CN101620166A - Manganese ion diagnosis/ determination kit and method for determining manganese ion concentration - Google Patents
Manganese ion diagnosis/ determination kit and method for determining manganese ion concentration Download PDFInfo
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- CN101620166A CN101620166A CN200810124236A CN200810124236A CN101620166A CN 101620166 A CN101620166 A CN 101620166A CN 200810124236 A CN200810124236 A CN 200810124236A CN 200810124236 A CN200810124236 A CN 200810124236A CN 101620166 A CN101620166 A CN 101620166A
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- reagent
- dehydrogenase
- manganese ion
- oxaloacetic
- coenzyme
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- WAEMQWOKJMHJLA-UHFFFAOYSA-N Manganese(2+) Chemical compound [Mn+2] WAEMQWOKJMHJLA-UHFFFAOYSA-N 0.000 title claims abstract description 38
- 229910001437 manganese ion Inorganic materials 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 18
- 238000003745 diagnosis Methods 0.000 title claims abstract description 15
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 68
- ZSLZBFCDCINBPY-ZSJPKINUSA-N acetyl-CoA Chemical compound O[C@@H]1[C@H](OP(O)(O)=O)[C@@H](COP(O)(=O)OP(O)(=O)OCC(C)(C)[C@@H](O)C(=O)NCCC(=O)NCCSC(=O)C)O[C@H]1N1C2=NC=NC(N)=C2N=C1 ZSLZBFCDCINBPY-ZSJPKINUSA-N 0.000 claims abstract description 40
- 239000005515 coenzyme Substances 0.000 claims abstract description 32
- 239000003381 stabilizer Substances 0.000 claims abstract description 25
- 108010066133 D-octopine dehydrogenase Proteins 0.000 claims abstract description 23
- 101710088194 Dehydrogenase Proteins 0.000 claims abstract description 21
- KHPXUQMNIQBQEV-UHFFFAOYSA-N oxaloacetic acid Chemical compound OC(=O)CC(=O)C(O)=O KHPXUQMNIQBQEV-UHFFFAOYSA-N 0.000 claims abstract description 21
- 239000004475 Arginine Substances 0.000 claims abstract description 20
- 229940100228 acetyl coenzyme a Drugs 0.000 claims abstract description 20
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims abstract description 20
- 238000006243 chemical reaction Methods 0.000 claims abstract description 16
- 238000002835 absorbance Methods 0.000 claims abstract description 13
- 108010011939 Pyruvate Decarboxylase Proteins 0.000 claims abstract description 11
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 claims abstract description 9
- 108090000790 Enzymes Proteins 0.000 claims abstract description 8
- 102000004190 Enzymes Human genes 0.000 claims abstract description 8
- 238000005516 engineering process Methods 0.000 claims abstract description 4
- 239000000203 mixture Substances 0.000 claims abstract description 4
- 238000013016 damping Methods 0.000 claims description 16
- 239000012530 fluid Substances 0.000 claims description 16
- 108090000489 Carboxy-Lyases Proteins 0.000 claims description 12
- LCTONWCANYUPML-UHFFFAOYSA-N Pyruvic acid Chemical compound CC(=O)C(O)=O LCTONWCANYUPML-UHFFFAOYSA-N 0.000 claims description 12
- 239000003795 chemical substances by application Substances 0.000 claims description 11
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- 239000007788 liquid Substances 0.000 claims description 6
- 239000011572 manganese Substances 0.000 claims description 6
- 238000003556 assay Methods 0.000 claims description 5
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 5
- 239000001569 carbon dioxide Substances 0.000 claims description 5
- 229940107700 pyruvic acid Drugs 0.000 claims description 5
- 125000000637 arginyl group Chemical group N[C@@H](CCCNC(N)=N)C(=O)* 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 230000000694 effects Effects 0.000 claims description 3
- 238000005259 measurement Methods 0.000 claims description 3
- IMXSCCDUAFEIOE-RITPCOANSA-N D-octopine Chemical compound [O-]C(=O)[C@@H](C)[NH2+][C@H](C([O-])=O)CCCNC(N)=[NH2+] IMXSCCDUAFEIOE-RITPCOANSA-N 0.000 claims description 2
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 claims 6
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims 5
- 235000011187 glycerol Nutrition 0.000 claims 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims 1
- 235000011130 ammonium sulphate Nutrition 0.000 claims 1
- 239000004615 ingredient Substances 0.000 claims 1
- 239000003755 preservative agent Substances 0.000 claims 1
- 238000012360 testing method Methods 0.000 abstract description 11
- 239000000376 reactant Substances 0.000 abstract description 4
- 238000004737 colorimetric analysis Methods 0.000 abstract description 2
- 238000002965 ELISA Methods 0.000 abstract 1
- 239000007853 buffer solution Substances 0.000 abstract 1
- NQMRYBIKMRVZLB-UHFFFAOYSA-N methylamine hydrochloride Chemical compound [Cl-].[NH3+]C NQMRYBIKMRVZLB-UHFFFAOYSA-N 0.000 description 6
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 229910052748 manganese Inorganic materials 0.000 description 3
- 238000012856 packing Methods 0.000 description 3
- 230000035484 reaction time Effects 0.000 description 3
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 108010017192 4-hydroxy-4-methyl-2-oxoglutarate aldolase Proteins 0.000 description 1
- 102100029589 Acylpyruvase FAHD1, mitochondrial Human genes 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- IMXSCCDUAFEIOE-UHFFFAOYSA-N D-Octopin Natural products OC(=O)C(C)NC(C(O)=O)CCCN=C(N)N IMXSCCDUAFEIOE-UHFFFAOYSA-N 0.000 description 1
- 108700033374 EC 4.1.1.3 Proteins 0.000 description 1
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- ACFIXJIJDZMPPO-NNYOXOHSSA-N NADPH Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](OP(O)(O)=O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NNYOXOHSSA-N 0.000 description 1
- 108010069823 Oxaloacetate decarboxylase Proteins 0.000 description 1
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000003321 atomic absorption spectrophotometry Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 229910002804 graphite Inorganic materials 0.000 description 1
- 239000010439 graphite Substances 0.000 description 1
- 238000002354 inductively-coupled plasma atomic emission spectroscopy Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229940076788 pyruvate Drugs 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention relates to a manganese ion diagnosis/ determination kit using the technology of an enzyme multiplication method, an enzyme colorimetric method and an ELISA method, a method for determining manganese ion concentration, and composition and components of a reagent, and belongs to the technical field of test and determination of medical science, food and environment. The kit comprises the following main components: buffer solution, reduced coenzyme, oxaloacetic acid, acetylcoenzyme A, arginine, oxaloacetic decarboxylase, pyruvate dehydrogenase, octopine dehydrogenase and a stabilizer. The method for determining manganese ion concentration comprises the following steps: mixing a sample and the reagent according to a certain volume ratio to perform a series of enzymic reaction, and then placing reactants under a UV/ visible light analyzer to test the descending speed of absorbance at the position where the dominant wave length is 340nm to determine the concentration of manganese ion.
Description
Technical field
The present invention relates to a kind of manganese ion diagnosis/mensuration kit, the invention still further relates to the method for measuring manganese ion concentration simultaneously, belong to medical science/food/environmental test determination techniques field.
Background technology
Manganese is indispensable essential composition in human body as a kind of trace element, also is the material poisonous to human body simultaneously.Under state of nature, exist with the form of two valencys, trivalent and tetravalence, degree of oxidation is low more, and toxicity is high more.Usually manganese has eight kinds of different states of oxidation.The biological significance of manganese is as accessory factor in the human body diversified function to be arranged.Yet, in many enzyme activating reactions, Mn
2+And Mg
2+Can substitute mutually.
Sampling Graphite Furnace Atomic Absorption spectrophotometry commonly used, atomic emissions ICP-OES.It is the best way that neutron excites analysis (NAA), but only can could measure in the good laboratory of some conditions.
Summary of the invention
The technical problem to be solved in the present invention is: propose a kind of enzyme multiplication method (Enzymatic DoublingMethod) that utilizes, enzymic colorimetric (Enzymatic Colorimetric Method) and enzyme (even) united method (CoupleReaction) technology, monitoring reduced form nicotinamide coenzyme (reduced coenzyme) is in the variation of 340nm wavelength place absorbance, measured the method for manganese ion concentration, simultaneously, the present invention also will provide in order to realize the manganese ion diagnosis/mensuration kit of this method, adopt this reagent not only can be ultraviolet analyser or half, carrying out manganese ion concentration on the automatic clinical chemistry analyzer measures, and finding speed is fast, the accuracy height, thereby can obtain practical applying.
Manganese ion concentration assay method principle of the present invention is as follows:
Oxaloacetic acid
Oxaloacetic decarboxylase/Mn 2+ Pyruvic acid+carbon dioxide
Carbon dioxide+acetyl coenzyme A+reduced coenzyme
Pyruvic dehydrogenasePyruvic acid+coacetylase+coenzyme
Pyruvic acid+arginine+reduced coenzyme
Octopine dehydrogenaseN2-(D-1-carboxyethyl)-L-arginine+coenzyme+water
This method application need manganese ion intensifies active oxaloacetic decarboxylase (oxaloacetatedecarboxylase; EC 4.1.1.3) enzyme (idol) connection pyruvic dehydrogenase (pyruvate dehydrogenase; EC1.2.1.51), octopine dehydrogenase (Octopine dehydrogenase; EC 1.5.1.11) enzymatic reaction continuous monitoring method/speed ratio color method.The activation degree of oxaloacetic decarboxylase is directly proportional with manganese ion concentration.The oxaloacetic decarboxylase enzymolysis oxaloacetic acid reaction that manganese ion intensifies under the activity produces pyruvic acid and carbon dioxide, the double action of uniting pyruvic dehydrogenase, octopine dehydrogenase again by (idol), final secondary is oxidized into coenzyme (not having absorption peak at the 340nm place) with reduced coenzyme (absorption peak being arranged at the 340nm place), thereby measured the speed that reduced coenzyme descends in 340nm place absorbance, by measuring the speed that 340nm place absorbance descends, can calculate the concentration of manganese ion.
Experiment shows, takes all factors into consideration from the accuracy of measurement result and economy two aspects of preparation cost, no matter is single agent, two agent or three doses, and the manganese ion diagnosis/mensuration kit of the present invention of following composition relation is ideal comparatively:
Damping fluid 100mmol/L
Stabilizing agent 500mmol/L
Reduced coenzyme 0.25mmol/L
Oxaloacetic decarboxylase 6000U/L
Pyruvic dehydrogenase 8000U/L
Octopine dehydrogenase 10000U/L
Oxaloacetic acid 12mmol/L
Acetyl coenzyme A 6mmol/L
Arginine 9mmol/L
Manganese ion diagnosis/mensuration kit of the present invention can be single agent, comprising:
Damping fluid, stabilizing agent, reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, arginine.
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, stabilizing agent, reduced coenzyme, oxaloacetic acid, acetyl coenzyme A, arginine.
Reagent 2
Damping fluid, stabilizing agent, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase.
Reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, the position of arginine 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.
Above-mentioned single agent reagent can also be made into following three doses of reagent:
Reagent 1
Damping fluid, stabilizing agent, reduced coenzyme, oxaloacetic acid, acetyl coenzyme A, arginine.
Reagent 2
Damping fluid, stabilizing agent, pyruvic dehydrogenase, octopine dehydrogenase.
Reagent 3
Damping fluid, stabilizing agent, oxaloacetic decarboxylase.
Reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, the position of arginine in reagent 1, reagent 2 or reagent 3 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.
No matter be single agent, two agent or three doses, the present invention measures the method for manganese ion concentration, and its reduced coenzyme can be a kind of among NADPH, NADH or the thio-NADH.
Embodiment
The present invention is further illustrated below in conjunction with examples of implementation.
Embodiment one
Manganese ion diagnosis/mensuration the reagent of present embodiment is single reagent, comprising:
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Reduced coenzyme 0.25mmol/L
Oxaloacetic decarboxylase 6000U/L
Pyruvic dehydrogenase 8000U/L
Octopine dehydrogenase 10000U/L
Oxaloacetic acid 12mmol/L
Acetyl coenzyme A 6mmol/L
Arginine 9mmol/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 1.8 ± 0.7, test predominant wavelength 340nm, test commplementary wave length 405nm, the volume ratio of tested manganese ion sample and reagent is 1/25, the Direction of Reaction is negative reaction (reaction descends), 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 340nm absorbance descends, thereby calculates the concentration of manganese ion.
Embodiment two
Manganese ion diagnosis/mensuration the reagent of present embodiment is double reagent, comprising:
Reagent 1
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 50mmol/L
Reduced coenzyme 0.25mmol/L
Oxaloacetic acid 12mmol/L
Acetyl coenzyme A 6mmol/L
Arginine 9mmol/L
Reagent 2
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Oxaloacetic decarboxylase 6000U/L
Pyruvic dehydrogenase 8000U/L
Octopine dehydrogenase 10000U/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 1.8 ± 0.7, test predominant wavelength 340nm, test commplementary wave length 405nm, the volume ratio of tested manganese ion sample and reagent 1, reagent 2 is 2/20/5, the Direction of Reaction is negative reaction (reaction descends), 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 340nm absorbance descends, thereby calculates the concentration of manganese ion.
Embodiment three
Manganese ion diagnosis/mensuration the reagent of present embodiment is three reagent, comprising:
Reagent 1
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 50mmol/L
Reduced coenzyme 0.25mmol/L
Oxaloacetic acid 12mmol/L
Acetyl coenzyme A 6mmol/L
Arginine 9mmol/L
Reagent 2
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Pyruvic dehydrogenase 8000U/L
Octopine dehydrogenase 10000U/L
Reagent 3
Three (ethyloic) aminomethane-hydrochloride buffer 100mmol/L
Stabilizing agent 500mmol/L
Oxaloacetic decarboxylase 6000U/L
Reagent divides the bottle of packing into after all dissolving and preparing, and makes liquid three reagent, can directly use.
When measuring manganese ion concentration, on automatic clinical chemistry analyzer, set: 37 ℃ of temperature, 10 minutes reaction time, initial absorbance 1.8 ± 0.7, test predominant wavelength 340nm, test commplementary wave length 405nm, the volume ratio of tested manganese ion sample and reagent 1, reagent 2, reagent 3 is 4/40/5/5, the Direction of Reaction is negative reaction (reaction descends), and 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 340nm absorbance descends, thereby calculates the concentration of manganese ion.
The applicant adopts other assay methods 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, experimental results show that: adopt assay method of the present invention can draw required measurement result by general biochemical analyzer fully---the blank reagent absorbance changes (Δ A/min)≤0.001; Absorbance time response curve should be decline curve until terminal point; Reagent can be surveyed effectively, and (R 〉=0.99) linear range can reach 5mmol/L; The inaccuracy of reagent test, its relative deviation be no more than ± and 5%; The coefficient of variation (CV)≤2% of the precision of reagent test (repeatability); The sensitivity of reagent can reach 0.18 ± 0.009 Δ A/mmol/L; Reagent is preserved down at 2-8 ℃, and activity can be stablized 1 year;---the present invention is highly sensitive, degree of accuracy good, the linear range broadness, and stationary phase is long, is enough to easy to utilize.
Claims (6)
1. manganese ion concentration assay method that utilizes enzyme multiplication method, enzymic colorimetric and enzyme-linked method technology, its method principle is as follows:
Oxaloacetic acid
Oxaloacetic decarboxylase/Mn 2+ Pyruvic acid+carbon dioxide
Carbon dioxide+acetyl coenzyme A+reduced coenzyme
Pyruvic dehydrogenasePyruvic acid+
Coacetylase+coenzyme
Pyruvic acid+arginine+reduced coenzyme
Octopine dehydrogenase
N2-(D-1-carboxyethyl)-L-arginine+coenzyme+water
The end reaction thing is placed under ultraviolet analyser or half, the automatic clinical chemistry analyzer, detect the speed that predominant wavelength 340nm absorbance descends, calculate the concentration measurement result of manganese ion.
2. manganese ion diagnosis/mensuration kit, principal ingredient comprises:
Damping fluid 20---500mmol/L
Stabilizing agent 1---4000mmol/L
Reduced coenzyme 0.1---0.35mmol/L
Oxaloacetic decarboxylase 1000---80000U/L
Pyruvic dehydrogenase 1000---80000U/L
Octopine dehydrogenase 1000---80000U/L
Oxaloacetic acid 1---50mmol/L
Acetyl coenzyme A 1---50mmol/L
Arginine 1---50mmol/L
The concentration of reagent composition not necessarily is only limited to above-mentioned scope; Effect is better in this scope, and outside this scope, reagent still can reagentia.
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 manganese ion diagnosis/mensuration of claim 2 kit, it is characterized in that:
Form single agent reagent by damping fluid, stabilizing agent, reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, arginine.
4. according to the described manganese ion diagnosis/mensuration of claim 2 kit, it is characterized in that:
Form two agent reagent by damping fluid, stabilizing agent, reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, arginine; Reagent 1 is made up of damping fluid, stabilizing agent, reduced coenzyme, oxaloacetic acid, acetyl coenzyme A, arginine; Reagent 2 is made up of damping fluid, stabilizing agent, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase.Reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, the position of arginine in reagent 1 or reagent 2 can not limit.
5. according to the described manganese ion diagnosis/mensuration of claim 2 kit, it is characterized in that:
Form multi-agent reagent by damping fluid, stabilizing agent, reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, arginine; Reagent 1 is made up of damping fluid, stabilizing agent, reduced coenzyme, oxaloacetic acid, acetyl coenzyme A, arginine; Reagent 2 is made up of damping fluid, stabilizing agent, pyruvic dehydrogenase, octopine dehydrogenase; Reagent 3 is made up of damping fluid, stabilizing agent, oxaloacetic decarboxylase.Reduced coenzyme, oxaloacetic decarboxylase, pyruvic dehydrogenase, octopine dehydrogenase, oxaloacetic acid, acetyl coenzyme A, the position of arginine in reagent 1, reagent 2 or reagent 3 can not limit.
6. according to the described manganese ion diagnosis/mensuration of claim 2 kit, it is characterized in that: also comprise stabilizing agent 1-4000mmol/L or 0.1%-100% volume ratio.Described stabilizing agent is: ammonium sulfate (AmmoniaSulfate), 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 |
|---|---|---|---|
| CN200810124236A CN101620166A (en) | 2008-06-30 | 2008-06-30 | Manganese ion diagnosis/ determination kit and method for determining manganese ion concentration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810124236A CN101620166A (en) | 2008-06-30 | 2008-06-30 | Manganese ion diagnosis/ determination kit and method for determining manganese ion concentration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101620166A true CN101620166A (en) | 2010-01-06 |
Family
ID=41513512
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200810124236A Pending CN101620166A (en) | 2008-06-30 | 2008-06-30 | Manganese ion diagnosis/ determination kit and method for determining manganese ion concentration |
Country Status (1)
| Country | Link |
|---|---|
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2008
- 2008-06-30 CN CN200810124236A patent/CN101620166A/en active Pending
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Open date: 20100106 |