CN1778942A - Determination of inorganic phosphorus and its diagnostic reagent kit - Google Patents

Abstract

The invention is about the measuring method of Inorganic Phosphates and its diagnosis reagent box. Producing hypoxanthine by reacting nucleoside phosphorylase with carnine under the existence of Inorganic Phosphates , then producing hydroperoxide by reactinghypoxanthine with xanthine oxidase, and then reacting hydroperoxide with peroxidaseand oxidating the colorless reduced chromogen combination to quinoneimine chromogen or indamide chromogen dyer with color, testing the variation of dominant wave-length400-600nmabsorbance during the reaction and finally measuring the content of Inorganic Phosphates . This method has high specificity and would not be contaminated by material of internal and exogenous sources, and the result is precise and accurate. Diving the diagnosis reagent box into double-dose or three-dose can reduces the cross interaction of each element, keeps the stability of the reagent and deposits chronically. Using this method can realize the fast testing in common ultraviolet/ visible light analyzer or semiautomatic/automatic analyzer and doesn't require special or additional apparatus, so the cost is low. Thus, this method can be easily promoted and applied in the whole industry.

Description

The measuring method of inorganic phosphorus and diagnostic kit thereof

Technical field

The present invention relates to measure the method for inorganic phosphorus, and use the formulated diagnostic kit of this method, belong to medical test determination techniques field.

Background technology

The phosphoric of inside of human body has 87% to be present in the bone approximately, and phosphorus in the blood and the phosphorus in the bone keep dynamic equilibrium relation.On the other hand, certain relation is arranged between the concentration of calcium and phosphorus in the blood, both products keep within limits, are unit with the milligram, and approximately the amassing of the content of calcium and phosphorus is 35～40 in per 100 milliliters of blood plasma.Normal people's blood calcium serium inorganic phosphorus that raises then reduces, and vice versa.When above-mentioned product greater than 40 the time, calcium and phosphorus are deposited on osseous tissue with the bone salts form; Greater than 70 o'clock, metastatic calcification can take place; Less than 35 o'clock, then will hinder the calcification of osseous tissue, even bone salts is dissolved again, influence osteogenesis, cause rickets or richets.Usually, can wait the concentration of regulating calcium and phosphorus in the blood plasma by vitamins D, Rat parathyroid hormone 1-34, thyrocalcitonin.

At present, the phosphorous total amount of human body can't directly be measured, and is that phosphorus acid ion concentration reflects in detection blood plasma or the serum mostly.Because H ₂PO ₄ ^-Can stable existence under natural condition such as blood, be the main existence form of inorganic phosphorus, thereby measure H ₂PO ₄ ^-Concentration just can represent or know by inference the total amount of inorganic phosphorus indirectly.Detection method commonly used has: phospho-molybdic acid reduction method/non-reduced method, dye binding method, enzyme process, isotopic dilution mass spectrometry, atomic absorption spectrophotometry, flow injection analysis or the like.Wherein, conclusive method is an isotopic dilution mass spectrometry, and the ordinary method that medium laboratory recommends to use is the phospho-molybdic acid colorimetry.

The phospho-molybdic acid reduction method is divided " no proteinemia filtrate method " and " not deproteinate method " again, and the former needs the Deproteinization with blood elder generation, and the latter needs to add nonionic surface active agent to avoid muddy in reagent.The kind of used reductive agent and tensio-active agent is a lot, and the more stable reductive agent of performance has ferrous sulfate, ferrous ammonium sulphate, hydrazonium sulfate, N-methyl p-aminophenol sulfate etc., and tensio-active agent is then the most desirable with polyethylene glycol groups phenyl ether (TritonX-100).

The non-reduced method of phospho-molybdic acid claims " direct ultraviolet method " again, is directly to measure the phospho-molybdic acid complex poly compounds at 340nm or 325nm wavelength, and method is easy, is convenient to automatization.But jaundice, haemolysis, the turbid serum of fat have absorption at 340nm, must do the sample blank, otherwise the result are higher.

Than other detection method, enzyme process is good, highly sensitive with its selectivity, can realize advantage such as automated analysis and enjoy people to favor, and is present people's research focus with the enzymatic assays content of inorganic phosphorus, also is development trend in the future.But, adopting different reagent and test route, the sensitivity of detection, the accuracy of detected result have very big difference, directly have influence on the use and the popularization of this method.

Summary of the invention

The objective of the invention is: a kind of method of enzymatic assays content of inorganic phosphorus is provided, and uses the formulated inorganic phosphorus diagnosis kit of this method.Adopt the reagent in this test kit, can utilize ultraviolet analyser or semi-automatic/automatic clinical chemistry analyzer that the content of inorganic phosphorus in the samples such as blood plasma, serum is carried out fast, accurately measures, so that promote the use of this detection method and diagnostic reagent, make in this area the medical test means of testing obtain abundant and development.

For realizing purpose of the present invention, a kind of method of measuring inorganic phosphorus, undertaken by following steps:

At first,, make it to take place following reaction with needs such as blood plasma, serum sample of measuring and the reagent mixing that contains inosine, Phosphatase, nucleotide, XOD, peroxidase and the combination of reduced form chromogen,

Then, reaction mixture is placed under ultraviolet analyser or half, the automatic clinical chemistry analyzer, detecting predominant wavelength is the lift velocity of the absorbancy of 400～600nm, and then calculates the content of inorganic phosphorus in the sample.

Above-mentioned " combination of reduced form chromogen " is to be made of reduced form chromogen A and another B component.Reduced form chromogen A is: " 3-methyl-2-Proxel hydrazone " (English MBTH that is called for short, full name is 3-methyl-2-benzothiazolinone-hydrszone) or " the amino anti-arsenic Yun of 4-" (English full name is 4-aminoantipyrine); Another B component is one of following 15 kinds of materials:

1. PHENOL 99.8 MIN ((CARBOLIC ACID)) (phenol),

2. N-ethyl-N-(3-thiopropyl)-m-thialdine amine (N-ethyl-N-(3-sulfopropyl)-m-anisidine, be called for short ESPAS),

3. N, the two ethyls of N--m-Tolylamine (N, N-Diethyl-m-toluidine),

4. 2, the two chlorine PHENOL 99.8 MIN ((CARBOLIC ACID)) of 4-(2,4-Dichloriphenol),

5. 2,4,6-three bromo-3-hydroxyl-Phenylsulfonic acid (2,4,6-Tribromo-3-hydroxy-benzenesulfonic acid is called for short TBHB),

6. 3, the two chlorine PHENOL 99.8 MIN ((CARBOLIC ACID)) sulfonic acid (3,5-Dichlorophenolsulfonic acid) of 5-,

7. 3, the two chloro-2-hydroxyl-Phenylsulfonic acids of 5-(3,5-Dichloro-2-hydroxy-benzenesulfonicacid is called for short DHBS),

8. N-ethyl-N-(2-hydroxyl-3-thiopropyl)-m-Tolylamine sodium salt (N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine sodium, be called for short TOOS),

9. three bromine hydroxy-benzoic acid (Tribromohydroxybenzoic acid),

10. two monomethylanilines (Dimethylaniline is called for short DMA),

(11) N-ethyl-N-(2-hydroxyl-3-thiopropyl)-m-Tolylamine (be called for short EHSPT, full name is N-Ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine),

(12) 2,2 '-Lian nitrogen-two (3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) di-ammonium salts (2,2 '-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid), be called for short ABTS),

(13) 2,2 '-azino-two (3-ethyl benzo thiophene pyrrolines-6-sulfonic acid), English name is 2,2 '-azino-bis-(3-ethylbenztbiozoline-6-sulfonic acid),

(14) vanillic acid (Vanillic acid (4-hydroxy-3-methoxybenzoicacid) is called for short HMB),

(15) 3-methyl-ethyl-hydroxyanilines (3-methyl-ethyl-hydroxyaniline is called for short MEHA).

In the mensuration process, the usage ratio of sample and reagent by volume was controlled at 1: 10～1: 500, and temperature of reaction is controlled at 20 ℃～50 ℃, and the reaction times was controlled at 2～30 minutes, set commplementary wave length during detection more than 500～700nm.

Realize that it can be single agent that the inventive method is measured the diagnostic kit of content of inorganic phosphorus, is grouped into by following one-tenth:

Damping fluid 40～200mmol/l,

Inosine 0.1～10mmol/l,

Phosphatase, nucleotide 500～50000U/l,

XOD 500～50000U/l,

Peroxidase 500～50000U/l,

Reduced form chromogen combination 0.1～20mmol/l,

Stablizer/reagent cumulative volume 10～80%.

Wherein the implication of " combination of reduced form chromogen " as previously mentioned, the concentration of two kinds of component A and B is 0.1～20mmol/l.

Also the various compositions in above single agent can be made up, be mixed with two agent, such as:

	Composition	Content range
			Reagent I	Damping fluid	40～200mmol/l
Inosine	0.1～10mmol/l
		Reduced form chromogen combination (one)		0.1～20mmol/l
Stablizer	10～50mmol/l
		Reagent II		Damping fluid	40～200mmol/l
Phosphatase, nucleotide	500～50000U/l
			XOD	500～50000U/l

	Peroxidase	500～50000U/l
			Reduced form chromogen combination (two)	0.1～20mmol/l
	Stablizer/reagent II cumulative volume	10～80％

Wherein, reduced form chromogen combination () can be: the amino anti-arsenic Yun of 3-methyl-2-Proxel hydrazone or 4-; Reduced form chromogen combination (two) can be one of aforementioned 15 kinds of materials.

The prescription of two agent is not limited only to above-mentioned prescription, reduced form chromogen combination (one) and (two) can transposition, and, inosine among the reagent I can be placed on reagent II, composition among the reagent II---Phosphatase, nucleotide, XOD, peroxidase etc. also can be put into reagent I, so can form multiple formulations, not enumerate one by one.

For making reagent more stable, can also be mixed with three doses, such as:

	Composition	Content range
			Reagent I	Damping fluid	40～200mmol/l
Inosine	0.1～10mmol/l
		Reduced form chromogen combination (one)		0.1～20mmol/l
Stablizer	10～50mmol/l
		Reagent II		Damping fluid	40～200mmol/l
XOD	500～50000U/l
			Peroxidase	500～50000U/l
Stablizer/reagent II cumulative volume	10～80％
			Reagent III	Damping fluid	40～200mmol/l
Phosphatase, nucleotide	500～50000U/l
		Reduced form chromogen combination (two)		0.1～20mmol/l
Stablizer/reagent III cumulative volume	10～80％

Similar with two agent, reduced form chromogen combination () can be the amino anti-arsenic Yun of 3-methyl-2-Proxel hydrazone or 4-in three doses, and combination (two) can be one of aforementioned 15 kinds of materials.

Three doses prescription also is not limited only to above-mentioned prescription, reduced form chromogen combination (one) and (two) can divide to come and is placed on any position of reagent I, II or III, the composition of reagent I---inosine, can be placed among reagent II or the reagent III, composition among the reagent II---XOD, peroxidase, can be placed among reagent I or the reagent III, composition among the reagent III---Phosphatase, nucleotide, also can be placed in the middle of reagent I or the reagent II, so can form multiple formulations, enumerate no longer one by one.

In the middle of the composition of above-mentioned diagnostic kit, selecting the basic demand of damping fluid is that pH value is within 6.0～11.0 scopes, can be: " Tutofusin tris～hydrochloric acid (Tris-HCl) damping fluid ", " trolamine (Triethanolamine) damping fluid ", " 2-amino-2-methyl-1-propanol (2-Amino-2-methyl-1-propanol) damping fluid ", " imidazoles～hydrochloric acid (Imidazole-HCl) damping fluid ", " glycylglycine (Glycylglycine) damping fluid ", " citric acid～sodium citrate buffer solution ", " Veronal sodium～hydrochloride buffer ", " boric acid～borate buffer solution ", " glycine～sodium hydrate buffer solution ", at least a in " borax～sodium hydrate buffer solution " or " yellow soda ash～sodium bicarbonate buffer liquid ", but range of choice be not subjected to these enumerate limit.

In addition, for reducing the cross influence between each reagent composition, the stability of maintenance reagent, so that standing storage, usually add stablizer in the middle of the reagent I/ reagent II of above single agent, two agent, three doses the reagent I/ reagent II/ reagent III, its consumption accounts for 10～80% (perhaps concentration is within 10～50mmol/l scopes) of place reagent cumulative volume.

Material with used as stabilizers can be: at least a in ethylene glycol, propylene glycol, glycerine, ammonium sulfate, thioglycol, adenosine diphosphate (ADP), bovine serum albumin, carbonate, cholate, dextran, ethylenediamine tetraacetic acid (EDTA), flavin adenine dinucleotide, vitamin B2 phosphate, glucose, glutaminate, reduced glutathion, lactose, N.F,USP MANNITOL, succinate or the sodium-chlor.

No matter studies show that, take all factors into consideration from the accuracy of measurement result and economy two aspects of preparation cost, be single agent, two agent or three doses, and the inorganic phosphorus diagnosis kit of following system component relation is comparatively desirable, also is preferred version of the present invention:

Damping fluid 80～120mmol/l,

Inosine 1～5mmol/l,

Phosphatase, nucleotide 5000～12000U/l,

XOD 5000～12000U/l,

Peroxidase 5000～12000U/l,

Reduced form chromogen combination 0.1～10mmol/l,

Stablizer/reagent cumulative volume 20～50%.

The present invention uses Phosphatase, nucleotide, in samples such as blood plasma, serum inorganic phosphorus in the presence of, produce xanthoglobulin with the inosine effect, xanthoglobulin produces hydrogen peroxide with the XOD effect again, hydrogen peroxide and peroxidase react, and colourless reduced form chromogen (Chromogen) combined system is oxidized to coloured quinone-imine chromogen (Quioneimine) or indoleamine chromogen (Indamine) dyestuff.Because the absorbing wavelength difference of different dyes in 400～600nm scope has well quantitatively corresponding relation between dyestuff content and the absorbancy, therefore, the absorbancy of measuring between the reaction period changes, and just can reflect the content of inorganic phosphorus in the sample well.

The outstanding substantive distinguishing features and the obvious improvement of technical solution of the present invention mainly shows:

(1) the present invention utilizes Enzymology method fully, enzyme digestion reaction has the high characteristics of specificity, by colourless reduced form chromogen combination is oxidized to coloured quinone-imine chromogen or indoleamine chromogen dyestuff, quantitative response goes out the content of inorganic phosphorus in the sample, and test result is accurate;

(2) reacted constituent of participation enzyme linked reaction system all adds, and is not subjected to the pollution of inside and outside source material, test process tolerance range height;

(3) this method is easy, easy to operate, can obtain detected result fast, and the reaction be under buffer conditions, to carry out, do not pollute the environment;

(4) this method just can detect on general ultraviolet/visible light analysis instrument or half, automatic clinical chemistry analyzer, does not need special or additional instruments, and testing cost is cheap, is convenient to apply in the industry;

(5) use the reagent that measuring method provided by the invention can be made various ways such as liquid reagent, powdered reagent, dried reagent, be mainly used in the content of measuring serium inorganic phosphorus in human body and other animal body, also can be, the content that supplementary means is measured inorganic phosphorus in other sample such as concentrate in conjunction with dilution;

(6) liquid inorganic phosphorus diagnosis kit provided by the invention, good stability, the three-D space structure that is present in wherein each kind of enzyme is kept perfectly, and has guaranteed the application testing effect well.Make after two agent or three doses, can further reduce the cross influence between the various compositions, improve the stability of reagent, be convenient to standing storage.

Embodiment

Below in conjunction with specific embodiment technical solution of the present invention is described further.These examples only are some exemplary applications, can not be interpreted as a kind of restriction to claim protection domain of the present invention.

Embodiment one (single agent)

Prepare inorganic phosphorus diagnosis kit in the serum by following composition and consumption:

Glycylglycine damping fluid 80mmol/l,

Inosine 1mmol/l,

Phosphatase, nucleotide 5000U/l,

XOD 5000U/l,

Peroxidase 5000U/l,

The amino anti-arsenic Yun 2mmol/l of 4-,

PHENOL 99.8 MIN ((CARBOLIC ACID)) 10mmol/l,

Ethylene glycol 50% (accounting for the reagent cumulative volume).

Go up to set at automatic clinical chemistry analyzer (Hitachi-7080): 37 ℃ of temperature, 10 minutes reaction times, test predominant wavelength 495～505nm, more than the test commplementary wave length 600nm, the volume ratio of sample and reagent is 1: 25, the Direction of Reaction is positive reaction (absorbancy rises, down together).

Adding serum sample and the single agent that is made into, both detect, write down the rising situation of 495～505nm absorbancy at the inner mixing automatically of analyser.Detect altogether and read a little 31 times, approximately every 20 seconds record one secondary data, according to methods such as rate method, end-point methods, the contrast corresponding standard curve calculates the content of inorganic phosphorus in the serum sample.Same batch sample is repeatedly repeated above process, test result good reproducibility, tolerance range height.

Embodiment two (two agent)

Prepare inorganic phosphorus diagnosis kit in the serum by following composition and consumption:

Reagent I---

Imidazoles～hydrochloride buffer 100mmol/l,

Inosine 3mmol/l,

The amino anti-arsenic Yun 1mmol/l of 4-,

Glycerine 50% (accounting for reagent I cumulative volume);

Reagent II---

Imidazoles～hydrochloride buffer 100mmol/l,

Phosphatase, nucleotide 9000U/l,

XOD 9000U/l,

Peroxidase 9 000U/l,

2,4,6-three bromo-3-hydroxyl-Phenylsulfonic acid 1mmol/l,

Dextran 50% (accounting for reagent II cumulative volume).

Go up to set at automatic clinical chemistry analyzer (Hitachi-7080): 30 ℃ of temperature, 15 minutes reaction times, test predominant wavelength 546nm, more than the test commplementary wave length 630nm, sample is 1: 25 with the ratio of the cumulative volume of reagent I and reagent II, reagent I is 4: 1 with the ratio of the consumption of reagent II, and the Direction of Reaction is positive reaction.

Adding serum sample and reagent I add reagent II after 5 minutes earlier, and the three detects, writes down the rising situation of 546nm absorbancy at the inner mixing automatically of analyser.According to methods such as rate method, end-point methods, the contrast corresponding standard curve calculates the content of inorganic phosphorus in the serum sample.Same batch sample is repeatedly repeated above process, test result good reproducibility, tolerance range height.

Embodiment three (three doses)

Prepare inorganic phosphorus diagnosis kit in the serum by following composition and consumption:

Reagent I---

Trolamine damping fluid 120mmol/l,

Inosine 5mmol/l,

3-methyl-2-Proxel hydrazone 2mmol/l,

Propylene glycol 20mmol/l;

Reagent II---

Trolamine damping fluid 120mmol/l,

XOD 12000U/l,

Peroxidase 12000U/l,

Propylene glycol 50% (accounting for reagent II cumulative volume);

Reagent III---

Trolamine damping fluid 120mmol/l,

Phosphatase, nucleotide 12000U/l,

Two monomethylaniline 2mmol/l,

N.F,USP MANNITOL 50mmol/l.

Go up to set at automatic clinical chemistry analyzer (Hitachi-7080): 25 ℃ of temperature, 20 minutes reaction times, test predominant wavelength 578nm, more than the test commplementary wave length 700nm, sample is 1: 25 with the ratio of the cumulative volume of reagent I, reagent II and reagent III, the consumption of reagent I, reagent II and reagent III is 8: 1: 1, and the Direction of Reaction is positive reaction.

Adding serum sample and reagent I, reagent II add reagent III after 5 minutes earlier, and they detect, write down the rising situation of 578nm absorbancy at the inner mixing automatically of analyser.According to methods such as rate method, end-point methods, the contrast corresponding standard curve calculates the content of inorganic phosphorus in the serum sample.Same batch sample is repeatedly repeated above process, test result good reproducibility, tolerance range height.

Embodiment four (two agent preference)

Prepare the serium inorganic phosphorus diagnostic kit by following composition and consumption:

Reagent I---

Tris-HCl damping fluid 100mmol/l,

XOD 12000U/l,

Peroxidase 12000U/l,

The amino anti-arsenic Yun 1mmol/l of 4-,

Glycerine 50% (accounting for reagent I cumulative volume);

Reagent II---

Tris-HCl damping fluid 120mmol/l,

Inosine 1mmol/l,

Phosphatase, nucleotide 12000U/l,

Two monomethylaniline 2mmol/l,

Glycerine 50% (accounting for reagent II cumulative volume).

Go up to set at automatic clinical chemistry analyzer (Hitachi-7080): 30 ℃ of temperature, 10 minutes reaction times, test predominant wavelength 578nm, more than the test commplementary wave length 700nm, sample is 1: 25 with the ratio of the cumulative volume of reagent I and reagent II, reagent I is 4: 1 with the ratio of the consumption of reagent II, and the Direction of Reaction is positive reaction.

Add serum sample and reagent I earlier, add reagent II after 5 minutes, following principle reaction takes place at the inner mixing automatically of analyser in the three:

Detect, write down the rising situation of 578nm absorbancy.According to methods such as rate method, end-point methods, the contrast corresponding standard curve calculates the content of inorganic phosphorus in the serum sample.Same batch sample is repeatedly repeated above process, test result good reproducibility, tolerance range height.

In a word, experimental results show that and adopt measuring method of the present invention, can pass through ultraviolet analyser or semi-automatic/automatic clinical chemistry analyzer device fully, determine the content of inorganic phosphorus in the samples such as blood plasma, serum, measurement sensitivity height, tolerance range are good, are not subjected to the pollution of inside and outside source material.And, inorganic phosphorus diagnosis kit provided by the invention, good stability still can accurately detect the content of inorganic phosphorus in all kinds sample after the long storage time.

Claims (9)

1. method of measuring inorganic phosphorus may further comprise the steps:

1. with testing sample and the reagent mixing that contains inosine, Phosphatase, nucleotide, XOD, peroxidase and the combination of reduced form chromogen, make it to take place following reaction,

2. reaction mixture is placed under ultraviolet analyser or the semi-automatic/automatic clinical chemistry analyzer, detecting predominant wavelength is the ascensional range of the absorbancy of 400～600nm, calculates the content of inorganic phosphorus in the sample.

2. the method for mensuration inorganic phosphorus according to claim 1, it is characterized in that: described " combination of reduced form chromogen " is to be made of reduced form chromogen A and another B component, reduced form chromogen A is " a 3-methyl-2-Proxel hydrazone " or " the amino anti-arsenic Yun of 4-", another B component is one of following 15 kinds of materials: PHENOL 99.8 MIN ((CARBOLIC ACID)), N-ethyl-N-(3-thiopropyl)-m-thialdine amine, N, the two ethyls of N--m-Tolylamine, 2, the two chlorine PHENOL 99.8 MIN ((CARBOLIC ACID)) of 4-, 2,4,6-three bromo-3-hydroxyl-Phenylsulfonic acid, 3, the two chlorine PHENOL 99.8 MIN ((CARBOLIC ACID)) sulfonic acid of 5-, 3, the two chloro-2-hydroxyl-Phenylsulfonic acids of 5-, N-ethyl-N-(2-hydroxyl-3-thiopropyl)-m-Tolylamine sodium salt, three bromine hydroxy-benzoic acid, two monomethylanilines, N-ethyl-N-(2-hydroxyl-3-thiopropyl)-m-Tolylamine, 2,2 '-Lian nitrogen-two (3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) di-ammonium salts, 2,2 '-azino-two (3-ethyl benzo thiophene pyrrolines-6-sulfonic acid), vanillic acid, 3-methyl-ethyl-hydroxyanilines.

3. the method for mensuration inorganic phosphorus according to claim 1 and 2, it is characterized in that: the usage ratio of testing sample and reagent by volume was controlled at 1: 10～1: 500, temperature of reaction is controlled at 20 ℃～50 ℃, reaction times was controlled at 2～30 minutes, set commplementary wave length during detection more than 500～700nm.

4. inorganic phosphorus diagnosis kit, reagent is grouped into by following one-tenth in the box:

Damping fluid 40～200mmol/l,

Inosine 0.1～10mmol/l,

Phosphatase, nucleotide 500～50000U/l,

XOD 500～50000U/l,

Peroxidase 500～50000U/l,

Reduced form chromogen combination 0.1～20mmol/l,

Stablizer/reagent cumulative volume 10～80%.

5. inorganic phosphorus diagnosis kit according to claim 4 is characterized in that:

Described " combination of reduced form chromogen " is to be made of reduced form chromogen A and another B component, and the concentration of A and B is 0.1～20mmol/l;

Reduced form chromogen A is " a 3-methyl-2-Proxel hydrazone " or " the amino anti-arsenic Yun of 4-";

Another B component is one of following 15 kinds of materials: PHENOL 99.8 MIN ((CARBOLIC ACID)), N-ethyl-N-(3-thiopropyl)-m-thialdine amine, N, the two ethyls of N--m-Tolylamine, 2, the two chlorine PHENOL 99.8 MIN ((CARBOLIC ACID)) of 4-, 2,4,6-three bromo-3-hydroxyl-Phenylsulfonic acid, 3, the two chlorine PHENOL 99.8 MIN ((CARBOLIC ACID)) sulfonic acid of 5-, 3, the two chloro-2-hydroxyl-Phenylsulfonic acids of 5-, N-ethyl-N-(2-hydroxyl-3-thiopropyl)-m-Tolylamine sodium salt, three bromine hydroxy-benzoic acid, two monomethylanilines, N-ethyl-N-(2-hydroxyl-3-thiopropyl)-m-Tolylamine, 2,2 '-Lian nitrogen-two (3-ethyl benzo thiazole phenanthroline-6-sulfonic acid) di-ammonium salts, 2,2 '-azino-two (3-ethyl benzo thiophene pyrrolines-6-sulfonic acid), vanillic acid, 3-methyl-ethyl-hydroxyanilines.

6. according to claim 4 or 5 described inorganic phosphorus diagnosis kits, it is characterized in that: the PH scope of described damping fluid is 6.0～11.0.

7. inorganic phosphorus diagnosis kit according to claim 6 is characterized in that: described damping fluid is " Tutofusin tris～hydrochloride buffer ", " trolamine damping fluid ", " 2-amino-2-methyl-1-propanol damping fluid ", " imidazoles～hydrochloride buffer ", " glycylglycine damping fluid ", " citric acid～sodium citrate buffer solution ", " Veronal sodium～hydrochloride buffer ", " boric acid～borate buffer solution ", " glycine～sodium hydrate buffer solution ", at least a in " borax～sodium hydrate buffer solution " or " yellow soda ash～sodium bicarbonate buffer liquid ".

8. according to claim 4 or 5 described inorganic phosphorus diagnosis kits, it is characterized in that: described stablizer is at least a in ethylene glycol, propylene glycol, glycerine, ammonium sulfate, thioglycol, adenosine diphosphate (ADP), bovine serum albumin, carbonate, cholate, dextran, ethylenediamine tetraacetic acid (EDTA), flavin adenine dinucleotide, vitamin B2 phosphate, glucose, glutaminate, reduced glutathion, lactose, N.F,USP MANNITOL, succinate or the sodium-chlor.

9. according to claim 4 or 5 described inorganic phosphorus diagnosis kits, it is characterized in that: described reagent is made into single agent, two agent or three doses.