CN104330392B - Hydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probe - Google Patents
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Abstract
The present invention discloses a kind of hydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probe, and it relates to, with the gold nano cluster Catalase determination method as fluorescent probe of N acetyl L cysteine protection, it is characterized in that utilizing Fe2+Catalysis H2O2Produce hydroxy radical and make the fluorescence generation quencher of gold nano cluster, and catalase can be catalyzed H2O2Decompose and generate H2O and O2, suppress gold nano cluster Quenching of fluorescence, thus show the change of fluorescence emission spectrum signature, may be used for catalatic detection.Fluorescence intensity change value Δ F in the range of 0.01 ~ 0.3 U/mL650Linear with catalase concentration, detection is limited to 0.002 U/mL.The present invention is highly sensitive, favorable reproducibility, can be used for catalatic mensuration in food, industry, environment and life system.
Description
Technical field
The present invention relates to, with the gold nano cluster catalatic assay method as fluorescent probe of N-acetyl-L-cysteine protection, belong to analytical chemistry and field of nanometer technology.
Background technology
Catalase (has another name called catalase), is a kind of oxidoreductase being widely present in biological tissue.Catalase is a kind of tetramer hemin enzyme, is made up of the subunit of four identical tetrahedral arrangement, and each subunit is 60000
G/mol, each molecule includes four protoferriheme groups, and molecular weight is about 240000.The accumulation of the reactive oxygen species and free radicals in organism, can cause Lipid peroxidation metabolism, thus cause the disorder of the metabolism of organism own.Many enzymatic reactions and non-enzymatic reaction can produce hydrogen peroxide (H2O2), H2O2It it is the precursor of the active oxygen of toxic effect.Catalatic major function is catalysis H2O2Being decomposed into water and oxygen, the hydrogen peroxide in purged body, so that cell protects against H2O2Murder by poisoning, this enzyme has scavenging action to hydroxy radical simultaneously.To this end, the most in succession establish some methods measuring catalase activity both at home and abroad, such as ultraviolet spectrophotometry, titrimetry, oxygen electrode method, polarography and photoelectric colorimetry etc..For these methods, some needing valuable instrument, have needs special reagent, and some operations are complicated, some methods itself with regard to poor repeatability, precision is inadequate.Therefore, the assay method setting up new catalase activity is extremely necessary.
Gold nano cluster (gold nanoclusters, Au NCs) as a kind of novel fluorescent nano material, it has that size is little, nontoxic, good water solubility, good light stability, Stokes displacement is big, specific surface area is big, preparation condition is gentle, surface is prone to modify and photoluminescent property is with outstanding advantages such as size adjustable, being study hotspot in recent years, it has been widely used in the fields such as catalysis, sensing detection, Nanoparticle labeling, medical imaging and optoelectronics.
The gold nano cluster that the present invention protects using N-acetyl-L-cysteine is as fluorescent probe, it is provided that the new method of a kind of simplicity, sensitive catalase detection.
Summary of the invention
It is an object of the invention to provide a kind of gold nano cluster protected with N-acetyl-L-cysteine catalatic assay method as fluorescent probe.
To achieve these goals, the present invention is by the following technical solutions: of the present inventionHydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probe, it is characterized in that utilizing Fe2+Catalysis H2O2Produce hydroxy radical and make the fluorescence generation quencher of gold nano cluster, and catalase can be catalyzed H2O2Decompose and generate H2O and O2, suppress gold nano cluster Quenching of fluorescence, thus show the change of fluorescence emission spectrum signature, catalatic detection can be used for.
The gold nano cluster of the N-acetyl-L-cysteine protection used uses the method for N-acetyl-L-cysteine reduction gold chloride to prepare: N-acetyl-L-cysteine solution that concentration is 0.02 ~ 0.18 mol/L and sodium hydroxide solution that concentration is 0.1 ~ 0.8 mol/L are joined concentration is 0.01 ~ 0.1
In the chlorauric acid solution of g/L, mixing, being placed in 20 ~ 70 ° of C constant temperature water baths to react 0 ~ 3.5 hour, reaction carries out, to reactant liquor, purification process of dialysing with the bag filter that molecular cut off is 3500 after terminating, and obtains N-acetyl-L-cysteine-fluorescent au nanocluster material aqueous solution.
The gold nano cluster of the N-acetyl-L-cysteine protection used preferably employs the method for N-acetyl-L-cysteine reduction gold chloride to be prepared: N-acetyl-L-cysteine solution that concentration is 0.08 mol/L and sodium hydroxide solution that concentration is 0.5 mol/L are joined concentration is 0.02
In the chlorauric acid solution of g/L, mixing, it is placed in 37 ° of C constant temperature water baths and reacts 2.5 hours, reaction carries out, to reactant liquor, purification process of dialysing with the bag filter that molecular cut off is 3500 after terminating, and obtains N-acetyl-L-cysteine-fluorescent au nanocluster material aqueous solution.
The present invention utilizes gold nano cluster that N-acetyl-L-cysteine the protects fluorescence intensity level (F at 650 nm650) to judge catalase content, the excitation wavelength used is 355 nm.
Of the present inventionHydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probeComprise the steps: 1) by the Catalase solution containing variable concentrations, containing the HEPES that concentration is 20 mmol/L, pH=7.4 in described Catalase solution, add in hydrogenperoxide steam generator, containing the HEPES that concentration is 20 mmol/L, pH=7.4 in described hydrogenperoxide steam generator, it is subsequently placed in 25 ° of C temperature bath reactions 15 ~ 150 minutes；2) by containing the Fe that concentration is 40 mmol/L sulphuric acid2+Solution and gold nano cluster solution are sequentially added in the reactant liquor of step 1), shake up and are placed on 25 ° of C water-baths reactions 1 ~ 15 minute, after reaction terminates, measure the fluorescence intensity level (F at 650 nm650), along with the increase of catalase concentration, F650It is gradually increased, 0.01 ~ 0.3
Fluorescence intensity change value Δ F in the range of U/mL650Linear with catalase concentration, detection is limited to 0.002 U/mL.
Above-mentioned steps 1) concentration of hydrogenperoxide steam generator that used is 10 μm ol/L, the response time is 90 minutes；Step 2) Fe that used2+Final concentration of 100 μm ol/L of solution, the response time is 10 minutes.
Described Catalase solution, hydrogenperoxide steam generator, Fe2+Solution, gold nano cluster solution 1:7:2:8 by volume mixes, and reaction cumulative volume is 0.45 mL.
Of the present inventionBased on catalatic method in gold nano cluster probe assay people's saliva, it is characterized in that comprising the steps: 1) and take Freshman saliva, after centrifugal 10 minutes, dilute with the buffer of pH=7.4 through 6000 revs/min, take 0.025 mL diluent and join the H that 0.175 milliliter of concentration is 10 μm ol/L2O2In solution, it is placed in 25 ° of C and reacts 90 minutes；2) by the Fe containing 40 mmol/L sulphuric acid that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution and 0.2 milliliter of gold nano cluster solution are sequentially added into above-mentioned steps 1) reactant liquor in, shake up and be placed on 25 ° of C water-baths and react 10 minutes, measure fluorescence intensity level F650, carry out quantitatively by standard curve, it is thus achieved thatCatalase content in saliva。
The gold nano cluster of the N-acetyl-L-cysteine protection used uses the method for N-acetyl-L-cysteine reduction gold chloride to prepare: N-acetyl-L-cysteine solution that concentration is 0.02 ~ 0.18 mol/L and sodium hydroxide solution that concentration is 0.1 ~ 0.8 mol/L are joined concentration is 0.01 ~ 0.1
In the chlorauric acid solution of g/L, mixing, being placed in 20 ~ 70 ° of C constant temperature water baths to react 0 ~ 3.5 hour, reaction carries out, to reactant liquor, purification process of dialysing with the bag filter that molecular cut off is 3500 after terminating, and obtains N-acetyl-L-cysteine-fluorescent au nanocluster material aqueous solution.
The gold nano cluster of the N-acetyl-L-cysteine protection used preferably employs the method for N-acetyl-L-cysteine reduction gold chloride to be prepared: join in the N-acetyl-L-cysteine solution that 4 mL concentration are 0.08 mol/L by sodium hydroxide solution and the chlorauric acid solution that 0.4 mL concentration is 0.02 g/L that 0.6 mL concentration is 0.5 mol/L, mixing, it is placed in 37 ° of C constant temperature water baths and reacts 2.5 h, reactant liquor is become colorless by light yellow, reaction is purified process with the bag filter that molecular cut off is 3500 to reactant liquor after terminating, gold nano cluster solution after purification is positioned over 4 ° of C refrigerators and keeps in Dark Place.
Specifically, the technical solution used in the present invention is:
(1) preparation of fluorescent au nanocluster material
The all glass drying ovens used in procedure below all soak through chloroazotic acid, and thoroughly clean with distilled water, dry.The preparation method of fluorescent au nanocluster material is as follows: join in the N-acetyl-L-cysteine solution that 4 mL concentration are 0.08 mol/L by sodium hydroxide solution and the chlorauric acid solution that 0.4 mL concentration is 0.02 g/L that 0.6 mL concentration is 0.5 mol/L, mixing, being placed in 37 ° of C constant temperature water baths reaction 2.5 hours, reactant liquor is become colorless by light yellow.Reaction is purified process with the bag filter that molecular cut off is 3500 to reactant liquor after terminating, and gold nano cluster solution after purification is positioned over 4 ° of C refrigerators and keeps in Dark Place.
(2) catalatic mensuration
Catalatic mensuration is undertaken in two steps: (1) 0.025 milliliter of sample solution (HEPES, 20 mmol/L pH=7.4) adds in the hydrogenperoxide steam generator that 0.175 milliliter of concentration is 10 μm ol/L, is placed in 25 ° of C and reacts 90 minutes；(2) by ferrous ion (Fe that 0.05 milliliter of concentration is 0.9 mmol/L2+) the gold nano cluster solution prepared with 0.2 milliliter of step () of solution (containing 40 mmol/L sulphuric acid) is sequentially added in above-mentioned reactant liquor, shake up and be placed on 25 ° of C water-baths and react 10 minutes.After reaction terminates, with 355 nm as excitation wavelength, measure 650
Fluorescence intensity level (F at nm650), pass through F650Standard curve carries out catalatic mensuration.
Advantages of the present invention:
(1) present invention is based on Fe2+Catalysis H2O2Produce hydroxy radical (Fenton reaction) and make the fluorescence generation quencher of gold nano cluster, and catalase can be catalyzed H2O2Decompose and generate H2O and O2, suppress gold nano cluster Quenching of fluorescence, thus show the change of fluorescence emission spectrum signature, may be used for catalatic detection.
(2) its preparation process of gold nano cluster material of the N-acetyl-L-cysteine protection that the present invention uses is quick, easy, it is not necessary to any front modification step.
(3) the method mensuration specificity constructed by the present invention is good, highly sensitive, and detection is limited to 0.002
U/mL。
(4) method constructed by the present invention can realize the catalatic mensuration of people's saliva without complicated sample pretreatment process.
Accompanying drawing explanation
Fig. 1 is gold nano cluster solution outward appearance comparison figure under uviol lamp.In figure: (A) gold nano cluster solution+10 μm ol/L H2O2
+ 100 μmol/L Fe2+；(B) gold nano cluster solution+10 μm ol/L H2O2 + 100 μmol/L Fe2+ + 0.5 U/mL
Catalase.
Fig. 2 is the emission spectrum figure of gold nano cluster solution.In figure: (A) gold nano cluster solution+10 μm ol/L H2O2
+ 100 μmol/L Fe2+；(B) gold nano cluster solution+10 μm ol/L H2O2 + 100 μmol/L Fe2+ + 0.5 U/mL
Catalase.
Fig. 3 is the catalase catalyst system and catalyzing response time gold nano cluster solution fluorescence intensity to be affected figure.
Fig. 4 is that disturbance material affects figure to gold nano cluster solution fluorescence intensity.(black post in figure: chaff interference+gold nano cluster solution+10 μm ol/L H2O2
+ 100 μmol/L Fe2+；Bai Zhu: chaff interference+catalase+gold nano cluster solution+10 μm ol/L H2O2
+ 100 μmol/L Fe2+).
Fig. 5 is fluorescence intensity change value (the Δ F of gold nano cluster solution650) and catalase concentration between linear relationship chart.
Detailed description of the invention
Of the present inventionHEPES refers to N-(2-ethoxy) piperazine-N'-2-ethane sulfonic acid.Fe used by present example2+Solution is Fe disclosed in any prior art2+Solution, it is preferably ferrous chloride and is dissolved in the Fe that 40 mM sulphuric acid are formulated2+Solution.
Example
1
:
Sodium hydroxide solution and the chlorauric acid solution that 0.4 mL concentration is 0.02 g/L that 0.6 mL concentration is 0.5 mol/L are joined in the N-acetyl-L-cysteine solution that 4 mL concentration are 0.08 mol/L, mixing, it is placed in 37 ° of C constant temperature water baths and reacts 2.5 h.Reaction is purified process with the bag filter that molecular cut off is 3500 to reactant liquor after terminating.It is colourless under obtained gold nano cluster solution visible ray, under ultra violet lamp, produces strong red fluorescence.4 ° of C dark places preserve, and can keep the most stable of at least one month.
Example
2
:
0.025 milliliter of concentration is that the Catalase solution of 0.5 U/mL is (containing HEPES N-(2-ethoxy) piperazine-N'-2-ethane sulfonic acid, concentration 20 mmol/L, pH=7.4) to add 0.175 milliliter of concentration be that the hydrogenperoxide steam generator of 10 μm ol/L is (containing HEPES, concentration 20 mmol/L, pH=7.4) in, it is placed in 25 ° of C and reacts 90 minutes.By the Fe that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution (containing 40 mmol/L sulphuric acid) is sequentially added in above-mentioned reactant liquor with the gold nano cluster solution of 0.2 milliliter of example 1 preparation, shakes up and is placed on 25 ° of C water-baths reactions 10 minutes.Observe under uviol lamp, be added without the red fluorescence generation quencher (A in Fig. 1) after Fenton reacts of catalatic gold nano cluster solution, and after adding catalase reaction, gold nano cluster solution recovers red fluorescence (B in Fig. 1).Fig. 2 is for being added without catalatic gold nano cluster solution (A in Fig. 2) and adding the catalatic gold nano cluster solution (B in Fig. 2) fluorescence emission spectrogram after Fenton reacts.
Example
3
:
0.025 milliliter of concentration is the Catalase solution (HEPES of 0.5 U/mL, 20 mmol/L pH=7.4) add the hydrogenperoxide steam generator (HEPES that 0.175 milliliter of concentration is 10 μm ol/L, 20 mmol/L pH=7.4) in, it is placed in 25 ° of C and reacts 15 ~ 130 minutes.By the Fe that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution (containing 40 mmol/L sulphuric acid) is sequentially added in above-mentioned reactant liquor with the gold nano cluster solution of 0.2 milliliter of example 1 preparation, shakes up and is placed on 25 ° of C water-baths reactions 10 minutes.As it is shown on figure 3, hydrogen peroxide enzymic catalytic reaction is after 90 minutes, fluorescence intensity level F650Change basically reaches steadily, therefore selecting the hydrogen peroxide enzymic catalytic reaction time is 90 minutes.
Example
4
:
Interference experiment: 0.025 milliliter of solution (HEPES, 20 mmol/L pH=7.4) containing disturbance material adds in the hydrogenperoxide steam generator (HEPES, 20 mmol/L pH=7.4) that 0.175 milliliter of concentration is 10 μm ol/L, is placed in 25 ° of C and reacts 90 minutes.By the Fe that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution (containing 40 mmol/L sulphuric acid) is sequentially added in above-mentioned reactant liquor with the gold nano cluster solution of 0.2 milliliter of example 1 preparation, shakes up and is placed on 25 ° of C water-baths reactions 10 minutes, measures fluorescence intensity level F650.As shown in Figure 4, the method capacity of resisting disturbance constructed by the present invention is strong.(0 ~ 17 is respectively blank, calcium chloride, zinc sulfate, potassium chloride, sodium bicarbonate, sodium dihydrogen phosphate, magnesium chloride, glucose, lactic acid, creatine, creatinine, carbamide, acetylcholine, glutathion, S-adenosylmethionine transferring enzyme, urase, carboxy-lesterase and acetylcholinesterase, wherein the concentration of catalase, S-adenosylmethionine transferring enzyme, urase, carboxy-lesterase and acetylcholinesterase is 0.5 U/mL, and the concentration of other interfering material is 100 μm ol/L)
Example
5
:
0.025 milliliter of Catalase solution (HEPES, 20 mmol/L pH=7.4) containing variable concentrations adds in the hydrogenperoxide steam generator (HEPES, 20 mmol/L pH=7.4) that 0.175 milliliter of concentration is 10 μm ol/L, is placed in 25 ° of C and reacts 90 minutes.By the Fe that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution (containing 40 mmol/L sulphuric acid) is sequentially added in above-mentioned reactant liquor with the gold nano cluster solution of 0.2 milliliter of example 1 preparation, shakes up and is placed on 25 ° of C water-baths reactions 10 minutes, measures fluorescence intensity level F650.As it is shown in figure 5, along with the increase of catalase concentration, fluorescence intensity change value Δ F650It is gradually increased, Δ F in the range of 0.01 ~ 0.3 U/mL650Linear with catalase concentration, detection is limited to 0.002
U/mL。
Example
6
:
0.025 milliliter of concentration is that 0.05 U/mL Catalase solution (HEPES, 20 mmol/L pH=7.4) adds in the hydrogenperoxide steam generator (HEPES, 20 mmol/L pH=7.4) that 0.175 milliliter of concentration is 10 μm ol/L, is placed in 25 ° of C and reacts 90 minutes.By the Fe that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution (containing 40 mmol/L sulphuric acid) is sequentially added in above-mentioned reactant liquor with the gold nano cluster solution of 0.2 milliliter of example 1 preparation, shakes up and is placed on 25 ° of C water-baths reactions 10 minutes, measures fluorescence intensity level F650.Repeat the above steps 12 times, obtaining relative standard deviation (RSD) is 2.7%, shows that this method repeatability is good.
Example
7
:
Take Freshman saliva, it is centrifuged 10 minutes with 6000 revs/min, the HEPES buffer (20 mmol/L) taking supernatant pH=7.4 dilutes 10 times, takes 0.025 mL diluent and joins in the hydrogenperoxide steam generator that 0.175 milliliter of concentration is 25 μm ol/L, is placed in 25 ° of C and reacts 90 minutes.By the Fe that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution (containing 40 mmol/L sulphuric acid) is sequentially added in above-mentioned reactant liquor with the gold nano cluster solution of 0.2 milliliter of example 1 preparation, shakes up and is placed on 25 ° of C water-baths reactions 10 minutes, measures fluorescence intensity level F650.Carry out quantitatively by standard curve, it is thus achieved that the catalase content in saliva.Comparing with standard method measurement result, result shows that method used in the present invention and ultraviolet method there was no significant difference (table 1).
Table 1
F0.05, 2, 2=19.00, t0.05, 4=2.776。
Claims (9)
1. one kindHydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probe, it is characterized in that utilizing Fe2+Catalysis H2O2Produce hydroxy radical and make the fluorescence generation quencher of gold nano cluster, and catalase can be catalyzed H2O2Decompose and generate H2O and O2, suppress gold nano cluster Quenching of fluorescence, thus show the change of fluorescence emission spectrum signature, catalatic detection can be used for；The gold nano cluster used uses the method for N-acetyl-L-cysteine reduction gold chloride to prepare: join in the chlorauric acid solution that concentration is 0.01 ~ 0.1 g/L by N-acetyl-L-cysteine solution and the sodium hydroxide solution that concentration is 0.1 ~ 0.8 mol/L that concentration is 0.02 ~ 0.18 mol/L; mixing; it is placed in 20 ~ 70 DEG C of constant temperature water baths to react 2.5 hours; reaction carries out, to reactant liquor, purification process of dialysing with the bag filter that molecular cut off is 3500 after terminating, and obtains the fluorescent au nanocluster material aqueous solution of N-acetyl-L-cysteine protection.
The most according to claim 1Hydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probeIt is characterized in that used gold nano cluster uses the method for N-acetyl-L-cysteine reduction gold chloride to prepare: join in the chlorauric acid solution that concentration is 0.02 g/L by N-acetyl-L-cysteine solution and the sodium hydroxide solution that concentration is 0.5 mol/L that concentration is 0.08 mol/L; mixing; it is placed in 37 DEG C of constant temperature water baths to react 2.5 hours; reaction carries out, to reactant liquor, purification process of dialysing with the bag filter that molecular cut off is 3500 after terminating, and obtains the fluorescent au nanocluster material aqueous solution of N-acetyl-L-cysteine protection.
The most according to claim 1Hydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probe, it is characterized in that the gold nano cluster utilizing N-acetyl-L-cysteine to protect fluorescence intensity level F at 650 nm650To judge catalase content, the excitation wavelength used is 355 nm.
4. one kindHydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probeComprise the steps: 1) by the Catalase solution containing variable concentrations, containing the HEPES that concentration is 20 mmol/L, pH=7.4 in described Catalase solution, add in hydrogenperoxide steam generator, containing the HEPES that concentration is 20 mmol/L, pH=7.4 in described hydrogenperoxide steam generator, it is subsequently placed in 25 DEG C of temperature bath reactions 15 ~ 150 minutes；2) by containing the Fe that concentration is 40 mmol/L sulphuric acid2+Solution and gold nano cluster solution are sequentially added in the reactant liquor of step 1), shake up and are placed on 25 DEG C of water-baths reactions 1 ~ 15 minute, after reaction terminates, measure the fluorescence intensity level F at 650 nm650, along with the increase of catalase concentration, F650It is gradually increased, fluorescence intensity change value Δ F in the range of 0.01 ~ 0.3 U/mL650Linear with catalase concentration, detection is limited to 0.002 U/mL.
The most according to claim 4Hydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probe, it is characterized in that the concentration of the hydrogenperoxide steam generator that step 1) used is 10 μm ol/L, the response time is 90 minutes；Step 2) Fe that used2+Final concentration of 100 μm ol/L of solution, the response time is 10 minutes.
6. according to described in claim 4 or 5Hydrogen peroxide enzymatic fluorimetric assay based on gold nano cluster probe, it is characterized in that Catalase solution, hydrogenperoxide steam generator, Fe2+Solution, gold nano cluster solution 1:7:2:8 by volume mixes, and cumulative volume is 0.45 mL.
7. one kindBased on catalatic method in gold nano cluster probe assay people's saliva, it is characterized in that comprising the steps: 1) and take Freshman saliva, after centrifugal 10 minutes, dilute with the buffer of pH=7.4 through 6000 revs/min, take 0.025 mL diluent and join the H that 0.175 milliliter of concentration is 10 μm ol/L2O2In solution, it is placed in 25 DEG C and reacts 90 minutes；2) by the Fe containing 40 mmol/L sulphuric acid that 0.05 milliliter of concentration is 0.9 mmol/L2+Solution and 0.2 milliliter of gold nano cluster solution are sequentially added into above-mentioned steps 1) reactant liquor in, shake up and be placed on 25 DEG C of water-baths and react 10 minutes, measure the fluorescence intensity level F at 650 nm650, carry out quantitatively by standard curve, it is thus achieved thatCatalase content in saliva。
The most according to claim 7Based on catalatic method in gold nano cluster probe assay people's salivaIt is characterized in that used gold nano cluster uses the method for N-acetyl-L-cysteine reduction gold chloride to prepare: join in the chlorauric acid solution that concentration is 0.01 ~ 0.1 g/L by N-acetyl-L-cysteine solution and the sodium hydroxide solution that concentration is 0.1 ~ 0.8 mol/L that concentration is 0.02 ~ 0.18 mol/L; mixing; it is placed in 20 ~ 70 DEG C of constant temperature water baths to react 2.5 hours; reaction carries out, to reactant liquor, purification process of dialysing with the bag filter that molecular cut off is 3500 after terminating, and obtains the fluorescent au nanocluster material aqueous solution of N-acetyl-L-cysteine protection.
The most according to claim 8Based on catalatic method in gold nano cluster probe assay people's saliva, it is characterized in that the gold nano cluster of used N-acetyl-L-cysteine protection uses the method for N-acetyl-L-cysteine reduction gold chloride to prepare: join in the chlorauric acid solution that 0.4 mL concentration is 0.02 g/L by N-acetyl-L-cysteine solution and the sodium hydroxide solution that 0.6 mL concentration is 0.5 mol/L that 4 mL concentration are 0.08 mol/L, mixing, it is placed in 37 DEG C of constant temperature water baths and reacts 2.5 h, reactant liquor is become colorless by light yellow, reaction is purified process with the bag filter that molecular cut off is 3500 to reactant liquor after terminating, gold nano cluster solution after purification is positioned over 4 DEG C of refrigerators and keeps in Dark Place.
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| CN106872546B (en) * | 2017-02-07 | 2018-12-04 | 福建医科大学 | Electrochemical reducing prepares high quantum production rate electrochemical luminescence gold nano cluster probe |
| CN108982462A (en) * | 2018-09-12 | 2018-12-11 | 福建医科大学 | Sulfatase measuring method based on gold nano cluster Ratiometric fluorescent probe |
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