CN101169447A - Ferrous ion diagnosis/determination reagent kit and ferrous ion concentration determination method - Google Patents
Ferrous ion diagnosis/determination reagent kit and ferrous ion concentration determination method Download PDFInfo
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- CN101169447A CN101169447A CNA2006100968893A CN200610096889A CN101169447A CN 101169447 A CN101169447 A CN 101169447A CN A2006100968893 A CNA2006100968893 A CN A2006100968893A CN 200610096889 A CN200610096889 A CN 200610096889A CN 101169447 A CN101169447 A CN 101169447A
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- reagent
- ferrous ion
- ferrous
- apotransferrin
- stabilizing agent
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Abstract
The invention relates to a ferrous ion diagnosing/testing reagent box which utilizes the enzyme color method technology, and also relates to a method for testing the consistence of ferrous ion, and the elements and the components of reagent. The invention belongs to the technical field of the medicine/food/environment check. The reagent box of the invention mainly comprises cushion fluid, apoferritin, ferrous ion and stabilizer. A series of enzyme reactions occur by mixing the sample and the reagent according to a certain volume rate. Then the reactant is arranged under an ultraviolet and visible light analysis instrument to test the lifting speed of the absorbency at the 460nm of the main wavelength, thereby testing the consistence of the ferrous ion. By adopting the invention, the testing result can be obtained by the ultraviolet and visible light analysis instrument.
Description
Technical field
The present invention relates to a kind of ferrous ion diagnosis/determination reagent kit, the invention still further relates to the method for measuring ferrous ion concentration simultaneously, belong to medical science/food/environmental test determination techniques field.
Background technology
Iron and total iron binding capacity are very important in the body metabolism process.Iron is indispensable material in the human body normal physiological processes.About 3～the 5g of human body iron content, wherein 70% is present in the corpuscular hemoglobin; 25% is distributed in mononuclear phagocyte system (being reticuloendothelial system, ferritin and hemosiderin in liver, spleen, the marrow); 4.9% is distributed in the enzyme of myoglobins, cytochrome, iron content; 0.1% is the iron in the circulating plasma.
Iron in the human body derives from food, mainly is absorbed at duodenum and upper part of small intestine.Iron forms with ferritin and hemosiderin and is stored in marrow, spleen and the liver in the body.There is small amounts of iron to combine, is delivered to each tissue, be utilized or stored by blood plasma with plasma transferrins.
The mensuration of serum levels of iron mainly is to use spectrophotometric method, secondly is atomic absorption method.The latter is because sensitivity is not high and the interference of substrate, than beam split photometry poor reliability.Spectrophotometric method uses a class and contains-the N=C-C=N-structure add lustre to formerly, with ferrous formation five-membered ring, produce aubergine.That early uses has dipyridine and a red shift phenanthroline; Recently the higher ferrous piperazines of sensitivity of recommending more.In addition, the reaction of using chrome azurol S and high ferro is still arranged.
Summary of the invention
The technical problem to be solved in the present invention is: propose a kind of enzymic colorimetric (EnzymaticColorimetric Method) technology of utilizing, the continuous monitoring transferrins is in the variation of 460nm wavelength place absorbance, measured the method for ferrous ion concentration, simultaneously, the present invention also will provide in order to realize the ferrous ion diagnosis/determination reagent kit of this method, adopt this reagent not only can be ultraviolet analyser or half, carrying out ferrous ion concentration on the automatic clinical chemistry analyzer measures, and finding speed is fast, the accuracy height, thereby can obtain practical applying.
Ferrous ion concentration assay method principle of the present invention is as follows:
Ferrous oxidase (the ferroxidase that this method application need ferrous ion activates; EC EC1.16.3.1) enzyme ' s reaction speeding colourimetry.The ferrous oxidase enzymatic reaction is with apotransferrin (apotransferrin; Do not have absorption peak at the 460nm place) be oxidized into transferrins (transferrin; At the 460nm place absorption peak is arranged), thus the speed that transferrins rises in 460nm place absorbance measured, by measuring the speed that 460nm place absorbance rises, can calculate the concentration of ferrous ion.
Experiment shows, takes all factors into consideration from the accuracy of measurement result and economy two aspects of preparation cost, and no matter be single agent or two agent, the ferrous ion diagnosis/determination reagent kit of the present invention of following composition relation is comparatively desirable:
Damping fluid 200mmol/L
Stabilizing agent 50mmol/L
Apotransferrin 30mmol/L
Ferrous oxidase 12000U/L
Ferrous ion diagnosis/determination reagent kit of the present invention can be single agent, comprising:
Damping fluid, stabilizing agent, apotransferrin, ferrous oxidase.
Kit can be a dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
Also above-mentioned single agent reagent can be made into following pair of agent reagent:
Reagent 1
Damping fluid, apotransferrin.
Reagent 2
Damping fluid, stabilizing agent, ferrous oxidase.
Apotransferrin, the position of ferrous oxidase in reagent 1 or reagent 2 can not limit.Kit can be a dry powder, and use the back that is dissolved in water before use; Also can be mixed with liquid reagent, directly use.
Embodiment
The present invention is further illustrated below in conjunction with examples of implementation.
Embodiment one
The ferrous ion diagnosis/determination reagent of present embodiment is single reagent, comprising:
Acetate buffer solution 200mmol/L
Stabilizing agent 50mmol/L
Apotransferrin 30mmol/L
Ferrous oxidase 12000U/L
Reagent divides the bottle of packing into after all dissolving and preparing, and carries out freeze drying, makes powdered reagent; Before the use, add pure water, use after redissolving.
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, initial absorbance≤0.1, test predominant wavelength 460nm, test commplementary wave length 550nm, the volume ratio of tested ferrous ion sample and reagent is 1/25, the Direction of Reaction is positive reaction (reaction of rising), about about 1 minute of time delay is about 2 minutes detection times.
After adding sample and reagent, make them mixed and have reaction, reactant places under the Biochemical Analyzer the most at last, detects the speed that predominant wavelength 460nm absorbance rises, thereby calculates the concentration of ferrous ion.
Embodiment two
The ferrous ion diagnosis/determination reagent of present embodiment is double reagent, comprising:
Reagent 1
Acetate buffer solution 200mmol/L
Apotransferrin 20mmol/L
Reagent 2
Acetate buffer solution 200mmol/L
Stabilizing agent 50mmol/L
Ferrous oxidase 16000U/L
Reagent divides the bottle of packing into after all dissolving and preparing, and makes liquid double reagent, can directly use.
On automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, initial absorbance≤0.1, test predominant wavelength 460nm, test commplementary wave length 550nm, the volume ratio of tested ferrous ion sample and reagent 1, reagent 2 is 2/20/5, the Direction of Reaction is positive reaction (reaction of rising), about about 1 minute of time delay is about 2 minutes detection times.
After adding sample and reagent, make them mixed and have reaction, reactant places under the Biochemical Analyzer the most at last, detects the speed that predominant wavelength 460nm absorbance rises, thereby calculates the concentration of ferrous ion.
The applicant adopts other various reduced form chromogens combinations of putting down in writing in the above summary of the invention all can reach purpose of the present invention through experimental verification, in view of situation such as determination step and above embodiment roughly the same, do not separately enumerate.
In a word, experiment showed, and adopt assay method of the present invention can draw required measurement result by general biochemical analyzer fully, and highly sensitive, degree of accuracy good, and is easy to utilize.
Claims (5)
1. the method for measurement of concentration of the ferrous ion of an enzymic colorimetric, its method principle is as follows:
The end reaction thing is placed under visible light analysis instrument or half, the automatic clinical chemistry analyzer, detect the speed that predominant wavelength 460nm absorbance rises, calculate the concentration measurement result of ferrous ion.
2. ferrous ion diagnosis/determination reagent kit, principal ingredient comprises:
Damping fluid 20---500mmol/L
Stabilizing agent 1---50mmol/L
Apotransferrin 1---50mmol/L
Ferrous oxidase 1000---80000U/L
It is characterized in that: kit can be a dry powder, and use the back that is dissolved in water before use;
Also can be mixed with liquid reagent, directly use.
3. according to the described ferrous ion diagnosis/determination reagent kit of claim 2, it is characterized in that:
Form single agent reagent by damping fluid, stabilizing agent, apotransferrin, ferrous oxidase.
4. according to the described ferrous ion diagnosis/determination reagent kit of claim 2, it is characterized in that:
Form two agent reagent by damping fluid, stabilizing agent, apotransferrin, ferrous oxidase; Reagent 1 is made up of damping fluid, apotransferrin; Reagent 2 is made up of damping fluid, stabilizing agent, ferrous oxidase.Apotransferrin, the position of ferrous oxidase in reagent 1 or reagent 2 can not limit.
5. according to the described ferrous ion diagnosis/determination reagent kit of claim 2, it is characterized in that: also comprise stabilizing agent 1-4000mmol/L or 0.1%-100% volume ratio.Described stabilizing agent is: ammonium sulfate (Ammonia Sulfate), glycerine (Glycerol), propylene glycol (Propylene Glycol), ethylene glycol (Ethylene glycol) and at least one of the preservatives.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006100968893A CN101169447A (en) | 2006-10-24 | 2006-10-24 | Ferrous ion diagnosis/determination reagent kit and ferrous ion concentration determination method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006100968893A CN101169447A (en) | 2006-10-24 | 2006-10-24 | Ferrous ion diagnosis/determination reagent kit and ferrous ion concentration determination method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101169447A true CN101169447A (en) | 2008-04-30 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006100968893A Pending CN101169447A (en) | 2006-10-24 | 2006-10-24 | Ferrous ion diagnosis/determination reagent kit and ferrous ion concentration determination method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105466869A (en) * | 2015-11-13 | 2016-04-06 | 李彬先 | Determination method of rate of ferrous oxidase |
-
2006
- 2006-10-24 CN CNA2006100968893A patent/CN101169447A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105466869A (en) * | 2015-11-13 | 2016-04-06 | 李彬先 | Determination method of rate of ferrous oxidase |
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Open date: 20080430 |