CN102716719A - Heavy metal adsorbent for extracorporeal perfusion method and preparation method of heavy metal absorbent - Google Patents
Heavy metal adsorbent for extracorporeal perfusion method and preparation method of heavy metal absorbent Download PDFInfo
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Abstract
The invention discloses an adsorbent for removing heavy metals through extracorporeal blood perfusion and a preparation method of the absorbent. Activating agents are used activating resin microsphere carriers with good biocompatibility, then sulihydryl reagents are fixed through bonding or coupling to form the heavy metal adsorbent, the surface of the heavy metal adsorbent is provided with a great quantity of sulihydryl functional groups and an adsorption effect through complexing or chelation between adsorption target liquid compositions and heavy metals and ions thereof is facilitated; the biocompatibility and the adsorption capacity are good; and the adsorbent has the characteristics of low preparation cost, simple synthesis process, strong adsorption capacity, rapid adsorption speed and the like, and can be used for extracorporeal whole-blood perfusion to remove in-vivo heavy metals and treat heavy metal poisoning.
Description
Technical field
The present invention relates to a kind of heavy metal absorbent that is used for external perfusion method and preparation method thereof, belong to field of biomedical materials.Relating in particular to a kind of being used for removes sorbing material of heavy metal and ion thereof and preparation method thereof with external perfusion method from blood of human body, serum, blood plasma, plasma component, bone marrow fluid, celiolymph, cell culture fluid, cell homogenized liquid, animal organ's extract, plant extraction liquid or other liquid component absorption (complexing, chelating).
Background of invention
Human in contact or use the air that polluted by various pollution sources such as industrial waste gas, waste water, waste residue, waste material, vehicle exhaust, agricultural chemicals, chemical fertilizer, chemical reagent, additive, paint, soil, river, lake, aquatic products, animal, plant, mineral, food, drinking water, domestic water, medicine, cosmetics, health products, toy etc., will receive each heavy metal species harm.For example, heavy metal Hg harmful to human nervous system makes brain impaired, causes mercury poisoning brain disease, the numb limb that causes, and symptoms such as ataxia, the visual field narrow down, hard of hearing, in addition in heart failure; Heavy metal cadmium can accumulate in human body and cause poisoning, and acute cadmium poisoning can make people's spitting blood, stomachache, cause death at last, and chronic cadium poisoning can make renal dysfunction, destroys skeleton, causes ostalgia, osteomalacia, paralysis; Heavy metal lead harm nervous system and kidney organ, infringement skeleton hemopoietic system causes anaemia, motion and cacesthesia appear in cerebral anoxia, encephaledema; The plumbous also neural plate of the brain cell of direct harm people, particularly fetus, it is shallow to cause congenital cerebral sulci to return, feeblemindedness; The elderly is caused dementia, brain death etc.
But the human method that generally lacks effective eliminating heavy metal of body first because in a single day heavy metal and ion thereof get into human body, stockpiles in the rapid blood at human body of meeting, bone, spinal cord, organ, the tissue, is difficult to eliminate; Second the body innerlich anwenden is got rid of heavy metal, the still kidney organ who relies at last, and speed is slow, and the infringement kidney; When three body innerlich anwendens were got rid of heavy metal, side effect was a lot.For example, up to the present, the human interior usefulness that especially effectively has no side effect that also do not find is driven plumbous medicine.The interior usefulness that clinical in history tradition (particularly labour health and occupational disease) is used is driven plumbous medicine, is used for severe lead poisoning patient's treatment more; Both be not suitable for morning, lead poisoning in the mid-term patient of subclinical pathology, and had serious problems such as side effect is big, can cause complexing syndrome, allergic reaction, kidney damage etc.; Lead discharging medicine calcium disodium edetate can only drug administration by injection, can produce general reaction such as shiver with cold, heating, tired, thirsty, headache, glycosuria, blood pressure decline temporarily, anaemia, dermatitis, the necrosis of kidney pipe etc.; In addition, the body innerlich anwenden drives lead, need to use continuously 40-60 talent reluctantly with Pb-B be reduced to poison below horizontal.
The hydrophilic gel microballoon of the preferred good biocompatibility of the present invention is as medium, and the heavy metal absorbent surface of processing has a large amount of sulfydryls, helps producing absorption (complexing, chelating) effect with the heavy metal that adsorbs in the object liquid component; Biocompatibility and adsorption capacity are good, and it is cheap to have a preparation cost, and building-up process is simple; Characteristics such as adsorbance is big, the rate of adsorption is fast; Can be used for the outer whole blood perfusion of human body, get rid of heavy metal in the human body, the treatment heavy metal poisoning.Be used for human body and remove blood lead, can exclusion process be reduced to 1-3 hour by 40-60 days of conventional method, effect very significantly.
Summary of the invention
The objective of the invention is, the problems to heavy metal, treatment heavy metal poisoning method in the present eliminating human body propose a kind of heavy metal absorbent that is used for external perfusion method and preparation method thereof.This method has following concrete characteristic:
1, a kind of heavy metal absorbent that is used for external perfusion method of the present invention and preparation method thereof; It is characterized in that: use the activator resin microsphere carrier activation that biocompatibility is good; Fixing then (bonding, coupling) SR; Form heavy metal absorbent, make the heavy metal absorbent surface have a large amount of sulfydryl functional groups, help producing absorption (complexing, chelating) effect with the heavy metal and the ion thereof that adsorb in the object liquid component.
2, heavy metal absorbent of the present invention; It is characterized in that; The good resin microsphere carrier of biocompatibility that is used to prepare heavy metal absorbent is the hydrophilic gel microballoon, includes but not limited to hydrophily agarose gel microsphere, hydrophily sephadex microsphere, HPAG microballoon, hydrophilic polyethene alcohol microballoon; But the preferred hydrophilic agarose gel microsphere is as the carrier matrix material; The particle diameter of matrix is the 50-160 micron, and the degree of cross linking is the 4-10% scope.
3, heavy metal absorbent of the present invention; It is characterized in that; The activator that is used to prepare heavy metal absorbent includes but not limited to epoxychloropropane, hexamethylene diamine, 1, and 1 '-carbonyl dimidazoles, two vinyl sulfone(Remzaol, ketene dimer, cyanogen bromide, allyl glycidyl ether, paratoluensulfonyl chloride, s-triazine, Cyanuric Chloride etc. can further connect groups such as amino on the aglucon, amido, hydroxyl, sulfydryl, silicon hydroxyl, boron hydroxyl, carboxyl, aldehyde radical.
4, heavy metal absorbent of the present invention; It is characterized in that; The SR that is used to prepare heavy metal absorbent be have free sulfhydryl groups (sulfhydryl compound SH) maybe can produce the reagent of free sulfhydryl groups through subsequent chemical reaction, includes but not limited to one or more composition of following reagent:
Metallothionein, sulfoprotein, sodium dimercaptosuccinate, dimercaptosuccinic acid, dimercaptosuccinic acid potassium, dimercaprol dimercaptopropanol, L-cysteine, thiocarbamide, TGA, 2 mercapto ethanol, vulcanized sodium, NaHS, potassium bisulfide, alkyl sulfide ortho acid sodium.
5, heavy metal absorbent of the present invention is used for external perfusion method absorption and gets rid of heavy metal, it is characterized in that said heavy metal includes but not limited to lead, cadmium, mercury, chromium, bismuth, cobalt, vanadium, tin, thallium and ion thereof.
6, absorption object liquid component of the present invention; It is characterized in that described absorption object liquid component is blood of human body, serum, blood plasma, plasma component, bone marrow fluid, celiolymph, cell culture fluid, cell homogenized liquid, animal organ's extract, plant extraction liquid or other liquid component.
The hydrophilic gel microballoon of the preferred good biocompatibility of the present invention is as medium, and the heavy metal absorbent surface of processing has a large amount of sulfydryls, helps producing absorption (complexing, chelating) effect with the heavy metal that adsorbs in the object liquid component; Biocompatibility and adsorption capacity are good, and it is cheap to have a preparation cost, and building-up process is simple; Characteristics such as adsorbance is big, the rate of adsorption is fast; Can be used for the outer whole blood perfusion of human body, get rid of heavy metal in the human body, the treatment heavy metal poisoning.Be used for human body and remove blood lead, can exclusion process be reduced to 1-3 hour by 40-60 days of conventional method, effect very significantly.The technology of heavy metal and ion thereof is not seen any report in the body fluid components such as the external perfusion method absorption of this use blood of human body.
The specific embodiment
Below through instance the present invention is further described:
Embodiment 1:
Measure the Ago-Gel Sepharose FF medium microsphere 20ml (settling volume) that is stored in 20% ethanol water, with excessive deionized water cyclic washing, to remove ethanol; Place G3 sintered glass funnel vacuum to drain 5min the gel after the washing; After clean the medium behind suction filtration with each 40ml of 20%, 50%, 70% the methyl-sulfoxide aqueous solution successively; Then, add a certain amount of methyl-sulfoxide (30%), dichloroethanes, epoxychloropropane (10%), NaOH (0.8M), water successively in the medium after handling; The medium suspension was in the following 40 ℃ of reactions of concussion condition 2 hours.The reaction back obtains epoxychloropropane activated agarose gel micro-ball with noresidue epoxide group in excessive deionized water cyclic washing to the cleaning fluid, and agarose epoxy-modified density is 60 μ mol/ml gels.
Embodiment 2
The K2HPO4 solution of the 0.2M of 8.5 volumes is mixed with the KH2PO4 solution of the 0.2M of 1.5 volumes, process coupling buffer.Get the epoxychloropropane activated agarose gel micro-ball among the 5g embodiment 1, be suspended from the 10ml coupling buffer.Get a certain amount of metallothionein and be dissolved in the coupling cushioning liquid, it is dissolved fully.With gel suspension and coupling cushioning liquid mixed suspension reaction with 1: 0.5 to 1: 1, in 20 ℃-45 ℃ water-bath, seal, continued to stir (or concussion, rock) 20 hours.Remove unnecessary part with the coupling buffer washing.To react washed gel and be suspended in the solution of 1mol/L monoethanolamine (pH 8.0), spend the night, to eliminate dielectric surface residual activity group in 40 ℃ of-50 ℃ of minimum reactions 4 hours or room temperature reaction.After having reacted, use the alternately washing of coupling buffer (pH 8.3) that contains the 0.1M acetate buffer solution (pH 4.0) of 0.5M NaCl and contain 0.5M NaCl, repeat above circulation cleaning 3 times at least.Gained medium ligand density 50 μ mol/ml.With its be stored in the SAS that contains 0.02% sodium azide (w/v) (0.2M, for use in pH5.4).
Embodiment 3:
Measure the Ago-Gel Sepharose FF medium microsphere 20ml (settling volume) that is stored in 20% ethanol water, with excessive deionized water cyclic washing, to remove ethanol; Place G3 sintered glass funnel vacuum to drain 5min the gel after the washing; After clean the medium behind the suction filtration with 30%, 70% aqueous acetone solution of 5 times of volumes successively; Wash 5 times with 100% dioxy, six alkane of 5 times of volumes the back, drains, and transfers in the 200ml flask; Add a certain amount of allyl glycidyl ether, boron trifluoride-ether successively in the medium after handling; The medium suspension was in the following 35 ℃ of reactions of concussion condition 1-2 hour.Reacted medium washs with 30%, 70% aqueous acetone solution of 5 times of volumes successively; The back is with noresidue epoxide group in excessive deionized water cyclic washing to the cleaning fluid; Obtain allyl glycidyl ether activated agarose gel micro-ball, it is 20-150 μ mol/ml gel that pi-allyl is modified density.
Embodiment 4:
Get the allyl glycidyl ether activated agarose gel micro-ball 10ml among the embodiment 3; Add the 1.20ml TGA, 10ml deionized water, 0.25g ammonium persulfate; The medium suspension under the concussion condition 50-70 ℃ the reaction 8-12 hour, the TGA agarose gel microsphere.
Embodiment 5:
Get the TGA agarose gel microsphere 10ml among the embodiment 4, successively with the medium behind 30%, 70%, the 100% aqueous acetone solution cleaning suction filtration of 5 times of volumes; Wash 5 times with 100% dioxy, six alkane of 5 times of volumes the back, transfers in the 200ml flask; Add a certain amount of dioxy six alkane, N-hydroxy-succinamide, N successively, N-dicyclohexyl carbimide in the medium after handling; The medium suspension was in the following 25 ℃ of reactions of concussion condition 8-12 hour.Reacted medium is used a large amount of dioxies six alkane, methyl alcohol, washing with acetone successively, obtains N-hydroxy-succinamide and modifies agarose gel microsphere, and it is 20-150 μ mol/ml gel that N-hydroxy-succinamide is modified density, and it is for use to be stored in the isopropyl alcohol lucifuge.
Embodiment 6:
The N-hydroxy-succinamide of getting among the embodiment 5 is modified agarose gel microsphere 10ml, uses 70%, 30% aqueous acetone solution of 5 times of volumes, the medium behind the washed with de-ionized water suction filtration successively; Transfer in the 200ml flask; Add a certain amount of metallothionein solution successively in the medium after handling; The medium suspension was in the following 25 ℃ of reactions of concussion condition 12-24 hour.Reacted medium is removed unnecessary part with the washing of the coupling buffer described in a large amount of embodiment 2.To react washed gel and be suspended in the solution of 1mol/L monoethanolamine (PH 8), room temperature reaction spends the night, to eliminate dielectric surface residual activity group.After having reacted, use the alternately washing of coupling buffer (pH 8.3) that contains the 0.1M acetate buffer solution (pH 4.0) of 0.5M NaCl and contain 0.5M NaCl, repeat above circulation cleaning 3 times at least, obtain metallothionein and modify agarose gel microsphere.With its be stored in the SAS that contains 0.02% sodium azide (w/v) (0.2M, for use in pH5.4).
Claims (6)
1. one kind is used for heavy metal absorbent of external perfusion method and preparation method thereof; It is characterized in that: use the activator resin microsphere carrier activation that biocompatibility is good; Fixing then (bonding, coupling) SR; Form heavy metal absorbent, make the heavy metal absorbent surface have a large amount of sulfydryl functional groups, help producing absorption (complexing, chelating) effect with the heavy metal and the ion thereof that adsorb in the object liquid component.
2. heavy metal absorbent according to claim 1; It is characterized in that; The good resin microsphere carrier of biocompatibility that is used to prepare heavy metal absorbent is the hydrophilic gel microballoon, includes but not limited to hydrophily agarose gel microsphere, hydrophily sephadex microsphere, HPAG microballoon, hydrophilic polyethene alcohol microballoon; But the preferred hydrophilic agarose gel microsphere is as the carrier matrix material; The particle diameter of matrix is the 50-160 micron, and the degree of cross linking is the 4-10% scope.
3. heavy metal absorbent according to claim 1; It is characterized in that; The activator that is used to prepare heavy metal absorbent includes but not limited to epoxychloropropane, hexamethylene diamine, 1, and 1 '-carbonyl dimidazoles, two vinyl sulfone(Remzaol, ketene dimer, cyanogen bromide, allyl glycidyl ether, paratoluensulfonyl chloride, s-triazine, Cyanuric Chloride etc. can further connect groups such as amino on the aglucon, amido, hydroxyl, sulfydryl, silicon hydroxyl, boron hydroxyl, carboxyl, aldehyde radical.
4. heavy metal absorbent according to claim 1; It is characterized in that; The SR that is used to prepare heavy metal absorbent is to have free sulfhydryl groups (sulfhydryl compound SH) maybe can produce the reagent of free sulfhydryl groups through subsequent chemical reaction, includes but not limited to one or more composition of following reagent: metallothionein, sulfoprotein, sodium dimercaptosuccinate, dimercaptosuccinic acid, dimercaptosuccinic acid potassium, dimercaprol dimercaptopropanol, L-cysteine, thiocarbamide, TGA, 2 mercapto ethanol, vulcanized sodium, NaHS, potassium bisulfide, alkyl sulfide ortho acid sodium.
5. be used for external perfusion method absorption according to the said heavy metal absorbent of claim 1 and get rid of heavy metal, it is characterized in that said heavy metal includes but not limited to lead, cadmium, mercury, chromium, bismuth, cobalt, vanadium, tin, thallium and ion thereof.
6. absorption object liquid component according to claim 1; It is characterized in that described absorption object liquid component is blood of human body, serum, blood plasma, plasma component, bone marrow fluid, celiolymph, cell culture fluid, cell homogenized liquid, animal organ's extract, plant extraction liquid or other liquid component.
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