CN111830019A - Method for detecting superoxide anion free radical generated by photosensitive organic dye - Google Patents
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Abstract
The invention discloses a method for detecting superoxide anion free radicals generated by photosensitive organic dye under the irradiation of visible light by taking hydroxylamine hydrochloride as a probe. A photosensitive organic dye containing hydroxylamine hydrochloride is irradiated under visible light, and then a ferric ion solution is added to the sample solution. The remaining hydroxylamine hydrochloride after reaction with the superoxide anion radical reduces the ferric ions in solution to ferrous ions. Then, adding 1, 10-phenanthroline solution, performing spectrophotometry measurement at 510nm, and indirectly indicating the residual amount of hydroxylamine hydrochloride by using an absorbance value at 510 nm. The production of superoxide anion radicals can be indirectly expressed as a reduction in hydroxylamine hydrochloride. Under the best experimental conditions, the linear range is 0.0-1.6X 10‑5M, the detection limit and the quantification limit of the method are respectively 8.23 multiplied by 10‑7And 2.49X 10‑6M，R2Is 0.9995. The method is simple and feasible, and can be used for stably measuring superoxide anion free radicals generated by photosensitive organic dyes under the irradiation of visible light.
Description
Technical Field
The invention belongs to the technical field of detection and analysis, and particularly relates to a simple, convenient and sensitive method for detecting superoxide anion radicals generated by photosensitive organic dye under the irradiation of visible light.
Background
The generation of reactive oxygen species by light is the primary mechanism for various chemical effects in photochemical processes. Free radical, in particular superoxide anion free radical (O)2·-) Has been widely regarded and applied in photochemical processes. Therefore, there is a need for a reliable and fast detection of O in an illumination system2·-The method of (1). Is commonly used for detecting O2·-The methods of (2) are Electron Spin Resonance (ESR), High Performance Liquid Chromatography (HPLC) and Chemiluminescence (CL). The above-described method has a number of disadvantages: for example, high-end instruments are used, which cannot be achieved in a laboratory, and the operation is complicated, and various instruments and medicines are used. Compared with the method, the spectrophotometry is the fastest and most convenient method, is also applicable to trace samples, and has the advantages of low cost and easy miniaturization. Nitro Blue Tetrazole (NBT) and 2- (4-iodophenyl) -3- (4-nitrophenyl) -5- (2, 4-disulfophenyl) -2H-tetrazole sodium salt (WST-1) are used for determining O2·-The most commonly used spectrophotometric probes. NBT by O2·-The formazan is oxidized into NBT formazan, and the formazan can be measured at 560nm by a spectrophotometry. However, NBT is unstable under visible light irradiation and NBT formazan is insoluble in water, which makes NBT unusable in aqueous solutions. In addition, another primary probe WST-1 was coupled with O2·-The WST-1 formazan can be generated by reaction, and the maximum absorbance is achieved at 438 nm. WST-1 formazan has high water solubility, so WST-1 is widely applied to various biological and chemical experiments. Nevertheless, WST-1 is very expensive and the optimal pH for the process is 8.0, which makes the process unsuitable for use under acidic conditions.
NH2OH & HCl can also be used as O2·-The spectrophotometric probe of (1). NH (NH)2OH & HCl by O2·-Oxidation to NO2 -。
NH2OH+2O2·-+H+→NO2 -+H2O2+H2O (1)
Conventional methods utilize NO2 -Diazotizing with sulfanilic acid to obtain diazonium salt, reacting with N-1-naphthyl ethylenediamine hydrochloride to obtain red azo dye, measuring at 540nm with spectrophotometer, and indirectly representing O by variation of absorbance value2·-The amount of production of (c). However, this method is not suitable for O in an illumination system2·-Measured because the photosensitizer can generate hydrated electrons (e) with strong reducibility under illuminationaq -). The hydrated electrons can react with nitrate ions (NO)3 -) And nitrite ion (NO)2 -) Reactions occur, such as reduction reactions (2) and (3). Thereafter, (NO)3·)2-And (NO)2·)2-Further conversion to N2O, finally reduced to N by hydrated electrons2(4)：
NO3 -+eaq -→(NO3·)2-(2)
NO2 -+eaq -→(NO2·)2-(3)
N2O+eaq -→N2+OH·+OH-(4)
Disclosure of Invention
Aiming at the problems, the invention discloses a method for detecting superoxide anion free radicals generated by photosensitive organic dye under the irradiation of visible light by taking hydroxylamine hydrochloride as a probe, which is efficient, convenient and stable to use under the illumination condition.
In an illumination system, NH2OH HCl due to O2·-Is consumed by oxidation, so that only NH is determined2The amount of OH HCl consumed can be based on NH2OH HCl and O2·-Calculating O from the reaction equation of (1)2·-The yield of (a).
In this method, the sample solution is treated by adding an auxiliary metal ion Fe3+To measure NH2OH HCl, residual NH in solution2OH HCl to react Fe3+Reduction to Fe2+. Then, 1,10-Adding the phenanthroline solution into the sample solution. Using 1, 10-phenanthroline in Fe3+/Fe2+1, 10-phenanthroline and Fe as chromogens in ionic colorimetric and spectrophotometric methods2+A water-soluble orange-red metal complex is formed. The resulting orange-red complex can then be measured spectrophotometrically at 510 nm. The absorbance at 510nm may be an indirect indication of NH2OH & HCl content, then with NH2Indirect calculation of O from OH & HCl consumption2·-The amount of production.
Optimizing the illumination time, the solution pH and the NH2After the influence factors such as the concentration of OH & HCl, the concentration of 1, 10-phenanthroline and the like, the interference of various coexisting ions on the method is considered, and the method for accurately and efficiently detecting the superoxide anion free radical generated by the photosensitive organic dye under the irradiation of visible light is obtained.
The technical scheme is as follows:
(1) establishment of a standard curve: configuring NH with different concentrations2OH HCl standard solution, the NH2The pH of the OH HCl standard solution is adjusted by adding a buffer solution, after which the NH is added2Adding Fe into OH & HCl standard solution3+Then adding 1, 10-phenanthroline solution to generate water-soluble orange-red metal complex, and measuring NH with different concentrations at 510nm by spectrophotometry2Orange-red complex obtained from OH & HCl solution to obtain NH2OH & HCl consumption and spectrophotometry DeltaA510A linear relationship therebetween; then according to the equation: NH (NH)2OH+2O2·-+H+→NO2 -+H2O2+H2The stoichiometric ratio of O obtains the concentration and the spectrophotometry delta of the superoxide anion free radicalA510A linear relationship therebetween; the different concentrations of NH2In OH & HCl solution, Fe3+Is added in an amount corresponding to NH2OH & HCl is in excess to ensure NH2OH & HCl can be consumed up;
(2) preparing aqueous solution of photosensitive organic dye, adding buffer solution into the aqueous solution of photosensitive organic dye to adjust pH, and then adding NH2OH & HCl solution to obtain a photosensitive organic dyeTaking out two identical parts of the superoxide anion radical reaction solution from the reaction solution, wherein one part is a reference sample 1, the other part is an experimental sample 1, and FeCl is sequentially added into the experimental sample 13The solution and the 1, 10-phenanthroline solution are developed, and then the reference sample 1 is used as a reference, the absorbance at 510nm is measured and is marked as A0；
(3) Placing the reaction solution of the rest superoxide anion free radicals under a xenon lamp light source for irradiation to generate superoxide anion free radicals O2·-Taking out two parts from the reaction solution after irradiation, wherein one part is a reference sample 2, one part is an experimental sample 2, and FeCl is added into the experimental sample 23After the solution and the 1, 10-phenanthroline solution are developed, taking a reference sample 2 as a reference, determining the absorbance of the experimental sample at 510nm, and recording as At；
(4) Analysis of the signal ΔA510＝A0-AtThe concentration and the degree of spectral distribution Delta of the superoxide anion radicals obtained in step (1)A510The linear relationship between the two groups determines the generation amount of superoxide anion free radicals;
the pH value of the buffer solution added in the step (1) and the step (2) is the same.
Preferably, the buffer solution is H3PO4–Na2HPO4Buffer solution or NaH2PO4–Na2HPO4A buffer solution; said H3PO4–Na2HPO4The pH value of the buffer solution is 2.0-5.0; the NaH2PO4–Na2HPO4The pH of the buffer solution is 6.0-7.0.
Preferably, the photosensitive organic dye is azure i.
Preferably, the step (2) includes the steps of:
step one, preparing 0.0153-0.2295 g of azure I solid into azure I solution with the concentration of 0.1-1.5 mM; taking 0.0035-0.0525 g of NH2OH & HCl solid is prepared into NH with the concentration of 0.1-1.5 mM2OH & HCl solution; taking 0.0081-0.1215 g FeCl3The solid is prepared into FeCl with the concentration of 0.1-1.5 mM3A solution; get0.57-1.72 g of 1, 10-phenanthroline solid is prepared into a 1, 10-phenanthroline solution with the concentration of 5.8-17.4 mM;
step two, taking 2.5mL of the azure I solution prepared in the step one, adding 37.5mL of deionized water, and then adding 5mL of H with pH of 5.03PO4–Na2HPO4Buffer solution, add 5mL of NH prepared in step one2OH & HCl solution, preparing to obtain superoxide anion free radical reaction solution of the photosensitive organic dye; 4.5mL of the reference sample 1 and the experimental sample 1 are taken; 0.5mL of FeCl prepared in the first step is added into the experimental sample 1 respectively3After the solution and 0.3mL of 1, 10-phenanthroline solution prepared in the first step are developed for 15-20 min, a spectrophotometer is used for measuring the absorbance of the sample at 510nm, and the absorbance is marked as A0。
Preferably, the step (3) includes the steps of:
irradiating the reaction solution prepared in the second step for 60-80 min under a xenon lamp light source, taking two 4.5mL samples from the irradiated superoxide anion radical reaction solution, taking one sample as a reference sample 2, taking the other sample as an experimental sample 2, and respectively adding 0.5mL FeCl prepared in the first step into the experimental sample 23After the solution and 0.3mL of 1, 10-phenanthroline solution prepared in the first step are developed for 15-20 min, a spectrophotometer is used for measuring the absorbance of the sample at 510nm, and the absorbance is marked as At。
Preferably, the xenon lamp (PLS-SXE300) is equipped with infrared and ultraviolet filters (broadband filtered infrared 400-1200 nm: diameter 63mm, UV CUT 400: diameter 63mm), the visible light output wavelength is 400nm-780nm, the xenon lamp is placed 10cm above the liquid surface, the diameter of the main spot is 3cm, the energy density is 150mW/cm2。
The invention has the following advantages:
(1) the method has the advantages of simple operation, easy mastering and high sensitivity, and can measure the superoxide anion free radical generated by the photosensitive organic dye under the irradiation of visible light in a short time.
(2) The reagents adopted by the invention are all water-soluble, so that the method is more favorable for measuring O in various water environments under illumination2·-The yield of (2). In addition to this, the present invention is,all reagents of the method are stable under visible light, and O can be measured in real time in a visible light system2·-。
(3) The method has wide practicability, is also suitable for other photosensitive organic wastewater, has good anti-interference capability, and can be applied to the detection of superoxide anion free radicals in acidic wastewater.
Drawings
Fig. 1 is a working curve of the present invention.
FIG. 2 is pH vs. assay O2·-The experimental conditions: [ azure I]＝0.05mM,[NH2OH·HCl]＝0.08mM,[Fe3+]0.08mM, [1, 10-phenanthroline]0.9mM, pH 2.0-7.0, and light irradiation at 25 deg.C for 60 min.
FIG. 3 is a graph of light exposure time versus assay O2·-The experimental conditions: [ azure I]＝0.05mM,[NH2OH·HCl]＝0.08mM,[Fe3+]0.08mM, [1, 10-phenanthroline]0.9mM, pH5.0, light at 25 deg.C for 10-80 min.
FIG. 4 is NH2OH & HCl concentration vs. determination of O2·-The experimental conditions: [ Azurie I ]]＝0.05mM,[NH2OH·HCl]＝0.02–0.10mM,[Fe3+]0.02-0.10 mM, [1, 10-phenanthroline]0.9mM, pH5.0, 60min at 25 ℃.
FIG. 5 is a graph of 1, 10-phenanthroline concentration versus assay O2·-The experimental conditions: [ Azurie I ]]＝0.05mM,[NH2OH·HCl]＝0.08mM,[Fe3+]0.08mM, [1, 10-phenanthroline]0.3-1.5 mM, pH5.0, and light at 25 deg.C for 60 min.
FIG. 6 shows the detection process of the superoxide anion radical of the present invention.
Detailed Description
Example 1
(1) Creation of the standard curve: 10 50mL volumetric flasks were pipetted with 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0mL of 0.1mM NH2OH HCl solution, said NH2OH & HCl solution was added by adding 5mLH3PO4–Na2HPO4The buffer solution adjusts the pH to 5Then, separately to the NH20.6, 1.1, 1.6, 2.1, 2.6, 3.1, 3.6, 4.1, 4.6, 5.1mL of Fe with a concentration of 0.1mM was added to the OH & HCl solution3+Solution, at which point the following reaction takes place:
2Fe3++2NH2OH·HCl＝2Fe2++N2+4H++2H2O+2Cl-(5)
then respectively adding 2.6mL of 17.4mM 1, 10-phenanthroline solution, and diluting to 50mL of 1, 10-phenanthroline and Fe with deionized water to constant volume2+A water-soluble orange-red metal complex is formed. After development for 15min, the NH concentrations at 510nm were measured spectrophotometrically2Orange-red complex obtained from OH & HCl solution to obtain NH2OH & HCl consumption and spectrophotometry DeltaA510The linear relationship between: delta A510＝8.0500C(NH2OH) + 0.0002; then according to the equation: NH (NH)2OH+2O2·-+H+→NO2 -+H2O2+H2The stoichiometric ratio of O obtains the concentration and the spectrophotometry delta of the superoxide anion free radicalA510The linear relationship between: delta A510＝4.0250C(O2·-) +0.0002(C units are mM). The linear relationship indicates that as the concentration of superoxide anion radicals increases, NH2Increased consumption of OH & HCl,. DELTA.A510The value of (A) increases with the linear range of the superoxide anion radical concentration from 0.0 to 1.6X 10-5M, the detection limit and the quantification limit of the method are respectively 8.23 multiplied by 10-7And 2.49X 10-6M，R2Is 0.9995.
(2) Take 7mL, 85% H3PO4Putting the solution into a 500mL volumetric flask, adding deionized water to constant volume, and preparing into H with the concentration of 0.2M3PO4Solution (solution a); collecting 35.8g of Na2HPO4·12H2Putting the O solid in a 500mL volumetric flask, using deionized water to fix the volume, and preparing Na with the concentration of 0.2M2HPO4Solution (solution B); 13.8g of NaH was taken2PO4·H2Placing the O solid in a 500mL volumetric flask, using deionized water to fix the volume, and preparing NaH with the concentration of 0.2M2PO4Solutions of(solution C), then H at pH 2.0, 3.0, 4.0, 5.0 was prepared as in Table 13PO4–Na2HPO4Buffer solution, NaH at pH 6.0, 7.0 prepared according to Table 22PO4–Na2HPO4And (4) buffer solution.
TABLE 1
TABLE 2
(3) Taking 0.153g of azure I solid in a 500mL volumetric flask, and preparing into 1mM azure I solution by using deionized water for constant volume.
(4) Take 0.028g NH2OH & HCl solid is put into a 500mL volumetric flask, the volume is determined by deionized water, and NH with the concentration of 0.8mM is prepared2OH & HCl solution; 0.065g of FeCl was taken3Putting the solid into a 500mL volumetric flask, diluting to constant volume with deionized water, and preparing FeCl with the concentration of 0.8mM3A solution; 1.72g of 1, 10-phenanthroline solid is taken and placed in a 500mL volumetric flask, the volume is determined by deionized water, and a 1, 10-phenanthroline solution with the concentration of 17.4mM is prepared.
(5) Taking 2.5mL of the azure I solution prepared in the step (3), adding 37.5mL of deionized water, and then adding 5mL of H with pH of 5.0 prepared in the step (2)3PO4–Na2HPO4Adding 5mL of NH prepared in the step (4) into the buffer solution2OH & HCl solution to prepare the superoxide anion free radical reaction solution with photosensitive organic dye azure I concentration of 0.05 mM.
(6) Taking two 4.5mL samples from the solution prepared in the step (5), one sample is used as a reference sample 1, the other sample is used as an experimental sample 1, and 0.5mL FeCl prepared in the step (4) is respectively added into the experimental sample 13And (3) after the solution and 0.3mL of the 1, 10-phenanthroline solution prepared in the step (4) are developed for 15min, taking the reference sample 1 as a reference, and measuring the absorbance of the sample at 510nm by using a spectrophotometer to be recorded as A0＝0.595。
(7) Placing the superoxide anion reaction solution prepared in the step (5) under a xenon lamp light source for irradiating for 60min, irradiating 0.05mM azure I solution through a xenon lamp to generate superoxide anion free radicals, wherein the xenon lamp (PLS-SXE300) is provided with an infrared and ultraviolet filter (broadband filtering infrared 400 + 1200 nm: diameter 63mM, UV CUT 400: diameter 63mM), the visible light output wavelength is 400nm-780nm, the xenon lamp is placed at 10cm above the liquid level, the diameter of a main light spot is 3cm, and the energy density is 150mW/cm2。
(8) After 60min of irradiation, two 4.5mL samples were taken from the irradiated superoxide anion radical reaction solution of step (7), one was used as a reference sample 2, and the other was used as an experimental sample 2, and 0.5mL of FeCl prepared in step (4) was added to the experimental sample 23And (3) after the solution and 0.3mL of the 1, 10-phenanthroline solution prepared in the step (4) are developed for 15min, taking a reference sample 2 as a reference, and measuring the absorbance of the sample at 510nm by using a spectrophotometer to be recorded as At＝0.542。
(9) Analyzing the signal
The concentration and the spectrophotometry delta of the superoxide anion free radicals obtained in the step (1)A510The linear relationship between the two radicals obtains the generation amount of superoxide anion free radicals under the illumination condition, and the generation amount of superoxide anion free radicals is 0.013 mM;
example 2
The detection procedure was the same as in example 1, except that: the pH of the buffer solution was 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, respectively. The results shown in FIG. 2 were obtained. Separately adding H with pH of 2.0-5.03PO4–Na2HPO4Buffer solution and NaH with pH of 6.0-7.02PO4–Na2HPO4And (4) buffering the solution, adjusting the pH value of the test solution, and analyzing according to an analysis program. At pH5.0 there is a maximum signal (. DELTA.)A510) Is 0.053. Therefore, pH5.0 was selected as the optimum pH.
Example 3
The detection procedure was the same as in example 1, except that: the illumination time is 10, 20, 30, 40, 50, 60, 70 and 80min respectively. To obtainTo the results shown in fig. 3. Xenon lamp exposure time pair O2·-The effect of the yield indicates that within 0-60min, the signal value (. DELTA.A510) And the linear relation with the irradiation time of the xenon lamp. Therefore, the irradiation time was selected to be 60 min.
Example 4
The detection procedure was the same as in example 1, except that: NH (NH)2OH HCl and Fe3+The concentrations were 0.02, 0.04, 0.06, 0.08, 0.10mM, respectively. The results shown in FIG. 4 were obtained. NH at various concentrations2OH & HCl reaction Signal value (. DELTA.)A510) The time curve shows that the oxidation reaction occurs rapidly and then gradually decreases over time. 0.08 and 0.10mM NH2OH HCl gave similar results. Therefore, an initial concentration of 0.08mM NH was selected in subsequent experiments2OH & HCl was used as the optimum initial concentration for the reaction. According to reaction equation (5), Fe in the experiment3+Is equal to NH2Concentration of OH & HCl.
2Fe3++2NH2OH·HCl＝2Fe2++N2+4H++2H2O+2Cl-(5)
Example 5
The detection procedure was the same as in example 1, except that: the concentrations of 1, 10-phenanthroline are respectively 0.3, 0.6, 0.9, 1.2 and 1.5 mM. The results shown in FIG. 5 were obtained. The results showed that there was a maximum signal value (Δ) with 0.9mM 1, 10-phenanthrolineA510) Was 0.054.
Example 6
The detection procedure was the same as in example 1, except that in step (8): after 60min of irradiation, three samples of 4.0mL were taken from the irradiated superoxide anion radical reaction solution of step (7), one as reference, one as run 1 and the other as run 2. Adding 0.5mL of deionized water into the experimental group 1, and then respectively adding 0.5mL of FeCl prepared in the step (4)3And (3) after the solution and 0.3mL of the 1, 10-phenanthroline solution prepared in the step (4) are developed for 15min, measuring the absorbance of the sample at 510nm by using a spectrophotometer, and recording the absorbance as At1. Adding 0.5mL of interference ion solution into the experimental group 2, shaking up, and then respectively adding 0.5mL of FeCl prepared in the step (4)3And (3) after the solution and 0.3mL of the 1, 10-phenanthroline solution prepared in the step (4) are developed for 15min, measuring the absorbance of the sample at 510nm by using a spectrophotometer, and recording the absorbance as At2. Analysis signal is DeltaA510(1)＝A0-At1And ΔA510(2)＝A0-At2. 20 coexisting ions were systematically detected as ΔA510(1)And ΔA510(2)Less than ± 5% of the error between (n ═ 3) is undisturbed. As shown in Table 3, the concentration of the ions in the table is within the range shown in the table and does not interfere with the present invention (mg. L)-1)。
TABLE 3
Claims (5)
1. A method for detecting superoxide anion radicals generated by a photosensitive organic dye, said method comprising:
(1) establishment of a standard curve: configuring NH with different concentrations2OH & HCl standard solutions, and sequentially adding buffer solution and Fe into each standard solution3+And 1, 10-phenanthroline solution to generate water-soluble orange-red metal complex, measuring the absorbance of each standard solution at 510nm by using a spectrophotometry, and calculating to obtain NH2OH & HCl consumption and spectrophotometry DeltaA510A linear relationship therebetween; then according to the equation: NH (NH)2OH+2O2·-+H+→NO2 -+H2O2+H2Obtaining the concentration and the spectrophotometry delta of the superoxide anion free radical by the stoichiometric ratio in OA510A linear relationship therebetween;
(2) preparing aqueous solution of photosensitive organic dye, and sequentially adding the buffer solution and NH into the aqueous solution2OH & HCl solution to obtain a reaction solution, and taking out two identical parts from the reaction solution, wherein one part is a reference sample1, one part is an experimental sample 1, and FeCl is added into the experimental sample 1 in sequence3After the solution and the 1, 10-phenanthroline solution are developed, taking a reference sample 1 as a reference, determining the absorbance of the experimental sample 1 at 510nm, and recording as A0；
(3) Irradiating the rest reaction solution under xenon lamp to generate superoxide anion free radical O2·-Taking out two identical parts from the reaction solution after irradiation, wherein one part is a reference sample 2, the other part is an experimental sample 2, and FeCl is sequentially added into the experimental sample 23After the solution and the 1, 10-phenanthroline solution are developed, taking a reference sample 2 as a reference, determining the absorbance of the experimental sample 2 at 510nm, and recording as At；
(4) Analysis of the signal ΔA510＝A0-AtThe concentration and the spectrophotometry delta of the superoxide anion free radicals obtained in the step (1)A510The linear relationship between them determines the amount of superoxide anion radicals generated.
2. The method of claim 1, wherein the buffer solution is H3PO4–Na2HPO4Buffer solution or NaH2PO4–Na2HPO4A buffer solution; said H3PO4–Na2HPO4The pH value of the buffer solution is 2.0-5.0; the NaH2PO4–Na2HPO4The pH of the buffer solution is 6.0-7.0.
3. The method of claim 1, wherein the photosensitive organic dye is azure i.
4. The method for detecting the generation of superoxide anion radicals from photosensitive organic dyes of claim 3, wherein the step (2) comprises the steps of:
step one, 0.0153-0.2295 g of azure I solid is prepared into the solid with the concentration of 0.1-1.5 mMAzure I solution; taking 0.0035-0.0525 g of NH2OH & HCl solid is prepared into NH with the concentration of 0.1-1.5 mM2OH & HCl solution; taking 0.0081-0.1215 g FeCl3The solid is prepared into FeCl with the concentration of 0.1-1.5 mM3A solution; preparing 0.57-1.72 g of 1, 10-phenanthroline solid into a 1, 10-phenanthroline solution with the concentration of 5.8-17.4 mM;
step two, taking 2.5mL of the azure I solution prepared in the step one, adding 37.5mL of deionized water, and then adding 5mL of H with pH of 5.03PO4–Na2HPO4Buffer solution, add 5mL of NH prepared in step one2OH & HCl solution, preparing to obtain superoxide anion free radical reaction solution of the photosensitive organic dye; 4.5mL of the reference sample 1 and the experimental sample 1 are taken; 0.5mL of FeCl prepared in the first step is added into the experimental sample 1 respectively3After the solution and 0.3mL of 1, 10-phenanthroline solution prepared in the first step are developed for 15-20 min, a spectrophotometer is used for measuring the absorbance of the sample at 510nm, and the absorbance is marked as A0。
5. The method for detecting the generation of superoxide anion radicals from photosensitive organic dyes of claim 4, wherein the step (3) comprises the steps of:
irradiating the reaction solution prepared in the second step in claim 4 under a xenon lamp light source for 60-80 min, then taking two 4.5mL samples from the irradiated superoxide anion radical reaction solution, wherein one sample is used as a reference sample 2, the other sample is used as an experimental sample 2, and 0.5mL FeCl prepared in the first step is added into the experimental sample 2 respectively3After the solution and 0.3mL of 1, 10-phenanthroline solution prepared in the first step are developed for 15-20 min, a spectrophotometer is used for measuring the absorbance of the sample at 510nm, and the absorbance is marked as At。
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Application publication date: 20201027 |