CN1313818C - Method for detecting phosphate radical in urine - Google Patents

Method for detecting phosphate radical in urine Download PDF

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CN1313818C
CN1313818C CNB2004100124481A CN200410012448A CN1313818C CN 1313818 C CN1313818 C CN 1313818C CN B2004100124481 A CNB2004100124481 A CN B2004100124481A CN 200410012448 A CN200410012448 A CN 200410012448A CN 1313818 C CN1313818 C CN 1313818C
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urine
solution
phosphate radical
visible light
cuvette
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CN1598539A (en
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杨频
阴彩霞
霍方俊
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Shanxi University
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Shanxi University
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Abstract

The present invention directly measures the content of phosphate radicals in undiluted urine of human bodies in an HEPES buffer solution with the pH of 7.0 more or less in a qualitative and quantitative method by using ytterbium ions and using pyrocatechol violet as a developer. The process of measurement is simple and convenient, and measurement results are accurate. The method has the advantages of high sensibility and no interference by other components in urine, and can be applied to clinics.

Description

A kind of method that detects phosphate radical in the urine
Technical field:
The present invention relates to a kind of detection method of phosphate radical, specifically belong to a kind of method that detects phosphate radical in the urine.
Background technology:
Phosphate radical participates in various important biomineralization processes, such as the formation of bone and the pathologic processes such as origin of kidney stone.Therefore, the detection of phosphate radical is very important in the biofluid.We know that urine has very important diagnostic value in clinical events, thus in the human body urine freely phosphate anion be necessary.Scientific result of study shows: contain too much ammonium magnesium phosphate in the urine, and carbapatite, hydroxyapatite, dicalcium phosphate dihydrate, crystal such as tricalcium phosphate belong to one of reason that causes urinary calculus.We know that lithiasis is attended by many miseries such as renal colic, n and V, and blood urine, heating etc., even cause other bigger disease.Therefore it is very crucial to keep in the urine certain density phosphate radical.Phosphate radical in the in view of this regular quantitative measurement urine is absolutely necessary.We can understand metabolism status and the relevant pathology state of biosome by the content of the phosphate radical in the periodic measurement urine.
About measuring the phosphate radical in the urine, can reduce following several to the method that detects the phosphate radical in the urine both at home and abroad in the past:
1, the use chromatography of ions detects phosphate radical (L.Opliti, R.Chiaraluce, V.Consalvi, N.Cetulli and R.Scandurra, Clinica Chimica Acta, 184 (1989) 155-166 in the urine; Journal ofChromatography, 433 (1988) 373-376;
2,, measure the phosphate radical (F.Grases andJ.G.March, Analytica Chimica Acta, 229 (1990) 249-254) in the urine based on the strong inhibition of phosphate radical to the lime carbonate crystal seed;
3, use single on-line calibration method and photometric means by the phosphate radical (D.G.Themelis in the sequence injection assay determination urine, A.Economou, a.Tsiomlektsis and p.D.Tzanavaras, AnalyticalBiochemistry 330 (2004) 193-198).
But there are some problems respectively in these above methods:
1, many instruments are measured the phosphate radical in the urine clinically because the narrow range of linearity need be diluted urine sample significantly, just can go to measure.
2, the method for the phosphate radical in many measurement urines is to come indirect measurement by some other composition of surveying in the urine.Such as the content of growing to determine phosphate radical by the calcium carbonate crystal of surveying urinary calculi.So both loaded down with trivial details, time-consuming again, bring many inconvenience clinically.
3, the method for phosphate radical in many detection urines, sensitivity is lower, and deviation is bigger, perhaps can not give an accurate quantitative results.
Summary of the invention:
The objective of the invention is to break through the method for phosphate radical in traditional in the past detection urine, invent a kind of simply, sensitive, fast, operable clinically, detect the method for phosphate radical in the undiluted urine.
The invention provides a kind of method that detects phosphate radical in the urine, this method be developer based on ytterbium ion, with the pyrocatechol violet pH be in the HEPES buffer solution of 6.5-7.5 directly and detect phosphate radical in the urine quantitatively, it is as follows that it specifically detects step:
1, HEPES (10mM) buffer solution of preparation pH6.5-7.5, and prepare 2 * 10 -3The Yb of M 3+Solution and 2 * 10 -3The pyrocatechol violet solution of M;
2, the HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank -3The pyrocatechol violet solution 10-100 μ l of M is added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 443nm has absorption maximum;
3, in above-mentioned cuvette, add 2 times of molal quantitys to the Yb of pyrocatechol violet solution 3+Solution, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, finds that maximum absorption band becomes 623nm by above-mentioned 443nm;
4, get urine sample, be added in this cuvette with microsyringe gradually, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with urine sample, maximum absorption band is become again to 443nm gradually by 623nm again, and the color of solution is also returned yellow by blue stain gradually, when absorption peak reaches maximum at 443nm, during the complete flavescence of the color of solution, stop to add urine sample;
5, by [Yb 3+] * 2ml * 95/m Urine* 100% calculates that phosphate radical gets content in the urine, m in the formula Urine=V Urine
Described Yb 3+Solution is YbCl 3Solution or Yb (ClO 4) 3Solution.
Described Yb 3+Can use Nd 3+, Sm 3+, Gd 3+, Er 3+Or Ho 3+Replace.
The present invention has following advantage: 1, our detected object is undiluted urine sample, saves preprocessor, bring clinically very big easily; 2, detect on the UV, visible light spectrophotometer, testing process is simple, and can with the naked eye obviously observe this effect; Although contain many other compositions in 3 urines, our detection method has shown good selectivity to phosphate radical, that is to say, our testing result is not disturbed in the existence of other composition; 4, our detection is quantitative, the very big range of linearity is arranged, and error is very little; Obviously, this invention can be applied to and detect urine sample clinically.
Description of drawings:
Fig. 1 to Fig. 9 is the UV, visible light absorption figure of phosphate radical in the detection urine of corresponding embodiment 1 to embodiment 9.
Embodiment:
HEPES (10mM) buffer solution of embodiment 1. preparation pH7.0, and prepare 2 * 10 -3The YbCl of M 3Solution and 2 * 10 -3The pyrocatechol violet solution of M; The HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette,, draws 2 * 10 with microsyringe as blank -3The pyrocatechol violet solution 50 μ l of M are added in this cuvette, and solution is detected on HP8453 UV, visible light spectrophotometer by colourless flavescence at this moment, and 443nm has absorption maximum; In above-mentioned cuvette, add 2 * 10 -3The Yb of M 3+Solution 100 μ l, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, finds that maximum absorption band becomes 623nm by above-mentioned 443nm; Get adult's urine sample, be added in this cuvette with microsyringe gradually, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with urine sample, maximum absorption band is become again to 443nm gradually by 623nm again, and the color of solution is also returned yellow by blue stain gradually, when absorption peak reaches maximum at 443nm, during the complete flavescence of the color of solution, this moment, the urine addition was 20 μ l; Calculate: 100 μ l * 2ml * 95/20g * 100%, in the urine phosphate content be 0.095%, UV, visible light absorption figure sees Fig. 1.
Press embodiment 1 and survey 10 health adult's urine samples, in the urine phosphate content be 0.0863-0.119%.If the discharge rate by adult's twenty-four-hour urine liquid is 1L, the discharge rate scope that draws adult's urine phosphorus hydrochlorate so is 0.863-1.19g, surely closes with the normal discharge rate (0.5～1.5g/24 hour) of the inorganic phosphate of statistics in the urine medically.
HEPES (10mM) buffer solution of embodiment 2. preparation pH6.5, all the other are with embodiment 1, in the urine phosphate content be 0.095%, UV, visible light absorption figure sees Fig. 2.
Embodiment 3. gets healthy child's urine of an one full year of life, and all the other calculate with embodiment 1: 100 μ l * 2ml * 95/4g * 100%, in the urine phosphate content be 0.38%, UV, visible light absorption figure sees Fig. 3.From the foregoing description, we find out that the phosphate content in child's urine is higher, this be because child and adult Comparatively speaking, owing to child is in the skeletonization animated period, the higher and accretion rate of alkaline phosphatase activities is quite fast etc., and reason causes.
Embodiment 4. usefulness Yb (ClO 4) 3Solution, get another adult's urine, all the other are with embodiment 1,100 μ l * 2ml * 95/16g * 100%, in the urine phosphate content be 0.119%, UV, visible light absorption figure sees Fig. 4.
Embodiment 5. uses ErCl 3Solution, all the other all the other with embodiment 1,100 μ l * 2ml * 95/20g * 100%, in the urine phosphate content be 0.095%, UV, visible light absorption figure sees Fig. 5.
Embodiment 6. uses NdClO 4) 3Solution, all the other are with embodiment 1,100 μ l * 2ml * 95/20g * 100%, in the urine phosphate content be 0.095%, UV, visible light absorption figure sees Fig. 6.
Embodiment 7. uses SmClO 4) 3Solution, all the other same EXAMPLE l, 100 μ l * 2ml * 95/20g * 100%, in the urine phosphate content be 0.095%, UV, visible light absorption figure sees Fig. 7.
Embodiment 8. uses GdCl 3Solution, all the other same EXAMPLE l, 100 μ l * 2ml * 95/16g * 100%, in the urine phosphate content be 0.095%, UV, visible light absorption figure sees Fig. 8.
Embodiment 9. uses HoCl 3Solution, all the other are with embodiment 1,100 μ l * 2ml * 95/16g * 100%, in the urine phosphate content be 0.095%, UV, visible light absorption figure sees Fig. 9.

Claims (3)

1, a kind of method that detects phosphate radical in the urine is characterized in that, this method comprises following detection step:
(1), preparation pH6.5-7.5 concentration is the HEPES buffer solution of 10mM, and prepares 2 * 10 -3The Yb of M 3+Solution and 2 * 10 -3The pyrocatechol violet solution of M;
(2), the HEPES buffer solution of 2ml is added in the clean ultraviolet cuvette, as blank, with microsyringe absorption 2 * 10 -3The pyrocatechol violet solution 10-100 μ l of M is added in this cuvette, and solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 443nm has absorption maximum;
(3), in above-mentioned cuvette, add 2 times of molal quantitys to the Yb of pyrocatechol violet solution 3+Solution, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, finds that maximum absorption band becomes 623nm by above-mentioned 443nm;
(4), get urine sample, be added in this cuvette with microsyringe gradually, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with urine sample, maximum absorption band is become again to 443nm gradually by 623nm again, and the color of solution is also returned yellow by blue stain gradually, when absorption peak reaches maximum at 443nm, during the complete flavescence of the color of solution, stop to add urine sample;
(5), by [Yb 3+] * 2ml * 95/m Urine* 100% calculates the content of phosphate radical in the urine, m in the formula Urine=V Urine
2, by the described a kind of method that detects phosphate radical in the urine of claim 1, it is characterized in that described Yb 3+Solution is YbCl 3Solution or Yb (ClO 4) 3Solution.
3, by claim 1 or 2 described a kind of methods that detect phosphate radical in the urine, it is characterized in that described Yb 3+Can use Nd 3+, Sm 3+, Gd 3+, Er 3+Or Ho 3+Replace.
CNB2004100124481A 2004-07-26 2004-07-26 Method for detecting phosphate radical in urine Expired - Fee Related CN1313818C (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100594373C (en) * 2008-04-25 2010-03-17 山西大学 Method for detecting aqueous solution oxalate

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1317564C (en) * 2005-04-08 2007-05-23 山西大学 Method for visual detection of inorganic phosphorus in urine and kit thereof
CN104267029B (en) * 2014-10-23 2017-05-17 攀枝花钢企欣宇化工有限公司 Quantitative analysis method for phosphate radical

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5991367A (en) * 1982-11-16 1984-05-26 Fuaintetsuku Japan:Kk Highly sensitive analysis of phosphate ion
US5117370A (en) * 1988-12-22 1992-05-26 Ford Motor Company Detection system for chemical analysis of zinc phosphate coating solutions
CN2288442Y (en) * 1997-03-31 1998-08-19 陈义新 Automatic detector for phosphate content in water
CN1044744C (en) * 1996-11-15 1999-08-18 衡水市环境保护局 Preparation method for phosphorus indicator paper

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5991367A (en) * 1982-11-16 1984-05-26 Fuaintetsuku Japan:Kk Highly sensitive analysis of phosphate ion
US5117370A (en) * 1988-12-22 1992-05-26 Ford Motor Company Detection system for chemical analysis of zinc phosphate coating solutions
CN1044744C (en) * 1996-11-15 1999-08-18 衡水市环境保护局 Preparation method for phosphorus indicator paper
CN2288442Y (en) * 1997-03-31 1998-08-19 陈义新 Automatic detector for phosphate content in water

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
氧化铝微柱预富集分光光度法测定天然水体中微量磷酸根 丁建华等，分析科学学报，第18卷第3期 2002 *
焦磷酸铜镀液中磷酸根和焦磷酸根的光度测定 邱星初等，材料保护，第33卷第12期 2000 *
焦磷酸铜镀液中磷酸根和焦磷酸根的光度测定 邱星初等，材料保护，第33卷第12期 2000;饲料中无机磷和有机磷的测定方法 钟国清，饲料检测，第24卷第7期 2003;氧化铝微柱预富集分光光度法测定天然水体中微量磷酸根 丁建华等，分析科学学报，第18卷第3期 2002 *
饲料中无机磷和有机磷的测定方法 钟国清，饲料检测，第24卷第7期 2003 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100594373C (en) * 2008-04-25 2010-03-17 山西大学 Method for detecting aqueous solution oxalate

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