CN105928918B - Application of the salicylhydroxamic acid in detecting hypochlorite - Google Patents
Application of the salicylhydroxamic acid in detecting hypochlorite Download PDFInfo
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- CN105928918B CN105928918B CN201610260595.3A CN201610260595A CN105928918B CN 105928918 B CN105928918 B CN 105928918B CN 201610260595 A CN201610260595 A CN 201610260595A CN 105928918 B CN105928918 B CN 105928918B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6428—Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes"
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Abstract
The present invention provides application of the salicylhydroxamic acid in detecting hypochlorite, the method for specifically providing fluoroscopic examination hypochlorite.The detection method is to be based on salicylhydroxamic acid (salicylhydroxamic acid), hypochlorous content is quantitatively detected in the HEPES solution that pH is 7.4, the detection method, high sensitivity and selectivity are shown to hypochlorite, detection reagent is cheap, detection process is easy, sensitive, quick, and testing result is accurate.
Description
Technical field
The present invention relates to hypochlorous acid to test and analyze technology, and in particular to salicylhydroxamic acid (Salicylhydroxamic
Acid) the application in detecting hypochlorite.
Background technology
The concentration range of sodium hypochlorite is from 10-5To 10-2Mol/L is widely used as in bleaching agent and disinfectant.However, high
There is greatly harm to the health of human and animal in the sodium hypochlorite of concentration.On the other hand, hypochlorite ion is biology weight
One of active oxygen (ROS) species wanted, it plays key player in immune system.Endogenous CLO-Be life base substance and
With anti-microbial property.However, the exception of hypochlorite can lead to tissue damage and disease, for example, hepatic ischemia-reperfusion damages
Wound, atherosclerosis, injury of lungs, rheumatoid arthritis, angiocardiopathy, neuronal degeneration, arthritis and cancer.Cause
This hypochlorous detection in environmental and biological samples is most important.However, there are process complexity, detections for most of detection methods
Reagent need organic building-up process, it is of high cost the problems such as.
Invention content
The application and fluoroscopic examination hypochlorous acid that the object of the present invention is to provide salicylhydroxamic acids in detecting hypochlorite
The method of root.The present invention should use commercially available and cheap reagent, realize hypochlorous quick detection by Fluorescence Spectrometer, also may be used
For cell imaging.
The present invention provides application of the salicylhydroxamic acid in detecting hypochlorite.
The present invention provides a kind of method of fluoroscopic examination hypochlorite, and step is：
(1), the salicylhydroxamic acid methanol solution of 2mM is prepared；
(2), by volume 800:Methanol solution and salicylhydroxamic acid methanol solution are added to clean fluorescence cuvette by 1
In, it is detected on Fluorescence spectrophotometer, with the addition for waiting for test sample, the fluorescence intensity of 415nm gradually increases；
(3), the NaClO solution that 2mM is prepared with distilled water, the salicylhydroxamic acid first of the methanol solution and 2.5 μ L of 2mL
Alcoholic solution is added in fluorescence cuvette, be gradually added into NaClO solution volume be 0.17,0.35,0.50,0.60,0.70,
0.95,1.0,1.25,1.36,1.54,1.64uL, while the corresponding fluorescence intensity F for measuring in Fluorescence Spectrometer 415nm is
65,156,233,284,325,457,496,589,650,736,787, with a concentration of abscissas of NaClO, with relative intensity of fluorescence
F-F0It draws and schemes for ordinate, F0﹦ 24 obtains the working curve of NaClO concentration；Equation of linear regression is：F-F0=
The unit of 489.232c-39.2948, c are 10-6mol/L；
(4), methanol solution 2000uL and salicylhydroxamic acid methanol solution 2.5uL are added to clean fluorescence cuvette
In, V ul testing sample solutions are drawn with microsyringe, are added in this clean fluorescence cuvette, in fluorescence spectrophotometry
It is detected on instrument, the fluorescence intensity measured is substituted into the equation of linear regression of (3), obtains concentration c, sample to be tested CWait for test sample=2000uL
×c×10-6/ VuL can acquire the concentration of NaClO.
The experiment proved that other analytes not measurement of the interference system to NaClO.
Compared with prior art, the invention has the advantages that and effect：The present invention uses commercially available and cheap reagent, leads to
Fluorescence Spectrometer is crossed to can be realized to hypochlorous quick detection and cell imaging.The detection method, shows hypochlorite
High sensitivity and selectivity, detection reagent is cheap, detection process is easy, sensitive, quick, and testing result is accurate.
Description of the drawings：
The fluorescent emission figure of 1 salicylhydroxamic acid of Fig. 1 embodiments and NaClO effects
The fluorescence block diagram of 2 salicylhydroxamic acid of Fig. 2 embodiments and various analytes
3 working curve of Fig. 3 embodiments
4 cell imaging figure of Fig. 4 embodiments
Specific implementation mode：
Embodiment 1
The salicylhydroxamic acid solution of 2mM is prepared with methanol；The methanol solution of 2mL and the salicylhydroxamic acid of 2.5 μ L
Methanol solution is added in clean fluorescence cuvette, takes the solution of NaClO, is gradually added in this cuvette with microsyringe,
It is detected on Fluorescence spectrophotometer in sample-adding, with the addition of NaClO, fluorescence intensity gradually increases at 415nm.Fluorescence is sent out
It penetrates figure and sees Fig. 1.
Embodiment 2
The NaClO solution of 2mM is prepared with methanol；In 12 fluorescence cuvettes, each methanol solution and 2.5 μ that 2mL is added
The salicylhydroxamic acid methanol solution of L, then be separately added into 1 molar equivalent NaClO and other various points of 50 molar equivalents
Analyse object (ClO-,H2O2,1O2,·O2 -, ROO, NO, OH), it is detected on Fluorescence spectrophotometer, draws different analytes
The block diagram of corresponding 415nm fluorescence intensities obtains fluorescent emission figure and sees that (NaClO makes the fluorescence of salicylhydroxamic acid strong to Fig. 2
Degree changes to 787 or so by 24, and other analytes do not cause the variation of salicylhydroxamic acid fluorescence intensity substantially).
The experiment proved that other analytes not measurement of the interference system to NaClO.
Embodiment 3
The NaClO solution of 2mM is prepared with distilled water, the methanol solution of 2mL and the salicylhydroxamic acid methanol of 2.5 μ L is molten
Liquid is added in fluorescence cuvette, be gradually added into NaClO solution volume be 0.17,0.35,0.50,0.60,0.70,0.95,
1.0,1.25,1.36,1.54,1.64uL, at the same the corresponding fluorescence intensity F for measuring in Fluorescence Spectrometer 415nm be 65,
156,233,284,325,457,496,589,650,736,787, with a concentration of abscissas of NaClO, with relative intensity of fluorescence F-
F0It draws and schemes for ordinate, F0﹦ 24 obtains the working curve of NaClO concentration；Equation of linear regression is：F-F0=489.232c-
39.2948, c unit is 10-6mol/L；See Fig. 3.
Embodiment 4
The salicylhydroxamic acid methanol solution of 2mM is prepared with methanol；It is thin that liver cancer is added in salicylhydroxamic acid methanol solution
In born of the same parents' culture solution so that its a concentration of 2.5 μM, with hepatocellular carcinoma H22 at 37 DEG C, react 30min, system is copolymerized in fluorescence
Unstressed configuration under burnt imager；Salicylhydroxamic acid methanol solution is added in liver cancer cells culture solution so that its a concentration of 2.5 μ
M reacts 30min, adds the NaClO of external source, make its a concentration of 5 μM with hepatocellular carcinoma H22 at 37 DEG C, at 37 DEG C,
30min is reacted again, and system is displayed in blue fluorescent emission under fluorescence co-focusing imager；It is introduced into the salicyl hydroxyl oxime of cell
Acid is acted on the NaClO for subsequently entering cell, makes its Fluorescence Increasing.Fig. 4 be fluorescence co-focusing imager under with salicyl hydroxyl oxime
What cell a (unstressed configuration) and its light field c and the effect of first and salicylhydroxamic acid after sour effect were acted on the NaClO of external source again
Cell b (blue-fluorescence) images and its light field d.
Claims (2)
1. application of the salicylhydroxamic acid in detecting hypochlorite, the environment and method of the application are by salicylhydroxamic acid
Methanol solution is added in detection reagent, passes through the variation detection wherein hypochlorite content of fluorescence intensity.
2. a kind of method of fluoroscopic examination hypochlorite, which is characterized in that step is：
(1), the salicylhydroxamic acid methanol solution of 2mM is prepared；
(2), by volume 800:1 is added to methanol solution and salicylhydroxamic acid methanol solution in clean fluorescence cuvette,
It is detected on Fluorescence spectrophotometer, with the addition for waiting for test sample, the fluorescence intensity of 415nm gradually increases；
(3), the NaClO solution of 2mM is prepared with distilled water, the methanol solution of 2mL and the salicylhydroxamic acid methanol of 2.5 μ L is molten
Liquid is added in fluorescence cuvette, be gradually added into NaClO solution volume be 0.17,0.35,0.50,0.60,0.70,0.95,
1.0,1.25,1.36,1.54,1.64uL, at the same the corresponding fluorescence intensity F for measuring in Fluorescence Spectrometer 415nm be 65,
156,233,284,325,457,496,589,650,736,787, with a concentration of abscissas of NaClO, with relative intensity of fluorescence F-
F0It draws and schemes for ordinate, F0﹦ 24 obtains the working curve of NaClO concentration；Equation of linear regression is：F-F0=489.232c-
39.2948, c unit is 10-6mol/L；
(4), methanol solution 2000uL and salicylhydroxamic acid methanol solution 2.5uL are added in clean fluorescence cuvette, are used
Microsyringe draws V ul testing sample solutions, is added in this clean fluorescence cuvette, on Fluorescence spectrophotometer
The fluorescence intensity measured is substituted into the equation of linear regression of step (3), obtains concentration c, sample to be tested C by detectionWait for test sample=2000uL
×c×10-6/ VuL can acquire the concentration of NaClO.
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| CN201610260595.3A CN105928918B (en) | 2016-04-25 | 2016-04-25 | Application of the salicylhydroxamic acid in detecting hypochlorite |
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| CN201610260595.3A CN105928918B (en) | 2016-04-25 | 2016-04-25 | Application of the salicylhydroxamic acid in detecting hypochlorite |
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| CN108169197B (en) * | 2017-12-19 | 2021-01-22 | 河南农业大学 | Method for detecting hypochlorite through near infrared |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103149184A (en) * | 2013-02-04 | 2013-06-12 | 山西大学 | Fluorescence detection method for hypochlorite |
| CN103926230A (en) * | 2014-05-08 | 2014-07-16 | 山西大学 | Application of curcumin to detection of hypochlorous acid |
| CN105203514A (en) * | 2015-09-23 | 2015-12-30 | 山西大学 | Reagent and method for fluorescence detection of hypochlorous acid (HClO) |
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2016
- 2016-04-25 CN CN201610260595.3A patent/CN105928918B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103149184A (en) * | 2013-02-04 | 2013-06-12 | 山西大学 | Fluorescence detection method for hypochlorite |
| CN103926230A (en) * | 2014-05-08 | 2014-07-16 | 山西大学 | Application of curcumin to detection of hypochlorous acid |
| CN105203514A (en) * | 2015-09-23 | 2015-12-30 | 山西大学 | Reagent and method for fluorescence detection of hypochlorous acid (HClO) |
Non-Patent Citations (2)
| Title |
|---|
| A highly specific fluorescent probe for hypochlorite based on fluorescein derivative and its endogenous imaging in living cells;BihuiWang et. al.;《Dyes and Pigments》;20150328;22-29 * |
| A reversible fluorescent probe for detecting hypochloric acid in living cells and animals: utilizing a novel strategy for effectively modulating the fluorescence of selenide and selenoxide;Zhangrong Lou et. al.;《Chem. Comm.》;20131231;2445-2447 * |
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