WO2020042200A1 - Blood lead remover for removing blood lead in vitro - Google Patents

Blood lead remover for removing blood lead in vitro Download PDF

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WO2020042200A1
WO2020042200A1 PCT/CN2018/103733 CN2018103733W WO2020042200A1 WO 2020042200 A1 WO2020042200 A1 WO 2020042200A1 CN 2018103733 W CN2018103733 W CN 2018103733W WO 2020042200 A1 WO2020042200 A1 WO 2020042200A1
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blood
lead
adsorbent
blood lead
magnetic
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PCT/CN2018/103733
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French (fr)
Chinese (zh)
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毛春
万密密
沈健
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南京师范大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61M—DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY; DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY; DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR
    • A61M1/00—Suction or pumping devices for medical purposes; Devices for carrying-off, for treatment of, or for carrying-over, body-liquids; Drainage systems
    • A61M1/36—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits
    • A61M1/3679—Other treatment of blood in a by-pass of the natural circulatory system, e.g. temperature adaptation, irradiation ; Extra-corporeal blood circuits by absorption
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/02—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material
    • B01J20/10—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate
    • B01J20/103—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising silica or silicate comprising silica
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/28—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties
    • B01J20/28002—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof characterised by their form or physical properties characterised by their physical properties
    • B01J20/28009—Magnetic properties
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
    • B01J20/30—Processes for preparing, regenerating, or reactivating
    • B01J20/305—Addition of material, later completely removed, e.g. as result of heat treatment, leaching or washing, e.g. for forming pores
    • B01J20/3057—Use of a templating or imprinting material ; filling pores of a substrate or matrix followed by the removal of the substrate or matrix

Definitions

  • the invention belongs to a biomedical instrument, and particularly relates to a blood lead-clearing instrument for removing blood lead in vitro.
  • the clinical treatment methods for blood lead poisoning can be divided into taking lead-driving drugs and blood perfusion;
  • the treatment methods for mild blood lead poisoning are taking small-molecule lead-driving drugs such as disodium calcium edetate, dimercaptopropanol, two Mercaptosuccinic acid, long treatment cycle (40-60 days), large side effects and poor treatment effect;
  • the treatment method for severe blood lead poisoning is hemoperfusion, which uses blood perfusion device containing resin or activated carbon with adsorption function to treat blood. Wash to absorb toxic substances in the serum. Because the size of the adsorbent in the blood perfusion device is large and fixed inside the perfusion device, it cannot enter the red blood cells to remove lead ions in the red blood cells.
  • the blood perfusion generally removes toxic substances in the serum, and it is almost impossible to remove lead ions in the red blood cells carry out. Therefore, there is an urgent need for a new type of blood lead-clearing instrument whose adsorbent can freely enter and exit red blood cells to capture lead ions in the red blood cells.
  • the blood lead-clearing instrument needs to provide sufficient space and time for the contact between the adsorbent and blood, and can quickly The adsorbent is separated from the blood to achieve safe and efficient removal of lead ions from the blood (especially lead ions bound to red blood cells).
  • the hemoperfusion adsorbent that is currently used to remove blood lead is immobilized, the size of the adsorbent is large (micron level), and it cannot enter red blood cells to take away lead ions in red blood cells. , And can not be in full contact with blood.
  • the design of the present invention highlights a contact device and a magnetic separation device, which provide sufficient space and time (more than 10 minutes) for the adsorbent to freely enter and exit the red blood cells.
  • the adsorbent can freely enter and exit the red blood cells, capture lead ions in the red blood cells, and then magnetically separate them. Achieve separation of blood and adsorbent.
  • the blood lead-clearing instrument of the present invention realizes safe and efficient removal of lead ions in blood, especially lead ions bound to red blood cells.
  • a blood lead-clearing instrument which can be used to remove blood lead in vitro according to the present invention includes an extracorporeal circulation machine, an adsorbent sampling device, a contact device, and a magnetic separation device connected in sequence;
  • the adsorbent injection device contains an adsorbent, and the injection amount of the adsorbent can be controlled by adjusting the injection rate;
  • the composition of the adsorbent is a magnetic trioxide with a regular mesoporous channel using amine-rich organic compounds as a template agent Iron / aminated mesoporous silica composite.
  • the extracorporeal circulation machine and the adsorbent sampling device are connected to the contact device through a Y-shaped tube.
  • the contact device is composed of a blood perfusion tube, and the aspect ratio of the blood perfusion tube is 300: 1-1000: 1.
  • the magnetic separation device is composed of an external magnetic field and a separation tube, and the aspect ratio of the separation tube is 50: 1-200: 1.
  • an external magnetic field is formed by a magnet surrounded by a separation tube.
  • the diameter of the separation tube is larger than the diameter of the blood perfusion tube to separate the adsorbent from the blood.
  • the size of the adsorbent is 50-500 nm, and the ratio of the adsorbent to blood volume is 1: 10-10: 1g / L.
  • the blood lead-clearing instrument for removing blood lead in vitro includes the following steps:
  • step (2) Mix the Fe 3 O 4 solution prepared in step (1) with hydrazine hydrate uniformly, sonicate it, add tetraethyl orthosilicate, collect the obtained product, and re-disperse it to contain hydrazine hydrate and cetyltrimethyl Ammonium bromide in deionized water, add tetraethyl orthosilicate after ultrasonication again;
  • step (3) The product obtained in the last reaction of step (2) is dried and calcined to remove cetyltrimethylammonium bromide, and the product obtained is collected and dispersed in deionized water containing hydrazine hydrate and amine-rich organic template, and ultrasonic After the treatment, tetraethyl orthosilicate was added to the reaction, and the reaction was continued to obtain a product magnetic iron trioxide / aminated mesoporous silica composite blood lead ion adsorbent.
  • the reaction temperature in step (1) is 150-220 ° C, and the reaction time is 10-24 h.
  • reaction temperature for each addition of tetraethyl orthosilicate described in step (2) and step (3) is 60-90 ° C, and the reaction time is 1 to 5 hours.
  • the molar ratio of the tetraethyl orthosilicate to the amine-rich organic template is 5: 1 to 50: 1.
  • the amine-rich organic substance used as a template for the synthesis of the adsorbent includes one of hyperbranched polyamide, polylysine, or chitosan.
  • the process of steps (2) and (3) is: dissolving sodium lauroyl sarcosinate in citric acid / sodium citrate (0.1M, pH 5.2) In the buffer solution, add ⁇ -polylysine to the solution to generate polylysine / sodium lauroyl sarcosinate complex micelles; add Fe 3 O 4 prepared in step (1) to citric acid / In a sodium citrate buffer solution, add a polylysine / sodium lauroyl sarcosinate complex micelle solution; then add tetraethyl orthosilicate and APTES (3-aminopropyltriethoxysilane) in sequence, and continue The product was stirred and transferred to a high-temperature reaction kettle at 80 ° C. for 8 hours to obtain a magnetic iron trioxide / polylysine mesoporous silica composite blood lead ion adsorbent.
  • a blood lead-clearing instrument that can be used to remove blood lead in vitro, wherein the mechanism of removing the lead ions in the blood by the adsorbent in the whole blood lead-clearing instrument is that the composite material enters and exits red blood cells freely and passes through mesoporous silica
  • the complexation of lead ions by the amino group in the structure captures lead-containing hemoglobin, and fixes the lead-containing hemoglobin through the mesoporous channels to achieve the removal of lead ions in red blood cells.
  • the present invention can be used for a blood lead removal instrument for removing blood lead in vitro, including an extracorporeal circulation machine, an adsorbent sampling device, a contact device (blood perfusion tube), and a magnetic separation device (plus a magnetic field and a separation tube).
  • a blood lead removal instrument for removing blood lead in vitro, including an extracorporeal circulation machine, an adsorbent sampling device, a contact device (blood perfusion tube), and a magnetic separation device (plus a magnetic field and a separation tube).
  • the blood lead clearer first connects the venous vessels of lead poisoning model pigs, and the blood enters the extracorporeal circulation machine 1 through the catheter to drive the blood circulation throughout the treatment process.
  • the extracorporeal circulation machine 1 and the adsorbent injection device 2 use Y
  • the catheter is connected to the blood perfusion tube 3 in the contact device, and then the blood is contacted and mixed with the adsorbent pushed out by the adsorbent injection device 2 through the Y-shaped catheter, and the adsorbent and blood are fully contacted in the blood perfusion tube (more than 10 minutes)
  • the diameter of the separation tube is larger than the diameter of the blood perfusion tube to achieve the separation of the adsorbent from the blood.
  • the blood flows back through the artery to the lead poisoning model pig and is completed once. Blood lead removal process, continue to repeat the cycle to clear.
  • the present invention has the following advantages:
  • the use of the blood lead-clearing instrument of the present invention can directly remove lead ions in the blood in a short time through extracorporeal blood circulation, quickly alleviate the condition of patients with blood lead poisoning, and reduce the patients' daily medication.
  • the side-by-side blood lead-clearing instrument of the present invention for removing blood lead in vitro has a simple composition, is convenient to use, and can flexibly change its working position, which solves the problem that the blood perfusion device cannot remove lead ions in red blood cells in the blood, and improves work efficiency. It is suitable for widespread promotion and use, and has broad application prospects in the medical field.
  • the design of the blood lead-clearing device of the present invention highlights a contact device and a magnetic separation device, which provide sufficient space and time (over 10 minutes) for the free passage of the adsorbent into and out of the red blood cells.
  • the adsorbent can freely enter and exit the red blood cells and capture the lead in the red blood cells.
  • the ions are then separated from the adsorbent by magnetic separation.
  • the blood lead-clearing instrument of the present invention can provide sufficient space and time for the free passage of the adsorbent into and out of the red blood cells.
  • the lead-containing hemoglobin is captured by the complexation of the lead with the amino group in the mesoporous silica structure, and passes through the mesoporous channels. Fixing leaded hemoglobin to achieve the purpose of removing lead ions in red blood cells. It overcomes the shortcomings of unclear principle and low efficiency of blood lead removal in current clinical and research.
  • the blood lead-clearing instrument of the present invention can be truly used to remove lead ions complexed with hemoglobin in the content of up to 95% of red blood cells, and can safely enter red blood cells and effectively capture blood lead and hemoglobin contaminated by blood lead, and then safely leave the red blood cells. And blood.
  • the sorbent in the sorbent sampling device of the blood lead meter for blood lead removal in vitro utilizes the self-assembly performance of the amine-rich organic compounds, and the self-assembly of the amine-rich organic compounds is used to form micelles.
  • the behavior is to realize the dehydration condensation of silicon source on the surface of magnetic iron trioxide nanoparticles, and to obtain a magnetic iron trioxide / rich aminated mesoporous silica composite material using amine-rich organic compounds as a template and having regular mesoporous channels. That is, in the process of material synthesis, amine-rich organic compounds are used as template agents to realize material synthesis and functional group modification through a "one-step method".
  • the formation process of the ferric oxide core mesoporous molecular sieve and the organic functional group modification process are combined to obtain rich Aminated magnetic triiron tetroxide / aminated mesoporous silica composite material has good biocompatibility, simple preparation process, short production cycle, and effectively overcomes the complicated steps of the current technology (post-modification method) Disadvantages of energy waste and uneven distribution of functional groups.
  • the invention proposes a new blood lead-expulsion mechanism.
  • the blood lead-clearing instrument and the attached adsorbent material are used to make full contact with blood red blood cells to capture most of the lead ions in the red blood cells, thereby achieving the purpose of efficiently removing blood lead. It overcomes the shortcomings of the current clinical lead-expulsion methods, such as long cycle, large side effects, and poor effect, and has broad application prospects in the medical field.
  • the adsorbent material supported by the blood lead-clearing instrument of the present invention is magnetic iron trioxide / rich amine
  • the mesoporous silica composite material has a regular mesoporous structure and highly dispersed organic functional groups, and can freely enter and exit red blood cells.
  • the lead-containing hemoglobin is captured by the complexation of lead ions by the amino group in the mesoporous silica structure, and Fixation of leaded hemoglobin through mesoporous channels to achieve the purpose of removing lead ions from red blood cells. It overcomes the shortcomings of unclear principle and low efficiency of blood lead removal in current clinical and research.
  • Example 1 is a transmission electron microscope image of (a) Fe 3 O 4 NPs and (b) MMS / P NPs prepared in Example 1;
  • Fig. 2 is a scanning electron microscope image of (a) MMS / PNPs, and (b) an elemental energy spectrum analysis chart of MMS / PNPs;
  • Figure 3 is the infrared spectrum of (a) Fe 3 O 4 NPs and (b) MMS / P NPs;
  • Figure 4 is the nitrogen adsorption and desorption isotherm and pore size distribution of MMS / P NPs before (a) and after (b) template removal (inside);
  • FIG. 5 is a schematic diagram of the in vitro coagulation time of MMS / P NPs before and after anticoagulation treatment
  • Figure 6 is a schematic diagram of the hemolysis rate of MMS / P NPs before and after heparin loading
  • Figure 7 is a schematic diagram of the relative content of common ions in blood before and after MMS / P NPs adsorption
  • Figure 9 is (a) nitrogen adsorption and desorption curve of MMS / H NPs; (b) schematic diagram of pore size distribution;
  • Figure 10 is a schematic diagram of the adsorption effect of MMS / H NPs on lead ions in real blood (rabbit and human blood);
  • Figure 11 shows the process of MMS / H NPs entering and exiting red blood cells: (a) pure red blood cells, (b) MMS / H NPs entering red blood cells, and (c) TEM images of red blood cells after MMS / H NPs are separated from red blood cells;
  • FIG. 12 is a schematic structural diagram of a device that can be used for a blood lead remover for removing blood lead in vitro, an extracorporeal circulation machine 1, an adsorbent sampling device 2, a contact device 3 (a blood perfusion tube), a magnetic separation device (an external magnetic field 4 and separation) Tube 5);
  • FIG. 13 is a schematic diagram of a blood lead-clearing apparatus that can be used to remove blood lead in vitro;
  • FIG. 14 is a process of removing lead from blood in vitro using a blood lead-clearing instrument for removing lead from lead poisoning model pigs in vitro (i) intravenous intubation and (ii, iii) extracorporeal circulation;
  • FIG. 15 is a routine blood test of lead poisoning model pigs before and after surgery (blood lead removal).
  • ferric trichloride (1.35 g, 5 mmol) and dissolve it in 30 mL of ethylene glycol solution.
  • sodium acetate (NaAc, 3.6 g) and polyethylene glycol-2000 (PEG-2000 1.0 g) with stirring. Stirring was continued for 30 min, and the solution was transferred to a high-temperature reaction kettle at 200 ° C for 72 hours. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
  • 100 mg of sodium lauroyl sarcosinate was dissolved into 10 mL of a citric acid / sodium citrate buffer solution (0.1 M, pH 5.2) at room temperature, and 150 ⁇ L of ⁇ -polylysine (20 wt%) was added to the solution. At this time, the solution immediately became an emulsion, and a polylysine / sodium lauroyl sarcosinate complex micelle was formed.
  • Fe 3 O 4 NPs are uniformly dispersed with a size of about 120 nm. Generally, the size of Fe 3 O 4 NPs is about 50-300 nm. The size of the obtained MMS / P NPs is about 170 nm, and the size of the obtained adsorbent is usually 50-500 nm. Scanning electron microscopy was used to characterize the size and morphology of the composite.
  • Nitrogen adsorption and desorption curves are often used to reflect the pore structure of mesoporous materials. As shown in Figure 4, the pore size of MMS / PNPs is approximately 5.1 nm.
  • the composite material is a biocompatible material with a hemolysis rate of 0.5 to 5%, an activated partial thromboplastin time of 12 to 30 s, a plasma prothrombin time of 8 to 20 s, and a thrombin time of 8 to 20 s.
  • Quantitative magnetic triiron tetroxide / aminated mesoporous silica composite was placed in a centrifuge tube, 3 mL of lead-containing blood (0.6 ppm) was added, and the centrifuge tube was placed on a constant temperature shaker at 37 ° C for a fixed time. Take 2mL of the supernatant and digest the solution by high temperature digestion. After cooling, make up to 5mL.
  • Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reactor for 200 h at 200 ° C. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
  • the Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 90 ° C for 2 h.
  • the obtained product was naturally cooled, and then washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C. The dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB.
  • the obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg (0.45 mmol) of PAMAM. Sonicate for 30 min, add 0.6 mL (2.88 mmol) of TEOS, and continue stirring at 90 ° C for 2 h.
  • the particle size of MMS / H NPs is about 300 nm.
  • a nitrogen adsorption desorption curve is used to characterize the pore structure of the material.
  • the BET surface area is approximately 248 m 2 ⁇ g -1 and the pore size is approximately 24 nm.
  • Quantitative magnetic triiron tetroxide / aminated mesoporous silica composite was placed in a centrifuge tube, 3 mL of lead-containing blood (0.6 ppm) was added, and the centrifuge tube was placed on a constant temperature shaker at 37 ° C for a fixed time. Take 2mL of the supernatant and digest the solution by high temperature digestion. After cooling, make up to 5mL.
  • MMS / H NPs can enter red blood cells and can be magnetically separated without affecting the red blood cell morphology.
  • Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reactor for 200 h at 200 ° C. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
  • the Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 90 ° C for 2 h.
  • the product obtained was naturally cooled, it was washed with deionized water and ethanol solution several times in that order, and dried under vacuum at 60 ° C. The dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB.
  • the obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 3.4 g (0.0576 mmol) of chitosan. Sonicate for 30 min, add 0.6 mL (2.88 mmol) of TEOS, and continue stirring at 90 ° C for 2 h.
  • Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reaction kettle at 150 ° C. for 24 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
  • the Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 60 ° C. for 5 h.
  • the obtained product was naturally cooled, and then washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C.
  • the dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB.
  • the obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 576 mg (0.576 mmol) of PAMAM. Sonicate for 30 min, add 0.6 mL (2.88 mmol) of TEOS, and continue stirring at 90 ° C for 2 h.
  • Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reaction kettle at 220 ° C. for 10 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
  • the Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 90 ° C for 1 h.
  • the obtained product was naturally cooled, and then washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C.
  • the dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB.
  • the obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of PAMAM. Sonicate for 30 min, add 0.6 mL of TEOS, and continue stirring at 90 ° C for 2 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
  • the product was immersed in a 10 mL heparin solution (10 mg / mL) for 12 h, and dried under vacuum at 60 ° C. to obtain a magnetic lead iron ion scavenger of ferric iron tetraoxide / hyperbranched polyamide mesoporous silica composite material.
  • Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reactor for 200 h at 200 ° C. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
  • a blood lead-clearing instrument which can be used for extracorporeal blood lead removal includes an extracorporeal circulation machine 1, an adsorbent sampling device 2, a contact device 3, and a magnetic separation device connected in sequence; an extracorporeal circulation machine 1 and an adsorption device
  • the agent injection device 2 is connected to the contact device 3 through a Y-shaped tube.
  • the composition of the contact device 3 is a blood perfusion tube.
  • the aspect ratio of the blood perfusion tube is 600: 1, and the aspect ratio of the blood perfusion tube can be 300: 1- Adjusting between 1000: 1, the magnetic separation device consists of an external magnetic field 4 (magnets can be used) and the separation tube 5, that is, the separation tube 5 has a magnet, and the length-to-diameter ratio of the separation tube is 100: 1. The ratio can be adjusted between 50: 1-200: 1. The diameter of the separation tube is larger than the diameter of the blood perfusion tube to achieve the separation of the adsorbent from the blood.
  • the composition of the adsorbent in the adsorbent injection device 3 is a magnetic triiron tetroxide / aminated mesoporous silica composite material using amine-rich organics as a template and having regular mesoporous channels.
  • This embodiment specifically uses
  • the magnetic lead iron ion adsorbent prepared in Example 2 is a magnetic iron trioxide / hyperbranched polyamide mesoporous silica composite material.
  • the blood lead clearer When used, the blood lead clearer first connects the venous vessels of lead poisoning model pigs, and the blood enters the extracorporeal circulation machine 1 through the catheter to drive the blood circulation throughout the treatment process.
  • the extracorporeal circulation machine 1 and the adsorbent injection device 2 use Y
  • the catheter is connected to the blood perfusion tube in the contact device 3, and then the blood is contacted and mixed with the adsorbent introduced by the adsorbent injection device 2 through the Y-type catheter, and the adsorbent and blood are fully contacted in the blood perfusion tube (more than 10 minutes)
  • the diameter of the separation tube 5 is larger than the diameter of the blood perfusion tube to separate the adsorbent from the blood, and the blood flows back through the artery to lead poisoning
  • the model pig complete the blood lead removal process once and continue to repeat the steps to remove lead.
  • a model of lead poisoning of about 0.5 ppm was established for pigs by the food poisoning method.
  • an extracorporeal circulation machine 1 for removing blood lead in vitro from Example 7, first, an extracorporeal circulation machine 1, an adsorbent sampling device 2, a contact device 3, and a magnetic separation device were connected to construct a blood lead cleaning device. Then establish the extracorporeal circulation pathway of the pigs ( Figure 13-14) as follows: Anesthetize the pigs by injecting 3% sodium pentobarbital (1ml / kg) in the abdominal cavity, and keep anesthesia during the blood lead cleaning, and continuously inject 0.9% NaCl And 2.5% sodium pentobarbital.
  • the venous vessels are connected to a blood lead cleaning device and return to arterial blood after passing through a magnetic separation device.
  • the blood flow speed is 25ml / min
  • the circulation time is 50min
  • the amount of adsorbent is 1g
  • the blood volume of the lead poisoning model pig is about 3L
  • the ratio of adsorbent to blood volume can be between 1: 10-10: 1g / L
  • the wound was sutured after the operation, and the blood routine and blood lead removal efficiency of the model pigs with lead poisoning before and after the operation were examined.
  • the experimental results show that when the blood lead content in pigs is 500ppb, after one cleaning process, the blood lead removal efficiency can reach 75%.
  • the secondary treatment can continue to improve the blood lead removal rate (the calculation method of the removal rate is:
  • the blood before and after the operation was detected with an inductively coupled plasma spectrometer before and after the operation.
  • E is the blood lead removal efficiency (%)
  • C 0 is the blood lead concentration (ppb) before the operation
  • C 1 is the blood lead concentration (ppb) after the operation).
  • the adsorbent has a better separation effect, which shows the higher removal efficiency of the adsorbent and the blood lead remover.
  • the blood routine before and after cleaning shows that the blood inflammation is reduced after the removal of lead ions ( Figure 15), which reflects the blood The safety and efficiency of the adsorbent and blood lead remover in the process of removing blood lead.

Abstract

A blood lead remover for removing blood lead in vitro. The blood lead remover comprises: an extracorporeal circulation unit (1), an adsorbent sample feeding device (2), a contact device (3), and a magnetic separation device which are sequentially connected; an adsorbent is loaded in the adsorbent sample feeding device (2), and the composition of the adsorbent is a magnetic ferroferric oxide/rich-aminated mesoporous silica composite material which takes an amine-rich organic matter as a template agent and has a regular mesoporous channel. Compared with an oral medication, use of the blood lead remover can directly remove lead ions in the blood in a short time by means of extracorporeal blood circulation, quickly alleviate the condition of a patient suffering from blood lead poisoning, and reduce side effects caused by daily medications. The blood lead remover has the beneficial effects that the structure is simple, the use is convenient, and a working position can be flexibly changed, thereby solving the problem that a blood perfusion device cannot remove lead ions in red blood cells in the blood, improving working efficiency, being suitable for wide application, and having a wide application prospect in the medical field.

Description

一种用于体外去除血铅的血液清铅仪Blood lead-clearing instrument for removing blood lead in vitro 技术领域Technical field
本发明属于生物医学仪器，具体涉及一种用于体外去除血铅的血液清铅仪。The invention belongs to a biomedical instrument, and particularly relates to a blood lead-clearing instrument for removing blood lead in vitro.
背景技术Background technique
人体健康受到铅离子暴露的极大威胁，铅离子会引起严重的毒性作用和疾病(如心脏病和肾病等)。当铅离子进入人体时，它主要沉积在骨骼、软组织和血液中。基于当今的生物医学水平，很难去除沉积在人体骨骼和软组织中的铅离子。因此，去除血液中痕量的铅离子(血铅)具有重要意义。然而，95％以上的血液铅离子位于红细胞中与血红蛋白结合，这造成了血铅去除的一大困难。目前临床上对血铅中毒的治疗方法可分为服用驱铅药物和血液灌流；轻度血铅中毒的治疗方法为服用小分子驱铅药物如依地酸二钠钙、二巯基丙醇、二巯基丁二酸，治疗周期长(40-60天)、副作用大且治疗效果差；重度血铅中毒的治疗方法为血液灌流，血液灌流是使用含有吸附功能的树脂或活性炭的血液灌流器对血液进行清洗，吸附血清中的有毒物质。由于血液灌流器中吸附剂尺寸较大且固定在灌流器内部，无法进入红细胞中去除红细胞中的铅离子，因此血液灌流一般是去除血清中的有毒物质，对去除红细胞中的铅离子几乎不可能完成。因此，亟需一种新型血液清铅仪，其吸附剂能够自由进出红细胞，捕获红细胞中的铅离子，且血液清铅仪需为吸附剂和血液接触提供足够的空间和时间，并能快速将吸附剂与血液进行分离，以实现安全高效地去除血液中的铅离子(尤其是与红细胞结合的铅离子)。Human health is greatly threatened by exposure to lead ions, which can cause serious toxic effects and diseases (such as heart disease and kidney disease). When lead ions enter the body, it is mainly deposited in bones, soft tissues and blood. At today's biomedical level, it is difficult to remove lead ions deposited in human bones and soft tissues. Therefore, it is of great significance to remove trace amounts of lead ions (blood lead) in the blood. However, more than 95% of blood lead ions are located in red blood cells and bind to hemoglobin, which causes a major difficulty in removing blood lead. At present, the clinical treatment methods for blood lead poisoning can be divided into taking lead-driving drugs and blood perfusion; the treatment methods for mild blood lead poisoning are taking small-molecule lead-driving drugs such as disodium calcium edetate, dimercaptopropanol, two Mercaptosuccinic acid, long treatment cycle (40-60 days), large side effects and poor treatment effect; The treatment method for severe blood lead poisoning is hemoperfusion, which uses blood perfusion device containing resin or activated carbon with adsorption function to treat blood. Wash to absorb toxic substances in the serum. Because the size of the adsorbent in the blood perfusion device is large and fixed inside the perfusion device, it cannot enter the red blood cells to remove lead ions in the red blood cells. Therefore, the blood perfusion generally removes toxic substances in the serum, and it is almost impossible to remove lead ions in the red blood cells carry out. Therefore, there is an urgent need for a new type of blood lead-clearing instrument whose adsorbent can freely enter and exit red blood cells to capture lead ions in the red blood cells. The blood lead-clearing instrument needs to provide sufficient space and time for the contact between the adsorbent and blood, and can quickly The adsorbent is separated from the blood to achieve safe and efficient removal of lead ions from the blood (especially lead ions bound to red blood cells).
发明内容Summary of the Invention
发明目的：针对现有技术存在的问题，即目前常用于去除血铅的血液灌流器吸附剂是固定不动的，吸附剂尺寸较大(微米级)，无法进入红细胞带走红细胞中的铅离子，也不能与血液充分接触。本发明设计中突出了一个接触装置和磁性分离装置，为吸附剂自由进出红细胞提供足够的空间和时间(10分钟以上)，吸附剂能够自由进出红细胞，捕获红细胞中的铅离子，然后通过磁性分离实现血液与吸附剂的分离。本发明的血液清铅仪实现安全高效地去除血液中的铅离子，尤其是与红细胞结合的铅离子。Object of the invention: In view of the problems existing in the prior art, the hemoperfusion adsorbent that is currently used to remove blood lead is immobilized, the size of the adsorbent is large (micron level), and it cannot enter red blood cells to take away lead ions in red blood cells. , And can not be in full contact with blood. The design of the present invention highlights a contact device and a magnetic separation device, which provide sufficient space and time (more than 10 minutes) for the adsorbent to freely enter and exit the red blood cells. The adsorbent can freely enter and exit the red blood cells, capture lead ions in the red blood cells, and then magnetically separate them. Achieve separation of blood and adsorbent. The blood lead-clearing instrument of the present invention realizes safe and efficient removal of lead ions in blood, especially lead ions bound to red blood cells.
技术方案：为了实现上述目的，如本发明所述一种可用于体外去除血铅的血液清铅仪，包括依次连接的体外循环机、吸附剂进样装置、接触装置、磁性分离装置；所述吸附剂进样装置中含有吸附剂，可通过调节进样速率控制吸附剂的进样量；所述吸附剂的成分为以富胺类有机物为模板剂且具有规整介孔孔道的磁性 四氧化三铁/富胺化介孔二氧化硅复合材料。Technical solution: In order to achieve the above-mentioned purpose, a blood lead-clearing instrument which can be used to remove blood lead in vitro according to the present invention includes an extracorporeal circulation machine, an adsorbent sampling device, a contact device, and a magnetic separation device connected in sequence; The adsorbent injection device contains an adsorbent, and the injection amount of the adsorbent can be controlled by adjusting the injection rate; the composition of the adsorbent is a magnetic trioxide with a regular mesoporous channel using amine-rich organic compounds as a template agent Iron / aminated mesoporous silica composite.
其中，所述体外循环机和吸附剂进样装置通过Y型管与接触装置连接。Wherein, the extracorporeal circulation machine and the adsorbent sampling device are connected to the contact device through a Y-shaped tube.
其中，所述接触装置的组成为血液灌注管，血液灌注管的长径比为300:1-1000：1。The contact device is composed of a blood perfusion tube, and the aspect ratio of the blood perfusion tube is 300: 1-1000: 1.
其中，所述磁性分离装置由外加磁场和分离管组成，分离管的长径比为50:1-200:1。所述磁性分离装置是由分离管外套有磁铁形成外加磁场，分离管口径大于血液灌注管的口径以实现吸附剂与血液的分离。The magnetic separation device is composed of an external magnetic field and a separation tube, and the aspect ratio of the separation tube is 50: 1-200: 1. In the magnetic separation device, an external magnetic field is formed by a magnet surrounded by a separation tube. The diameter of the separation tube is larger than the diameter of the blood perfusion tube to separate the adsorbent from the blood.
其中，所述吸附剂尺寸为50-500nm，吸附剂与血液体积比例为1:10-10:1g/L。Wherein, the size of the adsorbent is 50-500 nm, and the ratio of the adsorbent to blood volume is 1: 10-10: 1g / L.
本发明所述的可用于体外去除血铅的血液清铅仪，所述吸附剂的制备方法，包括以下步骤：The blood lead-clearing instrument for removing blood lead in vitro according to the present invention, and the preparation method of the adsorbent includes the following steps:
(1)称取六水合三氯化铁溶解于乙二醇溶液中，加入醋酸钠和三水合柠檬酸钠或聚乙二醇继续反应，得到Fe 3O 4溶液； (1) Weigh the ferric trichloride hexahydrate dissolved in the ethylene glycol solution, add sodium acetate and sodium citrate trihydrate or polyethylene glycol to continue the reaction to obtain a Fe 3 O 4 solution;
(2)将步骤(1)制得的Fe 3O 4溶液与水合肼混合均匀，超声处理，加入正硅酸四乙酯，收集所得产物，重新分散到包含水合肼和十六烷基三甲基溴化铵的去离子水中，再次超声后加入正硅酸四乙酯反应； (2) Mix the Fe 3 O 4 solution prepared in step (1) with hydrazine hydrate uniformly, sonicate it, add tetraethyl orthosilicate, collect the obtained product, and re-disperse it to contain hydrazine hydrate and cetyltrimethyl Ammonium bromide in deionized water, add tetraethyl orthosilicate after ultrasonication again;
(3)将步骤(2)最后反应得到的产物干燥后煅烧去除十六烷基三甲基溴化铵，收集获得的产物分散到包含水合肼和富胺类有机物模板剂的去离子水中，超声处理后加入正硅酸四乙酯反应，继续反应，得到产物磁性四氧化三铁/富胺化介孔二氧化硅复合材料血液铅离子吸附剂。(3) The product obtained in the last reaction of step (2) is dried and calcined to remove cetyltrimethylammonium bromide, and the product obtained is collected and dispersed in deionized water containing hydrazine hydrate and amine-rich organic template, and ultrasonic After the treatment, tetraethyl orthosilicate was added to the reaction, and the reaction was continued to obtain a product magnetic iron trioxide / aminated mesoporous silica composite blood lead ion adsorbent.
作为优选，步骤(1)所述反应温度为150-220℃，反应时间为10-24h。Preferably, the reaction temperature in step (1) is 150-220 ° C, and the reaction time is 10-24 h.
其中，步骤(2)和步骤(3)所述每次加入正硅酸四乙酯的反应温度为60-90℃，反应时间为1～5h。Wherein, the reaction temperature for each addition of tetraethyl orthosilicate described in step (2) and step (3) is 60-90 ° C, and the reaction time is 1 to 5 hours.
作为优选，步骤(3)所述正硅酸四乙酯与富胺类有机物模板剂的摩尔比为5：1～50：1。Preferably, the molar ratio of the tetraethyl orthosilicate to the amine-rich organic template is 5: 1 to 50: 1.
进一步地，所述吸附剂合成所用的以富胺类有机物为模板剂包含超支化聚酰胺、聚赖氨酸或壳聚糖中的一种。Further, the amine-rich organic substance used as a template for the synthesis of the adsorbent includes one of hyperbranched polyamide, polylysine, or chitosan.
其中，所述富胺类有机物模板剂为聚赖氨酸时，所述步骤(2)和(3)过程为：将月桂酰肌氨酸钠溶解在柠檬酸/柠檬酸钠(0.1M,pH 5.2)缓冲溶液中，向溶液中加入ε-聚赖氨酸，生成聚赖氨酸/月桂酰肌氨酸钠复合胶束；将步骤(1)所制备的Fe 3O 4加入到柠檬酸/柠檬酸钠缓冲溶液中，并加入聚赖氨酸/月桂酰肌氨酸钠复合胶束溶液；再依次加入正硅酸四乙酯和APTES(3-氨丙基三乙氧基硅烷)，继续搅拌，将产物转移到高温反应釜中80℃反应8h，得到磁性四氧化三铁/聚赖氨酸介孔二氧化硅复合材料血液铅离子吸附剂。 Wherein, when the amine-rich organic template is polylysine, the process of steps (2) and (3) is: dissolving sodium lauroyl sarcosinate in citric acid / sodium citrate (0.1M, pH 5.2) In the buffer solution, add ε-polylysine to the solution to generate polylysine / sodium lauroyl sarcosinate complex micelles; add Fe 3 O 4 prepared in step (1) to citric acid / In a sodium citrate buffer solution, add a polylysine / sodium lauroyl sarcosinate complex micelle solution; then add tetraethyl orthosilicate and APTES (3-aminopropyltriethoxysilane) in sequence, and continue The product was stirred and transferred to a high-temperature reaction kettle at 80 ° C. for 8 hours to obtain a magnetic iron trioxide / polylysine mesoporous silica composite blood lead ion adsorbent.
本发明所述的可用于体外去除血铅的血液清铅仪，其中，所述吸附剂在整个血液清铅仪中去除血液中铅离子机制过程为复合材料自由进出红细胞，通过介孔二氧化硅结构中的氨基对铅离子的络合作用捕获含铅血红蛋白，并通过介孔孔道对含铅血红蛋白进行固定，实现去除红细胞中铅离子。According to the present invention, a blood lead-clearing instrument that can be used to remove blood lead in vitro, wherein the mechanism of removing the lead ions in the blood by the adsorbent in the whole blood lead-clearing instrument is that the composite material enters and exits red blood cells freely and passes through mesoporous silica The complexation of lead ions by the amino group in the structure captures lead-containing hemoglobin, and fixes the lead-containing hemoglobin through the mesoporous channels to achieve the removal of lead ions in red blood cells.
工作原理：本发明可用于体外去除血铅的血液清铅仪，包括体外循环机、吸附剂进样装置、接触装置(血液灌注管)、磁性分离装置(外加磁场和分离管)。使用时血液清铅仪通过首先连接铅中毒模型猪的静脉血管，血液经过导管进入体外循环机1用以带动整个治疗过程的血液循环，体外循环机1和吸附剂进样装置2通过将使用Y型导管连接至接触装置中的血液灌注管3，随后血液通过Y型导管与吸附剂进样装置2推出的吸附剂接触混合，吸附剂和血液在血液灌注管中进行充分接触(10分钟以上)，后血液过血液灌注管3至套有磁铁4的分离管5中，分离管口径大于血液灌注管的口径以实现吸附剂与血液的分离，血液通过动脉流回铅中毒模型猪上，完成一次血液除铅过程，继续重复循环清除。Working principle: The present invention can be used for a blood lead removal instrument for removing blood lead in vitro, including an extracorporeal circulation machine, an adsorbent sampling device, a contact device (blood perfusion tube), and a magnetic separation device (plus a magnetic field and a separation tube). When used, the blood lead clearer first connects the venous vessels of lead poisoning model pigs, and the blood enters the extracorporeal circulation machine 1 through the catheter to drive the blood circulation throughout the treatment process. The extracorporeal circulation machine 1 and the adsorbent injection device 2 use Y The catheter is connected to the blood perfusion tube 3 in the contact device, and then the blood is contacted and mixed with the adsorbent pushed out by the adsorbent injection device 2 through the Y-shaped catheter, and the adsorbent and blood are fully contacted in the blood perfusion tube (more than 10 minutes) After the blood passes through the blood perfusion tube 3 to the separation tube 5 with the magnet 4, the diameter of the separation tube is larger than the diameter of the blood perfusion tube to achieve the separation of the adsorbent from the blood. The blood flows back through the artery to the lead poisoning model pig and is completed once. Blood lead removal process, continue to repeat the cycle to clear.
有益效果：与现有技术相比，本发明具有以下优点：Beneficial effect: Compared with the prior art, the present invention has the following advantages:
(1)本发明的血液清铅仪的使用相比于口服药物，通过体外血液循环可在短时间内直接清除血液中的铅离子，快速缓解血铅中毒病人病情，减少病人每天服用药物带来的副作用，本发明的用于体外去除血铅的血液清铅仪，组成简单，使用方便，可灵活变动其工作位置，解决了血液灌流器无法去除血液中红细胞内铅离子的问题，提高工作效率，适用于广泛推广使用，在医学领域有广阔的应用前景。本发明的血液清铅仪装置设计中突出了一个接触装置和磁性分离装置，为吸附剂自由进出红细胞提供足够的空间和时间(10分钟以上)，吸附剂能够自由进出红细胞，捕获红细胞中的铅离子，然后通过磁性分离实现血液与吸附剂的分离。(1) Compared with oral drugs, the use of the blood lead-clearing instrument of the present invention can directly remove lead ions in the blood in a short time through extracorporeal blood circulation, quickly alleviate the condition of patients with blood lead poisoning, and reduce the patients' daily medication. The side-by-side blood lead-clearing instrument of the present invention for removing blood lead in vitro has a simple composition, is convenient to use, and can flexibly change its working position, which solves the problem that the blood perfusion device cannot remove lead ions in red blood cells in the blood, and improves work efficiency. It is suitable for widespread promotion and use, and has broad application prospects in the medical field. The design of the blood lead-clearing device of the present invention highlights a contact device and a magnetic separation device, which provide sufficient space and time (over 10 minutes) for the free passage of the adsorbent into and out of the red blood cells. The adsorbent can freely enter and exit the red blood cells and capture the lead in the red blood cells. The ions are then separated from the adsorbent by magnetic separation.
此外，本发明的血液清铅仪能够为吸附剂自由进出红细胞提供足够的空间和时间，通过介孔二氧化硅结构中的氨基对铅离子的络合作用捕获含铅血红蛋白，并通过介孔孔道对含铅血红蛋白进行固定，实现去除红细胞中铅离子的目的。克服了当前临床和研究中血铅去除原理不清、效率不高的缺点。本发明的血液清铅仪可以真正用来去除红细胞中含量高达95％的与血红蛋白络合的铅离子，并且能够安全进入红细胞并有效捕获血铅和被血铅污染的血红蛋白，然后安全地离开红细胞和血液。In addition, the blood lead-clearing instrument of the present invention can provide sufficient space and time for the free passage of the adsorbent into and out of the red blood cells. The lead-containing hemoglobin is captured by the complexation of the lead with the amino group in the mesoporous silica structure, and passes through the mesoporous channels. Fixing leaded hemoglobin to achieve the purpose of removing lead ions in red blood cells. It overcomes the shortcomings of unclear principle and low efficiency of blood lead removal in current clinical and research. The blood lead-clearing instrument of the present invention can be truly used to remove lead ions complexed with hemoglobin in the content of up to 95% of red blood cells, and can safely enter red blood cells and effectively capture blood lead and hemoglobin contaminated by blood lead, and then safely leave the red blood cells. And blood.
(2)本发明的用于体外去除血铅的血液清铅仪的吸附剂进样装置中的吸附剂利用富胺类有机物的自组装性能，制备时利用富胺类有机物自组装形成胶束的行为实现硅源在磁性四氧化三铁纳米粒子表面的脱水缩合，得到以富胺类有机物 为模板剂且具有规整介孔孔道的磁性四氧化三铁/富胺化介孔二氧化硅复合材料，即在材料合成过程中富胺类有机物作为模板剂，通过“一步法”实现材料合成、官能团修饰等过程，将四氧化三铁核外介孔分子筛形成过程与有机官能团改性过程相融合，获得富胺化的磁性四氧化三铁/富胺化介孔二氧化硅复合材料，得到的复合材料生物相容性好，制备工艺简单，生产周期短，有效克服当前技术(后修饰法)的步骤繁琐、能源浪费且官能团分布不均匀的缺点。(2) The sorbent in the sorbent sampling device of the blood lead meter for blood lead removal in vitro according to the present invention utilizes the self-assembly performance of the amine-rich organic compounds, and the self-assembly of the amine-rich organic compounds is used to form micelles. The behavior is to realize the dehydration condensation of silicon source on the surface of magnetic iron trioxide nanoparticles, and to obtain a magnetic iron trioxide / rich aminated mesoporous silica composite material using amine-rich organic compounds as a template and having regular mesoporous channels. That is, in the process of material synthesis, amine-rich organic compounds are used as template agents to realize material synthesis and functional group modification through a "one-step method". The formation process of the ferric oxide core mesoporous molecular sieve and the organic functional group modification process are combined to obtain rich Aminated magnetic triiron tetroxide / aminated mesoporous silica composite material has good biocompatibility, simple preparation process, short production cycle, and effectively overcomes the complicated steps of the current technology (post-modification method) Disadvantages of energy waste and uneven distribution of functional groups.
(3)本发明提出新型血液驱铅机制，利用血液清铅仪及其附带的吸附剂材料与血液红细胞充分接触，捕获血液红细胞中的大部分铅离子，从而实现高效去除血铅的目的，有效克服当前临床上驱铅方法存在的周期长、副作用大、效果差的缺点，在医学领域有广阔的应用前景；此外，本发明血液清铅仪负载的吸附剂材料磁性四氧化三铁/富胺化介孔二氧化硅复合材料，具有规整的介孔结构和高度分散的有机官能团，能够自由进出红细胞，通过介孔二氧化硅结构中的氨基对铅离子的络合作用捕获含铅血红蛋白，并通过介孔孔道对含铅血红蛋白进行固定，实现去除红细胞中铅离子的目的。克服了当前临床和研究中血铅去除原理不清、效率不高的缺点。(3) The invention proposes a new blood lead-expulsion mechanism. The blood lead-clearing instrument and the attached adsorbent material are used to make full contact with blood red blood cells to capture most of the lead ions in the red blood cells, thereby achieving the purpose of efficiently removing blood lead. It overcomes the shortcomings of the current clinical lead-expulsion methods, such as long cycle, large side effects, and poor effect, and has broad application prospects in the medical field. In addition, the adsorbent material supported by the blood lead-clearing instrument of the present invention is magnetic iron trioxide / rich amine The mesoporous silica composite material has a regular mesoporous structure and highly dispersed organic functional groups, and can freely enter and exit red blood cells. The lead-containing hemoglobin is captured by the complexation of lead ions by the amino group in the mesoporous silica structure, and Fixation of leaded hemoglobin through mesoporous channels to achieve the purpose of removing lead ions from red blood cells. It overcomes the shortcomings of unclear principle and low efficiency of blood lead removal in current clinical and research.
附图说明BRIEF DESCRIPTION OF THE DRAWINGS
图1为实施例1中制备的(a)Fe 3O 4NPs，(b)MMS/P NPs的透射电镜图； 1 is a transmission electron microscope image of (a) Fe 3 O 4 NPs and (b) MMS / P NPs prepared in Example 1;
图2为(a)MMS/P NPs的扫描电镜图，(b)MMS/P NPs的元素能谱分析图；Fig. 2 is a scanning electron microscope image of (a) MMS / PNPs, and (b) an elemental energy spectrum analysis chart of MMS / PNPs;
图3为(a)Fe 3O 4NPs和(b)MMS/P NPs的红外光谱图； Figure 3 is the infrared spectrum of (a) Fe 3 O 4 NPs and (b) MMS / P NPs;
图4为模板去除(a)前、(b)后MMS/P NPs的氮气吸附脱附等温线和孔尺寸分布图(内附)；Figure 4 is the nitrogen adsorption and desorption isotherm and pore size distribution of MMS / P NPs before (a) and after (b) template removal (inside);
图5为抗凝血处理前后MMS/P NPs的体外凝血时间示意图；5 is a schematic diagram of the in vitro coagulation time of MMS / P NPs before and after anticoagulation treatment;
图6为负载肝素前后MMS/P NPs的溶血率示意图；Figure 6 is a schematic diagram of the hemolysis rate of MMS / P NPs before and after heparin loading;
图7为MMS/P NPs吸附前后血液中常见离子的相对含量比较示意图；Figure 7 is a schematic diagram of the relative content of common ions in blood before and after MMS / P NPs adsorption;
图8为实施例2制备的MMS/H NPs的TEM图；8 is a TEM image of MMS / H NPs prepared in Example 2;
图9为MMS/H NPs的(a)氮气吸附脱附曲线；(b)孔径分布示意图；Figure 9 is (a) nitrogen adsorption and desorption curve of MMS / H NPs; (b) schematic diagram of pore size distribution;
图10为MMS/H NPs对真实血液中铅离子的吸附效果(兔血和人血)示意图；Figure 10 is a schematic diagram of the adsorption effect of MMS / H NPs on lead ions in real blood (rabbit and human blood);
图11位MMS/H NPs进出红细胞的过程图示：(a)单纯红细胞，(b)MMS/H NPs进入红细胞，(c)MMS/H NPs与红细胞分离后的红细胞TEM图；Figure 11 shows the process of MMS / H NPs entering and exiting red blood cells: (a) pure red blood cells, (b) MMS / H NPs entering red blood cells, and (c) TEM images of red blood cells after MMS / H NPs are separated from red blood cells;
图12为可用于体外去除血铅的血液清铅仪的装置的结构示意图，体外循环机1、吸附剂进样装置2、接触装置3(血液灌注管)、磁性分离装置(外加磁场4和分离管5)；FIG. 12 is a schematic structural diagram of a device that can be used for a blood lead remover for removing blood lead in vitro, an extracorporeal circulation machine 1, an adsorbent sampling device 2, a contact device 3 (a blood perfusion tube), a magnetic separation device (an external magnetic field 4 and separation) Tube 5);
图13为可用于体外去除血铅的血液清铅仪的装置原理图；FIG. 13 is a schematic diagram of a blood lead-clearing apparatus that can be used to remove blood lead in vitro;
图14为可用于体外去除血铅的血液清铅仪用于铅中毒模型猪体外除铅的过程(i)静脉插管和(ii，iii)体外循环过程；FIG. 14 is a process of removing lead from blood in vitro using a blood lead-clearing instrument for removing lead from lead poisoning model pigs in vitro (i) intravenous intubation and (ii, iii) extracorporeal circulation;
图15为铅中毒模型猪在手术(血铅清除)前后的血常规检测。FIG. 15 is a routine blood test of lead poisoning model pigs before and after surgery (blood lead removal).
具体实施方式detailed description
以下结合附图和实施例对本发明作进一步说明。The invention is further described below with reference to the drawings and embodiments.
实施例1Example 1
1)磁性四氧化三铁的制备(Fe 3O 4NPs) 1) Preparation of magnetic ferric oxide (Fe 3 O 4 NPs)
称取六水合三氯化铁(1.35g,5mmol)，充分溶解于30mL乙二醇溶液中。搅拌条件下加入醋酸钠(NaAc,3.6g)和聚乙二醇-2000(PEG-2000 1.0g)。继续搅拌30min，将溶液转移至高温反应釜中200℃反应72h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it in 30 mL of ethylene glycol solution. Add sodium acetate (NaAc, 3.6 g) and polyethylene glycol-2000 (PEG-2000 1.0 g) with stirring. Stirring was continued for 30 min, and the solution was transferred to a high-temperature reaction kettle at 200 ° C for 72 hours. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
2)以聚赖氨酸为模板剂的磁性四氧化三铁/富胺化介孔二氧化硅复合材料的制备(MMS/P NPs)2) Preparation of magnetic Fe3O4 / aminated mesoporous silica composites using polylysine as a template (MMS / P NPs)
室温下溶解100mg月桂酰肌氨酸钠至10mL柠檬酸/柠檬酸钠缓冲溶液(0.1M,pH 5.2)中，向溶液中加入150μLε-聚赖氨酸(20wt％)。此时溶液立刻变为乳浊液，生成聚赖氨酸/月桂酰肌氨酸钠复合胶束。100 mg of sodium lauroyl sarcosinate was dissolved into 10 mL of a citric acid / sodium citrate buffer solution (0.1 M, pH 5.2) at room temperature, and 150 μL of ε-polylysine (20 wt%) was added to the solution. At this time, the solution immediately became an emulsion, and a polylysine / sodium lauroyl sarcosinate complex micelle was formed.
室温下将150mg上文所制备的Fe 3O 4NPs加入到50mL柠檬酸/柠檬酸钠缓冲溶液(0.1M,pH 5.2)中，并加入10mL聚赖氨酸/月桂酰肌氨酸钠复合胶束溶液。搅拌20min后，依次加入500μL的TEOS和50μL的APTES，继续室温搅拌2h，将产物转移到高温反应釜中80℃反应8h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥，得到磁性四氧化三铁/聚赖氨酸介孔二氧化硅复合材料血液铅离子清除剂。 Add 150 mg of Fe 3 O 4 NPs prepared above to 50 mL of citric acid / sodium citrate buffer solution (0.1M, pH 5.2) at room temperature, and add 10 mL of polylysine / lauroyl sarcosinate composite gel Beam solution. After stirring for 20 min, 500 μL of TEOS and 50 μL of APTES were sequentially added, and stirring was continued at room temperature for 2 h. The product was transferred to a high-temperature reaction kettle at 80 ° C. for 8 h. After being naturally cooled, it was sequentially washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C. to obtain a magnetic lead iron ion scavenger of polyiron methoxide / polylysine silica composite material.
利用透射电镜图来表征复合材料合成过程中纳米粒子的尺寸和形貌，如图1所示。Fe 3O 4NPs分散均匀，尺寸大约为120nm，一般情况下得到Fe 3O 4NPs尺寸大约为50-300nm。得到的MMS/P NPs尺寸约为170nm，制得吸附剂尺寸通常为50-500nm。进一步采用扫描电镜图表征复合材料的尺寸和形貌，如图2所示，能谱分析结果表明铁(Fe)，氧(O)，硅(Si)，碳(C)，氮(N)元素的含量依次为51.66％、25.20％、4.65％、17.23％、1.25％。以上结果均证实了MMS/P NPs的成功构建。 TEM images were used to characterize the size and morphology of the nanoparticles during composite synthesis, as shown in Figure 1. Fe 3 O 4 NPs are uniformly dispersed with a size of about 120 nm. Generally, the size of Fe 3 O 4 NPs is about 50-300 nm. The size of the obtained MMS / P NPs is about 170 nm, and the size of the obtained adsorbent is usually 50-500 nm. Scanning electron microscopy was used to characterize the size and morphology of the composite. As shown in Figure 2, the results of energy spectrum analysis showed that iron (Fe), oxygen (O), silicon (Si), carbon (C), and nitrogen (N) elements The content was 51.66%, 25.20%, 4.65%, 17.23%, and 1.25%, respectively. The above results confirm the successful construction of MMS / P NPs.
利用红外光谱图对MMS/P NPs的合成过程进行表征，如图3所示。修饰之前，如图(a)中，在587cm -1处有明显的特征吸收峰，对应于Fe-O峰。如图(b)所 示，随着进一步反应，在1608cm -1和1409cm -1处出现明显的特征吸收峰，表明了酰胺键的存在。此外，在3000～3500cm -1之间有一个宽峰，对应于氨基基团(-NH 2,-NH-)的伸缩振动峰。酰胺键和氨基基团的存在反映了反应过程中ε-聚赖氨酸得以保留。此外，在1094cm -1和802cm -1处也同时出现了特征峰，分别对应于Si-O-Si和Si-O峰，证明了二氧化硅壳层的成功构建。 The infrared spectrum was used to characterize the synthesis of MMS / P NPs, as shown in Figure 3. Before the modification, as shown in Figure (a), there is a distinct characteristic absorption peak at 587 cm -1 , which corresponds to the Fe-O peak. As shown in Figure (b), with further reaction, obvious characteristic absorption peaks appeared at 1608 cm -1 and 1409 cm -1 , indicating the existence of amide bonds. In addition, there is a broad peak between 3000 and 3500 cm -1 , which corresponds to the stretching vibration peak of the amino group (-NH 2 , -NH-). The presence of amide bonds and amino groups reflects the retention of ε-polylysine during the reaction. In addition, characteristic peaks also appeared at 1094 cm -1 and 802 cm -1 , corresponding to the Si-O-Si and Si-O peaks respectively, which proved the successful construction of the silica shell.
氮气吸附脱附曲线常被用来反映介孔材料的孔道结构。如图4所示，MMS/P NPs的孔径大小约为5.1nm。Nitrogen adsorption and desorption curves are often used to reflect the pore structure of mesoporous materials. As shown in Figure 4, the pore size of MMS / PNPs is approximately 5.1 nm.
通过比较如图5所示的抗凝血处理前后的凝血时间可以看出，与空白组相比由于材料中带有大量氨基基团可以与表面带负点的蛋白结合，未经抗凝处理的MMS/P NPs APTT、PT、TT时间均有所缩短。抗凝血处理后，APTT、PT、TT时间恢复到正常水平，证明修饰后的材料不会对凝血***产生影响。如图6所示，肝素的负载可有效降低溶血率。复合材料为生物相容性材料，溶血率为0.5～5％，活化部分凝血活酶时间为12～30s，血浆凝血酶原时间为8～20s，凝血酶时间为8～20s。By comparing the coagulation time before and after the anticoagulation treatment shown in FIG. 5, it can be seen that compared with the blank group, the material with a large number of amino groups can bind to proteins with negative points on the surface compared with the blank group. MMS / P, NPs, APTT, PT, and TT times have all been shortened. After anticoagulant treatment, the APTT, PT, and TT times returned to normal levels, proving that the modified materials will not affect the coagulation system. As shown in Figure 6, heparin loading can effectively reduce the hemolysis rate. The composite material is a biocompatible material with a hemolysis rate of 0.5 to 5%, an activated partial thromboplastin time of 12 to 30 s, a plasma prothrombin time of 8 to 20 s, and a thrombin time of 8 to 20 s.
3)步骤2)制得的磁性四氧化三铁/富胺化介孔二氧化硅复合材料对血铅的吸附行为研究：3) Step 2) Study on the adsorption of blood lead by the magnetic iron trioxide / aminated mesoporous silica composite:
取定量磁性四氧化三铁/富胺化介孔二氧化硅复合材料放置于离心管中，加入3mL的含铅血液(0.6ppm)，将离心管置于恒温振荡器上37℃振荡固定时间，取2mL上清液，采用高温消解法消解溶液，冷却后定容到5mL。Quantitative magnetic triiron tetroxide / aminated mesoporous silica composite was placed in a centrifuge tube, 3 mL of lead-containing blood (0.6 ppm) was added, and the centrifuge tube was placed on a constant temperature shaker at 37 ° C for a fixed time. Take 2mL of the supernatant and digest the solution by high temperature digestion. After cooling, make up to 5mL.
为了解MMS/P NPs对真实血液中铅的吸附情况，采用真实血液进行吸附测试。如图7所示，吸附30min后，吸附效率达到52.64％。In order to understand the adsorption of lead by MMS / P NPs in real blood, real blood was used for the adsorption test. As shown in Figure 7, after 30 minutes of adsorption, the adsorption efficiency reached 52.64%.
实施例2Example 2
1)磁性四氧化三铁的制备1) Preparation of magnetic iron oxide
采用水热法合成四氧化三铁纳米粒子(Fe 3O 4NPs)。具体方法如下：称取六水合三氯化铁(1.35g,5mmol)，充分溶解于40mL乙二醇溶液中。搅拌条件下加入醋酸钠(NaAc,1.8g)和三水合柠檬酸钠(Na 3Cit 0.2g)。继续搅拌1h，将溶液转移至高温反应釜中200℃反应20h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。 Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reactor for 200 h at 200 ° C. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
2)以超支化聚酰胺(PAMAM)为模板剂的磁性四氧化三铁/富胺化介孔二氧化硅复合材料的制备(MMS/H NPs)2) Preparation of magnetic triiron tetroxide / aminated mesoporous silica composites using hyperbranched polyamide (PAMAM) as a template (MMS / H NPs)
将Fe 3O 4NPs溶液(3mg/mL,50mL)与9mL水合肼混合均匀，超声处理30min。混合物转移到三口烧瓶中，加入70mL去离子水。随后加入90mg TEOS，90℃继续搅拌2h。磁性收集所得产物，重新分散到150mL去离子水中，其中 包含2.1mL水合肼和450mg CTAB。再次超声30min后加入0.6mL TEOS于90℃搅拌2h。获得的产物自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。干燥后的产物于空气气氛下550℃煅烧5h去除模板剂CTAB。收集获得的产物(100mg)分散到150mL去离子水中，其中包含2.1mL水合肼和450mg(0.45mmol)PAMAM。超声处理30min，加入0.6mL(2.88mmol)的TEOS，继续于90℃搅拌2h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。最后将产物于10mL肝素溶液中(10mg/mL)浸泡12h，60℃真空干燥，得到磁性四氧化三铁/超支化聚酰胺介孔二氧化硅复合材料血液铅离子清除剂。 The Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 90 ° C for 2 h. The obtained product was naturally cooled, and then washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C. The dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB. The obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg (0.45 mmol) of PAMAM. Sonicate for 30 min, add 0.6 mL (2.88 mmol) of TEOS, and continue stirring at 90 ° C for 2 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C. Finally, the product was immersed in a 10 mL heparin solution (10 mg / mL) for 12 h, and dried under vacuum at 60 ° C. to obtain a magnetic lead iron ion scavenger of ferric iron tetraoxide / hyperbranched polyamide mesoporous silica composite material.
如图8所示，MMS/H NPs的粒子尺寸约为300nm。如图9所示，采用氮气吸附脱附曲线来表征材料的孔道结构，BET表面积约为248m 2·g -1，孔径大小约为24nm。 As shown in Figure 8, the particle size of MMS / H NPs is about 300 nm. As shown in FIG. 9, a nitrogen adsorption desorption curve is used to characterize the pore structure of the material. The BET surface area is approximately 248 m 2 · g -1 and the pore size is approximately 24 nm.
3)步骤2)制得的磁性四氧化三铁/富胺化介孔二氧化硅复合材料对血铅的吸附行为研究：3) Step 2) Study on the adsorption of blood lead by the magnetic iron trioxide / aminated mesoporous silica composite:
取定量磁性四氧化三铁/富胺化介孔二氧化硅复合材料放置于离心管中，加入3mL的含铅血液(0.6ppm)，将离心管置于恒温振荡器上37℃振荡固定时间，取2mL上清液，采用高温消解法消解溶液，冷却后定容到5mL。Quantitative magnetic triiron tetroxide / aminated mesoporous silica composite was placed in a centrifuge tube, 3 mL of lead-containing blood (0.6 ppm) was added, and the centrifuge tube was placed on a constant temperature shaker at 37 ° C for a fixed time. Take 2mL of the supernatant and digest the solution by high temperature digestion. After cooling, make up to 5mL.
如图10所示，在吸附剂添加量达到5mg/mL时，吸附效率为53％，当吸附剂添加量扩大到10mg/mL时，吸附效率增加到78％。如图11所示，MMS/H NPs可以进入到红细胞当中，并可以通过磁性分离，不会对红细胞形态产生影响。As shown in FIG. 10, when the amount of the adsorbent added reaches 5 mg / mL, the adsorption efficiency is 53%, and when the amount of the adsorbent is expanded to 10 mg / mL, the adsorption efficiency increases to 78%. As shown in Figure 11, MMS / H NPs can enter red blood cells and can be magnetically separated without affecting the red blood cell morphology.
实施例3Example 3
1)磁性四氧化三铁的制备1) Preparation of magnetic iron oxide
采用水热法合成四氧化三铁纳米粒子(Fe 3O 4NPs)。具体方法如下：称取六水合三氯化铁(1.35g,5mmol)，充分溶解于40mL乙二醇溶液中。搅拌条件下加入醋酸钠(NaAc,1.8g)和三水合柠檬酸钠(Na 3Cit 0.2g)。继续搅拌1h，将溶液转移至高温反应釜中200℃反应20h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。 Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reactor for 200 h at 200 ° C. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
2)以壳聚糖为模板剂的磁性四氧化三铁/富胺化介孔二氧化硅复合材料的制备2) Preparation of magnetic triiron tetroxide / aminated mesoporous silica composite with chitosan as template
将Fe 3O 4NPs溶液(3mg/mL,50mL)与9mL水合肼混合均匀，超声处理30min。混合物转移到三口烧瓶中，加入70mL去离子水。随后加入90mg TEOS，90℃继续搅拌2h。磁性收集所得产物，重新分散到150mL去离子水中，其中包含2.1mL水合肼和450mg CTAB。再次超声30min后加入0.6mL TEOS于90℃搅拌2h。获得的产物自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃ 真空干燥。干燥后的产物于空气气氛下550℃煅烧5h去除模板剂CTAB。收集获得的产物(100mg)分散到150mL去离子水中，其中包含2.1mL水合肼和3.4g(0.0576mmol)壳聚糖。超声处理30min，加入0.6mL(2.88mmol)的TEOS，继续于90℃搅拌2h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。最后将产物于10mL肝素溶液中(10mg/mL)浸泡12h，60℃真空干燥，得到磁性四氧化三铁/壳聚糖介孔二氧化硅复合材料血液铅离子清除剂。 The Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 90 ° C for 2 h. After the product obtained was naturally cooled, it was washed with deionized water and ethanol solution several times in that order, and dried under vacuum at 60 ° C. The dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB. The obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 3.4 g (0.0576 mmol) of chitosan. Sonicate for 30 min, add 0.6 mL (2.88 mmol) of TEOS, and continue stirring at 90 ° C for 2 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C. Finally, the product was immersed in a 10 mL heparin solution (10 mg / mL) for 12 h, and dried under vacuum at 60 ° C. to obtain a blood lead ion scavenger of a magnetic iron trioxide / chitosan mesoporous silica composite material.
实施例4Example 4
1)磁性四氧化三铁的制备1) Preparation of magnetic iron oxide
采用水热法合成四氧化三铁纳米粒子(Fe 3O 4NPs)。具体方法如下：称取六水合三氯化铁(1.35g,5mmol)，充分溶解于40mL乙二醇溶液中。搅拌条件下加入醋酸钠(NaAc,1.8g)和三水合柠檬酸钠(Na 3Cit 0.2g)。继续搅拌1h，将溶液转移至高温反应釜中150℃反应24h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。 Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reaction kettle at 150 ° C. for 24 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
2)以超支化聚酰胺为模板剂的磁性四氧化三铁/富胺化介孔二氧化硅复合材料的制备(MMS/H NPs)2) Preparation of magnetic triiron tetroxide / aminated mesoporous silica composites using hyperbranched polyamide as a template (MMS / H NPs)
将Fe 3O 4NPs溶液(3mg/mL,50mL)与9mL水合肼混合均匀，超声处理30min。混合物转移到三口烧瓶中，加入70mL去离子水。随后加入90mg TEOS，90℃继续搅拌2h。磁性收集所得产物，重新分散到150mL去离子水中，其中包含2.1mL水合肼和450mg CTAB。再次超声30min后加入0.6mL TEOS于60℃搅拌5h。获得的产物自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。干燥后的产物于空气气氛下550℃煅烧5h去除模板剂CTAB。收集获得的产物(100mg)分散到150mL去离子水中，其中包含2.1mL水合肼和576mg(0.576mmol)PAMAM。超声处理30min，加入0.6mL(2.88mmol)的TEOS，继续于90℃搅拌2h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。最后将产物于10mL肝素溶液中(10mg/mL)浸泡12h，60℃真空干燥，得到磁性四氧化三铁/超支化聚酰胺介孔二氧化硅复合材料血液铅离子清除剂。 The Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 60 ° C. for 5 h. The obtained product was naturally cooled, and then washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C. The dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB. The obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 576 mg (0.576 mmol) of PAMAM. Sonicate for 30 min, add 0.6 mL (2.88 mmol) of TEOS, and continue stirring at 90 ° C for 2 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C. Finally, the product was immersed in a 10 mL heparin solution (10 mg / mL) for 12 h, and dried under vacuum at 60 ° C. to obtain a magnetic lead iron ion scavenger of ferric iron tetraoxide / hyperbranched polyamide mesoporous silica composite material.
实施例5Example 5
1)磁性四氧化三铁的制备1) Preparation of magnetic iron oxide
采用水热法合成四氧化三铁纳米粒子(Fe 3O 4NPs)。具体方法如下：称取六水合三氯化铁(1.35g,5mmol)，充分溶解于40mL乙二醇溶液中。搅拌条件下加入醋酸钠(NaAc,1.8g)和三水合柠檬酸钠(Na 3Cit 0.2g)。继续搅拌1h，将溶液转移至高温反应釜中220℃反应10h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。 Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reaction kettle at 220 ° C. for 10 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
2)以超支化聚酰胺为模板剂的磁性四氧化三铁/富胺化介孔二氧化硅复合材料的制备2) Preparation of magnetic triiron tetroxide / aminated mesoporous silica composite using hyperbranched polyamide as a template
将Fe 3O 4NPs溶液(3mg/mL,50mL)与9mL水合肼混合均匀，超声处理30min。混合物转移到三口烧瓶中，加入70mL去离子水。随后加入90mg TEOS，90℃继续搅拌2h。磁性收集所得产物，重新分散到150mL去离子水中，其中包含2.1mL水合肼和450mg CTAB。再次超声30min后加入0.6mL TEOS于90℃搅拌1h。获得的产物自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。干燥后的产物于空气气氛下550℃煅烧5h去除模板剂CTAB。收集获得的产物(100mg)分散到150mL去离子水中，其中包含2.1mL水合肼和450mg PAMAM。超声处理30min，加入0.6mL的TEOS，继续于90℃搅拌2h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。最后将产物于10mL肝素溶液中(10mg/mL)浸泡12h，60℃真空干燥，得到磁性四氧化三铁/超支化聚酰胺介孔二氧化硅复合材料血液铅离子清除剂。 The Fe 3 O 4 NPs solution (3 mg / mL, 50 mL) was mixed with 9 mL of hydrazine hydrate, and sonicated for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 90 ° C for 1 h. The obtained product was naturally cooled, and then washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C. The dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB. The obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of PAMAM. Sonicate for 30 min, add 0.6 mL of TEOS, and continue stirring at 90 ° C for 2 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C. Finally, the product was immersed in a 10 mL heparin solution (10 mg / mL) for 12 h, and dried under vacuum at 60 ° C. to obtain a magnetic lead iron ion scavenger of ferric iron tetraoxide / hyperbranched polyamide mesoporous silica composite material.
实施例6Example 6
1)磁性四氧化三铁的制备1) Preparation of magnetic iron oxide
采用水热法合成四氧化三铁纳米粒子(Fe 3O 4NPs)。具体方法如下：称取六水合三氯化铁(1.35g,5mmol)，充分溶解于40mL乙二醇溶液中。搅拌条件下加入醋酸钠(NaAc,1.8g)和三水合柠檬酸钠(Na 3Cit 0.2g)。继续搅拌1h，将溶液转移至高温反应釜中200℃反应20h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。 Fe 3 O 4 NPs were synthesized by hydrothermal method. The specific method is as follows: Weigh ferric trichloride (1.35 g, 5 mmol) and dissolve it fully in 40 mL of ethylene glycol solution. Add sodium acetate (NaAc, 1.8 g) and sodium citrate trihydrate (Na 3 Cit 0.2 g) with stirring. Stirring was continued for 1 h, and the solution was transferred to a high-temperature reactor for 200 h at 200 ° C. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C.
2)以超支化聚酰胺为模板剂的磁性四氧化三铁/富胺化介孔二氧化硅复合材料的制备将Fe 3O 4NPs溶液(3mg/mL,50mL)与9mL水合肼混合均匀，超声处理30min。混合物转移到三口烧瓶中，加入70mL去离子水。随后加入90mg TEOS，90℃继续搅拌2h。磁性收集所得产物，重新分散到150mL去离子水中，其中包含2.1mL水合肼和450mg CTAB。再次超声30min后加入0.6mL TEOS于90℃搅拌2h。获得的产物自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。干燥后的产物于空气气氛下550℃煅烧5h去除模板剂CTAB。收集获得的产物(100mg)分散到150mL去离子水中，其中包含2.1mL水合肼和57.6mg(0.0576mmol)PAMAM。超声处理30min，加入0.6mL(2.88mmol)的TEOS，继续于90℃搅拌2h。自然冷却后依次用去离子水和乙醇溶液多次洗涤，60℃真空干燥。最后将产物于10mL肝素溶液中(10mg/mL)浸泡12h，60℃真空干燥，得到磁性四氧化三铁/超支化聚酰胺介孔二氧化硅复合材料血液铅离子清除剂。 2) Preparation of magnetic triiron tetroxide / aminated mesoporous silica composite using hyperbranched polyamide as a template. Fe 3 O 4 NPs solution (3mg / mL, 50mL) was mixed with 9mL of hydrazine hydrate, Sonicate for 30 min. The mixture was transferred to a three-necked flask, and 70 mL of deionized water was added. Subsequently, 90 mg of TEOS was added, and stirring was continued at 90 ° C for 2 h. The resulting product was magnetically collected and re-dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 450 mg of CTAB. After sonicating again for 30 min, 0.6 mL of TEOS was added and stirred at 90 ° C for 2 h. The obtained product was naturally cooled, and then washed with deionized water and an ethanol solution for several times, and dried under vacuum at 60 ° C. The dried product was calcined in an air atmosphere at 550 ° C for 5 hours to remove the template agent CTAB. The obtained product (100 mg) was collected and dispersed into 150 mL of deionized water, which contained 2.1 mL of hydrazine hydrate and 57.6 mg (0.0576 mmol) of PAMAM. Sonicate for 30 min, add 0.6 mL (2.88 mmol) of TEOS, and continue stirring at 90 ° C for 2 h. After leaving to cool, it was washed with deionized water and ethanol solution several times in sequence, and dried under vacuum at 60 ° C. Finally, the product was immersed in a 10 mL heparin solution (10 mg / mL) for 12 h, and dried under vacuum at 60 ° C. to obtain a magnetic lead iron ion scavenger of ferric iron tetraoxide / hyperbranched polyamide mesoporous silica composite.
实施例7Example 7
如图12所示，一种可用于体外去除血铅的血液清铅仪，包括依次连接的体外循环机1、吸附剂进样装置2、接触装置3、磁性分离装置；体外循环机1和吸附剂进样装置2通过Y型管与接触装置3连接，接触装置3的组成为血液灌注管，血液灌注管的长径比为600:1，血液灌注管的长径比可以在300:1-1000：1之间调整，磁性分离装置由有外加磁场4(可以使用磁铁)和分离管5组成，即分离管5外套有磁铁，分离管的长径比为100:1，分离管的长径比可以在50:1-200:1之间调整，分离管口径大于血液灌注管的口径以实现吸附剂与血液的分离。吸附剂进样装置3中的吸附剂的成分为以富胺类有机物为模板剂且具有规整介孔孔道的磁性四氧化三铁/富胺化介孔二氧化硅复合材料，本实施例具体采用实施例2制备得到的磁性四氧化三铁/超支化聚酰胺介孔二氧化硅复合材料血液铅离子吸附剂。As shown in FIG. 12, a blood lead-clearing instrument which can be used for extracorporeal blood lead removal includes an extracorporeal circulation machine 1, an adsorbent sampling device 2, a contact device 3, and a magnetic separation device connected in sequence; an extracorporeal circulation machine 1 and an adsorption device The agent injection device 2 is connected to the contact device 3 through a Y-shaped tube. The composition of the contact device 3 is a blood perfusion tube. The aspect ratio of the blood perfusion tube is 600: 1, and the aspect ratio of the blood perfusion tube can be 300: 1- Adjusting between 1000: 1, the magnetic separation device consists of an external magnetic field 4 (magnets can be used) and the separation tube 5, that is, the separation tube 5 has a magnet, and the length-to-diameter ratio of the separation tube is 100: 1. The ratio can be adjusted between 50: 1-200: 1. The diameter of the separation tube is larger than the diameter of the blood perfusion tube to achieve the separation of the adsorbent from the blood. The composition of the adsorbent in the adsorbent injection device 3 is a magnetic triiron tetroxide / aminated mesoporous silica composite material using amine-rich organics as a template and having regular mesoporous channels. This embodiment specifically uses The magnetic lead iron ion adsorbent prepared in Example 2 is a magnetic iron trioxide / hyperbranched polyamide mesoporous silica composite material.
使用时血液清铅仪通过首先连接铅中毒模型猪的静脉血管，血液经过导管进入体外循环机1用以带动整个治疗过程的血液循环，体外循环机1和吸附剂进样装置2通过将使用Y型导管连接至接触装置3中的血液灌注管，随后血液通过Y型导管与吸附剂进样装置2推出的吸附剂接触混合，吸附剂和血液在血液灌注管中进行充分接触(10分钟以上)，后血液过血液灌注管至套有磁铁(即有外加磁场4)的分离管5中，分离管5口径大于血液灌注管的口径以实现吸附剂与血液的分离，血液通过动脉流回铅中毒模型猪上，完成一次血液除铅过程，继续重复步骤循环除铅。When used, the blood lead clearer first connects the venous vessels of lead poisoning model pigs, and the blood enters the extracorporeal circulation machine 1 through the catheter to drive the blood circulation throughout the treatment process. The extracorporeal circulation machine 1 and the adsorbent injection device 2 use Y The catheter is connected to the blood perfusion tube in the contact device 3, and then the blood is contacted and mixed with the adsorbent introduced by the adsorbent injection device 2 through the Y-type catheter, and the adsorbent and blood are fully contacted in the blood perfusion tube (more than 10 minutes) After the blood passes through the blood perfusion tube to the separation tube 5 with a magnet (that is, an external magnetic field 4), the diameter of the separation tube 5 is larger than the diameter of the blood perfusion tube to separate the adsorbent from the blood, and the blood flows back through the artery to lead poisoning On the model pig, complete the blood lead removal process once and continue to repeat the steps to remove lead.
实施例8Example 8
将血液清铅仪用于铅中毒模型猪的血铅去除实验Blood lead removal instrument for blood lead removal experiment of lead poisoning model pigs
通过食物中毒方法对猪建立了约0.5ppm的铅中毒模型。采用实施例7的用于体外去除血铅的血液清铅仪，首先将体外循环机1、吸附剂进样装置2、接触装置3、磁力分离装置连接起来构建血铅清理装置。然后建立猪的体外循环途径(图13-14)如下：通过在腹腔中注射3％戊巴比妥钠(1ml/kg)麻醉猪，并且在血铅清洁过程中保持麻醉，连续注射0.9％NaCl和2.5％戊巴比妥钠。静脉血管与血铅清理装置连接，并在通过磁分离装置后返回动脉血。血流速度为25ml/min，循环时间为50min，吸附剂的量为1g，铅中毒模型猪的血液量约为3L，吸附剂与血液体积比例，可以在1:10-10:1g/L之间调整。手术后对伤口缝合，并检测了手术前后铅中毒模型猪的血常规和血铅去除效率。实验结果显示当猪体内血铅含量为500ppb时，经过一次清洗过程,血铅的去除效率可达75％，可通过二次处理，继续提高血铅去除率(去除率的计算方法为：分别抽取手术前后的血液， 用电感耦合等离子光谱仪检测手术前后的浓度分别标记为C 0，C 1，通过公式E＝(C 0-C 1)/C 0*100％计算出血铅的去除效率。公式中E为血铅去除效率(％)，C 0为手术前的血铅浓度(ppb)，C 1为手术后的血铅浓度(ppb))。且吸附剂分离效果较好，显示了该吸附剂和血液清铅仪较高的去除效率,同时清洗前后的血常规显示铅离子去除后血液炎症有所减小(图15)，体现出该血液吸附剂和血液清铅仪在去除血铅过程中的安全性和高效性。 A model of lead poisoning of about 0.5 ppm was established for pigs by the food poisoning method. Using the blood lead removal instrument for removing blood lead in vitro from Example 7, first, an extracorporeal circulation machine 1, an adsorbent sampling device 2, a contact device 3, and a magnetic separation device were connected to construct a blood lead cleaning device. Then establish the extracorporeal circulation pathway of the pigs (Figure 13-14) as follows: Anesthetize the pigs by injecting 3% sodium pentobarbital (1ml / kg) in the abdominal cavity, and keep anesthesia during the blood lead cleaning, and continuously inject 0.9% NaCl And 2.5% sodium pentobarbital. The venous vessels are connected to a blood lead cleaning device and return to arterial blood after passing through a magnetic separation device. The blood flow speed is 25ml / min, the circulation time is 50min, the amount of adsorbent is 1g, the blood volume of the lead poisoning model pig is about 3L, and the ratio of adsorbent to blood volume can be between 1: 10-10: 1g / L Between adjustments. The wound was sutured after the operation, and the blood routine and blood lead removal efficiency of the model pigs with lead poisoning before and after the operation were examined. The experimental results show that when the blood lead content in pigs is 500ppb, after one cleaning process, the blood lead removal efficiency can reach 75%. The secondary treatment can continue to improve the blood lead removal rate (the calculation method of the removal rate is: The blood before and after the operation was detected with an inductively coupled plasma spectrometer before and after the operation. The concentrations were marked as C 0 and C 1 , and the lead removal efficiency was calculated by the formula E = (C 0 -C 1 ) / C 0 * 100%. In the formula, E is the blood lead removal efficiency (%), C 0 is the blood lead concentration (ppb) before the operation, and C 1 is the blood lead concentration (ppb) after the operation). And the adsorbent has a better separation effect, which shows the higher removal efficiency of the adsorbent and the blood lead remover. At the same time, the blood routine before and after cleaning shows that the blood inflammation is reduced after the removal of lead ions (Figure 15), which reflects the blood The safety and efficiency of the adsorbent and blood lead remover in the process of removing blood lead.

Claims (10)

  1. 一种可用于体外去除血铅的血液清铅仪，其特征在于，包括依次连接的体外循环机、吸附剂进样装置、接触装置、磁性分离装置；所述吸附剂进样装置中负载有吸附剂，所述吸附剂的成分为以富胺类有机物为模板剂且具有介孔孔道的磁性四氧化三铁/富胺化介孔二氧化硅复合材料。A blood lead-clearing instrument which can be used to remove blood lead in vitro, which comprises an extracorporeal circulation machine, an adsorbent sampling device, a contact device, and a magnetic separation device connected in sequence; the adsorbent sampling device is loaded with adsorption The component of the adsorbent is a magnetic triiron tetroxide / aminated mesoporous silica composite material using amine-rich organic matter as a template and having mesoporous channels.
  2. 根据权利要求1所述的可用于体外去除血铅的血液清铅仪，其特征在于，所述体外循环机和吸附剂进样装置通过Y型管与接触装置连接。The blood lead removal instrument according to claim 1, wherein the extracorporeal circulation machine and the sorbent sampling device are connected to a contact device through a Y-shaped tube.
  3. 根据权利要求1所述的可用于体外去除血铅的血液清铅仪，其特征在于，所述接触装置的组成为血液灌注管，血液灌注管的长径比为300:1-1000：1。The blood lead-clearing instrument for removing blood lead in vitro according to claim 1, wherein the composition of the contacting device is a blood perfusion tube, and the aspect ratio of the blood perfusion tube is 300: 1-1000: 1.
  4. 根据权利要求1所述的可用于体外去除血铅的血液清铅仪，其特征在于，所述磁性分离装置由外加磁场和分离管组成，分离管的长径比为50:1-200:1。The blood lead removal instrument according to claim 1, wherein the magnetic separation device is composed of an external magnetic field and a separation tube, and the aspect ratio of the separation tube is 50: 1-200: 1 .
  5. 根据权利要求1所述的可用于体外去除血铅的血液清铅仪，其特征在于，所述吸附剂尺寸为50-500nm，吸附剂与血液体积比例为1:10-10:1g/L。The blood lead-clearing instrument for removing blood lead in vitro according to claim 1, wherein the size of the adsorbent is 50-500 nm, and the ratio of the adsorbent to the blood volume is 1: 10-10: 1g / L.
  6. 根据权利要求1所述的可用于体外去除血铅的血液清铅仪，其特征在于，所述吸附剂的制备方法，包括以下步骤：The blood lead-clearing instrument for removing blood lead in vitro according to claim 1, wherein the method for preparing the adsorbent comprises the following steps:
    (1)称取六水合三氯化铁溶解于乙二醇溶液中，加入醋酸钠和三水合柠檬酸钠或聚乙二醇继续反应，得到Fe 3O 4溶液； (1) Weigh the ferric trichloride hexahydrate dissolved in the ethylene glycol solution, add sodium acetate and sodium citrate trihydrate or polyethylene glycol to continue the reaction to obtain a Fe 3 O 4 solution;
    (2)将步骤(1)制得的Fe 3O 4溶液与水合肼混合均匀，超声处理，加入正硅酸四乙酯，收集所得产物，重新分散到包含水合肼和十六烷基三甲基溴化铵的去离子水中，再次超声后加入正硅酸四乙酯反应； (2) Mix the Fe 3 O 4 solution prepared in step (1) with hydrazine hydrate uniformly, sonicate it, add tetraethyl orthosilicate, collect the obtained product, and re-disperse it to contain hydrazine hydrate and cetyltrimethyl Ammonium bromide in deionized water, add tetraethyl orthosilicate after ultrasonication again;
    (3)将步骤(2)最后反应得到的产物干燥后煅烧去除十六烷基三甲基溴化铵，收集获得的产物分散到包含水合肼和富胺类有机物模板剂的去离子水中，超声处理后加入正硅酸四乙酯反应，继续反应，得到产物磁性四氧化三铁/富胺化介孔二氧化硅复合材料血液铅离子吸附剂。(3) The product obtained in the last reaction of step (2) is dried and calcined to remove cetyltrimethylammonium bromide, and the product obtained is collected and dispersed in deionized water containing hydrazine hydrate and amine-rich organic template, and ultrasonic After the treatment, tetraethyl orthosilicate was added to the reaction, and the reaction was continued to obtain a product magnetic iron trioxide / aminated mesoporous silica composite blood lead ion adsorbent.
  7. 根据权利要求6所述的可用于体外去除血铅的血液清铅仪，其特征在于，步骤(2)和步骤(3)所述每次加入正硅酸四乙酯的反应温度为60-90℃，反应时间为1～5h。The blood lead remover for removing blood lead in vitro according to claim 6, characterized in that the reaction temperature of each step of adding tetraethyl orthosilicate in step (2) and step (3) is 60-90 ℃, reaction time is 1 ~ 5h.
  8. 根据权利要求6所述的可用于体外去除血铅的血液清铅仪，其特征在于，步骤(3)所述正硅酸四乙酯与富胺类有机物模板剂的摩尔比为5：1～50：1。The blood lead-clearing instrument for removing blood lead in vitro according to claim 6, wherein the molar ratio of the tetraethyl orthosilicate to the amine-rich organic template is 5: 1 ～ 50: 1.
  9. 根据权利要求1-8任一所述的可用于体外去除血铅的血液清铅仪，其特征在于，所述吸附剂合成所用的以富胺类有机物为模板剂包含超支化聚酰胺、聚赖氨酸或壳聚糖中的一种。The blood lead clearing instrument for removing blood lead in vitro according to any one of claims 1 to 8, characterized in that the amine-rich organic matter used as a template for the synthesis of the adsorbent comprises hyperbranched polyamide, polypropylene One of amino acid or chitosan.
  10. 根据权利要求1-8任一所述的可用于体外去除血铅的血液清铅仪，其特征在于，所述吸附剂在整个血液清铅仪中去除血液中铅离子机制过程为复合材料 自由进出红细胞，通过介孔二氧化硅结构中的氨基对铅离子的络合作用捕获含铅血红蛋白，并通过介孔孔道对含铅血红蛋白进行固定，实现去除红细胞中铅离子。The blood lead-clearing instrument for removing blood lead in vitro according to any one of claims 1 to 8, characterized in that the mechanism of removing the lead ions in the blood by the adsorbent in the whole blood lead-clearing instrument is that the composite material enters and exits freely For red blood cells, lead-containing hemoglobin is captured by the complexation of lead ions by the amino group in the mesoporous silica structure, and lead-containing hemoglobin is fixed through the mesoporous channels to achieve the removal of lead ions in red blood cells.
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