CN1260564C - Quick determination method of trace lead being in blood of human body - Google Patents
Quick determination method of trace lead being in blood of human body Download PDFInfo
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- CN1260564C CN1260564C CN 03126945 CN03126945A CN1260564C CN 1260564 C CN1260564 C CN 1260564C CN 03126945 CN03126945 CN 03126945 CN 03126945 A CN03126945 A CN 03126945A CN 1260564 C CN1260564 C CN 1260564C
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Abstract
The present invention relates to a high-speed measuring method for a trace amount of lead in blood of a human body, which belongs to in-situ mercury plating stripping voltammetry or stripping voltammetry of hanging mercury electrodes or mercury film electrodes. Nafion or Teflon solution with the mass percentage of 3% is dropped onto the surface of a work electrode, and a film is formed by drying. Capillaries of a complexing agent are placed in a blood sample collector, wherein the complexing agent comprises one of tartrate, EGTA, DCTA, EDTA and citric acid or the mixture of some of tartrate, EGTA, DCTA, EDTA and citric acid. Base solution is prepared by adding metal ions to configured buffer solution, the stability constant of a complex compound of the metal ions and the complexing agent is larger than the stability constant of lead and the complexing agent, and the concentration rate of the metal ions is slightly higher than the concentration of lead in blood samples. In the present invention, a test can be completed by collecting a small amount of blood from the fingertip or the earlobe, testing work does not require professional laboratories or professional analysts, and the measurement of the content of lead in blood samples can be completed at high speed.
Description
(1) technical field
The present invention relates to the analysis test method technical field, specifically be meant the plumbous rapid assay methods of trace in a kind of blood of human body.
(2) background technology
Lead is the heavy metal contaminants that extensively distributes in the physical environment, and biosome is had the cumulative toxicity effect.Plumbous toxic action to human body shows as the physiological function that influence each organoid, and as nerve, hematopoiesis, digestion, uropoiesis, reproduction, cardiovascular, endocrine, immunity, skeleton etc., nervous system wherein, hemopoietic system are the main target organs that lead acts on.The World Health Organization (WHO) thinks, plumbous in the environmental poisonous substance children's's (comprising baby, child) threat likend to the National People's Congress, and this is because the former contact the approach that enters body etc. than the height of being grown up to neurological susceptibility, absorptivity and the property accumulated of lead, through life.The chronic lead poisoning meeting causes children's growth and behavior, intelligence development obstacle, brain functions such as damage recognizing ability, neurobehavioral and learning and memory.Light poisoner shows the attention deficient disorder of overacfivity, and serious poisoner then causes dementia.At present, lead content exceeds standard becomes a big factor that influences Chinese children health, realize the blood lead content crowd of exceeding standard is carried out finding morning early treatment, must set up a kind of easy, method fast that is used for that the human body blood lead content detects.
Lead content in the human body can obtain by the lead content of measuring in the blood, and whether normal standard general provision is 100 μ g/L to blood lead content.The mensuration of blood lead belongs to trace materials to be measured, and the method that generally adopt the measurement of blood lead at present the home and abroad mainly is graphite furnace atomic absorption spectrometry, ICP-MS method etc.Wherein, owing to exist a large amount of biomacromolecules in the blood sample, these big molecules generally all have certain suction-operated to lead, simultaneously, also there is interference in the existence of these materials to measurement result, so sample generally need be under the control of the condition of strictness digestion process repeatedly, to remove biomacromolecule.The digestion process of sample causes then that the sampling amount of these methods is big, the sample test time is long, sample recovery rate is lower, also needs the lab technician of analytical chemistry specialty to carry out simultaneously.In addition, method such as atomic absorption method and ICP-MS exist also that instrument costs an arm and a leg, analysis cost height, complicated operating process and be subjected to shortcomings such as matrix interference easily.And utilize lead in the organic extraction method matrix separation, then to use organic solvent, and operate loaded down with trivial details environment and human body harmful.Adopt stripping voltammetry directly to carry out mensuration plumbous in the blood report is also arranged, but because the biomacromolecule that exists in the blood pollutes to the suction-operated of lead with to electrode, though these methods can realize in that some Specialty Experiments are indoor, still can't be applied to clinical assays.
(3) summary of the invention
Purpose of the present invention is exactly in order to solve above-mentioned the deficiencies in the prior art part, and the plumbous rapid assay methods of trace in a kind of blood of human body is provided.This method is simple, does not need blood sample is carried out digestion process, need not the analyst of specialty, and the clinical express-analysis that promptly can carry out blood lead content detects.
The plumbous rapid assay methods of trace in a kind of blood of human body of the present invention, belong to coordination plating mercury stripping voltammetry or hanging mercury electrode or mercury film electrode stripping voltammetry, at first the blood sample in the blood specimen collection device is blown in the end liquid, adopt three-electrode system to measure then, it is characterized in that, the surface of the working electrode in the described three-electrode system drips that mass percent is arranged is 3% Nafion or Teflon solution, drying and forming-film; Described blood specimen collection device is the kapillary of built-in complexing agent, and described complexing agent comprises one or more potpourris in tartrate, ethylene glycol diethyl ether ethylenediamine tetraacetic acid (EDTA) (EGTA), CDTA (DCTA), ethylenediamine tetraacetic acid (EDTA), the citric acid; Liquid of the described end is to add metallic ion in the buffer solution that configures, and described metallic ion is bigger with the complexing agent stability constant than plumbous with the complex compound stability constant of complexing agent, and the concentration ratio of metallic ion wants the concentration of lead in the blood sample high slightly.
In order to realize the present invention better, described metallic ion comprises Fe
3+, Co
3-, Cr
3+
The present invention is the fast measuring that is used for blood lead content, also is applicable to the mensuration of lead content in other body fluid of human or animal, also is applicable to other metal (as Cu, Zn, Cd) Determination on content in the human or animal body.
The present invention compared with prior art has following advantage and beneficial effect:
1. the present invention does not need sample is carried out digestion process, do not need to extract a large amount of venous blood, only gather a spot of finger tip or ear-lobe blood just can be finished test, test job does not need specialized laboratory and specialty analysis personnel yet, can realize finishing fast the mensuration of lead content in the blood sample.
2. the kapillary that is coated with complexing agent in the present invention adopts is the blood specimen collection device, can when gathering, finish The pretreatment, promptly in blood specimen collection, the lead that is adsorbed by biomacromolecule in the blood sample is all discharged, and combine with complexing agent and to form complex compound, by controlling internal diameter control sampling amount capillaceous, the troublesome operation when reducing quantitative sampling.
3. the present invention adds in the end of stripping analysis liquid and the stronger metallic ion of complexing agent binding ability, as the lead ion releasing agent, can make lead ion dissociate out easily and carry out stripping voltammetry mensuration.The stabilizing solution that liquid of the employed end is made up of pH value buffering agent, mercury ion and lead ion releasing agent etc. only needs 1mL during test, can make kit easily and be used for clinical and on-site measurement.
4. the present invention once measures finger tip blood or the ear-lobe blood that only needs 25 μ L, without venous blood samples, more helps children and baby's blood lead generaI investigation.
5. the cleaning of electrode of the present invention only need scan 1 minute with constant potential method or cyclic voltammetric method with activation in the nitric acid of 0.1mol/L, and method is easy.
6. the making of standard working curve of the present invention, only need utilize the blood lead standard model, with the repeatedly identical mensuration of standard addition method, promptly can guarantee under the constant condition of matrix condition, finish rapidly, and only need make the mensuration that the one action curve can be finished 500 duplicate samples an electrode.
(4) embodiment
Below in conjunction with embodiment, the present invention is done detailed description further.
The employed electrode of embodiment can be commercialization disk electrode or the round loop electrode of directly buying, and electrode material can be gold, silver, glass carbon or carbon electrode.
After the polishing of electrode process, cleaning, drying, the surperficial mass percent that upwards drips 20 μ L is 3% Nafion or Teflon solution, knock gently and make the even branch of solution at whole electrode surfaces, perhaps electrode is directly immersed modification solution, take out also rapidly and vibrate 1 minute gently, just form one deck decorative layer at electrode surface after the air dry.Conventional stripping voltammetry generally is to adopt ruhende electrode or bare electrode, but for direct mensuration plumbous in the blood sample, owing to contain a large amount of biomacromolecules in the sample, electrode is produced pollute easily, general can the generation electrode in the preenrichment process polluted the reappearance variation that makes mensuration.By modification to electrode surface, adopt coordination plating mercury stripping voltammetry to measure blood content in the blood sample, can prevent electrode fouling effectively, and the electrode after modifying can reuse more than 500 times, can also keep good reappearance.
The sampling kapillary of blood sample can be to draw by the kapillary drawing machine, or directly adopts the commodity kapillary, and every batch of (the 50 or 100) internal diameter capillaceous that requires to use is wanted evenly, and relative deviation is less than 3%.After single batch of kapillary cleans simultaneously, vertically downward their tip is immersed in the plumbous enveloping agent solution, solution can be tartrate, EGTA, DCTA, EDTA, citric acid etc., and the concentration of solution is 1 * 10
-5Mol/L.Difference according to tested object also can be adjusted concentration, and as professional patient is measured, can suitably heighten concentration (can be to 5 * 10
-5Mol/L).Placed about 1 minute, treat to take out simultaneously after liquid level no longer raises in the kapillary and air dry standby.
Get the pure anhydrous sodium acetate 50g of top grade and be dissolved in the inferior boiling water, add the pure glacial acetic acid of 60mL top grade, be made into buffer solution, get the pure Hg (NO of top grade of 0.1623g again
3)
2Be dissolved in the inferior boiling water and and mix, in mixed solution, add a certain amount of metallic ion (Fe for example then with buffer solution
3+, Co
3+, Cr
3+), be diluted to 5000mL at last and feed high pure nitrogen deoxygenation in 10 minutes promptly can be used as the test end liquid.Wherein, the kind of the metallic ion that adds is looked different complexing agent and difference, the principle of selecting is, the metallic ion that is added is bigger than plumbous and corresponding complexing agent stability constant with the complex compound stability constant of use complexing agent, and the concentration ratio of metallic ion wants the concentration of the lead in the working sample high slightly.
Liquid is put into small beaker at the bottom of getting 1ml, puncture finger tip or ear-lobe then, small amounts of blood is flowed out naturally, drop of blood is aimed at the tip of blood specimen collection device, blood sample can be adopted into collector automatically by capillarity, and the blood sample in the blood specimen collection device is blown in the small beaker, leaves standstill after shaking up about 1 minute, adopt three-electrode system with plating Mo electrode then, carry out coordination plating mercury stripping voltammetry and measure.
After each sample determination is finished, electrode is placed the nitric acid of 0.1mol/L, locate to leave standstill cleaning and the activation of carrying out electrode in 1 minute at-0.1V (to the Ag/AgCl electrode).
Stripping peak current or peak area with lead are index, adopt standard addition method or standard working curve method to carry out quantitatively.Wherein the method for making of standard working curve is, adopt the blood lead standard sample of 30 μ g/L to measure by above-mentioned steps, record peak current or peak area adopt standard addition method to same sample then, measure successively and obtain one group of data, adopt linear regression production standard working curve.
The present invention adopts the method that plates Nafion film or Teflon film on disk electrode (also can be round loop electrode) to make electrode, and when kind electrode can prevent effectively that blood lead from measuring, the biomacromolecule in the blood sample was to the pollution of electrode; The kapillary that employing scribbles complexing agent carries out the collection of blood sample and since the complexing agent that is coated with stronger than the biomolecule in the blood to the complexing power of lead, so can in blood specimen collection, finish The pretreatment; Except containing the needed whole reagent of coordination plating mercury stripping voltammetry, added the excess metal ion stronger than lead in the test end liquid, can effectively the sampling back have been discharged fully by the lead ion of complexing, so that measure with the complexing agent complexing power; The acquisition method of blood sample is to gather finger tip blood or ear-lobe blood, utilization be that capillarity is sampled automatically, the collection capacity of blood can generally be controlled at 20～50 μ L by internal diameter control capillaceous; Adopt coordination plating mercury stripping voltammetry to measure, also can adopt hanging mercury electrode or mercury film electrode to measure, the stripping voltammetry that adopts can be single sweep stripping voltammetry, square wave stripping voltammetry, differentiated pulse stripping volt-ampere and continuous current current potential stripping voltammetry, and quantitative test can be the quantitative or peak area quantification of peak current; Behind each sample determination, use potentiostatic method that electrode is cleaned, also can clean electrode with cyclic voltammetry; Employing adds the method for standard lead and carries out the making of working curve in the blood lead standard, can guarantee the synchronism of matrix effect.
As mentioned above, can realize the present invention preferably.
Claims (2)
1. the plumbous rapid assay methods of trace in the blood of human body, belong to coordination plating mercury stripping voltammetry or hanging mercury electrode or mercury film electrode stripping voltammetry, at first the blood sample in the blood specimen collection device is blown in the end liquid, adopt three-electrode system to measure then, it is characterized in that, the surface of the working electrode in the described three-electrode system drips that mass percent is arranged is 3% Nafion or Teflon solution, drying and forming-film; Described blood specimen collection device is the kapillary of built-in complexing agent, and described complexing agent comprises one or more potpourris in tartrate, ethylene glycol diethyl ether ethylenediamine tetraacetic acid (EDTA), CDTA, ethylenediamine tetraacetic acid, the citric acid; Liquid of the described end is to add metallic ion in the buffer solution that configures, and described metallic ion is bigger with the complexing agent stability constant than plumbous with the complex compound stability constant of complexing agent, and the concentration of metallic ion is higher slightly than concentration plumbous in the blood sample.
2. the plumbous rapid assay methods of trace is characterized in that described metallic ion comprises Fe in a kind of blood of human body according to claim 1
3+, Co
3+, Cr
3+
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03126945 CN1260564C (en) | 2003-06-23 | 2003-06-23 | Quick determination method of trace lead being in blood of human body |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03126945 CN1260564C (en) | 2003-06-23 | 2003-06-23 | Quick determination method of trace lead being in blood of human body |
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| CN1460853A CN1460853A (en) | 2003-12-10 |
| CN1260564C true CN1260564C (en) | 2006-06-21 |
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| CN 03126945 Expired - Fee Related CN1260564C (en) | 2003-06-23 | 2003-06-23 | Quick determination method of trace lead being in blood of human body |
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Families Citing this family (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100346159C (en) * | 2004-05-03 | 2007-10-31 | 北京艾联联合科技发展有限公司 | Determination reagent for trace content of heavy metal |
| CN102495117B (en) * | 2011-12-19 | 2013-11-20 | 天津理工大学 | Method for improving repeatability of working electrode and application of method |
| CN103149255A (en) * | 2013-02-25 | 2013-06-12 | 长沙理工大学 | Method for quickly determining lead content in blood |
| CN103940756A (en) * | 2013-08-06 | 2014-07-23 | 江苏天瑞仪器股份有限公司 | Reagent pack for detection of lead ions in water |
| CN103472230B (en) * | 2013-09-28 | 2015-12-02 | 河南科技学院 | Detect indirect competitive enzyme-linked immunosorbent kit of lead ion and preparation method thereof |
| CN103592356B (en) * | 2013-11-15 | 2016-10-05 | 天津理工大学 | A kind of method using scan anode stripping voltammetry quickly to detect lead, cadmium |
| CN104950021B (en) * | 2014-03-28 | 2018-04-17 | 无锡市申瑞生物制品有限公司 | A kind of deserted silk-screen printing sensor for blood lead analysis and preparation method thereof |
| CN105628684B (en) * | 2015-12-30 | 2019-10-08 | 河南省有色金属地质勘查总院 | A method of utilizing high-content lead in ICP-AES method measurement Pb-Zn deposits |
| CN108627565B (en) * | 2018-05-14 | 2020-08-14 | 桂林理工大学 | Bismuth-copper mixed coating test strip and preparation method and application thereof |
| CN113237939A (en) * | 2021-05-10 | 2021-08-10 | 深圳市朗石科学仪器有限公司 | Method for measuring co-plating mercury film of multiple heavy metals in surface water |
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