CN100514040C - Rhodamine S association complex microparticles enzyme catalysis fluorescent method for detecting trace amount hydrogen peroxide - Google Patents
Rhodamine S association complex microparticles enzyme catalysis fluorescent method for detecting trace amount hydrogen peroxide Download PDFInfo
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- CN100514040C CN100514040C CNB2007100506032A CN200710050603A CN100514040C CN 100514040 C CN100514040 C CN 100514040C CN B2007100506032 A CNB2007100506032 A CN B2007100506032A CN 200710050603 A CN200710050603 A CN 200710050603A CN 100514040 C CN100514040 C CN 100514040C
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Abstract
The invention discloses a detecting method for rhodamine S associated matter atom enzyme catalysis fluorescence through trace the quantity of hydrogen peroxide. After adding KI, HRP, H<SUB>2</SUB>O<SUB>2</SUB> and rhodamine S into sub acidity HAc-NaAc solution, shaking to uniformity and constant volume, the detected hierarchy with given deepness is gained. Excitation and emission spectra of the hierarchy is gained through scanning of a fluorophotometer. The fluorescence intensity F and the reagent blank F<SUB>0</SUB> of the hierarchy are detected at a point where the length of exciting light wave is 500nm and the length of transmitting light wave is 556nm. The value of Delta F=F-F<SUB>0</SUB> is calculated and the Delta F of the detected matter is determined. The H<SUB>2</SUB>O<SUB>2</SUB> concentration of the detected matter with unknown concentration is reckoned up according to a working curve. The method has the advantages of mild reaction conditions, simple equipment, easy operation, available and low-cost reagent, high sensitivity, good selectivity, and handy and quick speed. The invention can be widely applied easily.
Description
Technical field:
The present invention relates to H
2O
2Detection method, particularly detect the rhodamine S association complex microparticles enzyme catalysis fluorescent method of trace amount hydrogen peroxide.
Background technology:
Hydrogen peroxide is a kind of strong oxidizer, can be used as that bleaching agent, sanitizer, deoxidizer, liquid propellant etc. are widely used in that chemicals is synthetic, papermaking, environmental protection, medicine, weaving, mining industry and agricultural residue processing and other fields.Long-term contacted hydrogen oxide can decolour by hair, symptoms such as respiratory tract and skin irritatin occur.The method that detects hydrogen peroxide both at home and abroad at present mainly contains conventional titrimetry, electrochemical process, spectrophotometric method, fluorescence method, chemoluminescence method, chromatography.It is many to have plenty of disturbing factor in these methods, has plenty of instrument than complex and expensive, and it is low to have plenty of sensitivity, and measuring has error.
Summary of the invention:
The objective of the invention is will be for overcoming the deficiencies in the prior art, and provide a kind of highly sensitive, detect the rhodamine S association complex microparticles enzyme catalysis fluorescent method that detects trace amount hydrogen peroxide fast and accurately.
The present invention realizes by following technical proposals:
A kind of rhodamine S association complex microparticles enzyme catalysis fluorescent method that detects trace amount hydrogen peroxide, it is to utilize horseradish peroxidase (HRP) that hydrogen peroxide is measured in the fluorescent quenching characteristic combination of hydrogen peroxide catalyzed selectivity and rhodamine S (RhS) association microparticle.
Under acid condition, HRP can catalysis H
2O
2With excessive I
-Reaction generates I
3 -, I
3 -Form association microparticle (RhS-I with RhS
3 -), thereby cause the fluorescent value of system to reduce, demonstrate the fluorescent quenching effect.Concentration of hydrogen peroxide is the alkalescence relation with the fluorescence intensity of rhodamine S respectively within the specific limits.Set up in view of the above a kind of highly sensitive, selectivity good, the fluorescence method of easy fast measuring trace amount hydrogen peroxide.
Key reaction is as follows:
Concrete detection method comprises the steps:
1. prepare known H
2O
2The test system of concentration pipettes the HAc-NaAc damping fluid in the 5mL test tube with microscale sampler; The H that in the solution of above-mentioned test tube, adds KI, horseradish peroxidase (HRP), known variable concentrations
2O
2, rhodamine S, shake up, be settled to 3ml;
2. the method according to step (1) prepares the reagent blank system, does not add H
2O
2
3. exciting and emitting fluorescence spectrum of the system that obtains with fluorophotometer scanning, at excitation wavelength 500nm, emission wavelength 556nm place, the fluorescence intensity of mensuration system is F and F
0, calculate Δ F=F-F
0Value;
According to Δ F to H
2O
2The concentration relationship workmanship make curve;
5. the method according to step (1) prepares detection architecture, wherein the H of Jia Ruing
2O
2For containing H
2O
2But the measured object of unknown concentration is obtained the Δ F value of measured object;
6. according to the working curve of (4), calculate the contained H of measured object
2O
2Concentration.
Wherein:
The pH value of the described HAc-NaAc damping fluid of step 1 is 3.0-6.0, and volume is 0.05-1.0mL;
KI concentration is 2 * 10
-3Mol/L, volume are 0.1-1mL;
HRP concentration is 0.2 μ g/mL, and volume is 0.02-0.2mL;
Rhodamine S concentration is 2 * 10
-5Mol/L, volume are 0.1-0.8mL.
The invention has the advantages that:
(1) reaction is at room temperature carried out, and the reaction conditions gentleness is beneficial to and popularizes;
(2) equipment is simple, simple to operate;
(3) reagent be easy to get, inexpensive;
(4) highly sensitive, quick, using value is preferably arranged.
Description of drawings:
Fig. 1 is the exciting light spectrogram that horseradish peroxidase enzyme catalytic hydrogen peroxide and iodide ion and rhodamine S associate in the embodiment of the present invention,
Wherein curve a to d represents:
a.HAc-NaAc(pH＝4.4)-3.33×10
-5mol/LKI-4×10
-3μg/mL?HRP-2×10
-5mol/LRh-S；
b.a-1.44×10
-6mol/L?H
2O
2；
c.a-2.52×10
-6mol/L?H
2O
2；
d.a-3.6×10
-6mol/L?H
2O
2。
Fig. 2 is the fluorescence spectrum figure that horseradish peroxidase enzyme catalytic hydrogen peroxide and iodide ion and rhodamine S associate in the embodiment of the present invention;
Wherein curve a to d represents: a.HAc-NaAc (pH=4.4)-3.33 * 10
-5Mol/L KI-4 * 10
-3μ g/mL HRP-2 * 10
-5Mol/L Rh-S, b.a-1.44 * 10
-6Mol/L H
2O
2, c.a-2.52 * 10
-6Mol/LH
2O
2, d.a-3.6 * 10
-6Mol/L H
2O
2
Embodiment:
Following embodiment has been described in weakly acidic HAC-NaAC damping fluid and H
2O
2Exist down, but HRP catalysis H
2O
2Generate OH, the oxidable I of OH generates I
2, excessive I and I
2Can be in conjunction with forming I
3, and RhS can with I
3In conjunction with forming (RhS-I
3)
nThe association particulate causes the fluorescent value of RhS system to descend.In view of the above, set up the fluorescence new method of indirect determination hydrogen peroxide.Its key reaction as previously mentioned.
A kind of rhodamine S association complex microparticles enzyme catalysis fluorescent method that detects trace amount hydrogen peroxide is described below:
(1) with microscale sampler pipette 0.08mL pH value be 4.4 HAc-NaAc damping fluid in the 5ml test tube, adding concentration in the solution of test tube respectively is 2 * 10
-3The KI0.5ml of mol/L, adding concentration again is the HRP 0.1ml of 0.2 μ g/mL, adds the H of known variable concentrations again
2O
20.1ml adding concentration again is 2 * 10
-5The rhodamine S 0.4ml of mol/L shakes up and is settled to 3ml, makes tested systems.
(2) method with step (1) prepares the reagent blank system;
(3) system that above-mentioned each solution is obtained with Cary Eclipse fluorospectrophotometer (U.S. Varian company) scanning excites and emitting fluorescence spectrum, at excitation wavelength 500nm place and emission wavelength is wavelength of fluorescence 556nm place, measures the fluorescence intensity F of system and the F of reagent blank
0, calculate Δ F=F-F
0Value;
(4) the Δ F and the H of the measured object of the known variable concentrations of foundation
2O
2The concentration relationship workmanship be Δ F=7.1444c+8.2037 as curve, c-H
2O
2Concentration, unit is 10
-7Mol/L, the range of linearity is 3.6 * 10
-8Mol/L to 3.6 * 10
-6Mol/L;
(5) set by step the method for (1) prepares detection architecture, and what wherein add is to contain H
2O
2But the measured object of unknown concentration, according to the method for step (3), the Δ F that records the measured object of certain unknown concentration is 52;
(6), calculate the H of the measured object of unknown concentration according to working curve
2O
2Concentration is 6.13 * 10
-7Mol/L.
Claims (1)
1. detect the rhodamine S association complex microparticles enzyme catalysis fluorescent method of trace amount hydrogen peroxide, it is characterized in that: assay method comprises that step is as follows:
(1) the known H of preparation
2O
2The test system of concentration pipettes the HAc-NaAc damping fluid in test tube with microscale sampler, adds the H of KI, horseradish peroxidase HRP, known variable concentrations in the solution of above-mentioned test tube
2O
2, rhodamine S, shake up, be settled to 3ml, the pH value of described HAc-NaAc damping fluid is 3.0-6.0, volume is 0.05-1.0mL; Described KI concentration is 2 * 10
-3Mol/L, volume are 0.1-1mL; Described HRP concentration is 0.2 μ g/mL, and volume is 0.02-0.2mL; Described rhodamine S concentration is 2 * 10
-5Mol/L, volume are 0.1-0.8mL;
(2) method according to step (1) prepares the reagent blank system, does not add H
2O
2
(3) exciting and emitting fluorescence spectrum of the system that obtains with fluorophotometer scanning, at excitation wavelength 500nm, emission wavelength 556nm place, the fluorescence intensity of mensuration system is the F of F and reagent blank
0, calculate △ F=F-F
0Value;
(4) according to △ F to H
2O
2The concentration relationship workmanship make curve;
(5) method according to step (1) prepares detection architecture, wherein the H of Jia Ruing
2O
2For containing H
2O
2But the measured object of unknown concentration is obtained the △ F value of measured object;
(6), calculate the contained H of measured object according to the typical curve of (4)
2O
2Concentration.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2007100506032A CN100514040C (en) | 2007-11-19 | 2007-11-19 | Rhodamine S association complex microparticles enzyme catalysis fluorescent method for detecting trace amount hydrogen peroxide |
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|---|---|---|---|
| CNB2007100506032A CN100514040C (en) | 2007-11-19 | 2007-11-19 | Rhodamine S association complex microparticles enzyme catalysis fluorescent method for detecting trace amount hydrogen peroxide |
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|---|---|
| CN101158642A CN101158642A (en) | 2008-04-09 |
| CN100514040C true CN100514040C (en) | 2009-07-15 |
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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102590164A (en) * | 2012-02-03 | 2012-07-18 | 桂林理工大学 | Method for determining hydrogen peroxide content |
| CN104330391A (en) * | 2014-11-04 | 2015-02-04 | 福建医科大学 | Hydrogen peroxide measurement method based on N-acetyl-L-cysteine-gold nanoclusters |
| CN105424659B (en) * | 2015-10-30 | 2019-07-05 | 重庆医科大学附属永川医院 | Utilize graphene oxide-rhodamine-uricase mixed solution detection uric acid method and the detection architecture |
| CN114397284A (en) * | 2022-01-20 | 2022-04-26 | 甘肃医学院 | Method for determining activity of serum catalase based on butyl rhodamine B fluorescence quenching method |
| CN114560839B (en) * | 2022-03-26 | 2023-09-05 | 甘肃医学院 | Enhanced fluorescent probe and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1160201A (en) * | 1996-03-18 | 1997-09-24 | 株式会社京都第一科学 | Method of and apparatus for determining hydrogen peroxide by Raman schattering |
| KR20020069834A (en) * | 2001-02-28 | 2002-09-05 | 광주과학기술원 | Methodology development of peroxy/superoxide radical determination |
| CN1844124A (en) * | 2006-04-11 | 2006-10-11 | 山东师范大学 | Fluorescent probe for detecting hydrogen peroxide and its synthesis method and use |
| CN101004387A (en) * | 2007-01-11 | 2007-07-25 | 山东新华医疗器械股份有限公司 | Method for detecting residual hydrogen dioxide on surface of sterilized object |
-
2007
- 2007-11-19 CN CNB2007100506032A patent/CN100514040C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1160201A (en) * | 1996-03-18 | 1997-09-24 | 株式会社京都第一科学 | Method of and apparatus for determining hydrogen peroxide by Raman schattering |
| KR20020069834A (en) * | 2001-02-28 | 2002-09-05 | 광주과학기술원 | Methodology development of peroxy/superoxide radical determination |
| CN1844124A (en) * | 2006-04-11 | 2006-10-11 | 山东师范大学 | Fluorescent probe for detecting hydrogen peroxide and its synthesis method and use |
| CN101004387A (en) * | 2007-01-11 | 2007-07-25 | 山东新华医疗器械股份有限公司 | Method for detecting residual hydrogen dioxide on surface of sterilized object |
Non-Patent Citations (6)
| Title |
|---|
| A simple and sensitive resonance scattering spectral methodfordetermination of hydroxyl radical in Fenton system usingrhodamine S and its application to screening the antioxidant. Ai-Hui Liang, Su-Mei Zhou et al.Science Direct Talanta,Vol.70 No.2. 2006 |
| A simple and sensitive resonance scattering spectral methodfordetermination of hydroxyl radical in Fenton system usingrhodamine S and its application to screening the antioxidant. Ai-Hui Liang, Su-Mei Zhou et al.Science Direct Talanta,Vol.70 No.2. 2006 * |
| 一个灵敏测定过氧化氢的吖啶红共振散射光谱新方法. 梁爱惠，蒋治良等.光谱学与光谱分析，第27卷第1期. 2007 |
| 辣根过氧化物酶分光光度法测定黄嘌呤氧化酶的活性. 李忠琴，许小平等.分析化学研究简报，第34卷第6期. 2006 |
| 辣根过氧化物酶分光光度法测定黄嘌呤氧化酶的活性. 李忠琴，许小平等.分析化学研究简报，第34卷第6期. 2006 * |
| 银纳米微粒荧光猝灭法测定水中痕量ClO2. 康彩艳，陈媛媛等.光谱学与光谱分析，第26卷第6期. 2006 |
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