CN112697735A - Glutathione detection method and related detection kit - Google Patents
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Abstract
The invention relates to the field of chemical analysis, in particular to a glutathione detection method and a related detection kit. The invention provides a method for detecting glutathione, which comprises the following steps: mixing Fe3O4Co-incubating the @ C nanoparticles with a sample to be tested to provide first nanoparticles; dispersing the first nanoparticles in a reaction system comprising a peroxide and a developer to provide second nanoparticles and a liquid phase; acquiring the absorbance of the liquid phase substance; and providing the content of the GSH in the sample to be detected according to the absorbance of the liquid phase substance. The invention provides a detection method and a related detection kit, and relates to the field of Fe detection according to GSH3O4Inhibition of catalytic Activity of the @ C nanoparticle, based on Fe3O4The catalytic activity of the @ C nanoparticles is similar to that of peroxidase, and specific binding can be achieved with the nanoparticlesThe qualitative and/or quantitative detection of the target substance is converted into the qualitative and/or quantitative determination of the peroxide, and has the characteristics of low detection limit, wide detection range, high specificity and the like.
Description
Technical Field
The invention relates to the field of chemical analysis, in particular to a glutathione detection method and a related detection kit.
Background
Glutathione (GSH), a thiolated tripeptide with multiple physiological functions and medical value, is composed of glutamic acid, cysteine and glycine. Present in almost every cell of the body. Glutathione can help maintain normal immune system function, and has antioxidant and antidotal effects. As important endogenous antioxidants, free radical scavengers and detoxifying substances, GSH can be widely involved in immune regulation, human metabolism, energy transport and other physiological processes. Therefore, glutathione has been widely used for protecting the liver and kidney, detoxifying and anticancer drugs. Various analytical methods for detecting glutathione have been developed, including fluorescence, High Performance Liquid Chromatography (HPLC), electrochemistry, capillary electrophoresis colorimetric assays, photoelectrochemical methods, and the like.
The application of the nano material brings a new opportunity for developing a high-sensitivity and high-selectivity biosensor.
Disclosure of Invention
In view of the above-mentioned disadvantages of the prior art, the present invention aims to provide a method for detecting glutathione and a related detection kit, which are used for solving the problems in the prior art.
In order to achieve the above and other related objects, the present invention provides a method for detecting glutathione, comprising:
1) mixing Fe3O4Co-incubating the @ C nanoparticles with a sample to be tested to provide first nanoparticles;
2) dispersing the first nanoparticles provided in the step 1) in a reaction system comprising peroxide and a color developing agent to provide second nanoparticles and a liquid phase substance;
3) acquiring the absorbance of the liquid phase substance provided in the step 2);
4) providing the content of the GSH in the sample to be detected according to the absorbance of the liquid phase substance provided by the step 3).
In some embodiments of the invention, in step 1), the content of GSH in the sample to be tested is 0.0002 to 500 μ g/ml.
In some embodiments of the invention, in step 1), Fe3O4The @ C nanoparticles are spherical;
and/or, in said step 1), Fe3O4The particle size of the @ C nano particle is 100 nm-220 nm.
In some embodiments of the invention, in the step 1), Fe is contained in the reaction system3O4The content of the @ C nano particles is 0.1-3 mg/ml, preferably 0.5-1.5 mg/ml.
In some embodiments of the invention, in the step 1), the incubation temperature of the co-incubation is 20-40 ℃, preferably 36-38 ℃, and the incubation time is not less than 10min, preferably 90-150 min;
and/or in the step 1), the reaction is carried out in the presence of a solvent, the solvent of the reaction system is one or a combination of more of acetic acid-sodium acetate buffer solution and PBS buffer solution, and the pH value of the reaction system is 4-8, preferably 7.2-7.6.
In some embodiments of the invention, in step 2), the peroxide is selected from hydrogen peroxide;
and/or, in the step 2), the concentration of the peroxide in the reaction system is 1-50 mM, preferably 4-6 mM;
and/or, in the step 2), the color developing agent is selected from TMB;
and/or in the step 2), the concentration of the color developing agent in the reaction system is 0.1-2 mM, preferably 0.8-1.2 mM.
In some embodiments of the present invention, in the step 2), the reaction temperature is 20 to 50 ℃, preferably 25 to 30 ℃;
and/or in the step 2), the reaction is carried out in the presence of a solvent, the solvent of the reaction system is one or a combination of more of acetic acid-sodium acetate buffer solution and PBS buffer solution, and the pH value of the reaction system is 3-4.5, preferably 3.8-4.2.
In some embodiments of the invention, in the step 3), the absorbance of the liquid phase is at 400-800 nm.
In some embodiments of the present invention, in the step 4), the method for providing the content of GSH in the sample to be tested according to the absorbance of the liquid phase provided in the step 3) comprises: and (3) providing the content of the GSH in the sample to be detected according to the relation between the absorbance of the liquid phase substance and the absorbance of the calibrated liquid phase substance and the concentration of the GSH.
In a second aspect, the invention provides a glutathione detection kit, which is suitable for the glutathione detection method, and comprises Fe3O4@ C nanoparticles.
Drawings
FIG. 1 is a schematic diagram showing the principle of the method for detecting glutathione in the present invention.
FIG. 2 is a TEM image of Fe3O4@ C nanoparticles as in example 1 of the present invention.
FIG. 3 is a graph showing an absorbance curve in example 2 of the present invention, in which a is a GSH solution having a concentration of 0 and b is a GSH solution having a concentration of 0.2 mg/ml.
FIG. 4 is a schematic diagram showing the absorbance curves in example 2 of the present invention, in which a to h are GSH solutions having concentrations of 0, 200pg/mL, 2ng/mL, 20ng/mL, 200ng/mL, 2. mu.g/mL, 20. mu.g/mL, and 200. mu.g/mL, respectively.
FIG. 5 is a graph showing the relationship between concentration and absorbance in example 2 of the present invention.
FIG. 6 is a graph showing the linear relationship between the concentration and the absorbance in example 2 of the present invention.
FIG. 7 shows a schematic representation of the specificity of GSH detection in example 3 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments, and other advantages and effects of the present invention will be apparent to those skilled in the art from the disclosure of the present specification.
The inventor of the invention provides a glutathione detection method through a great deal of practical research, and the detection method is implemented through Fe with a core-shell structure3O4Reaction of @ C nanoparticles with GSH to generate peroxidase catalytic activityThe present invention has been completed based on the finding that the detection of a target protein is converted into qualitative and/or quantitative detection based on a redox reaction of a peroxide (e.g., hydrogen peroxide), thereby effectively improving the detection limit and detection sensitivity and widening the detection range.
The first aspect of the present invention provides a method for detecting glutathione, comprising:
1) mixing Fe3O4Co-incubating the @ C nanoparticles with a sample to be tested to provide first nanoparticles;
2) dispersing the first nanoparticles provided in the step 1) in a reaction system comprising peroxide and a color developing agent to provide second nanoparticles and a liquid phase substance;
3) acquiring the absorbance of the liquid phase substance provided in the step 2);
4) providing the content of the GSH in the sample to be detected according to the absorbance of the liquid phase substance provided by the step 3).
The glutathione detection method provided by the invention can comprise the following steps: mixing Fe3O4The @ C nanoparticles are incubated with a sample to be tested to provide first nanoparticles. The sample to be tested may be generally a liquid and/or a gas, etc., and may thus be brought into contact with Fe3O4Full contact of @ C nanoparticles to realize Fe3O4Co-incubation of the @ C nanoparticles with the sample to be tested. If GSH is contained in the sample to be detected, then the GSH is added into Fe3O4The surface of the @ C nanoparticle will be coated with an amount of GSH, thereby providing a first nanoparticle whose surface, if coated with GSH, may correspond to the production of inhibition of peroxidase catalytic activity due to the surface enrichment effect.
In the method for detecting glutathione, the content of GSH in the sample to be detected may be usually 0.0002 to 500. mu.g/ml, 0.0002 to 0.0005. mu.g/ml, 0.0005 to 0.001. mu.g/ml, 0.001 to 0.002. mu.g/ml, 0.002 to 0.005. mu.g/ml, 0.005 to 0.01. mu.g/ml, 0.01 to 0.02. mu.g/ml, 0.02 to 0.05. mu.g/ml, 0.05 to 0.1. mu.g/ml, 0.1 to 0.2. mu.g/ml, 0.2 to 0.5. mu.g/ml, 0.5 to 1. mu.g/ml, 1 to 2. mu.g/ml, 2 to 5. mu.g/ml, 5 to 10. mu.g/ml, 10 to 20. mu.g/ml, 20 to 50. mu.g/ml, 50 to 100. mu.g/ml, 200. mu.g/ml, 300 to 300. mu.g/ml, 400 to 500. mu.g/ml, 500. mu.g/ml, 600-800 μ g/ml, 800-1000 μ g/ml, 1000-1500 μ g/ml, 1500-2000 μ g/ml, or higher.
In the method for detecting glutathione, Fe is suitably supplied3O4The method of @ C nanoparticles should be known to the person skilled in the art. For example, one can refer to Wang N, Zhang M, Liu L, et al, space-defined pyrolysis for simulation of peptides-like Fe3O4@ C-Ni nanostructures for analysis and protein adsorption [ J]Nanotechnology,2019,30 (41.) related methods provide Fe3O4@ C nanoparticles. As another example, Fe3O4The @ C nanoparticles may be spherical. As another example, Fe3O4The particle size of the @ C nanoparticle may be 100 nm-220 nm, 100 nm-110 nm, 110 nm-120 nm, 120 nm-130 nm, 140 nm-150 nm, 150 nm-160 nm, 150 nm-170 nm, 170 nm-180 nm, 180 nm-190 nm, 190 nm-200 nm, 200 nm-210 nm, or 210 nm-220 nm.
In the method for detecting glutathione described above, Fe3O4The using amount of the @ C nano-particles can be adjusted according to the possible GSH content in a sample to be detected, and Fe in a reaction system is generally used3O4Content of @ C nanoparticles (i.e., Fe)3O4Mixing the @ C nano particles with a sample to be detected to form a reaction system, and then mixing Fe in the reaction system before the reaction is not started3O4The content of the @ C nanoparticles) may be 0.1 to 5mg/ml, 0.1 to 0.2mg/ml, 0.2 to 0.3mg/ml, 0.3 to 0.4mg/ml, 0.4 to 0.5mg/ml, 0.5 to 0.6mg/ml, 0.6 to 0.7mg/ml, 0.7 to 0.8mg/ml, 0.8 to 0.9mg/ml, 0.9 to 1.0mg/ml, 1.0 to 1.2mg/ml, 1.2 to 1.5mg/ml, 1.5 to 2mg/ml, 2 to 2.5mg/ml, or 2.5 to 3 mg/ml.
In the method for detecting glutathione described above, one skilled in the art can select appropriate reaction conditions so that Fe is present3O4The @ C nanoparticles react well with possible GSH in the sample to be tested. For example, the incubation temperature for co-incubation can be 20-40 ℃, 36-38 ℃, 20-24 ℃, 24-28 ℃, 28-32 ℃, 32-36 ℃, 36-38 ℃, or 38-40 ℃. For another example, at the time of incubationThe time interval may be not less than 10min, 10-20 min, 20-30 min, 30-40 min, 40-60 min, 60-90 min, 90-120 min, 120-150 min, or 150-180 min.
In the method for detecting glutathione, the co-incubation may be performed in the presence of a solvent. Suitable solvents may be such that Fe3O4The @ C nano particles and a sample to be detected are fully dispersed and fully contacted in a reaction system, and the integral pH value of the reaction system can be kept stable. For example, the applicable solvent may be a buffer solution in general, and specifically, an acetic acid-sodium acetate buffer solution, a PBS buffer solution, or the like. For another example, the pH of the reaction system is 4 to 8, 7.2 to 7.6, 4 to 4.4, 4.4 to 4.8, 4.8 to 5.2, 5.2 to 5.6, 5.6 to 6, 6.4 to 6.8, 6.8 to 7.0, 7.0 to 7.2, 7.2 to 7.4, 7.4 to 7.6, 7.6 to 7.8, or 7.8 to 8.
The glutathione detection method provided by the invention can also comprise the following steps: dispersing the first nanoparticles provided in the step 1) in a reaction system comprising peroxide and a color developing agent to provide second nanoparticles and a liquid phase material. The first nanoparticles are dispersed in a reaction system containing peroxide and a color developing agent, namely, the first nanoparticles can be used for generating peroxidase catalytic activity inhibition, the inhibition generated by the first nanoparticles can be correspondingly different for the GSH content in different samples to be detected, and the introduction of the color developing agent can reflect the content of the residual peroxide in the reaction system.
In the method for detecting glutathione, the type and amount of peroxide are generally required to be equal to Fe3O4The activities of the @ C nanoparticles are matched so that Fe can pass through3O4The @ C nanoparticles accelerate the reaction between the peroxide and the developer. For example, the peroxide may be selected from hydrogen peroxide and the like. For another example, the concentration of the peroxide in the reaction system may be 1 to 50mM, 4 to 6mM, 1 to 2mM, 2 to 3mM, 3 to 4mM, 4 to 4.5mM, 4.5 to 5mM, 5 to 5.5mM, 5.5 to 6mM, 6 to 7mM, 7 to 8mM, 8 to 10mM, 10 to 15mM, 15 to 20mM, 20 to 30mM, 30 to 50mM, or 40 to 50 mM.
In the method for detecting glutathione, the type and the amount of the color developing agent are generally required to be matched with the activity of the peroxide, so that the color developing agent can be oxidized by the residual peroxide (relative to the reaction between the first nano-ion and the peroxide) to enable the reaction system to develop color, and the amount of the residual peroxide in the reaction system can be fully reflected. For example, the color developer may be selected from TMB (3,3',5,5' -tetramethylbenzidine) and the like. For another example, the concentration of the color-developing agent in the reaction system may be 0.1 to 2mM, 0.8 to 1.2mM, 0.1 to 0.2mM, 0.2 to 0.4mM, 0.4 to 0.6mM, 0.6 to 0.8mM, 0.8 to 1mM, 1 to 1.2mM, 1.2 to 1.5mM, or 1.5 to 2 mM.
In the method for detecting glutathione, one skilled in the art can select appropriate reaction conditions so that the first nanoparticles are sufficiently reacted with the peroxide and the remaining peroxide is sufficiently colored in the reaction system including the color-developing agent. For example, the reaction temperature may be room temperature, 20-50 ℃, 20-22 ℃, 22-24 ℃, 24-26 ℃, 26-28 ℃, 28-30 ℃, 30-35 ℃, 35-40 ℃, 40-45 ℃, or 45-50 ℃. For another example, the reaction time may be 5min or more, 5 to 6min, 6 to 8min, 8 to 10min, 10 to 15min or more.
In the method for detecting glutathione described above, the reaction between the first nanoparticle and the peroxide and the color-developing agent may be carried out in the presence of a solvent. The suitable solvent may allow the first nanoparticles, the peroxide, the color developer, and the like to be sufficiently dispersed and contacted in the reaction system, and may allow the overall pH of the reaction system to be maintained stable. For example, the applicable solvent may be a buffer solution in general, and specifically, an acetic acid-sodium acetate buffer solution, a PBS buffer solution, or the like. For another example, the pH of the reaction system may be 3 to 4.5, 3 to 3.2, 3.2 to 3.4, 3.4 to 3.6, 3.6 to 3.8, 3.8 to 3.9, 3.9 to 4, 4 to 4.1, 4.1 to 4.2, 4.2 to 4.3, or 4.3 to 4.5.
The glutathione detection method provided by the invention can also comprise the following steps: acquiring the absorbance of the liquid phase substance provided by the step 2). As mentioned above, the residual peroxide in the reaction system can oxidize the color developing agent, and the liquid phase substance provided by the reaction system can correspondingly develop color, so that the liquid phase substance has certain absorbance. The absorbance of the liquid phase is usually matched with the color developer and the reaction system. For example, the absorbance of the liquid phase may be 400 to 800nm, 400 to 450nm, 450 to 500nm, 500 to 550nm, 550 to 600nm, 600 to 650nm, 650 to 700nm, 700 to 750nm, or 750 to 800 nm.
The glutathione detection method provided by the invention can also comprise the following steps: providing the content of the GSH in the sample to be detected according to the absorbance of the liquid phase substance provided by the step 3). The liquid phase provided by the reaction system is developed correspondingly as described above, and thus has a certain absorbance, and the absorbance generally corresponds to (e.g., is linear with) the content of the sample to be tested.
In the method for detecting glutathione, the step of providing the content of GSH in the solution to be detected can be qualitative detection, namely providing whether the sample to be detected contains GSH, or can be quantitative detection, namely providing the specific content of GSH in the sample to be detected. For example, the method for providing the content of GSH in the sample to be tested according to the absorbance of the liquid phase substance provided in step 3) may specifically include: and (3) providing the content of the GSH in the sample to be detected according to the relation between the absorbance of the liquid phase substance and the absorbance of the calibrated liquid phase substance and the concentration of the GSH. After the absorbance of the liquid phase substance obtained by the corresponding treatment of the sample to be detected is obtained, the content of the GSH in the sample to be detected can be obtained according to the relationship between the calibrated absorbance of the liquid phase substance and the concentration of the GSH. Methods for providing a relationship between absorbance of a calibration liquid phase and GSH concentration should be known to those skilled in the art. For example, the relationship between the absorbance of the liquid phase corresponding to each of the GSH solutions at the calibration concentrations, i.e., the absorbance of the liquid phase corresponding to the GSH solution at the calibration concentrations and the GSH concentration, can be obtained by referring to the above glutathione detection method (i.e., replacing the sample to be detected with the GSH solution at the calibration concentrations) for a series of GSH solutions at the calibration concentrations (e.g., 200 μ g/mL, 20 μ g/mL, 2 μ g/mL, 200ng/mL, 20ng/mL, 2ng/mL, 200pg/mL, 0GSH concentration (PBS) buffer).
The second aspect of the invention provides a glutathione detection kit, which is suitable for the detection method of glutathione provided by the first aspect of the invention. The kit may typically comprise Fe3O4The @ C nano-particle can also comprise one or more of a peroxide, a color developing agent, a buffer solution and the like.
The invention provides a glutathione detection method and a related detection kit, and relates to Fe detection according to GSH3O4Inhibition of catalytic Activity of the @ C nanoparticle, based on Fe3O4The catalytic activity of the @ C nano particle is similar to that of peroxidase, qualitative and/or quantitative detection of a target substance specifically combined with the @ C nano particle can be converted into qualitative and/or quantitative determination of the peroxide, the detection range of the detection method can be 0.0002-200 mu g/mL, the detection limit can reach 0.02ng/mL, and the method has the characteristics of low detection limit, wide detection range, high specificity and the like, has wide application prospect in medical diagnosis, treatment and detection directions, and has good industrial prospect.
The present application is further illustrated by the following examples, which are not intended to limit the scope of the present application.
Example 1
Fe3O4Preparation of @ C nanoparticles: ferrocene solution was prepared by dissolving 0.3g ferrocene (spherical material) in 30mL acetone, after 30 minutes sonication. 1mL of hydrogen peroxide was added dropwise to the above ferrocene solution and reacted for 10 minutes with stirring, and the resulting solution was transferred to a total volume of 50mL of a Teflon-lined stainless autoclave and reacted at 200 ℃ for 24 hours. Centrifugally washing for multiple times (at least 3 times) and drying in an oven to obtain Fe3O4@ C nanoparticle powder, Fe obtained by preparation3O4TEM images of the @ C nanoparticles are shown in FIG. 2.
Example 2
Taking Fe3O4@ C nanoparticles 1mg (prepared in example 1), dispersed in 1mL1 XPBS solution (pH 7.4), 25 uL of GSH solution (200. mu.g/mL, 20. mu.g/mL, 2. mu.g/mL, 200ng/mL, 20ng/mL, 2ng/mL, 200pg/mL, 0, solvent 1 XPBS solution) at various concentrations and Fe as described above3O4@ C solution 100. mu.L was mixed well, incubated at 37 ℃ for 2 hours, and then the prepared particles were stored in PBS buffer.
258.75 μ l of 0.2M pH 4.0Acetic acid-sodium acetate buffer solution into a centrifuge tube, 11.25. mu.l of nanoparticle solution (obtained in the previous step) with a concentration of 0.8mg/mL was added, followed by 15. mu.L of TMB (20mM), 15. mu. L H2O2(0.1M), uniformly mixing the obtained solution, reacting for 10 minutes at room temperature, separating the nano particles from the reaction solution by an external magnetic field, and measuring the ultraviolet absorption spectrum of the mixed solution at 652nm by using an ultraviolet-visible absorption spectrophotometer.
The results of the absorbance curves obtained from the measurement of standards of different GSH concentrations are shown in fig. 3 to 6. As can be seen from fig. 3 to 6, there is a good linear relationship between the GSH concentration in the standard and the measured absorbance.
Example 3
Mixing Fe3O4@ C nanoparticles (1Mg/mL, prepared in example 1) were mixed with a solution containing aspartic acid (Asp), arginine (Arg), glutamic acid (Glu), serine (Ser), and Mg2+(MgCl2)、K+(KCl) (concentration of each substance was 40mg/mL) were mixed with each solution (solvent 1 XPBS buffer), and then shaken for 2 hours on a homomixer (37 ℃, 1500 rpm).
The mixture of the above steps (final concentration 30. mu.gmL)-1) With 20mM TMB, 15. mu.L (final concentration 1mM) and H2O2After incubation of 0.1M, 15. mu.L (final concentration 5mM) HAc-NaAc buffer (0.2M, pH 4.0) mixed at room temperature for 10 minutes, nanoenzymes were then rapidly removed by applying a magnetic field or centrifugation. Then, the solution was transferred to a quartz cuvette, and UV-Vis spectra were measured at 652nm, and the results are shown in FIG. 7, in which the absorbance of GSH, i.e., the absorbance of GSH obtained in example 2 at a concentration of 200. mu.g/mL, is shown in FIG. 7.
As can be seen from FIG. 7, Fe3O4The absorbance of the @ C nano particle is not obviously interfered after the reaction with other 6 interferents respectively, and the absorbance of oxTMB at 652nm is obviously reduced after the reaction with GSH, which indicates that the GSH has Fe to the GSH3O4The catalytic activity of the @ C nanoparticle produces significant inhibition, and this inhibition has very significant specificity.
In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (10)
1. A method for detecting glutathione, which comprises the following steps:
1) mixing Fe3O4Co-incubating the @ C nanoparticles with a sample to be tested to provide first nanoparticles;
2) dispersing the first nanoparticles provided in the step 1) in a reaction system comprising peroxide and a color developing agent to provide second nanoparticles and a liquid phase substance;
3) acquiring the absorbance of the liquid phase substance provided in the step 2);
4) providing the content of the GSH in the sample to be detected according to the absorbance of the liquid phase substance provided by the step 3).
2. The method for detecting glutathione according to claim 1, wherein in the step 1), the content of GSH in the sample to be detected is 0.0002-500 μ g/ml.
3. The method for detecting glutathione according to claim 1, wherein in the step 1), Fe3O4The @ C nanoparticles are spherical;
and/or, in said step 1), Fe3O4The particle size of the @ C nano particle is 100 nm-220 nm.
4. The method for detecting glutathione according to claim 1, wherein in the step 1), Fe is contained in the reaction system3O4The content of the @ C nano particles is 0.1-3 mg/ml, preferably 0.5-1.5 mg/ml。
5. The glutathione detection method according to claim 1, wherein in the step 1), the incubation temperature of the co-incubation is 20-40 ℃, preferably 36-38 ℃, and the incubation time is not less than 10min, preferably 90-150 min;
and/or in the step 1), the reaction is carried out in the presence of a solvent, the solvent of the reaction system is one or a combination of more of acetic acid-sodium acetate buffer solution and PBS buffer solution, and the pH value of the reaction system is 4-8, preferably 7.2-7.6.
6. The method for detecting glutathione according to claim 1, wherein in the step 2), the peroxide is selected from the group consisting of hydrogen peroxide;
and/or, in the step 2), the concentration of the peroxide in the reaction system is 1-50 mM, preferably 4-6 mM;
and/or, in the step 2), the color developing agent is selected from TMB;
and/or in the step 2), the concentration of the color developing agent in the reaction system is 0.1-2 mM, preferably 0.8-1.2 mM.
7. The method for detecting glutathione according to claim 1, wherein in the step 2), the reaction temperature is 20 to 50 ℃, preferably 25 to 30 ℃;
and/or in the step 2), the reaction is carried out in the presence of a solvent, the solvent of the reaction system is one or a combination of more of acetic acid-sodium acetate buffer solution and PBS buffer solution, and the pH value of the reaction system is 3-4.5, preferably 3.8-4.2.
8. The method for detecting glutathione according to claim 1, wherein in the step 3), the absorbance of the liquid phase is the absorbance of the liquid phase at 400 to 800 nm.
9. The method for detecting glutathione according to claim 1, wherein the step 4) of providing the content of GSH in the sample to be detected according to the absorbance of the liquid phase substance provided in the step 3) comprises the following steps: and (3) providing the content of the GSH in the sample to be detected according to the relation between the absorbance of the liquid phase substance and the absorbance of the calibrated liquid phase substance and the concentration of the GSH.
10. A glutathione detection kit, which is suitable for the glutathione detection method of any one of claims 1 to 9, and comprises Fe3O4@ C nanoparticles.
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| CN109342420A (en) * | 2018-12-07 | 2019-02-15 | 上海工程技术大学 | Fe3O4The application of@C one-dimensional nano line |
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| JP2004267067A (en) * | 2003-03-06 | 2004-09-30 | Osaka Industrial Promotion Organization | Method for alcohol analysis |
| CN109342420A (en) * | 2018-12-07 | 2019-02-15 | 上海工程技术大学 | Fe3O4The application of@C one-dimensional nano line |
| CN111024636A (en) * | 2019-10-16 | 2020-04-17 | 济南大学 | Colorimetric method for detecting glutathione based on CoOOH-TMB oxidation system |
| CN111307725A (en) * | 2020-03-10 | 2020-06-19 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Method for measuring content of glutathione |
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| CN116874556A (en) * | 2023-06-01 | 2023-10-13 | 四川农业大学 | Preparation method and application of selenium glutathione nano-enzyme with antioxidant activity |
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