CN101241076A - Hydroxy free radical concentration determination method - Google Patents
Hydroxy free radical concentration determination method Download PDFInfo
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- CN101241076A CN101241076A CNA2008100196933A CN200810019693A CN101241076A CN 101241076 A CN101241076 A CN 101241076A CN A2008100196933 A CNA2008100196933 A CN A2008100196933A CN 200810019693 A CN200810019693 A CN 200810019693A CN 101241076 A CN101241076 A CN 101241076A
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- hydroxyl
- free radical
- terephthalic acid
- tpa
- concentration
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Abstract
The present invention provides a method for detecting concentration of hydroxyl radical. In the system produced by adding capture agent terephthalic acid sodium salt (NaTA) into hydroxyl radical, 2-hydroxyl terephthalic acid (HTA) is produced by reaction of terephthalic acid sodium salt and hydroxyl radical, which has stable fluorescence and is detected by fluorescence spectrophotometer (excited wavelength lambada ex=315 nm, projecting wavelength lambada em=425 nm), and then the concentration of 2-hydroxyl terephthalic acid is conversed to the concentration of hydroxyl radical. The detecting method of present invention is easy to do, credibility and nicety, especial for used in the conditions of circumstance produced hydroxyl radical at the value of pH<12, or the solution of the hydroxyl radical produced system required the conductivity.
Description
Technical field
The present invention relates to a kind of assay method of hydroxyl radical free radical, use the terephthalic acid (TPA) sodium salt, generate 2-hydroxyl terephthalic acid (TPA), utilize fluorospectrophotometer method indirect determination hydroxyl radical free radical then as the hydroxyl radical free radical trapping agent.
Background technology
Because hydroxyl radical free radical be extremely active material, what the speed that it and majority of compounds are reacted made it very soon exists the life-span 10
-9The s order of magnitude, thus very difficult to the mensuration of hydroxyl radical free radical, also be scientific circles' research focuses.
Very many about the document of hydroxyl free based assays both at home and abroad at present, mainly be divided into direct mensuration and indirect determination.
It is representative that direct assay method mainly contains with electron spin resonance spectroscopy (ESR), but the shortcoming of this method is an insufficient sensitivity, and can not the quantitative measurement hydroxyl radical free radical.Directly the needed instrument of determination method all is very expensive in addition, and common laboratory can't meet the demands.
Indirect determination method is meant aromatic hydroxy, cardinal principle: use aromatic compounds (phenol, benzoic acid, salicylic acid etc.) as trapping agent and hydroxyl free radical reaction, generate metastable hydroxylation product, use various means to measure metastable hydroxylation product then, thus the indirect determination hydroxyl radical free radical.The indirect determination method exists the possibility that reduces cost of determination, so become the focus of research rapidly simultaneously because the selection of its trapping agent is ununified as yet at present.
Abroad the hydroxylation product that produces for hydroxyl radical free radical indirect determination method is measured the main instruments such as high performance liquid chromatography, gas chromatography that rely on, and the domestic document that then uses spectrophotometer method to measure hydroxyl radical free radical is many.The indirect determination method is because the mechanism of selected trapping agent and hydroxyl free radical reaction is very complicated, and the factor that influences accuracy of measurement is very many.Whether trapping agent and hydroxyl radical free radical reaction path be unique, and whether the hydroxylation product after the reaction is stable, all has to be determined.
Chinese invention patent discloses two kinds of hydroxyl radical free radical assay methods (CN1412553 and CN101042371), uses isopropyl alcohol and P-hydroxybenzoic acid to be trapping agent respectively, owing to the mechanism complexity, influence and easily cause systematic error more than the factor of accuracy of measurement; For the mensuration of intermediate product, the gas chromatography degree of accuracy is high but operation is comparatively loaded down with trivial details, by the response current potential measurement is simple to operate but degree of accuracy is then poor slightly.
The hydroxyl radical free radical widespread use makes the research of its method for measurement of concentration become focus, and the raising of the accuracy of hydroxy free radical concentration assay method and the reduction of cost will be the trend of future studies.
Summary of the invention
The object of the invention provides a kind of assay method of hydroxyl radical free radical, uses the terephthalic acid (TPA) sodium salt as trapping agent, the limitation that can only dissolve under the highly basic condition when overcoming terephthalic acid (TPA) as trapping agent.For environment pH＜12 that produce system at hydroxyl radical free radical, or the hydroxyl radical free radical solution that produces system has under the situation of requirement (as: Pulsed Discharge hydroxyl radical free radical system in the liquid) this assay method especially suitable to conductivity.Use fluorescence spectrophotometry, the 2-hydroxyl P-phthalic acid at concentration according to the crest area estimation is converted into hydroxy free radical concentration at last, and this assay method is row, reliable, accurate easily.
Technical scheme of the present invention is: preparation 2-hydroxyl terephthalic acid (TPA) standard solution with the spectrogram of each standard solution of fluorescent spectrophotometer assay, obtains the typical curve of 2-hydroxyl terephthalic acid (TPA) concentration of standard solution and spectrogram area corresponding relation; In the system that during practical measurement the excessive adding hydroxyl radical free radical of trapping agent terephthalic acid (TPA) sodium salt is produced, generate 2-hydroxyl terephthalic acid (TPA) with the hydroxyl free radical reaction that produces in the system, adopt fluorospectrophotometer to detect mixed solution, excited-emission spectrum characteristics figure; At last, the crest area that provides according to characteristic pattern, typical curve by crest area and 2-hydroxyl terephthalic acid (TPA) volumetric molar concentration relation, obtain 2-hydroxyl terephthalic acid (TPA) volumetric molar concentration corresponding under this crest area, the reduction formula that is obtained by the chemical reaction relational expression converses the volumetric molar concentration of hydroxyl radical free radical again.
In the said determination method mensuration process, to feed O in the system of terephthalic acid (TPA) sodium salt and hydroxyl free radical reaction
2, the feeding time is 30min, the solution under the reaction system reaches oxygen saturation at this moment; (0.2M, pH=7.4) buffer solution is regulated the pH value in 6～11 scopes to adopt the fluorospectrophotometer detection to use sodium phosphate before.
When above-mentioned fluorescence spectrophotometry detects, use the fluorescence of wavelength 315nm to excite, collect the fluorescence spectrum of sample in 340～625nm wavelength coverage.
The present invention utilizes trapping agent and hydroxyl free radical reaction, by the concentration of intermediate products indirect determination hydroxyl radical free radical of assaying reaction.The terephthalic acid (TPA) sodium salt is caught hydroxyl radical free radical, and to produce the reaction path of 2-hydroxyl terephthalic acid (TPA) as follows:
Catch the research of hydroxyl radical free radical reaction principle according to the terephthalic acid (TPA) sodium salt, when dissolved oxygen was saturated in system, the efficient of capture reaction was 35%, and the volumetric molar concentration of promptly known 2-hydroxyl terephthalic acid (TPA) can be calculated hydroxy free radical concentration according to following formula:
In the formula: C
OHBe hydroxy free radical concentration, M;
C
2-hydroxyl terephthalic acid (TPA)Be 2-hydroxyl P-phthalic acid at concentration, M
Use fluorescence spectrophotometry 2-hydroxyl P-phthalic acid at concentration,, obtain hydroxy free radical concentration indirectly by above-mentioned formula.
The trapping agent that the present invention uses and the intermediate product 2-hydroxyl terephthalic acid (TPA) of hydroxyl free radical reaction are 0～2.0 * 10
-3The M concentration range has linear response relationship to fluorescence spectrum crest area, so hydroxyl radical free radical sensing range of the present invention is 0～5.7 * 10
-3M.2-hydroxyl terephthalic acid (TPA) is 5.0 * 10
-7Can detect fluorescence spectrum during M concentration, lowest detection promptly of the present invention is limited to 1.5 * 10
-6M.
Embodiment
The key step that the present invention measures hydroxy free radical concentration is as follows:
1. drawing standard curve
(1) preparation 0～2.0 * 10
-3A series of product 2-hydroxyl terephthalic acid (TPA) standard solution of catching in the M scope, 6 normal concentration: 0M, 0.2 * 10 are got in suggestion
-3M, 0.5 * 10
-3M, 1.0 * 10
-3M, 1.5 * 10
-3M, 2.0 * 10
-3M.
(2) to get the 5.00ml standard solution at every turn, add the 5.00ml buffer solution of sodium phosphate, use fluorescence spectrophotometry, calculating excites-the crest area of emission spectrum characteristics figure;
(3) the crest area with exciting of recording-emission spectrum characteristics figure is an ordinate, is horizontal ordinate with 2-hydroxyl terephthalic acid (TPA) concentration of standard solution, the drawing standard curve;
2. measure 2-hydroxyl P-phthalic acid at concentration
Aerating oxygen is up to oxygen saturation in tested systems, with excessive terephthalic acid (TPA) sodium salt is trapping agent, reaction time 20～30min, take out the 5.00ml reaction solution and carry out the fluorescence method analysis according to (1) in the step 1, use wavelength 315nm to excite, collect the fluorescence generation spectrum of sample in 340～625nm wavelength coverage, excited-the crest area of emission spectrum characteristics figure; The typical curve of being drawn by (3) in the crest area of the exciting of the water sample that records-emission spectrum characteristics figure and the step 1 obtains 2-hydroxyl terephthalic acid (TPA) volumetric molar concentration in the solution.
3. pass through formula
Calculate the hydroxyl radical free radical volumetric molar concentration.
Embodiment
The hydroxy free radical concentration that liquid mesohigh pulsed discharge system generates is measured.In beaker, dispose 5 * 10 with distilled water
-4The terephthalic acid (TPA) sodium solution 1L of M concentration uses magnetic stirrer, spends the night, and is stand-by.The NaCl solution regulator solution conductivity of using 0.1mol/L is to 150 μ s/cm.The solution that configures is placed the reaction vessel of liquid mesohigh pulse discharge means, in reactor, feed O with the flow of 400ml/min
2, start-up system power supply after the 10min, crest voltage 60kV, 15min and 30min respectively get the 5.00ml reaction solution once after the reaction, add the buffer solution of sodium phosphate 5.00ml of pH=7.4.Use fluorescent spectrophotometer assay solution fluorescence, calculate and excite-the crest area of emission spectrum characteristics figure, 2-hydroxyl terephthalic acid (TPA) is carried out quantitative test by the typical curve of drawing.The result: behind the energising 15min, the solution hydroxy free radical concentration is 2.57 * 10
-5M, behind the energising 30min, the solution hydroxy free radical concentration is 4.96 * 10
-5M.
Claims (7)
1, a kind of assay method of hydroxy free radical concentration, it is characterized in that: preparation 2-hydroxyl terephthalic acid (TPA) standard solution, with the spectrogram of each standard solution of fluorescent spectrophotometer assay, obtain the typical curve of 2-hydroxyl terephthalic acid (TPA) concentration of standard solution and spectrogram area corresponding relation; In the system that during practical measurement the excessive adding hydroxyl radical free radical of trapping agent terephthalic acid (TPA) sodium salt is produced, generate 2-hydroxyl terephthalic acid (TPA) with the hydroxyl free radical reaction that produces in the system, adopt fluorospectrophotometer to detect mixed solution, excited-emission spectrum characteristics figure; At last, the crest area that provides according to characteristic pattern, typical curve by crest area and 2-hydroxyl terephthalic acid (TPA) volumetric molar concentration relation, obtain 2-hydroxyl terephthalic acid (TPA) volumetric molar concentration corresponding under this crest area, the reduction formula that is obtained by the chemical reaction relational expression converses the volumetric molar concentration of hydroxyl radical free radical again.
2, the assay method of a kind of hydroxy free radical concentration according to claim 1 is characterized in that: described reduction formula is:
In the formula: C
OHBe the hydroxyl radical free radical volumetric molar concentration;
C
2-hydroxyl terephthalic acid (TPA)Be 2-hydroxyl terephthalic acid (TPA) volumetric molar concentration.
3, the assay method of a kind of hydroxy free radical concentration according to claim 1 is characterized in that: feed O in the system of terephthalic acid (TPA) sodium salt and hydroxyl free radical reaction
2Until oxygen saturation; Adopting fluorospectrophotometer to detect mixed solution uses buffer solution of sodium phosphate to regulate mixed solution pH value in 6～11 scopes before.
4, the assay method of a kind of hydroxy free radical concentration according to claim 3 is characterized in that: O
2The feeding time is 30min, and buffer solution of sodium phosphate concentration is 0.2M, and the pH value is 7.4.
5, the assay method of a kind of hydroxy free radical concentration according to claim 1 is characterized in that: when above-mentioned fluorescence spectrophotometry detects, get the 5.00ml standard solution at every turn, add the 5.00ml buffer solution of sodium phosphate; Use the fluorescence of wavelength 315nm to excite, collect the fluorescence spectrum of sample in 340～625nm wavelength coverage.
6, the assay method of a kind of hydroxy free radical concentration according to claim 1 is characterized in that: during preparation 2-hydroxyl terephthalic acid (TPA) standard solution, getting 2-hydroxyl terephthaldehyde acid solutions scope is 0～2.0 * 10
-3M.
7, the assay method of a kind of hydroxy free radical concentration according to claim 1 is characterized in that: with the reaction time in the system of the excessive adding hydroxyl radical free radical generation of disodium terephthalate is 20～30min.
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|---|---|---|---|
| CNA2008100196933A CN101241076A (en) | 2008-03-12 | 2008-03-12 | Hydroxy free radical concentration determination method |
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Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101968437A (en) * | 2010-10-14 | 2011-02-09 | 中国科学院安徽光学精密机械研究所 | Atmosphere OH radical measuring system |
| CN101413896B (en) * | 2008-12-02 | 2011-05-18 | 上海理工大学 | Method for measuring hydroxy free radical |
| CN101718705B (en) * | 2009-11-13 | 2011-09-21 | 上海理工大学 | Method for detecting hydroxyl radicals |
| CN101587065B (en) * | 2009-06-25 | 2011-11-30 | 上海理工大学 | Method for detecting oxygen radical on surfaces of plasma functionalized carbon nanotubes |
| WO2013058676A1 (en) | 2011-10-21 | 2013-04-25 | ОБЩЕСТВО С ОГРАНИЧЕННОЙ ОТВЕТСТВЕННОСТЬЮ "ДИСофт" | Method for determining the quantity and/or activity of antioxidants |
| CN103149188A (en) * | 2013-03-05 | 2013-06-12 | 东华大学 | Fluorescent quantitative method for detecting hydroxyl radical |
| CN103674911A (en) * | 2013-12-02 | 2014-03-26 | 中国科学院安徽光学精密机械研究所 | Fluorescent pool for atmosphere Hox free radical real-time measurement |
| CN103728287A (en) * | 2014-01-10 | 2014-04-16 | 福建医科大学 | Fluorescence analysis method for determining glucose by employing nanometer copper oxide as simulated peroxide |
| CN103728288A (en) * | 2014-01-10 | 2014-04-16 | 福建医科大学 | Method for oxidizing terephthalic acid to enhance fluorescence by employing nanometer copper oxide to catalyze hydrogen peroxide |
| CN103760145A (en) * | 2014-01-27 | 2014-04-30 | 同济大学 | Ratio fluorescent probe for detecting hydroxyl radical and synthesis method and application of ratio fluorescent probe |
| CN103776808A (en) * | 2014-01-24 | 2014-05-07 | 鼎泰（湖北）生化科技设备制造有限公司 | Method for detecting hydroxyl radical concentration |
| CN106124463A (en) * | 2016-06-09 | 2016-11-16 | 天津城建大学 | A kind of micro-interface hydroxyl radical free radical characterizing method based on fluorescence imaging analysis |
| CN106248811A (en) * | 2016-06-20 | 2016-12-21 | 环境保护部华南环境科学研究所 | A kind of detect the method that airborne fine particulate matter produces reactive oxygen free radical in human body |
| CN108088820A (en) * | 2017-12-14 | 2018-05-29 | 大连理工大学 | A kind of method that hydroxyl radical free radical is quantitatively detected using laser flash photolysis technology |
| CN110291383A (en) * | 2017-02-23 | 2019-09-27 | 株式会社Ihi | OH free radical detects detector, OH determining free radicals device and OH determining free radicals method |
| CN110658163A (en) * | 2018-06-29 | 2020-01-07 | 成都先导药物开发股份有限公司 | Method for monitoring reaction in synthesis of DNA coding compound |
| CN112285100A (en) * | 2020-10-16 | 2021-01-29 | 天津工业大学 | Method for detecting concentration of free radicals in sodium hypochlorite aqueous solution |
| US11585761B2 (en) | 2018-08-23 | 2023-02-21 | Ihi Corporation | OH radical measuring device and OH radical measuring method |
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2008
- 2008-03-12 CN CNA2008100196933A patent/CN101241076A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101413896B (en) * | 2008-12-02 | 2011-05-18 | 上海理工大学 | Method for measuring hydroxy free radical |
| CN101587065B (en) * | 2009-06-25 | 2011-11-30 | 上海理工大学 | Method for detecting oxygen radical on surfaces of plasma functionalized carbon nanotubes |
| CN101718705B (en) * | 2009-11-13 | 2011-09-21 | 上海理工大学 | Method for detecting hydroxyl radicals |
| CN101968437A (en) * | 2010-10-14 | 2011-02-09 | 中国科学院安徽光学精密机械研究所 | Atmosphere OH radical measuring system |
| CN101968437B (en) * | 2010-10-14 | 2012-05-30 | 中国科学院安徽光学精密机械研究所 | Atmosphere OH radical measuring system |
| WO2013058676A1 (en) | 2011-10-21 | 2013-04-25 | ОБЩЕСТВО С ОГРАНИЧЕННОЙ ОТВЕТСТВЕННОСТЬЮ "ДИСофт" | Method for determining the quantity and/or activity of antioxidants |
| CN103149188B (en) * | 2013-03-05 | 2015-05-13 | 东华大学 | Fluorescent quantitative method for detecting hydroxyl radical |
| CN103149188A (en) * | 2013-03-05 | 2013-06-12 | 东华大学 | Fluorescent quantitative method for detecting hydroxyl radical |
| CN103674911A (en) * | 2013-12-02 | 2014-03-26 | 中国科学院安徽光学精密机械研究所 | Fluorescent pool for atmosphere Hox free radical real-time measurement |
| CN103728287A (en) * | 2014-01-10 | 2014-04-16 | 福建医科大学 | Fluorescence analysis method for determining glucose by employing nanometer copper oxide as simulated peroxide |
| CN103728288A (en) * | 2014-01-10 | 2014-04-16 | 福建医科大学 | Method for oxidizing terephthalic acid to enhance fluorescence by employing nanometer copper oxide to catalyze hydrogen peroxide |
| CN103728288B (en) * | 2014-01-10 | 2016-03-30 | 福建医科大学 | The method of fluorescence is strengthened with nano cupric oxide catalyzing hydrogen peroxide oxidation terephthalic acid (TPA) |
| CN103728287B (en) * | 2014-01-10 | 2016-03-30 | 福建医科大学 | Nano cupric oxide Mimetic enzyme measures the fluorescence analysis method of glucose |
| CN103776808A (en) * | 2014-01-24 | 2014-05-07 | 鼎泰（湖北）生化科技设备制造有限公司 | Method for detecting hydroxyl radical concentration |
| CN103760145A (en) * | 2014-01-27 | 2014-04-30 | 同济大学 | Ratio fluorescent probe for detecting hydroxyl radical and synthesis method and application of ratio fluorescent probe |
| CN106124463A (en) * | 2016-06-09 | 2016-11-16 | 天津城建大学 | A kind of micro-interface hydroxyl radical free radical characterizing method based on fluorescence imaging analysis |
| CN106124463B (en) * | 2016-06-09 | 2018-11-09 | 天津城建大学 | A kind of micro-interface hydroxyl radical free radical characterizing method based on fluorescence imaging analysis |
| CN106248811A (en) * | 2016-06-20 | 2016-12-21 | 环境保护部华南环境科学研究所 | A kind of detect the method that airborne fine particulate matter produces reactive oxygen free radical in human body |
| CN110291383A (en) * | 2017-02-23 | 2019-09-27 | 株式会社Ihi | OH free radical detects detector, OH determining free radicals device and OH determining free radicals method |
| CN110291383B (en) * | 2017-02-23 | 2021-12-28 | 株式会社Ihi | OH radical detection probe, OH radical measurement device, and OH radical measurement method |
| US11313803B2 (en) | 2017-02-23 | 2022-04-26 | Ihi Corporation | OH radical measurement device and method using an OH radical detection probe |
| CN108088820A (en) * | 2017-12-14 | 2018-05-29 | 大连理工大学 | A kind of method that hydroxyl radical free radical is quantitatively detected using laser flash photolysis technology |
| CN108088820B (en) * | 2017-12-14 | 2020-09-01 | 大连理工大学 | Method for quantitatively detecting hydroxyl free radicals by using laser flash photolysis technology |
| CN110658163A (en) * | 2018-06-29 | 2020-01-07 | 成都先导药物开发股份有限公司 | Method for monitoring reaction in synthesis of DNA coding compound |
| US11585761B2 (en) | 2018-08-23 | 2023-02-21 | Ihi Corporation | OH radical measuring device and OH radical measuring method |
| CN112285100A (en) * | 2020-10-16 | 2021-01-29 | 天津工业大学 | Method for detecting concentration of free radicals in sodium hypochlorite aqueous solution |
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