CN101477059A - Method for rapidly detecting inorganic phosphorus in water solution - Google Patents
Method for rapidly detecting inorganic phosphorus in water solution Download PDFInfo
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- CN101477059A CN101477059A CNA2009100736438A CN200910073643A CN101477059A CN 101477059 A CN101477059 A CN 101477059A CN A2009100736438 A CNA2009100736438 A CN A2009100736438A CN 200910073643 A CN200910073643 A CN 200910073643A CN 101477059 A CN101477059 A CN 101477059A
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Abstract
The invention provides a method for quickly detecting inorganic phosphorus in a water solution, which directly, qualitatively and quantificationally detects the inorganic phosphorus in a buffer solution with a pH value of 7.0 by taking 4-(2-pyridylazo)-resorcinol (PAR) as a visualization reagent based on divalent lead Pb<2+> ions. The method concretely comprises a spectrometry, a reagent kit method and a test paper method. The methods can be directly used for the determination of the inorganic phosphorus in environmental water sources, blood, urine, milk, cola, beer, and so on. The method has the advantages of simple detection process, intuitive and convenient discrimination, and easy popularization and promotion. The method has accurate measurement result and high sensitivity, is not interfered by other anions, can be promoted and applied to the determination of the inorganic phosphorus in clinical blood samples, urine samples, the environmental water sources, and food, and has larger market development value and application prospect.
Description
Technical field:
The present invention relates to the detection technique of Phos, specifically belong to the method for Phos in a kind of fast detecting aqueous solution.
Background technology:
Phosphorus is accumulation and the conduction and other the most basic key element of biological respinse of synthetic DNA and RNA in the life system, energy, is one of more element of content in the human body, is only second to calcium.80 percent and calcium in conjunction with and be stored in bone and the tooth, remaining 20 percent, be present in the cell or as the form of organophosphoruss such as phosphide, nucleic acid, ATP and be present in the extracellular as Phos.Phosphorus in the diet is considered to damage a factor suffering from chronic renal failure patient residual renal function.Phosphorus also participates in various important biomineralization processes, such as the formation of bone and the pathologic processes such as origin of kidney stone.It is reported, in the urine Phos can cause unusually lithiasis, prostatic disorders, hypoparathyroidism or hyperfunction, late period chronic nephritis, Huppert's disease, rickets or osteomalacia etc., if untimely healing even causes other bigger disease.In human serum, typical phosphate concentration scope is 0.81-2.26mmol/L.In the individuality, it is too much that unusual high phosphate concentration is diagnosed as high phosphate, show as acute or chronic kidney failure, simultaneously possible situation is: take place prostatic disorders, hypoparathyroidism, late period chronic nephritis, vitamin D picked-up too much, Huppert's disease.And the multiple disease that the individuality of low phosphate radical content will be caused by hypophosphoremia will in the blood: hyperparathyroidism, tubular degeneration pathology, rickets or osteomalacia, parathyroid secondary hyperplasia, long-term diarrhea or diseases such as malabsorption, Fanconi's syndrome.Therefore to keep certain density Phos very crucial for individuality.And we can understand metabolism status and the relevant pathology state of biosome by the content of periodic monitoring Phos.In view of this at food, environment, detect Phos clinically and be absolutely necessary.
At present, about measuring Phos, enzyme process, the synthetic acceptor of use go identification to measure Phos to have the chromatography of ions, sequence to inject fixedly, also have many colourimetrys or the like.It may be noted that many instruments are measured Phos clinically because the narrow range of linearity needs dilution significantly to treat test sample, just can go to measure; Use synthetic acceptor, need a large amount of synthetic works, and the water-soluble of acceptor can be made troubles also to surveying work; Many colourimetrys detect Phos, or further are reduced to assorted poly-molybdenum blue based on phosphomolybdic acid, or based on inferior molybdenum of direct mensuration and inferior vanadium phosphomolybdic acid, perhaps based on the complex of phosphomolybdic acid and basic-dyeable fibre.These chemical methodes have serious defective, that is: the speed that formed by slight pH variable effect, complex of the molybdate reduction pH acidity that had a strong impact on, need by the concentration of albumen will cause the hydrolysis of organophosphorus, so all can cause the concentration of the Phos that records higher.
Summary of the invention:
The object of the present invention is to provide a kind of simple, sensitive, fast, directly measure the method for Phos in the aqueous solution.
The invention provides the method for Phos in a kind of easy fast detecting aqueous solution, the total design of this method is based on the plumbous Pb of divalence
2+Ion, be developer at pH with 4-(2-pyridylazo) resorcinol (PAR) be direct qualitative, quantitative ground detection Phos in 7.0 the HEPES buffer solution, its concrete detection method comprises following several:
The method of Phos in a kind of fast detecting aqueous solution (spectroscopic methodology detection) comprises the steps:
(1) preparation pH=6.5-7.5, concentration is the HEPES buffer solution of 1-100mM, and preparation 2mM, the plumbous Pb of divalence
2+The PAR ethanolic solution of solution, 2mM;
(2) the HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws the PAR solution 20 μ l of 2mM with microsyringe as blank, be added in the above-mentioned cuvette, solution is detected on the UV, visible light spectrophotometer by colourless flavescence look at this moment, and there is absorption maximum at the 412nm place;
(3) in the cuvette of step (2), add Pb with the PAR equivalent
2+Solution, solution is detected on the UV, visible light spectrophotometer by yellow purpling look at this moment, finds that maximum absorption band becomes 520nm by above-mentioned 412nm, adds Pb
2+Liquor capacity is designated as VPb;
(4) get Phos aqueous solution to be measured, be added in the cuvette of step (3) with microsyringe gradually, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with the Phos aqueous solution, maximum absorption band is become again to 412nm gradually by 520nm again, and the color of solution also becomes yellow again by purple gradually, when absorption peak when 412nm reaches maximal value, stop to add the Phos aqueous solution, add Phos aqueous solution volume and count the V Phos this moment;
(5) by [Pb
2+] * V
Pb/ V
PhosPhos gets content in calculating.
The method (kit method) of Phos comprises the steps: in the another kind of fast detecting solution
(1), preparation pH=7.0, concentration is the HEPES buffer solution of 10mM, is labeled as R
1The PAR ethanolic solution of 2mM is labeled as R
2The Pb of 2mM
2+Solution is labeled as R
3
(2), according to R
1, R
2, R
3Volume ratio is 100:1:1, adds in the color comparison tube, and this moment, solution was purple;
(3), get R
3The Phos aqueous solution to be measured of volume 1/10 is added in the above-mentioned color comparison tube with injector, behind the shake 20-30s, contrasts with kit standard color range, draws the inorganic phosphorus concentration scope in the test sample for the treatment of.
The formulation of kit standard color range: at the plumbous Pb of the divalence of the PAR that contains 20 μ M, 20 μ M
2+Ion, pH=7.0 and concentration are in the HEPES buffer solution of 10mM, in five color comparison tubes, add phosphate radical solution respectively successively, make the concentration of phosphate radical be respectively 0,5,10,15,20 μ M, obtain the color range (see figure 3) of kit standard, that is: color from left to right is followed successively by, 0.0,2.5,5.0,7.5,10.0mM purple, aubergine, reddish yellow, dark brown, yellow are represented corresponding inorganic phosphorus concentration:.
The method of Phos (test paper colourimetry) comprises the steps: in another fast detecting aqueous solution
(1), preparation pH=6.5-7.5, concentration are the HEPES buffer solution of 1-100mM, and prepare the plumbous Pb of divalence of 2mM
2+The PAR ethanolic solution of solution, 2mM;
(2) with the Pb of 20 μ l
2+The PAR ethanolic solution of solution and 20 μ l is added in the HEPES buffer solution of 2ml, and after stirring, this moment, solution was purple;
(3) bulk filter paper is immersed 5-10 second in the purple solution of step (2), takes out and in air, dry, soak into 1-3 time repeatedly, dry afterwards the Phos test paper, color is purple, it is standby that this test paper is cut into strip;
(4) get step (3) strip Phos test paper and be infiltrated on that 2-5 takes out second in the Phos aqueous solution to be measured,, judge the content for the treatment of the test sample Phos according to test paper change color and the contrast of Phos test paper color range.
Phos test paper color range is formulated:
Get 5 of step (3) stick form test papers, be infiltrated on concentration respectively and be 0.0,2.5,5.0,7.5,2-5 second in the Phos aqueous solution of 10.0mM, take out, the color that promptly obtains from left to right is followed successively by: purple, pink, khaki, brassy and golden yellow, represent that inorganic phosphorus concentration is followed successively by 0,2.5,5,7.5, the 10mM (see figure 4).
Described Pb
2+Solion is all solubility divalence lead salt solution of lead ion, and is formulated as plumbi nitras, lead acetate, lead chloride etc.
Described HEPES buffer solution can use acetic acid-sodium acetate buffer solution, Tris-HCl, citric acid to replace.
Described Phos aqueous solution can be urine, blood, emulsion (as breast milk, milk or goat milk), beverage (as cola, coffee, tea), beer or environmental water systems water sample etc.
Compared with prior art, the present invention has following advantage and effect: 1, the detection architecture cost is low, is easy to extensively promote the use of, and helping Phos detected becomes a conventional sense, improves the people's livelihood, diet, medical condition; 2, can on the UV, visible light spectrophotometer, detect; Also can detect by visual colorimetric determination without any need for instrument; 3 can be prepared into the Phos test paper directly detects; 4, detected object is undressed sample, saves loaded down with trivial details in the past preprocessor, brings great convenience to the application on clinical, food, the environment; Although contain many other compositions in the 5 detection samples, the detection method of this invention has shown good selectivity to Phos, that is to say, our testing result is not disturbed in the existence of other composition; 6, detection architecture is to carry out in the physiological condition aqueous solution, has avoided the secondary pollution that with an organic solvent may bring; Obviously this invention can be widely applied to environment, food, detect Phos clinically.
Description of drawings:
Fig. 1 is containing 20 μ M PAR, and pH 7.0 HEPES (10mM) add Pb in the buffer solution gradually
2+The UV, visible light of (0-20 μ l) absorbs figure;
Fig. 2 is containing 20 μ M PAR and 20 μ M Pb
2+, pH 7.0 HEPES (10mM) add phosphate radical (0-20 μ l) gradually in the buffer solution UV, visible light absorbs figure;
Fig. 3 is a kit standard color range of the present invention;
Fig. 4 Phos test paper of the present invention color range;
Fig. 5 is containing 20 μ M PAR and 20 μ M Pb
2+, pH 7.0 HEPES (10mM) but add various anionic colorimetric views in the buffer solution.
Fig. 6 is containing 20 μ M PAR and 20 μ M Pb
2+, pH 7.0 HEPES (10mM) add serum (0-18 μ l) gradually in the buffer solution UV, visible light absorbs figure;
Fig. 7 kit method detects the color diagram of inorganic phosphorus concentration in the blood;
Fig. 8 test paper colourimetry detects Phos color diagram in the blood;
Fig. 9 is containing 20 μ M PAR and 20 μ M Pb
2+, pH 7.0 HEPES (10mM) add urine (0-4 μ l) gradually in the buffer solution UV, visible light absorbs figure;
Figure 10 kit method detects the color diagram of inorganic phosphorus concentration in the urine;
Figure 11 test paper colourimetry detects Phos color diagram in the urine;
Figure 12 is containing 20 μ M PAR and 20 μ M Pb
2+, the UV, visible light that adds laughable (0-20 μ l) in pH 7.0 HEPES (10mM) buffer solution gradually absorbs figure;
Figure 13 kit method detects the color diagram of inorganic phosphorus concentration in the cola.
Embodiment:
Embodiment 1: the method for Phos in the spectroscopic methodology fast detecting aqueous solution
HEPES (10mM) buffer solution of preparation pH=7.0, and prepare 2 * 10
-3The lead nitrate solution of M, 2 * 10
-3The PAR ethanolic solution and 2 * 10 of M
-3The sodium radio-phosphate,P-32 solution of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The PAR ethanolic solution 20 μ l of M are added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 412nm has absorption maximum; In above-mentioned cuvette, add 2 * 10
-3The plumbi nitras 20 μ l of M, solution is detected on the UV, visible light spectrophotometer by yellow purpling look at this moment, finds that maximum absorption band becomes 520nm by above-mentioned 412nm, and UV, visible light absorption figure sees Fig. 1; Get sodium radio-phosphate,P-32 solution, application of sample limit, limit is detected on HP8453 UV, visible light spectrophotometer, adding along with phosphate radical, maximum absorption band is become again to 412nm gradually by 520nm again, the color of solution also becomes yellow again by purple gradually, when absorption peak reaches maximum at 412nm, during the complete flavescence of the color of solution, this moment, the phosphate radical addition was V
Phosμ l; Calculate: 20 μ l * 2 * 10
-3/ V
Phosμ l calculates.UV, visible light absorption figure sees Fig. 2.
Embodiment 2: the method for Phos in solution colourimetry (kit method) the fast detecting aqueous solution
The kit preparation:
HEPES (10mM) buffer solution of preparation pH=7.0, and prepare 2 * 10
-3The lead nitrate solution of M, 2 * 10
-3The PAR ethanolic solution of M.Get six color comparison tubes, add the pH=7.0 of 2ml and the HEPES buffer solution, 2 * 10 that concentration is 10mM respectively
-3The PAR solution 20 μ l of M, this moment, 1-6 color comparison tube solution was yellow; In the 2-6 color comparison tube, add 2 * 10 respectively
-3The lead nitrate solution 20 μ l of M, this moment, 2-6 color comparison tube solution colour became purple by yellow; In the 2-6 color comparison tube, add different amount phosphate radical solution subsequently successively respectively, make the concentration of phosphate radical be respectively 0,5,10,15,20 μ M, obtain the color range figure of standard, that is: 2-6 color comparison tube color from left to right is followed successively by, 0,2.5,5.0,7.5, the 10mM (see figure 3) purple, aubergine, reddish yellow, dark brown, yellow, the corresponding test sample concentration range for the treatment of of representative is:.
The unknown concentration content of inorganic phosphorus is measured:
HEPES (10mM) buffer solution of preparation pH=7.0, and prepare 2 * 10
-3M lead nitrate solution, 2 * 10
-3The PAR ethanolic solution of M; Get color comparison tube, add HEPES buffer solution, the 20 μ l of 2ml, 2 * 10
-3The PAR ethanolic solution of M, solution adds 2 * 10 by colourless flavescence at this moment
-3The lead nitrate solution 20 μ l of M, this moment, solution was by yellow purpling look; In above-mentioned color comparison tube, get and treat test sample 2 μ l, be added in this color comparison tube with microsyringe, behind the shake, observe the color range contrast of solution colour variation and standard, promptly decidable is learnt inorganic phosphorus concentration in institute's test sample product.
Embodiment 3: the method for content of inorganic phosphorus in the test paper colourimetry fast detecting aqueous solution
HEPES (10mM) aqueous buffer solution of preparation pH=7.0, and prepare 2 * 10
-2The plumbi nitras aqueous solution, 2 * 10 of M
-2The PAR ethanolic solution of M and concentration are 0,2.5,5.0,7.5, the Phos aqueous solution of 10mM; Pb with 20 μ l
2+The PAR ethanolic solution of solution and 20 μ l is added in HEPES (10mM) buffer solution of 2ml, the back solution that stirs is purple, bulk filter paper was immersed in this purplish red solution 30 seconds, taking-up is dried in air, soak into repeatedly again dry 3 times available Phos test paper, it is stand-by that this test paper is cut into strip;
Get 5 of step stick form test papers, be infiltrated on concentration respectively and be 0,2.5,5.0,7.5, in the Phos aqueous solution of 10mM 5 seconds, take out, the Phos test paper concentration gradient color range figure that promptly obtains purple (blank), pink (2.5mM), khaki (5mM), brassy (7.5mM) and golden yellow (10mM) sees Fig. 4;
Get the Phos test paper and be infiltrated on and treat in the test sample to take out in 3 seconds, contrast with inorganic phosphorus concentration gradient color range figure according to the test paper change color, i.e. the content of decidable sample Phos to be measured
Embodiment 4: other negative ion do not disturb content of inorganic phosphorus to measure
HEPES (10mM) buffer solution of preparation pH=7.0, and prepare 2 * 10
-3The lead nitrate solution of M, 2 * 10
-3The PAR ethanolic solution and 2 * 10 of M
-3The sodium phosphate of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The PAR solution 20 μ l of M are added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 412nm has absorption maximum; In above-mentioned cuvette, add 2 * 10
-3The lead nitrate solution 20 μ l of M, solution is detected on the UV, visible light spectrophotometer by yellow purpling look at this moment, finds that maximum absorption band becomes 520nm by above-mentioned 412nm, and UV, visible light absorption figure sees Fig. 1; In the above-mentioned cuvette, add 10 times of acetate (AcO successively to the Phos amount
-), nitrate radical (NO
3 -), carbonate (CO
3 2-), oxalate (
-O0CC00
-), fluorine ion (F
-), chlorion (Cl
-), bromide ion (Br
-), sulfate ion (SO
4 2-), on the UV, visible light spectrophotometer, detecting, the adding of above-mentioned various ions does not influence the detection of system to phosphate anion, and ultraviolet spectrum does not change at the 520nm maximum absorption band, and the system color does not also have change (Fig. 5).
Embodiment 5: the mensuration of content of inorganic phosphorus in the blood
Spectrographic determination:
From healthy people's blood sampling, centrifugal that the yellowish clear liquid in upper strata was placed 2 hours on TGL-16B type high speed tabletop centrifuge; HEPES (10mM) buffer solution of preparation pH7.0, and prepare 2 * 10
-3The lead nitrate solution of M and 2 * 10
-3The PAR ethanolic solution of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The PAR solution 20 μ l of M are added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence look at this moment, and 412nm has absorption maximum; In above-mentioned cuvette, add 2 * 10
-3The Pb of M
2+Solution 20 μ l, solution is detected on the UV, visible light spectrophotometer by yellow purpling look at this moment, finds that maximum absorption band becomes 520nm by above-mentioned 412nm; Get above-mentioned limpid liquid, be added in this cuvette with microsyringe gradually, application of sample limit, limit is detected on HP8453 UV, visible light spectrophotometer, adding along with limpid liquid, maximum absorption band is become again to 412nm gradually by 520nm again, and the color of solution is also returned yellow by purple stain gradually, when absorption peak reaches maximum at 412nm, during the complete flavescence of the color of solution, this moment, limpid liquid addition was 18 μ l; Calculate: 20 μ l * 2 * 10
-3/ 18 μ l, in the blood content of inorganic phosphorus be 2.22mmol/L, UV, visible light absorption figure sees Fig. 6.
The kit method is measured:
Get centrifugal blood and handle the gained yellowish clear liquid sample 2 μ l in upper strata, be added in this color comparison tube, behind the shake, observe kit standard color range contrast among solution colour variation and the embodiment 2, find that this blood sample content of inorganic phosphorus is about 2.5mM with microsyringe.See Fig. 7 than chromatic graph
Phos test paper colorimetric method for determining:
Get volunteer's blood sample, after the centrifugal treating, the Phos test paper for preparing among the embodiment 3 is immersed in this blood sample, find that the test paper color becomes reddish yellow by purple, judges that this blood sample content of inorganic phosphorus is about 2.5mM.See Fig. 8 than chromatic graph
Embodiment 6: the mensuration of content of inorganic phosphorus in the urine
Spectrographic determination:
HEPES (10mM) buffer solution of preparation pH=7.0, and prepare 2 * 10
-3The lead nitrate solution of M, 2 * 10
-3The PAR ethanolic solution of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The PAR solution 50 μ l of M are added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 412nm has absorption maximum; In above-mentioned cuvette, add 2 * 10
-3The plumbi nitras 20 μ l of M, solution is detected on the UV, visible light spectrophotometer by yellow purpling look at this moment, finds that maximum absorption band becomes 520nm by above-mentioned 412nm; Get urine sample, application of sample limit, limit is detected on HP8453 UV, visible light spectrophotometer, adding along with urine sample, maximum absorption band is become again to 412nm gradually by 520nm again, the color of solution also becomes yellow again by purple gradually, when absorption peak reaches maximum at 412nm, during the complete flavescence of the color of solution, this moment, the urine sample addition was 4 μ l; Calculate: 20 μ l * 2 * 10
-3/ 4 μ l, the content that gets Phos in the urine sample is 10 * 10
-3M.UV, visible light absorption figure sees Fig. 9.
The kit method is measured:
Get volunteer's urine sample 2 μ l, be added in this color comparison tube, behind the shake, observe kit standard color range contrast among solution colour variation and the embodiment 2, find that this urine sample content of inorganic phosphorus is between 7.5-10mM with microsyringe.See Figure 10 than chromatic graph
Phos test paper colorimetric method for determining:
Get volunteer's urine sample, the Phos test paper for preparing among the embodiment 3 is immersed in this urine sample, find that the test paper color becomes brassy by purple, judges that this urine sample content of inorganic phosphorus is between 7.5-10mM.See Figure 11 than chromatic graph.
Embodiment 7: the mensuration of content of inorganic phosphorus in the milk
Spectrographic determination:
HEPES (10mM) buffer solution of preparation pH=7.0, and prepare 2 * 10
-3The lead nitrate solution of M, 2 * 10
-3The PAR ethanolic solution of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The PAR solution 50 μ l of M are added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 412nm has absorption maximum; In above-mentioned cuvette, add 2 * 10
-3The plumbi nitras 20 μ l of M, solution is detected on the UV, visible light spectrophotometer by yellow purpling look at this moment, finds that maximum absorption band becomes 520nm by above-mentioned 412nm; Get the milk sample, application of sample limit, limit is detected on HP8453 UV, visible light spectrophotometer, adding along with the milk sample, maximum absorption band is become again to 412nm gradually by 520nm again, the color of solution also becomes yellow again by purple gradually, when absorption peak reaches maximum at 412nm, during the complete flavescence of the color of solution, this moment, milk sample addition was 4 μ l; Calculate: 20 μ l * 2 * 10
-3/ 5 μ l, the content that gets Phos in the milk sample is 8 * 10
-3M.UV, visible light absorbs figure as Fig. 9.
The kit method is measured:
Get milk sample 2 μ l, be added in this color comparison tube, behind the shake, observe kit standard color range contrast among solution colour variation and the embodiment 2, find that this milk sample content of inorganic phosphorus is between 7.5-10mM with microsyringe.Phos test paper colorimetric method for determining:
Get the milk sample, the Phos test paper for preparing among the embodiment 3 is immersed in this milk sample, find that the test paper color becomes brassy by purple, judges that this milk sample content of inorganic phosphorus is between 7.5-10mM.
Embodiment 8: the mensuration of content of inorganic phosphorus in the cola
Spectrographic determination:
Buy laughable from the market; HEPES (10mM) buffer solution of preparation pH=7.0, and prepare 2 * 10
-3The lead nitrate solution of M, 2 * 10
-3The PAR ethanolic solution of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank
-3The PAR solution 20 μ l of M are added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 412nm has absorption maximum; In above-mentioned cuvette, add 2 * 10
-3Plumbi nitras 20 μ l, this moment solution by yellow purpling look, on the UV, visible light spectrophotometer, detect, find that maximum absorption band becomes 520nm by above-mentioned 412nm; Get laughable sample, application of sample limit, limit is detected on HP8453 UV, visible light spectrophotometer, adding along with laughable sample, maximum absorption band is become again to 412nm gradually by 520nm again, the color of solution also becomes yellow again by purple gradually, when absorption peak reaches maximum at 412nm, during the complete flavescence look of the color of solution, this moment, Coca-Cola addition was 7.2 μ l; Calculate: 20 μ l * 2 * 10
-3/ 7.2 μ l, the content that gets Phos in Coca-Cola is 5.56 * 10
-3M.UV, visible light absorption figure sees Figure 12.
The kit method is measured:
Get laughable sample 2 μ l, be added in this color comparison tube, behind the shake, observe kit standard color range contrast among solution colour variation and the embodiment 2, find that content of inorganic phosphorus is about 5mM in this laughable sample with microsyringe.See Figure 13 than chromatic graph
Phos test paper colorimetric method for determining:
Get laughable sample, the Phos test paper for preparing among the embodiment 3 is immersed in this laughable sample, find that the test paper color becomes brassy by purple, judges that this laughable sample content of inorganic phosphorus is about 5mM.
Embodiment 9: with acetic acid-sodium acetate (pH 7.0) is that buffer solution detects content of inorganic phosphorus
Acetic acid-sodium-acetate buffer of preparation pH7.0, with the HEPES buffer solution in its alternate embodiment 1, other solution preparation and detection step and embodiment 1, embodiment 2 is identical, and it is identical with Fig. 1, Fig. 2 that spectroscopic methodology detects ultraviolet-visible absorption spectroscopy; The colourimetry testing process is with embodiment 2, and is more identical with Fig. 3 than chromatic graph.
Embodiment 10: detect content of inorganic phosphorus with the lead acetate obtain solution
Preparation 2 * 10
-3The lead acetate solution of M, with the lead nitrate solution in its alternate embodiment 1, other solution preparation and detection step and embodiment 1, embodiment 2, embodiment 3 are identical, and it is identical with Fig. 1, Fig. 2 that spectroscopic methodology detects ultraviolet-visible absorption spectroscopy; The colourimetry testing process is with embodiment 2, and is more identical with Fig. 3 than chromatic graph.
Claims (6)
1, the method for Phos in a kind of fast detecting aqueous solution is characterized in that, comprises the steps:
(1) preparation pH=6.5-7.5, concentration is the HEPES buffer solution of 1-100mM, and preparation 2mM, the plumbous Pb of divalence
2+The PAR ethanolic solution of solution, 2mM;
(2) the HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws the PAR solution 20 μ l of 2mM with microsyringe as blank, be added in the above-mentioned cuvette, solution is detected on the UV, visible light spectrophotometer by colourless flavescence look at this moment, and there is absorption maximum at the 412nm place;
(3) in the cuvette of step (2), add Pb with the PAR equivalent
2+Solution, solution is detected on the UV, visible light spectrophotometer by yellow purpling look at this moment, finds that maximum absorption band becomes 520nm by above-mentioned 412nm, adds Pb
2+Liquor capacity is designated as V
Pb
(4) get Phos aqueous solution to be measured, be added in the cuvette of step (3) with microsyringe gradually, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with the Phos aqueous solution, maximum absorption band is become again to 412nm gradually by 520nm again, and the color of solution also becomes yellow again by purple gradually, when absorption peak when 412nm reaches maximal value, stop to add the Phos aqueous solution, add Phos aqueous solution volume and count V this moment
Phos
(5) by [Pb
2+] * V
Pb/ V
PhosPhos gets content in calculating.
2, the method for Phos in a kind of fast detecting solution is characterized in that, comprises the steps:
(1), preparation pH=7.0, concentration is the HEPES buffer solution of 10mM, is labeled as R
1The PAR ethanolic solution of 2mM is labeled as R
2The Pb of 2mM
2+Solution is labeled as R
3
(2), according to R
1, R
2, R
3Volume ratio is 100:1:1, adds in the color comparison tube, and this moment, solution was purple;
(3), get R
3The Phos aqueous solution to be measured of volume 1/10 is added in the above-mentioned color comparison tube with injector, behind the shake 20-30s, contrasts with the standard color range, draws and treats inorganic phosphorus concentration in the test sample.
0.0,2.5,5.0,7.5,10.0mM described standard color range: color from left to right is followed successively by, and purple, aubergine, reddish yellow, dark brown, yellow are represented corresponding inorganic phosphorus concentration:.
3, the method for Phos in a kind of fast detecting aqueous solution is characterized in that, comprises the steps:
(1), preparation pH=6.5-7.5, concentration are the HEPES buffer solution of 1-100mM, and prepare the plumbous Pb of divalence of 2mM
2+The PAR ethanolic solution of solution, 2mM;
(2) with the Pb of 20 μ l
2+The PAR ethanolic solution of solution and 20 μ l is added in the HEPES buffer solution of 2ml, and after stirring, this moment, solution was purple;
(3) filter paper is immersed 5-10 second in the purple solution of step (2), takes out and in air, dry, soak into 1-3 time repeatedly, dry afterwards color be the Phos test paper of purple;
(4) get step (3) strip Phos test paper and be infiltrated on that 2-5 takes out second in the Phos aqueous solution to be measured,, judge the content for the treatment of the test sample Phos according to test paper change color and the contrast of Phos test paper color range.
Described Phos test paper color range: color from left to right is followed successively by: purple, pink, khaki, brassy and golden yellow, represent that inorganic phosphorus concentration is followed successively by 0,2.5,5,7.5,10mM.
4, as the method for Phos in claim 1, the 2 or 3 described a kind of fast detecting aqueous solution, it is characterized in that described Pb
2+Solion is a solubility divalence lead salt solution.
As the method for Phos in claim 1, the 2 or 3 described a kind of fast detecting aqueous solution, it is characterized in that 5, described HEPES buffer solution replaces with acetic acid-sodium acetate buffer solution, Tris-HCl or citric acid.
6, as the method for Phos in claim 1, the 2 or 3 described a kind of fast detecting aqueous solution, it is characterized in that described Phos aqueous solution is urine, blood, emulsion, beverage, beer or environmental water systems.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102183472A (en) * | 2011-01-11 | 2011-09-14 | 山西大学 | Method and reagent kit for detecting mercury ions |
| CN102419310A (en) * | 2011-09-07 | 2012-04-18 | 山西大学 | Hg<2+> detection method |
| CN108267412A (en) * | 2018-03-20 | 2018-07-10 | 东北大学 | The quick repeatable hexavalent chromium detecting system and method utilized |
| CN110849869A (en) * | 2019-11-18 | 2020-02-28 | 南京江原安迪科正电子研究发展有限公司 | Color development method and kit for detecting copper ion concentration limit in radioactive injection |
| CN111141898A (en) * | 2019-12-31 | 2020-05-12 | 东北大学 | Urinary calculus risk factor detection paper chip and preparation method thereof |
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2009
- 2009-01-09 CN CN2009100736438A patent/CN101477059B/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102183472A (en) * | 2011-01-11 | 2011-09-14 | 山西大学 | Method and reagent kit for detecting mercury ions |
| CN102419310A (en) * | 2011-09-07 | 2012-04-18 | 山西大学 | Hg<2+> detection method |
| CN108267412A (en) * | 2018-03-20 | 2018-07-10 | 东北大学 | The quick repeatable hexavalent chromium detecting system and method utilized |
| CN108267412B (en) * | 2018-03-20 | 2020-09-29 | 东北大学 | Rapid and reusable hexavalent chromium ion detection system and method |
| CN110849869A (en) * | 2019-11-18 | 2020-02-28 | 南京江原安迪科正电子研究发展有限公司 | Color development method and kit for detecting copper ion concentration limit in radioactive injection |
| CN111141898A (en) * | 2019-12-31 | 2020-05-12 | 东北大学 | Urinary calculus risk factor detection paper chip and preparation method thereof |
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Inventor after: Huo Fangjun Inventor after: Liu Yanling Inventor after: Yin Caixia Inventor after: Sun Yuanqiang Inventor before: Yin Caixia Inventor before: Huo Fangjun Inventor before: Sun Yuanqiang |
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