrombin Time (PT) is improved, the hemostasis is promoted mainly by activating an endogenous blood coagulation path, the exogenous blood coagulation path is not obviously influenced, and the Y molecular sieve carrier-based calamine hemostatic composite has more efficient blood coagulation performance compared with the conventional hemostatic medicaments, and is an efficient and safe antibacterial hemostatic composite.

Drawings

Fig. 1 is an XRD pattern of samples prepared in example 1 and comparative example 1.

FIG. 2 is an SEM photograph of samples prepared in example 1 and comparative example 1.

Figure 3 is the XRD pattern of the example 1 preparation after firing.

Fig. 4 is the APTT and PT test results for different hemostatic materials.

Detailed Description

The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples. The following examples are only for more clearly illustrating the technical solutions of the present invention so as to enable those skilled in the art to better understand and utilize the present invention, and do not limit the scope of the present invention.

The names and abbreviations of the experimental methods, production processes, instruments and equipment involved in the examples and comparative examples of the present invention are those commonly known in the art and are clearly and clearly understood in the relevant fields of use, and those skilled in the art can understand the conventional process steps and apply the corresponding equipment according to the names and perform the operations according to the conventional conditions or conditions suggested by the manufacturers.

The various starting materials or reagents used in the examples of the present invention and comparative examples are not particularly limited in their sources, and are all conventional products commercially available. They may also be prepared according to conventional methods well known to those skilled in the art.

Example 1.

30mL of deionized water, 2.33g of sodium hydroxide, 0.2g of sodium chloride, 1.99g of sodium metaaluminate and 4.33mL of 40% silica sol are sequentially added into a container with a polytetrafluoroethylene lining, and the mixture is stirred at room temperature for 60 min.

And then placing the container in a water bath kettle at 25 ℃, stirring for 24h at constant temperature, raising the temperature of the water bath kettle to 45 ℃, stirring for 24h at constant temperature, finally raising the temperature to 60 ℃, stirring for 12h at constant temperature, and stopping stirring.

Adding 1.5g of medical absorbent cotton torn into fiber sheets and 0.2g of calamine into a container with a polytetrafluoroethylene lining in a mode of adding a layer of medical absorbent cotton and scattering a layer of calamine, putting the container into a stainless steel reaction kettle for sealing, and heating the container in a thermostat to 100 ℃ for constant-temperature crystallization reaction for 24 hours.

And taking out the reaction kettle, quickly cooling to room temperature in a quenching tank, taking out a reaction product, washing for 3 times by using deionized water, carrying out suction filtration, and drying for 12 hours at 70 ℃ in a vacuum drying oven to obtain a Y molecular sieve carrier-based calamine hemostatic composite sample.

Comparative example 1.

In the comparative example, a guide agent method is adopted to prepare the Y molecular sieve growing on the cotton fiber.

19.0mL of deionized water, 5.96g of sodium hydroxide, 0.56g of sodium metaaluminate and 5.93mL of 40% silica sol are sequentially added into a container, and the mixture is placed into a 35 ℃ water bath kettle to be stirred and reacted for 18 hours, so that the guiding agent for preparing the Y molecular sieve is obtained.

Then 25.6mL of deionized water, 3.94g of sodium hydroxide, 1.24g of sodium metaaluminate and 9.7mL of 40% silica sol are sequentially added into a container with a polytetrafluoroethylene lining, 1.44mL of guiding agent is dropwise added, and the mixture is stirred for 2 hours.

Adding 1.5g of medical absorbent cotton torn into fiber sheets and 0.2g of calamine into a container with a polytetrafluoroethylene lining in a mode of adding a layer of medical absorbent cotton and scattering a layer of calamine, putting the container into a stainless steel reaction kettle for sealing, and heating the container in a thermostat to 100 ℃ for constant-temperature crystallization reaction for 24 hours.

And taking out the reaction kettle, quickly cooling to room temperature in a quenching tank, taking out a reaction product, washing for 3 times by using deionized water, carrying out suction filtration, and drying for 12 hours at 70 ℃ in a vacuum drying oven to obtain a calamine hemostatic composite sample of the growth Y molecular sieve on the cotton fiber prepared by the guiding agent method.

The XRD patterns of the samples prepared as described above are shown in fig. 1, in which a is the sample of the complex prepared in example 1 and b is the sample of the complex prepared in comparative example 1.

In the figure, characteristic diffraction peaks of the Y molecular sieve appear at 2 theta =6.1 degrees, 10.0 degrees, 15.5 degrees, 18.6 degrees, 20.2 degrees, 23.5 degrees, 26.9 degrees, 30.6 degrees, 31.2 degrees, 32.3 degrees, 33.9 degrees and the like of a and b, which indicates that the added calamine and cotton fibers existing as a support carrier do not influence the crystallization of a sample, and the Y molecular sieve is successfully synthesized by the two methods.

However, the characteristic diffraction peak intensities of a at 2 theta =6.1 degrees, 15.5 degrees, 18.6 degrees and 26.9 degrees are higher than b, and the half-peak width is also smaller than b, which indicates that the crystallinity of the Y molecular sieve obtained by the method is higher than that of the Y molecular sieve obtained by the guide agent method, and the Y molecular sieve with better crystallinity can be obtained.

FIG. 2 further shows SEM images of samples prepared as described above, wherein a and b are surface topographies of the composite sample prepared in example 1, b is a partial magnified view of a, and c is the surface topography of the composite sample prepared in comparative example 1.

In the figure, the Y molecular sieves of both samples grew firmly on the rough surface and grooves of the cotton fiber. It can be seen from a and b that the Y molecular sieve particles of the sample prepared in example 1 have an agglomeration phenomenon on the surface of the cotton fiber, the particle size is small, about 300-450 nm, the Y molecular sieve growing on the cotton fiber is relatively more under the load of the calamine, a certain amount of particles are also on the surface of the smoother cotton fiber at rough and groove positions, and the agglomeration phenomenon in the groove is alleviated. In the comparative example 1 of c, the Y molecular sieve in the sample prepared by the guiding agent method is spherical, the particle size is uniformly distributed, and the particle size is about 550-750 nm.

The formation of molecular sieve crystals is divided into two steps of crystal nucleation and growth. The preparation method of the invention increases the temperature of the raw material solution from 25 ℃, 45 ℃ and 60 ℃ to 100 ℃ in a gradient way, provides a mild and slow forming environment for crystallization reaction, has enough time to form a large number of crystal nuclei, and generates the Y molecular sieve with smaller grain diameter. According to the invention, the cotton fiber is added after the crystal is nucleated and grown for a certain time, so that a growth substrate with more space is provided for the molecular sieve, and the active alcoholic hydroxyl on the cotton fiber can realize the firm growth of the Y molecular sieve, so that the compound with small particle size and good crystallinity is obtained.

To confirm that the calamine payload was on the composite sample, the XRD pattern of the calcined product after subjecting the calamine hemostatic composite sample based on Y molecular sieve support prepared in example 1 to calcination treatment is shown in fig. 3. In the figure, sharp diffraction peaks appearing at 2 theta = 5-35 degrees are characteristic peaks of a Y molecular sieve, and diffraction peaks appearing at 2 theta =30.9 degrees, 34.7 degrees, 36.2 degrees, 47.1 degrees, 62.7 degrees and 68.2 degrees are characteristic peaks of ZnO, and the effective component of calamine is analyzed to be ZnCO₃And ZnO, ZnCO after calcination₃Converted to ZnO, and the calamine drug was loaded on the composite sample at a characteristic peak surface of 2 θ = 30.9-68.2 °.

Example 2.

35mL of deionized water, 1.285g of sodium hydroxide, 0.2g of sodium chloride, 1.99g of sodium metaaluminate and 2.33g of white carbon black are sequentially added into a container with a polytetrafluoroethylene lining, and the mixture is stirred for 60min at room temperature.

And then placing the container in a water bath kettle at 25 ℃, stirring for 24h at constant temperature, raising the temperature of the water bath kettle to 45 ℃, stirring for 24h at constant temperature, finally raising the temperature to 60 ℃, stirring for 12h at constant temperature, and stopping stirring.

Adding 1.5g of medical absorbent cotton torn into fiber sheets and 0.27g of calamine into a container with a polytetrafluoroethylene lining in a mode of adding a layer of medical absorbent cotton and scattering a layer of calamine, putting the container into a stainless steel reaction kettle for sealing, and heating the container in a thermostat to 100 ℃ for constant-temperature crystallization reaction for 24 hours.

And taking out the reaction kettle, quickly cooling to room temperature in a quenching tank, taking out a reaction product, washing for 3 times by using deionized water, carrying out suction filtration, and drying for 12 hours at 70 ℃ in a vacuum drying oven to prepare the Y molecular sieve carrier-based calamine hemostatic compound.

Example 3.

32mL of deionized water, 1.8g of sodium hydroxide, 0.2g of sodium chloride, 3.41g of sodium metaaluminate and 9.7mL of 40% silica sol are sequentially added into a container with a polytetrafluoroethylene lining, and the mixture is stirred at room temperature for 60 min.

And then placing the container in a water bath kettle at 25 ℃, stirring for 24h at constant temperature, raising the temperature of the water bath kettle to 45 ℃, stirring for 24h at constant temperature, finally raising the temperature to 60 ℃, stirring for 12h at constant temperature, and stopping stirring.

Adding 1.0g of medical absorbent cotton torn into fiber sheets and 0.15g of calamine into a container with a polytetrafluoroethylene lining in a mode of adding a layer of medical absorbent cotton and scattering a layer of calamine, putting the container into a stainless steel reaction kettle for sealing, and heating the container in a thermostat to 100 ℃ for constant-temperature crystallization reaction for 24 hours.

And taking out the reaction kettle, quickly cooling to room temperature in a quenching tank, taking out a reaction product, washing for 3 times by using deionized water, carrying out suction filtration, and drying for 12 hours at 70 ℃ in a vacuum drying oven to prepare the Y molecular sieve carrier-based calamine hemostatic compound.

Application example 1: rabbit ear artery trauma hemostasis test.

The average weight of 15 rabbits for the experiment is 2.5 Kg.

After 15 white rabbits were adaptively raised in a single cage for 1 week, the animals were randomly divided into 3 groups of 5 animals each, including a blank group, an experimental group and a control group. Wherein the blank group adopts blank cotton to stop bleeding, the experimental group adopts the calamine hemostatic compound sample prepared in the example 1 to stop bleeding, and the control group adopts cotton directly soaked with calamine to stop bleeding.

Rabbits were fasted for 8h before the experiment, water was freely drunk, and anesthesia was performed by intravenous anesthesia injection at the ear margin with 10% chloral hydrate. Fixing on an operating table after anesthesia, disinfecting the central area of the back side of the rabbit ear with iodophor, transecting the ear artery with an operating knife but not cutting through the ear, and establishing an ear artery bleeding model.

Each group of cotton was accurately weighed in advance before the experiment. Immediately after the model is made, different groups of cotton with the same mass are respectively coated on the wound surfaces of each group, the cotton is pressurized by a weight of 100g, the timing is started, the observation time points are respectively 30s, 20s, 10s and 5s, then the wound surface bleeding condition is observed for 1 time every 5s, and the hemostasis time is recorded. So that bleeding does not occur any more 3min after the dressing and the weight are removed until the blood is successful.

Accurately weighing the wet weight of each group of cotton after hemostasis, and calculating the bleeding amount. Bleeding amount = weight of wet cotton after hemostasis-weight of dry cotton before hemostasis.

All data of this experiment were usedShowing that SPSS 24.0 software is used for processing data and comparing between groupstAnd (4) checking the test result,Pdifferences < 0.05 are statistically significant.

The experimental results show that the bleeding amount and bleeding time of the control group and the experimental group are obviously reduced compared with the blank group; compared with a control group, the bleeding amount and bleeding time of the experimental group are obviously reduced; all had statistical differences, see table 1.

Application example 2: activated Partial Thromboplastin Time (APTT) and Prothrombin Time (PT) tests.

The venous blood of the testee is extracted in vacuum in the early morning under the condition of empty stomach, and APTT and PT index detection is carried out by adopting a STA-R full-automatic coagulometer of French STATO company. Whether the material activates the intrinsic or extrinsic coagulation pathway is judged by APTT and PT values.

The sample prepared in example 1 was used as a test sample, and the test sample was mixed with plasma, incubated at 37 ℃ for 3min, allowed to stand for 10min, and tested on a computer. All blood samples are detected within 2h after collection, so that the influence of delayed detection on the authenticity of a detection result is avoided.

The detection results are shown in FIG. 4, wherein A is the APTT result of each sample; b is the PT result of each sample; in the figure, a represents calamine, b represents a Y molecular sieve, c represents a sample of example 1, and d represents Yunnan white drug hemostatic.

The APTT experimental result shown in figure 4A shows that the calamine has obviously increased APTT value compared with Yunnan Baiyao, and the calamine and Yunnan Baiyao have obvious difference, namely the calamine used alone has the effect of hindering the endogenous blood coagulation system; compared with Yunnan white drug powder, the hemostatic compound prepared by the invention effectively reduces the APTT value, obviously shortens the blood coagulation time of blood plasma, and shows that the hemostatic compound can accelerate the start of an endogenous blood coagulation system and has better blood coagulation effect compared with the Yunnan white drug powder.

The PT experimental result shown in FIG. 4B shows that the PT value of calamine is obviously increased compared with that of Yunnan white drug powder, and the significant difference is that the calamine used alone also has an inhibiting effect on an exogenous blood coagulation system; compared with Yunnan white drug powder, the hemostatic compound prepared by the invention has no significant difference, which shows that the hemostatic compound has no influence on exogenous blood coagulation.

APTT and PT experiments show that the hemostatic composite provided by the invention mainly activates an endogenous coagulation pathway to promote hemostasis, has no significant influence on the exogenous coagulation pathway, and has more efficient coagulation performance compared with Yunnan white drug.

The above embodiments of the present invention are not intended to be exhaustive or to limit the invention to the precise form disclosed. Various changes, modifications, substitutions and alterations to these embodiments will be apparent to those skilled in the art without departing from the principles and spirit of this invention.