CN100594373C

CN100594373C - Method for detecting aqueous solution oxalate

Info

Publication number: CN100594373C
Application number: CN200810054882A
Authority: CN
Inventors: 阴彩霞; 孙远强; 霍方俊; 刘艳林
Original assignee: Shanxi University
Current assignee: Shanxi University
Priority date: 2008-04-25
Filing date: 2008-04-25
Publication date: 2010-03-17
Anticipated expiration: 2028-04-25
Also published as: CN101285776A

Abstract

The invention provides a method for detecting oxalate in water solution. The method uses pyrocatechol violet as color developing reagent based on Cu<2+> ions and directly and quantificationally detects the oxalate in the water solution in HEPES buffer solution with a pH of 6.0, thereby the determining process is simple and convenient and the measuring result is accurate. The method has high sensibility, can not be interfered by other anions in the water solution, and can be applied to detection of the oxalate in clinic, foodstuff and environment.

Description

A kind of method that detects aqueous solution medium-height grass acid group

Technical field:

The present invention relates to a kind of detection method of oxalate, specifically belong to a kind of method that detects aqueous solution medium-height grass acid group.

Background technology:

Oxalic acid is a kind of high toxicity compound, extensively is present in plant, animal, the microorganism, and accumulating in vivo can cause multiple pathological state, as urolithiasis, hyperoxaluria and renal failure etc.It may be the symptom of diseases such as diabetes that urine mesoxalic acid salinity increases, and attenuating may be an acute and chronic nephritis.The diet mesoxalic acid is excessive, can form insolubles with calcium before absorption, hinders the absorption of calcium; Can form calculus with calcium and other materials again after the absorption, it is reported that 70%-80% kidney stone is is principal ingredient with the calcium oxalate.Utilize the beverage of ozone sterilization, when wherein ascorbic content is higher, can form oxalic acid, thereby promote the regeneration of microorganism, reduce bactericidal effect.Simultaneously oxalic acid is as the toxin of fungus secretion and participate in causing a disease, at present Sclerotinia, aspergillus, chestnut epidemic disease shell Pseudomonas, fall and report arranged in the genus such as a cup Agaricus, Rhizoctonia, sclerotium and Crystulariella.Oxalic acid all plays an important role in the pathogenic course of above-mentioned fungi.In view of this, the detection by quantitative at the aqueous solution mesoxalic acid is very important.

About measuring the oxalate in the aqueous solution, can reduce following several to the method that detects oxalate both at home and abroad in the past:

1, titrimetry is a titrant with the ceric sulfate, is indicator (military Xiao Ling etc., Xinjiang Medicine University's newspaper, 22 (1999) 148) with iron-adjacent luxuriant and rich with fragrance gong quinoline;

2, colourimetry, with oxalic acid oxidation or reduction, or direct and certain developer reaction generates coloring matter (Shen You etc., spectroscopy and spectral analysis, 18 (1998) 756-758);

3, the chromatography of ions (Del Nozal, M.J, J.Chromatograohy A, 881 (2000) 629-638);

4, capillary electrophoresis analysis method (Bryant, C.N, Chromatography A, 771 (1997) 285-299);

5, high performance liquid chromatography (Yu Le, Peng Xinxiang, Yang Chong etc., analytical chemistry research notes, 30 (2002) 1119-1122);

6, enzyme process (Thakue, M., Goyal, L., and Pundie, C.S, J.BiochemBiophys.Methods, 44 (2000) 77-88).

But there are some problems respectively in these above methods:

1, titrimetry sensitivity is low, operates more loaded down with trivial details, consuming time;

2, other colourimetrys need preformed precipitate, be difficult to fully oxalate be extracted, and complex operation and repeated relatively poor;

3, the Capillary Electrophoresis instrument is not popularized as yet;

4, high performance liquid chromatography requires oxalate is separated, in the detachment process interference of mineral acid more serious, and instrument costliness is had relatively high expectations to operating personnel;

5, the oxalate oxidase cost is higher, has limited the widespread adoption of this method.

Summary of the invention:

The object of the present invention is to provide the method for low, simple to operate, the highly sensitive detection aqueous solution medium-height grass acid group of a kind of cost.

The invention provides a kind of method that detects aqueous solution medium-height grass acid group, this method is based on Cu ²⁺Ion is a developer with the pyrocatechol violet, pH be in 6.0 the HEPES buffer solution directly and detect oxalate in the aqueous solution quantitatively, it specifically detects step and comprises:

1, the concentration of preparation pH6.0 is the HEPES buffer solution of 10mM, and compound concentration is the Cu of 2mM respectively ²⁺Solution and pyrocatechol violet solution;

2, the HEPES buffer solution with above-mentioned 2ml is added in the ultraviolet cuvette, as blank, to draw above-mentioned pyrocatechol violet solution 10-100 μ l with microsyringe and be added in the cuvette, this moment, solution was by colourless flavescence look, detect on the UV, visible light spectrophotometer, 443nm has absorption maximum;

3, in above-mentioned cuvette, add 2 times of molal quantitys again to the Cu of pyrocatechol violet solution ²⁺Solution, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, finds that maximum absorption band becomes 624nm by above-mentioned 443nm;

4, get tested oxalate solution, be added in the cuvette gradually with microsyringe, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with solution, maximum absorption band is become again to 443nm gradually by 624nm 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 solution;

5, by [Cu ²⁺] * 2 * V _Cu/ V _OxalateCalculate the content of solution medium-height grass acid group.

Described Cu ²⁺Solution is solution of water miscible cupric inorganic salt solution such as cupric chloride, copper nitrate, copper sulphate, Schweinfurt green, cupric perchlorate etc.

That the method that the present invention detects aqueous solution medium-height grass acid group can be applied to is clinical, the detection of food and environment medium-height grass acid group.

Detection method of the present invention is single-minded to relative other negative ion of the selectivity of oxalate, i.e. the mensuration of this method to oxalate is not disturbed in other anionic existence.Proof experiment: in the described cuvette of step 3, add 10 times of molal quantitys respectively or successively to acetate, nitrate radical, carbonate, phosphate radical, hydrogen phosphate, fluorine ion, chlorion or the bromide ion of step 4 medium-height grass acid group amount, on the UV, visible light spectrophotometer, detect, all less than changing, the system color does not change yet at the 624nm maximum absorption band.

The present invention has following advantage: 1, the detection architecture cost is low; 2, detect on the UV, visible light spectrophotometer, testing process is simple, and can with the naked eye obviously observe this effect and realize the naked eyes monitoring; 3, this method has shown good selectivity to oxalate, that is to say, other anionic existence is interference detection results not; 4, it is quantitative detecting, and the very big range of linearity is arranged, and error is very little; 5, testing environment is to carry out in aqueous solution, and therefore, this invention can be applied to the detection of environmental water systems or clinical body fluid medium-height grass acid group.

Description of drawings:

Fig. 1 is the UV, visible light absorption figure of embodiment 1

Fig. 2 is the UV, visible light absorption figure of embodiment 2

Fig. 3 is the UV, visible light absorption figure of embodiment 3

Fig. 4 is the UV, visible light absorption figure of embodiment 4

Fig. 5 is the change color figure of embodiment 5 testing processes

Embodiment:

Embodiment 1:

HEPES (10mM) buffer solution of preparation pH6.0, and the Cu (NO of preparation 2mM ₃) ₂3H ₂The pyrocatechol violet solution of O solution, 2mM and the sodium oxalate solution of 2mM; The HEPES buffer solution of 2ml is added in the ultraviolet cuvette,, draws the pyrocatechol violet solution 50 μ l of 2mM with microsyringe as blank, be added in this cuvette, solution is detected on the UV, visible light spectrophotometer by colourless flavescence at this moment, and 443nm has absorption maximum; In above-mentioned cuvette, add the Cu of 2mM ²⁺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 624nm by above-mentioned 443nm; Get sodium oxalate solution, application of sample limit, limit is detected on HP8453 UV, visible light spectrophotometer, adding along with oxalate, maximum absorption band is become again to 443nm gradually by 624nm again, 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 oxalate addition was 200 μ l; Calculate: 100 μ l * 2 * 10 ^-3* 2/200 μ l, the content that gets the solution mesoxalic acid is 2mM.UV, visible light absorption figure sees Fig. 1.

Embodiment 2:

The CuCl of preparation 2mM ₂3H ₂O solution, all the other are with embodiment 1, and uv absorption figure sees Fig. 2.

Embodiment 3:

The CuSO of preparation 2mM ₄5H ₂O solution, all the other are with embodiment 1, and uv absorption figure sees Fig. 3.

Embodiment 4:

Containing 50 μ M pyrocatechol violets and 100 μ M Cu (NO ₃) ₂3H ₂O, pH6.0HEPES (10mM) add 10 times of acetates to oxalate of molal quantity, nitrate radical, carbonate, phosphate radical, hydrogen phosphate, fluorine ion, chlorion and bromide ion successively in the buffer solution, uv absorption figure sees Fig. 4.

Embodiment 5

Containing 50 μ M pyrocatechol violets and 100 μ M Cu (NO ₃) ₂3H ₂O, pH6.0HEPES (10mM) add the oxalate solution that concentration is 2mM (0,50,100,150,200 μ l) gradually in the buffer solution.Change color figure sees Fig. 5.

Claims

1, a kind of method that detects aqueous solution medium-height grass acid group is characterized in that, comprises following detection step:

(1), the concentration of preparation pH6.0 is the HEPES buffer solution of 10mM, and compound concentration is the Cu of 2mM respectively ²⁺Solution and pyrocatechol violet solution;

(2) the HEPES buffer solution with above-mentioned 2ml is added in the ultraviolet cuvette, as blank, to draw above-mentioned pyrocatechol violet solution 10-100 μ l with microsyringe and be added in the cuvette, this moment, solution was by colourless flavescence look, detect on the UV, visible light spectrophotometer, 443nm has absorption maximum;

(3), in above-mentioned cuvette, add 2 times of molal quantitys again to the Cu of pyrocatechol violet solution ²⁺Solution, solution is detected on the UV, visible light spectrophotometer by xanthochromia indigo plant at this moment, finds that maximum absorption band becomes 624nm by above-mentioned 443nm;

(4), get tested oxalate solution, be added in the cuvette gradually with microsyringe, application of sample limit, limit is detected on the UV, visible light spectrophotometer, adding along with oxalate, maximum absorption band is become again to 443nm gradually by 624nm 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 oxalate;

(5), by [Cu ²⁺] * 2 * V _Cu/ V _OxalateCalculate the content of oxalate;

Described Cu ²⁺Solution is the solution of cupric chloride, copper nitrate, copper sulphate, Schweinfurt green or cupric perchlorate.