Carbon nanospheres, preparation method thereof and application of carbon nanospheres in preparation of organ / tissue toxin removal reagent

Abstract

The invention belongs to the technical field of adsorbents, and particularly relates to a carbon nanosphere, a preparation method of the carbon nanosphere and application of the carbon nanosphere in preparation of an organ / tissue toxin removal reagent. The nano carbon sphere provided by the invention is rich in micropore structure, large in specific surface area, smooth in surface and free of dust, and further can be used as or used for preparing an organ / tissue toxin removing reagent through a specific diameter, so that toxin and impurities in organs or tissues are directionally removed, specific toxin is accurately removed, the poisoning condition is corrected, and the toxic and side effects are reduced. The positive significance is realized on treating acute and chronic renal failure of animals. The carbon nanosphere provided by the invention can be used as or used for preparing an organ / tissue toxin removing reagent. When lipopolysaccharide in blood is removed, the removal rate is 80% or above, and the influence on other beneficial components in the blood is smaller than 10%.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of adsorbents, and particularly relates to a nanometer carbon sphere, a preparation method thereof and application of the nanometer carbon sphere in preparation of an organ / tissue toxin removal reagent. BACKGROUND

[0002] In the field of pet medical treatment, diseases such as acute and chronic renal failure and poisoning pose a serious threat to the health of pets. Blood perfusion as an important blood purification technology can effectively remove toxins and harmful substances in the blood. However, the adsorbents used in current blood perfusion devices have limitations in precisely adsorbing specific toxins.

[0003] Traditional adsorbents often cannot efficiently remove target toxins while minimizing the impact on beneficial components in the blood. For example, when removing toxins such as lipopolysaccharide (LPS), it is difficult to achieve the dual goals of high removal rate and minimal impact on beneficial components. SUMMARY

[0004] The application aims to provide a nanometer carbon sphere, a preparation method thereof and application of the nanometer carbon sphere in preparation of an organ / tissue toxin removal reagent. The nanometer carbon sphere provided by the application can remove toxins and impurities in organs / tissues in a targeted manner.

[0005] To achieve the above-mentioned purpose, the application provides the following technical solutions.

[0006] The application provides a nanometer carbon sphere with a diameter of 100-500 microns and a specific surface area of 300-800 m 2 / g. The surface of the nanometer carbon sphere has a microporous structure.

[0007] Preferably, the pore size of the micropores in the nanometer carbon sphere is 1-5 nanometers.

[0008] Preferably, the elemental composition of the nanometer carbon sphere includes C, N and O.

[0009] The application also provides a preparation method of the nanometer carbon sphere described in the above-mentioned solution, which includes the following steps.

[0010] (1) mixing a surfactant, water and a pH regulator to obtain a template solution;

[0011] (2) mixing a carbon source solution and the template solution to perform a hydrothermal carbonization reaction, and then performing solid-liquid separation to obtain a nanometer carbon sphere precursor;

[0012] (3) annealing the nanometer carbon sphere precursor to obtain the nanometer carbon sphere.

[0013] Preferably, the surfactant is an organic ammonium salt.

[0014] Preferably, the standard for adding the pH adjuster is to adjust the pH value of the template solution to 10-12.

[0015] Preferably, the equipment for the hydrothermal carbonization reaction is a reactor and an oven; the filling degree of the reactor is 60-80%; the temperature of the hydrothermal carbonization reaction is 180-220℃, and the holding time is 12-24 hours.

[0016] Preferably, the annealing temperature is 500–800°C, and the holding time is 1–3 hours; the annealing is carried out in a protective atmosphere.

[0017] The present invention also provides the application of the nanocarbon spheres described in the above-described scheme or the nanocarbon spheres obtained by the preparation method described in the above-described scheme in the preparation of organ / tissue toxin removal reagents.

[0018] Preferably, the method of application is as follows: the nanocarbon spheres are filled into the adsorption column of a hemoperfusion device, and animal blood is passed through the adsorption column at a flow rate of 50-200 mL / min.

[0019] This invention provides a carbon nanosphere. The carbon nanosphere provided by this invention has a rich microporous structure, a large specific surface area, and a smooth surface that does not generate dust. Furthermore, through a specific diameter, it can be used as or for preparing organ / tissue toxin removal reagents. Through differences in diffusion rates and the synergistic effect of functional groups and charges, it can selectively remove toxins (e.g., lipopolysaccharides) and impurities (e.g., cellular debris and byproducts) in organs / tissues, precisely removing specific toxins, correcting poisoning conditions, and has positive significance for treating acute and chronic renal failure in animals.

[0020] This invention also provides a method for preparing the carbon nanospheres described above. The preparation method provided by this invention, using a hydrothermal template method, prepares carbon nanospheres with excellent physical properties. The steps are simple, safe, and stable, and it has promising applications for large-scale industrial production.

[0021] This invention also provides the application of the carbon nanospheres described in the above-described scheme or the carbon nanospheres prepared by the above-described method in the preparation of organ / tissue toxin removal reagents. The carbon nanospheres provided by this invention can be used as or in the preparation of organ / tissue toxin removal reagents to achieve the removal of specific toxins. When removing lipopolysaccharides from the blood, the removal rate is above 80%, and the impact on other beneficial components in the blood is less than 10%. Attached Figure Description

[0022] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0023] Figure 1 The graph shows the functional test results of the nanospheres prepared in Example 1 for blood perfusion.

[0024] Figure 2 The image shows the pathological staining results of blood perfusion of the nanospheres prepared in Example 1.

[0025] Figure 3 X-ray photoelectron spectrum of the nanospheres prepared in Example 1;

[0026] Figure 4 The pore size distribution curve of the nanospheres prepared in Example 1;

[0027] Figure 5 Low-magnification scanning electron microscope image of the nanospheres prepared in Example 1;

[0028] Figure 6 This is a high-magnification scanning electron microscope image of the nanospheres prepared in Example 1. Detailed Implementation

[0029] This invention provides a carbon nanosphere with a diameter of 100–500 micrometers and a specific surface area of ​​300–800 m². 2 / g; the surface of the carbon nanospheres has a microporous structure.

[0030] In this invention, the diameter of the carbon nanospheres can be 200 to 400 micrometers, specifically 250 micrometers, 300 micrometers, or 350 micrometers.

[0031] In this invention, the specific surface area of ​​the carbon nanospheres can be 400–700 m². 2 / g, specifically 450m 2 / g、500m 2 / g、550m 2 / g、600m 2 / g or 650m 2 / g.

[0032] In this invention, the pore size of the micropores in the carbon nanospheres can be 1 to 5 nanometers, specifically 2 nanometers, 3 nanometers or 4 nanometers.

[0033] In this invention, the elemental composition of the carbon nanospheres may include C, N and O.

[0034] The application further provides a preparation method of the nanometer carbon sphere.

[0035] (1) mixing a surfactant, water and a pH regulator to obtain a template solution;

[0036] (2) mixing a carbon source solution and the template solution to perform a hydrothermal carbonization reaction, and then performing solid-liquid separation to obtain a nanometer carbon sphere precursor;

[0037] (3) annealing the nanometer carbon sphere precursor to obtain the nanometer carbon sphere.

[0038] In the application, the surfactant can be an organic ammonium salt; the organic ammonium salt can be cetyltrimethylammonium bromide (CTAB).

[0039] In the application, the water can be deionized water.

[0040] In the application, the ratio of the amount of substance of the surfactant to the volume of water can be (0.1-0.5) mol:1 L, and specifically can be 0.2 mol:1 L, 0.3 mol:1 L or 0.4 mol:1 L.

[0041] In the application, the pH regulator can be a base; the base can be a hydroxide; the hydroxide can be a sodium hydroxide solution; the concentration of the sodium hydroxide solution can be 0.1-1 mol / L, and specifically can be 0.5 mol / L; and the addition amount of the pH regulator can be such that the pH value of the template solution is adjusted to 10-12.

[0042] In the application, the first mixing can be: premixing the surfactant and the water to obtain a premixing solution, then adding the pH regulator dropwise to the premixing solution while stirring, and continuing to stir.

[0043] In the application, the premixing can be magnetic stirring mixing, and the rotating speed of the magnetic stirring mixing can be 200-400 rpm, and specifically can be 300 rpm. The application enables the surfactant to be uniformly dispersed in the water through premixing.

[0044] In the application, the rotating speed of the stirring while dropwise adding can be 500-800 rpm, and specifically can be 600 rpm or 700 rpm, and the dropwise adding rate can be 1-2 drops per second.

[0045] In the application, the time of the continued stirring can be 30-60 minutes, and specifically can be 40 minutes or 50 minutes. The application enables the surfactant to form a stable micellar structure as a template through the first mixing.

[0046] After obtaining the template solution, the present application mixes the carbon source solution and the template solution (denoted as second mixing) to perform a hydrothermal carbonization reaction, and then performs solid-liquid separation to obtain a nanocarbon sphere precursor. In the present application, the carbon source in the carbon source solution can be a monosaccharide; the monosaccharide can be glucose; the solvent of the carbon source solution can be deionized water; and the concentration of the carbon source solution can be 0.5-1.5 mol / L, and specifically can be 1 mol / L.

[0047] In the present application, the preparation method of the carbon source solution can be mixing the carbon source and deionized water (denoted as third mixing); and the temperature of the third mixing can be no higher than 60℃, and specifically can be 60℃, 50℃, 40℃, 30℃ or 20℃. The present application accelerates the dissolution of the carbon source by heating and mixing.

[0048] In the present application, the second mixing can be: using a constant-pressure dropping funnel to drop the carbon source solution into the template solution and continuing to stir; the dropping rate can be 40-60 mL / s, and specifically can be 50 mL / s; and the time of the continued stirring can be 1-2 hours.

[0049] In the present application, the equipment of the hydrothermal carbonization reaction can be a reaction kettle and an oven; the filling degree of the reaction kettle can be 60-80%, and specifically can be 70%; and the reaction kettle can be sealed before the hydrothermal carbonization reaction. The present application performs the hydrothermal carbonization reaction under the above filling degree to ensure that the reaction is performed in a suitable pressure environment.

[0050] In the present application, the temperature of the hydrothermal carbonization reaction can be 180-220℃, and specifically can be 190℃, 200℃ or 210℃, and the holding time can be 12-24 hours, and specifically can be 15 hours or 18 hours. The present application forms a nanocarbon sphere precursor by carbonization of the carbon source under the limitation of the template through the hydrothermal carbonization reaction.

[0051] In the present application, before the solid-liquid separation, the obtained reaction system can be cooled and then the product can be taken out; the cooling can be natural cooling; and the final temperature of the cooling can be room temperature.

[0052] In the present application, the solid-liquid separation can be discarding the supernatant after centrifugation; the rotation speed of the centrifugation can be 5000-10000 rpm, and specifically can be 6000 rpm, 7000 rpm, 8000 rpm or 9000 rpm; and the time of the centrifugation can be 10-15 minutes, and specifically can be 12 minutes.

[0053] In the present application, the solid-liquid separation can further include alternately washing and dispersing the obtained solid; the washing can include sequentially water washing and alcohol washing; the water used in the water washing can be deionized water; the alcohol used in the alcohol washing can be ethanol; the number of times of washing can be more than 1, and specifically can be 5 times; the dispersing can be ultrasonic; the time of ultrasonic can be 3 minutes. The present application removes templates and impurities in the product through washing and dispersing. During the solid-liquid separation and the washing process, the acceleration and deceleration process of the centrifuge is smooth, avoiding the agglomeration of the product caused by violent vibration.

[0054] After obtaining the nanocarbon sphere precursor, the present application anneals the nanocarbon sphere precursor to obtain the nanocarbon sphere. In the present application, the annealing can further include drying the nanocarbon sphere precursor before the annealing; the temperature of drying can be 60-80℃, and the holding time can be 12-24 hours; the drying equipment can be an oven with a gas-permeable partition. The gas-permeable partition can ensure air circulation when the oven is drying.

[0055] In the present application, the annealing equipment can be a tube furnace; the temperature of annealing can be 500-800℃, and specifically can be 600℃ or 700℃, and the holding time can be 1-3 hours, and specifically can be 2 hours; the annealing can be carried out in a protective atmosphere; the protective atmosphere can be an inert gas; the inert gas can be argon; the flow rate of the inert gas can be 50-150mL / min, and specifically can be 80mL / min, 100mL / min or 120mL / min.

[0056] The present application also provides the use of the nanocarbon sphere prepared by the method described in the above scheme or the nanocarbon sphere prepared by the method described in the above scheme in the preparation of an organ / tissue toxin removal reagent.

[0057] In the present application, the method of use can be: loading the nanocarbon sphere into an adsorption column of a hemoperfusion device, and making the blood of an animal pass through the adsorption column at a flow rate of 50-200mL / min.

[0058] In the present application, the loading mode can be series connection or parallel connection; the series connection can be series connection (type I).

[0059] The nanocarbon sphere provided by the present application can be used in a hemoperfusion device for rapid and convenient use, the nanocarbon sphere is loaded into an adsorption column of a hemoperfusion device, the bed pressure drop is small, the blood can pass through smoothly, and the removal of specific toxins can be achieved.

[0060] In order to further illustrate the present application, the schemes of the present application are described in detail below in combination with the drawings and examples, but they should not be understood as limiting the scope of protection of the present application.

[0061] Example 1

[0062] 1.6644 g of CTAB was weighed and dissolved in 50 mL of deionized water using a magnetic stirrer at a speed of 200 rpm to prepare a CTAB solution with a concentration of 0.1 mol / L. A 0.1 mol / L sodium hydroxide solution was added dropwise to the CTAB solution under stirring at 700 rpm, the dropping speed was controlled at 1 drop per second, the pH value of the CTAB solution was adjusted to 10, and the stirring was continued for 60 minutes to form a stable micelle structure of CTAB, thereby obtaining a template solution;

[0063] 4.5 g of glucose was heated to 40°C and dissolved in 30 mL of deionized water to prepare a carbon source solution with a concentration of 0.5 mol / L. The carbon source solution was then added to the template solution at a rate of 50 mL / s using a constant pressure dropping funnel, and the stirring was continued for 2 hours to fully disperse the glucose, thereby obtaining a mixed solution;

[0064] The mixed solution was transferred to a 100 mL reaction kettle with a filling degree of 60%, and then was placed in an oven after being sealed. The glucose was subjected to a hydrothermal carbonization reaction under the restriction of the CTAB template at 180°C for 24 hours. After the reaction, the product was naturally cooled to room temperature, and then was centrifuged at a speed of 5000 rpm for 15 minutes using a centrifuge. The supernatant was discarded, and the precipitate was washed with deionized water and ethanol for 5 times. After each washing, the precipitate was subjected to ultrasonic dispersion for 3 minutes, thereby obtaining a nanocarbon sphere precursor;

[0065] The nanocarbon sphere precursor was placed in an oven with a gas-permeable partition at 60°C for drying for 24 hours, and then was placed in a tube furnace. Under the protection of argon gas with a flow rate of 50 mL / min, the nanocarbon sphere precursor was annealed at 500°C for 3 hours, thereby obtaining nanocarbon spheres with a diameter of 100 microns, a specific surface area of 300 m 2 / g, and a surface with a rich microporous structure (micropore size of 1-5 nanometers, tested by the Brunauer-Emmett-Teller (BET) method).

[0066] Test Example 1

[0067] The nanocarbon spheres prepared in Example 1 were loaded in an adsorption column of a blood perfusion device, and the animal blood was passed through the column at a flow rate of 50 mL / min. The results showed that the nanocarbon spheres of Example 1 had a clearance rate of 82% for lipopolysaccharide (LPS) and had an influence of less than 10% on other beneficial components in the blood.

[0068] Preparation of a dialysis experiment object:

[0069] An adult male New Zealand rabbit was used as the experiment object, and the weight of the adult male New Zealand rabbit was 2.0-2.5 kg.

[0070] Grouping: The adult male New Zealand rabbits were divided into three groups, i.e. a control group, a sepsis model group, and a sepsis plus hemoperfusion group;

[0071] Sepsis model construction: The adult male New Zealand rabbits in the sepsis model group and the sepsis plus hemoperfusion group were injected with lipopolysaccharide (LPS) through the marginal ear vein to construct a sepsis model, and the injection dose of the lipopolysaccharide was 500 μg / kg BW;

[0072] Treatment operation of the sepsis plus hemoperfusion group:

[0073] Anesthesia and blood vessel treatment: Before the treatment started, the adult male New Zealand rabbits in the sepsis plus hemoperfusion group were weighed, and then anesthetized through intraperitoneal injection of tribromoethanol, and the injection dose of the tribromoethanol was 1 mL / kg. After the anesthesia was successful, the right femoral artery and vein of the adult male New Zealand rabbits were separated, punctured with a 24G indwelling needle, and fixed;

[0074] Pretreatment of the hemoperfusion device: The fixed indwelling needle was connected to the hemoperfusion device, and the entire extracorporeal circuit was pretreated with heparin solution (concentration of 125 IU / mL), specifically, first pretreated at a flow rate of 20 mL / min for 30 min, and then the flow rate was reduced to 5 mL / min for continued pretreatment for 30 min;

[0075] Hemoperfusion implementation: After the LPS treatment for 1.5 h, the hemoperfusion program was started. Heparin was used for anticoagulation, and an initial loading dose of 500 IU was given, and then 200 IU was intermittently injected every 30 min. During the hemoperfusion process, the time points of 30 min, 60 min and 120 min were recorded, and after the hemoperfusion ended, physiological saline was used for fluid replacement;

[0076] Postoperative treatment: The surgical incision was carefully sutured, and protamine was given to reverse the anticoagulation effect of heparin;

[0077] Sample collection and detection: After the LPS treatment for 24 h, the animals in all groups were anesthetized for sampling, and the serum was collected for functional detection, and the heart, lung, liver and kidney tissues were taken for pathological staining (hematoxylin-eosin staining). The results are shown in Figures 1-2 .

[0078] According to Figure 1It can be seen that compared with the control group, the level of lactate dehydrogenase (LDH) in the serum of the sepsis model group animals was significantly increased, indicating that LPS caused multiple organ damage such as heart, liver, kidney and the like in animals, but after blood perfusion, the level of LDH in the serum of sepsis animals was significantly reduced, indicating that blood perfusion can significantly reduce the damage of multiple organs such as heart, liver and kidney; further detection of liver function, kidney function indicators, such as Figure 1 As shown, compared with the control group, the content of blood urea nitrogen (BUN), serum creatinine (SCr), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the serum of the sepsis model group animals was significantly increased, and after blood perfusion, the above indicators were significantly reduced, fully indicating that blood perfusion can alleviate liver and kidney dysfunction caused by sepsis.

[0079] According to Figure 2 It can be seen that the hematoxylin-eosin staining results show that compared with the control group, the heart, lung, liver and kidney of the sepsis model animals have significant inflammatory reactions (as shown by the arrows in the figure), and after blood perfusion, the inflammation in the above organs of the animals is significantly alleviated.

[0080] Example 2

[0081] 8.322g of CTAB was weighed, and a magnetic stirrer was used to dissolve it uniformly in 50mL of deionized water at a speed of 400rpm to prepare a CTAB solution with a concentration of 0.5mol / L. While stirring, a 1mol / L sodium hydroxide solution was slowly added dropwise, and the pH value of the CTAB solution was adjusted to 12 at a speed of 2 drops per second. The stirring was continued for 30 minutes to form stable CTAB micelles, and a template solution was obtained.

[0082] 13.5g of glucose was heated to 60℃ and dissolved in 30mL of deionized water to prepare a carbon source solution with a concentration of 1.5mol / L. The carbon source solution was added to the template solution at a rate of 50mL / s through a constant pressure dropping funnel, and stirring was performed for 1 hour to fully disperse it, and a mixed solution was obtained.

[0083] The mixed solution was transferred to a 100mL reaction kettle with a filling degree of 80%, and after sealing, it was placed in an oven at 220℃ for 12 hours for hydrothermal carbonization reaction of glucose. After the reaction, it was naturally cooled to room temperature, and the product was centrifuged at a speed of 10000rpm for 10 minutes. After separation, the supernatant was discarded, and the precipitate was washed with deionized water and ethanol for 5 times, and after each washing, it was ultrasonically dispersed for 3 minutes to remove impurities, and a nanocarbon sphere precursor was obtained.

[0084] The nanometer carbon sphere precursor was dried in an oven with a gas-permeable partition at 80°C for 12 hours, and then was annealed in a tube furnace at 800°C for 1 hour under the protection of argon gas with a flow rate of 150 mL / min. The nanometer carbon spheres with a diameter of 500 microns and a specific surface area of 800 m 2 / g and abundant surface micropores (micropore diameter of 1-5 nm) were obtained.

[0085] Test Example 2

[0086] The nanometer microspheres prepared in Example 2 were filled in an adsorption column of a blood perfusion device, and the blood of an animal was passed through at a flow rate of 200 mL / min. The results showed that the clearance rate of lipopolysaccharide in the blood of the animal was 85%, and the influence on other beneficial components was less than 10%.

[0087] Test Example 3

[0088] The nanometer carbon spheres prepared in Example 1 were precisely controlled in pore size by a liquid-phase carbon deposition method, and then were analyzed by X-ray photoelectron spectroscopy. The X-ray photoelectron spectroscopy of O1s orbit was obtained, and the results are shown in Figure 3 .

[0089] According to Figure 3 The oxygen-related chemical states on the surface of the nanometer carbon spheres were analyzed. The blue peak at a binding energy of 531.01 eV was attributed to the oxygen in the C-OH group, the red peak at 532.70 eV corresponded to the oxygen in the C-O-C structure, and the green peak at 534.03 eV was attributed to the oxygen in the C=O group.

[0090] Test Example 4

[0091] The nanometer carbon spheres prepared in Example 1 were precisely controlled in pore size by a liquid-phase carbon deposition method, and then were analyzed by pore size measurement. The results are shown in Figure 4 .

[0092] According to Figure 4 It can be seen that the pore size distribution curve shows the pore size distribution of the nanometer carbon spheres of Example 1. The curve shows that there is a significant peak in the pore volume in the small pore size region, indicating that there are a large number of pore structures in the nanometer carbon spheres in this pore size range.

[0093] Test Example 5

[0094] The nanometer carbon spheres prepared in Example 1 were precisely controlled in pore size by a liquid-phase carbon deposition method, and then were analyzed by scanning electron microscopy (SEM). The results are shown in Figures 5-6 .

[0095] According to Figures 5-6It can be seen that a large number of spherical particles can be observed in the image, and the particle size distribution has certain difference, and the particle dispersion is relatively uniform when the magnification is 30kX; the image presents a highly porous microstructure when the magnification is 1000kX, the pores interweave with each other to form a complex network; the fine porous structure has important influence on specific surface area, adsorption performance and ion transmission of the material and the like.

[0096] From the above examples, the nanometer carbon sphere provided by the application can be used as or for preparing an organ / tissue toxin scavenging reagent to perform directional scavenging on toxins and impurities in animal blood.

[0097] Although the above examples make a detailed description of the application, it is only a part of the examples of the application, not all the examples, and other examples can be obtained under the premise of no creativity according to the examples, and these examples all belong to the protection scope of the application.

Claims

1. A type of carbon nanosphere, characterized in that, Diameter ranges from 100 to 500 micrometers, with a specific surface area of ​​300 to 800 m². 2 / g; the surface of the carbon nanospheres has a microporous structure.

2. The nanocarbon spheres according to claim 1, characterized in that, The pore size of the micropores in the carbon nanospheres is 1–5 nanometers.

3. The carbon nanospheres according to claim 1 or 2, characterized in that, The elemental composition of the carbon nanospheres includes C, N, and O.

4. The method for preparing the carbon nanospheres according to any one of claims 1 to 3, characterized in that, Includes the following steps: (1) Mix the surfactant, water and pH adjuster to obtain the template solution; (2) After mixing the carbon source solution and the template solution and carrying out a hydrothermal carbonization reaction, solid-liquid separation is performed to obtain the precursor of carbon nanospheres; (3) Anneal the precursor of the carbon nanospheres to obtain the carbon nanospheres.

5. The preparation method according to claim 4, characterized in that, The surfactant is an organic ammonium salt.

6. The preparation method according to claim 4, characterized in that, The standard for adding the pH adjuster is to adjust the pH value of the template solution to 10-12.

7. The preparation method according to claim 4 or 5, characterized in that, The equipment for the hydrothermal carbonization reaction includes a reactor and an oven; the filling degree of the reactor is 60-80%; the temperature of the hydrothermal carbonization reaction is 180-220℃, and the holding time is 12-24 hours.

8. The preparation method according to claim 4, characterized in that, The annealing temperature is 500–800°C, and the holding time is 1–3 hours; the annealing is carried out in a protective atmosphere.

9. The use of the carbon nanospheres according to any one of claims 1 to 3 or the carbon nanospheres obtained by the preparation method according to any one of claims 4 to 8 in the preparation of organ / tissue toxin removal reagents.

10. The application according to claim 9, characterized in that, The method of application is as follows: the nano-carbon spheres are filled into the adsorption column of a blood perfusion device, and animal blood is passed through the adsorption column at a flow rate of 50-200 mL / min.

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