CN109596580A - Method based on glutamic-pyruvic transaminase in copper nanocluster fluorescence probe quantitative detection solution - Google Patents
Method based on glutamic-pyruvic transaminase in copper nanocluster fluorescence probe quantitative detection solution Download PDFInfo
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Abstract
The invention discloses a kind of methods based on glutamic-pyruvic transaminase in copper nanocluster fluorescence probe quantitative detection solution; it is protectant copper nano-cluster as fluorescence probe using glutathione, passes through glutamic-pyruvic transaminase content in fluorescence " off-on " mode specific detection solution.It is realized by using fluorescence " off-on " mode and carries out label-free, highly sensitive, selective detection to glutamic-pyruvic transaminase, this method is easy, fast, the specific detection to glutamic-pyruvic transaminase may be implemented, it is wide to detect the range of linearity, detection limit is low, the present invention have preferable detection sensitivity and selectivity, disease detection and in terms of have a good application prospect.
Description
This patent obtains state natural sciences fund general project 21375095, Tianjin Natural Science Fund In The Light youth's project
(No.17JCQNJC05800), Tianjin Normal University doctor fund (No.52XB1510), Tianjin Normal University are " inorganic-organic
Key lab, hydridization functional material chemical education portion ", " Tianjin functional molecular structure and performance focus laboratory " open base
The support of golden project and Tianjin Normal University " the following thousand people plan " project (WLQR201711, WLQR201814).
Technical field
The invention belongs to application field of the metallic copper nano-cluster in terms of fluorescence sense, and in particular to one kind passes through fluorescence
" off-on " mode is using copper nano-cluster as fluorescence probe, Gu Bingzhuan ammonia in label-free, efficient, specific detection complex system
Method in terms of enzyme content.
Background technique
Glutamic-pyruvic transaminase (ALT) also known as glutamate transaminase are a kind of important transaminases in human body, it can be catalyzed L- third
It is amino acid converting for a-ketoglutaric acid in propylhomoserin.Glutamic-pyruvic transaminase is located in the most cells of body, the concentration in liver
Highest.Earlier studies have shown that glutamic-pyruvic transaminase is the marker of liver diseases, glutamic-pyruvic transaminase raising is that liver function is asked
One important indicator of topic, all kinds of hepatitis can all cause glutamic-pyruvic transaminase to increase, after liver cell is seriously damaged, Gu Bingzhuan ammonia
The content of enzyme may rise to 50 times of customary amount, the degree phase one that the raised degree of glutamic-pyruvic transaminase is damaged with liver cell
It causes, this is because caused by glutamic-pyruvic transaminase is released in blood after liver is destroyed, the higher master of glutamic-pyruvic transaminase
If causing damages to liver, liver cell is caused constantly to damage, at the same can metabolism to liver and detoxification ability reduce, thus
So that drug metabolism and toxins in body cannot be discharged in time, the burden of liver is further aggravated, glutamic-pyruvic transaminase rises for a long time
Height can cause lesion, and serious person but will cause liver cancer.Therefore, the detection of glutamic-pyruvic transaminase content be for medical diagnosis on disease to
It closes important.Currently, the method for assessment glutamic-pyruvic transaminase content routine is spectrophotometry.However, this result may be not
Accurately, and the usual detection limit of these methods is higher or to need enzyme or cofactors etc. expensive reagent, therefore, to understand
Certainly these problems, and there is the method for many new detection glutamic-pyruvic transaminase contents to be established, such as: colorimetric method, chromatography
Deng, but due to its detect pre-treatment harsh conditions, detect purity high requirement, detection sensitivity is low the problems such as all limit
The extensive use of these methods.It is well known that fluorimetric method process is quick, it is convenient to operate, without complicated pre-treatment
Process, high sensitivity have lower detection limit, therefore, establish a kind of utilization fluorescence spectrophotometry analysis detection Gu Bingzhuan
The method of adnosine deaminase content is of great significance in terms of practical application.The present invention is closed by protective agent and reducing agent of glutathione
At copper nano-cluster, and by fluorescence " off-on " mode, using Tiopronin (Trp) as quencher, glutamic-pyruvic transaminase conduct
Restorative realizes and carries out unmarked, highly sensitive detection to the content of glutamic-pyruvic transaminase in solution.
Summary of the invention
It is an object of the invention to overcome the detection method of traditional complexity, establishes and a kind of utilize fluorescence spectrophotometry, side
Just, quickly in selective enumeration method complex system glutamic-pyruvic transaminase content method, the present invention provides one kind be based on gluathione
Peptide is the copper nano-cluster of stabilizer as fluorescence probe, is realized by fluorescence " off-on " mode and is exempted to glutamic-pyruvic transaminase
Label, highly sensitive, selective detection, this method is simple, quick, and the detection range of linearity is wide, and detection limit is low, adopts in the present invention
Glutamic-pyruvic transaminase is detected with copper nano-cluster, there is preferable detection sensitivity and selectivity, in disease detection and clinical application etc.
Aspect has a good application prospect.
To achieve the above object, the invention discloses one kind based on paddy third in copper nanocluster fluorescence probe quantitative detection solution
The method of transaminase, it is protectant copper nano-cluster as fluorescence probe using glutathione, passes through fluorescence " off-on " mode
Glutamic-pyruvic transaminase content in specific detection solution, it is characterised in that carried out by following step:
(1) Tiopronin mother liquor configures: weighing 0.1000 g Tiopronin and is dissolved in 5 mL high purity waters；By the concentration of mother liquor dilution
For the low concentration of 1 mg/mL, cryo-conservation is stand-by；
(2) the glutamic-pyruvic transaminase solution that concentration is 1,5,50,100,500,1000 U/L is respectively configured, cryo-conservation is stand-by；
(3) it is evenly spread to what is prepared in high purity water based on the copper nano-cluster that glutathione is stabilizer, is configured to concentration
For 0.675 mM, volume is the detection architecture of 4 mL, and utilizes the fluorescence intensity of fluorescent spectrophotometer assay at this time, is being excited
Under the excitation of 354 nm of wavelength, which shows to emit more by force at 632 nm；
(4) 1.2 mL copper nano-clusters are evenly spread in 2.6 mL high purity waters, after mixing, is added into mixed solution
The Tiopronin solution of 0.1 mL, 1 mg/mL increases 0.1 mL of pure water, sufficiently acts on to Tiopronin and copper nano-cluster solution
The fluorescence intensity of test system afterwards, obvious quenching occurs for fluorescence intensity at this time, therefore Tiopronin can be used as the detection architecture
Quencher；
(5) it is added into centrifuge tube in 2.6 mL high purity waters, 1.2 mL copper nano-cluster solution, after mixing, to mixed solution
The middle Tiopronin solution that 0.1 mL, 1 mg/mL is added sufficiently adds 0.1 mL glutamic-pyruvic transaminase solution after reaction, and sulphur is general
Luo Ningyu glutamic-pyruvic transaminase sufficiently acts on so that its fluorescence emission spectrum is restored and detected to fluorescence intensity, by with fluorescence in (4)
Intensity contrast, the recovery value that can use fluorescence emission spectral intensity prove that the copper nano-cluster detects Gu Bingzhuan as fluorescence probe
The feasibility of adnosine deaminase；
(6) the linear measurement of the content of glutamic-pyruvic transaminase in solution is detected
0.2 ~ 3.4 mL high purity water, 0.4 ~ 3.6 mL copper nano-cluster solution, 0.1 general sieve of mL sulphur are separately added into centrifuge tube
Peaceful solution is separately added into the glutamic-pyruvic transaminase solution of 0.1 mL various concentration, 1 ~ 1000 U/L into mixed solution, sufficiently reacts 1
~ 15 min detect the fluorescence intensity before and after glutamic-pyruvic transaminase is added with sepectrophotofluorometer respectively；The copper nano-cluster is
Refer to the copper nano-cluster based on glutathione for stabilizer.The experimental results showed that in glutamic-pyruvic transaminase concentration in 1-1000 U/L model
In enclosing, the fluorescence intensity recovery value of copper nano-cluster and the concentration of glutamic-pyruvic transaminase are presented linear relationship, linear equation be DF=
116.90744+0.54101X(DF being the fluorescence intensity after glutamic-pyruvic transaminase is added and copper nano-cluster after only addition Tiopronin
Fluorescence intensity obtains difference, and X is the concentration of glutamic-pyruvic transaminase), linearly dependent coefficient is 0 .992, and detection is limited to 0.61 U/L.
Copper nano-cluster solution referred to above is the copper nano-cluster based on glutathione for stabilizer, specific to synthesize
Method is shown in embodiment 1.
In terms of copper nano-cluster disclosed by the invention is as glutamic-pyruvic transaminase content in fluorescence probe specific detection solution
It is as follows using possessed good effect:
(1) the copper nano-cluster synthesized has stable optical property, and synthetic material partial size is small, and fluorescence property is good, synthetic method letter
It is single, quick, the complicated processes such as heating, adjusting pH, functionalization are not needed in synthesis process, synthetic material luminous position is 632
Nm, it is in the UV lamp it can be seen that apparent red.
(2) fluorescence " off on " is used using the copper nano-cluster with unique optical properties of synthesis as fluorescence probe
Mode, efficient selective sense the content of glutamic-pyruvic transaminase, and process is simple, quick, can be directly realized by glutamic-pyruvic transaminase
Selective enumeration method.
(3) using Tiopronin as quencher, the fluorescence intensity of copper nano-cluster is made to drop to lower numerical value, recycles paddy
Pyruvic transaminase makes fluorescence intensity be restored to high value as restorative, and the special of glutamic-pyruvic transaminase may be implemented in this kind of method
Property detection, detection the range of linearity it is wide, detection limit is low.
Detailed description of the invention
Fig. 1 is to illustrate to close using glutathione as the transmission electron microscope picture (TEM) of protective agent and the copper nano-cluster of reducing agent
At copper nanocluster size size uniformity, partial size is smaller and is evenly distributed；
Fig. 2 is to show using glutathione as the fluorescence excitation spectrum and launching light spectrogram of protective agent and the copper nano-cluster of reducing agent
Its maximum excitation wavelength is 354 nm, and maximum emission wavelength is 632 nm；
Fig. 3 is the feasibility point that the glutamic-pyruvic transaminase in solution is detected by the copper nano-cluster of protective agent and reducing agent of glutathione
Analysis；
Fig. 4 be using glutathione as the copper nano-cluster of protective agent and reducing agent, copper nano-cluster in Tiopronin and copper be added receive
The ultraviolet lamp picture of Tiopronin and glutamic-pyruvic transaminase is added in rice cluster；Wherein 1 expression copper nano-cluster is presented red in the UV lamp
Color, 2 indicate that solution becomes cloudy after Tiopronin is added, and solution reverts to original clarification shape again after glutamic-pyruvic transaminase is added in 3
State；
Fig. 5 is the linear graph that the glutamic-pyruvic transaminase in solution is detected by the copper nano-cluster of protective agent and reducing agent of glutathione,
The range of linearity is 1-1000 U/L, and detection limit is 0.61 U/L.
Specific embodiment
The present invention is described below by specific embodiment.Unless stated otherwise, technological means used in the present invention
It is method known in those skilled in the art.In addition, embodiment is interpreted as illustrative, it is not intended to limit the present invention
Range, the spirit and scope of the invention are limited only by the claims that follow.To those skilled in the art, without departing substantially from this
Under the premise of invention spirit and scope, to the various changes or change of material component and dosage progress in these embodiments
It belongs to the scope of protection of the present invention.Agents useful for same is that analysis is pure, and agents useful for same and manufacturer are as follows: glutathione, Beijing
Prosperity Bioisystech Co., Ltd, ancient cooking vessel state；Ascorbic acid, Tianjin Kermel Chemical Reagent Co., Ltd.；Copper chloride (99%), day
Saliva recovers Fine Chemical Co., Ltd；Sodium hydroxide, Tianjin Ke Wei Co., Ltd；Tiopronin, the raw work bioengineering in Shanghai
Limited liability company；Glutamic-pyruvic transaminase pacifies resistance to Jilin Chemical Science and Technology Ltd..The preparation method of copper nano-cluster can refer to (Wang,
C.; Ling, L.; Yao, Y.; Song, Q. Nano Research 2015,8(6), 1975-1986) or see implementation
Example 1.
Embodiment 1
It is carried out in accordance with the following steps under room temperature using glutathione as the preparation of the copper nano-cluster of stabilizer:
The preparation of (1) 0.1 M copper chloride solution: 1.7048 g CuCl are weighed2∙2H2O is dissolved in 100 mL high purity waters, sufficiently
It is spare after dissolution；
(2) it the preparation of copper nano-cluster: under room temperature, weighs 0.28 g of glutathione and is dissolved in 15 mL H2In O, thereto plus
Enter 450 mL CuCl2(0.1M) is added 0.1 g ascorbic acid (AA), adds 1 mL NaOH (1M) sufficiently after reaction,
1 h is reacted, until white suspension, which is completely dissolved, becomes light yellow clear transparent solutions, it was demonstrated that the formation of copper nano-cluster.By saturating
Electron microscope (TEM) (Fig. 1) is penetrated it can be seen that copper nano-cluster is uniformly dispersed, partial size is smaller.
Embodiment 2
Method of the copper nano-cluster as fluorescence probe specific detection glutamic-pyruvic transaminase, it is characterised in that in accordance with the following steps into
Row:
(1) Tiopronin mother liquor configures: weighing 0.1000 g Tiopronin and is dissolved in 5 mL high purity waters；By the dense of mother liquor dilution
Degree is the low concentration of 1 mg/mL, and cryo-conservation is stand-by；
(2) a series of preparation of glutamic-pyruvic transaminase various concentration solution:
The glutamic-pyruvic transaminase solution that concentration is 1,5,50,100,500,1000 U/L is respectively configured, cryo-conservation is stand-by；
(3) it is evenly spread to what is prepared in high purity water based on the copper nano-cluster that glutathione is stabilizer, is configured to concentration
For 0.675 mM, volume is the detection architecture of 4 mL, and utilizes the fluorescence intensity of fluorescent spectrophotometer assay at this time, is being excited
Under the excitation of 354 nm of wavelength, which shows to emit more by force at 632 nm；
(4) 1.2 mL copper nano-clusters are evenly spread in 2.6 mL high purity waters, after mixing, is added into mixed solution
The Tiopronin solution of 0.1 mL, 1 mg/mL increases 0.1 mL of pure water, sufficiently acts on to Tiopronin and copper nano-cluster solution
The fluorescence intensity of test system afterwards, obvious quenching occurs for fluorescence intensity at this time, therefore Tiopronin can be used as the detection architecture
Quencher；
(5) it is added into centrifuge tube in 2.6 mL high purity waters, 1.2 mL copper nano-cluster solution, after mixing, to mixed solution
The middle Tiopronin solution that 0.1 mL, 1 mg/mL is added sufficiently adds 0.1 mL glutamic-pyruvic transaminase solution after reaction, and sulphur is general
Luo Ningyu glutamic-pyruvic transaminase sufficiently acts on so that fluorescence restores and detects its fluorescence emission spectrum, by with fluorescence intensity in (4)
Comparison, the recovery value that can use fluorescence emission spectral intensity prove that the copper nano-cluster detects glutamic-pyruvic transaminase as fluorescence probe
Feasibility；
(6) measurement that glutamic-pyruvic transaminase content is linear in solution is detected
It is separately added into 2.6 mL high purity waters into centrifuge tube, 1.2 bronze medal nano-cluster solution, 0.1 mL Tiopronin solution, to mixed
The glutamic-pyruvic transaminase solution that 0.1 mL various concentration, 1 ~ 1000 U/L is separately added into solution is closed, sufficiently 1 ~ 15 min of reaction, point
The fluorescence intensity before and after glutamic-pyruvic transaminase is added is not detected with sepectrophotofluorometer.The experimental results showed that in glutamic-pyruvic transaminase
For concentration within the scope of 1-1000 U/L, linear close is presented in the fluorescence intensity recovery value of copper nano-cluster and the concentration of glutamic-pyruvic transaminase
System, linear equation be DF=116.90744+0.54101X(DF be added glutamic-pyruvic transaminase after fluorescence intensity with only addition sulphur it is general
The fluorescence intensity of Luo Ninghou copper nano-cluster obtains difference, and X is the concentration of glutamic-pyruvic transaminase), linearly dependent coefficient 0.992, detection
It is limited to 0.61 U/L.
Embodiment 3
It 1, is the preparation reference embodiment 1 of protectant copper nano-cluster with glutathione；
2, using glutathione as the measurement of the excitation spectrum and emission spectrum of the copper nano-cluster of stabilizer:
Copper nano-cluster is distributed in high purity water, the fluorescence excitation spectrum of material and fluorescence emission spectrum are measured, such as Fig. 2
Shown, the maximum excitation wavelength of copper nano-cluster is 354 nm, under the excitation of maximum excitation wavelength, fluorescence emission wavelengths 632
nm。
Embodiment 4
It 1, is the preparation reference embodiment 1 of protectant copper nano-cluster with glutathione；
2, it uses fluorescence for means of testing, utilizes with glutathione as protectant copper nano-cluster specific detection Gu Bingzhuan ammonia
Enzyme:
Take 2 empty centrifuge tubes respectively, number 1., 2., pipette respectively 2.6 mL of high purity water be added to 1., 2. in number centrifuge tube, then
It pipettes 1.2 mL copper nano-cluster solution to be separately added into different centrifuge tubes, after mixing, continues to add into 1. number centrifuge tube
Enter 0.1 mL Tiopronin, then 0.1 mL high purity water is added thereto as blank control group, is added into 2. number centrifuge tube
0.1 mL Tiopronin, then 0.1 mL glutamic-pyruvic transaminase solution is added thereto, 10 min are sufficiently reacted, so that fluorescence occurs
Restore and detect fluorescent emission intensity with fluophotometer, excitation wavelength is 354 nm, and launch wavelength is 632 nm；Detection is limited to
0.61 U/L
Embodiment 5
It 1, is the preparation reference embodiment 1 of protectant copper nano-cluster with glutathione；
2, under ultraviolet lamp using glutathione as in the copper nano-cluster of protective agent and reducing agent, copper nano-cluster be added Tiopronin and
The sample luminous situation of Tiopronin and glutamic-pyruvic transaminase is added in copper nano-cluster:
3 empty centrifuge tubes are taken respectively, are numbered (1), (2), (3), are pipetted 2.6 mL of high purity water respectively and be added to (1), (2), (3) number
In centrifuge tube, then pipette 1.2 mL copper nano-cluster solution and be added in different centrifuge tubes, after mixing, continue to (1) number from
0.2 mL high purity water is added in heart pipe, 0.1 mL Tiopronin is added into (2) number centrifuge tube, then 0.1 mL is added thereto
0.1 mL Tiopronin is added into (3) number centrifuge tube for high purity water, then 0.1 mL glutamic-pyruvic transaminase solution is added thereto, fills
Divide reaction 10 min so that fluorescence occurs to restore simultaneously to detect its luminous situation with ultraviolet lamp box.As shown in figure 4,1 expression copper is received
Red clear state is presented in rice cluster in the UV lamp, and solution becomes cloudy (2) after Tiopronin is added, and glutamic-pyruvic transaminase is added in 3
Solution reverts to original clear state again afterwards.
Embodiment 6
It 1, is the preparation reference embodiment 1 of protectant copper nano-cluster with glutathione；
2, it using the means of testing of fluorescence, utilizes with glutathione as the paddy in protectant copper nano-cluster specific detection solution
Pyruvic transaminase feasibility: taking 2 empty centrifuge tubes respectively, 1., 2. number, pipettes 2.2 mL of high purity water respectively and is added to 1., 2. number
In centrifuge tube, then pipette 1.6 mL copper nano-cluster solution and be added in different centrifuge tubes, after mixing, continue to 1. number from
0.1 mL Tiopronin is added in heart pipe, 0.1 mL is added into 2. number centrifuge tube as blank control group in 0.1 mL high purity water
Tiopronin adds 0.1 mL glutamic-pyruvic transaminase solution, reacts 10 min, so that fluorescence restores and examined with fluophotometer
Survey fluorescent emission intensity；Detection is limited to 0.61 U/L.
Embodiment 7
The detection of glutamic-pyruvic transaminase: taking 2 empty centrifuge tubes respectively, 1., 2. number, pipettes 0.4 mL of high purity water respectively and is added to
1., 2. in number centrifuge tube, then pipette 3.2 mL copper nano-cluster solution and be added in different centrifuge tubes, after mixing, continue to
1. 0.1 mL Tiopronin is added in number centrifuge tube, 0.1 mL high purity water is added as blank control group into 2. number centrifuge tube
0.1 mL Tiopronin, adds 0.1 mL glutamic-pyruvic transaminase solution, reacts 10 min, so that fluorescence restores and with fluorescence light
Degree meter detection fluorescent emission intensity；Detection is limited to 0.61 U/L.
Embodiment 8
The detection of glutamic-pyruvic transaminase: taking 2 empty centrifuge tubes respectively, 1., 2. number, pipettes 2.8 mL of high purity water respectively and is added to
1., 2. in number centrifuge tube, then pipette 1.0 mL copper nano-cluster solution and be added in different centrifuge tubes, after mixing, continue to
1. 0.1 mL Tiopronin is added in number centrifuge tube, 0.1 mL high purity water is added as blank control group, to 2. number centrifuge tube
0.1 mL Tiopronin of middle addition adds 0.1 mL glutamic-pyruvic transaminase, reacts 10 min, so that fluorescence occurs quenching and is used in combination
Fluophotometer detects fluorescent emission intensity.Detection is limited to 0.61 U/L.
Claims (2)
1. a kind of method based on glutamic-pyruvic transaminase in copper nanocluster fluorescence probe quantitative detection solution, it is to protect with glutathione
The copper nano-cluster of agent is protected as fluorescence probe, is contained by glutamic-pyruvic transaminase in fluorescence " off-on " mode specific detection solution
Amount, it is characterised in that carried out by following step:
The configuration of Tiopronin mother liquor: it weighs 0.1000 g Tiopronin and is dissolved in 5 mL high purity waters；It is 1 by the concentration of mother liquor dilution
The low concentration of mg/mL, cryo-conservation are stand-by；
The glutamic-pyruvic transaminase solution that concentration is 1,5,50,100,500,1000 U/L is respectively configured, cryo-conservation is stand-by；
(3) it is evenly spread to what is prepared in high purity water based on the copper nano-cluster that glutathione is stabilizer, is configured to concentration
For 0.675 mM, volume is the detection architecture of 4 mL, and utilizes the fluorescence intensity of fluorescent spectrophotometer assay at this time, is being excited
Under the excitation of 354 nm of wavelength, which shows to emit more by force at 632 nm；
(4) 1.2 mL copper nano-clusters are evenly spread in 2.6 mL high purity waters, after mixing, is added into mixed solution
The Tiopronin solution of 0.1 mL, 1 mg/mL increases 0.1 mL of pure water, sufficiently acts on to Tiopronin and copper nano-cluster solution
The fluorescence intensity of test system afterwards, obvious quenching occurs for fluorescence intensity at this time, therefore Tiopronin can be used as the detection architecture
Quencher；
(5) it is added into centrifuge tube in 2.6 mL high purity waters, 1.2 mL copper nano-cluster solution, after mixing, to mixed solution
The middle Tiopronin solution that 0.1 mL, 1 mg/mL is added sufficiently adds 0.1 mL glutamic-pyruvic transaminase solution after reaction, and sulphur is general
Luo Ningyu glutamic-pyruvic transaminase sufficiently acts on so that its fluorescence emission spectrum is restored and detected to fluorescence intensity, by with fluorescence in (4)
Intensity contrast, the recovery value that can use fluorescence emission spectral intensity prove that the copper nano-cluster detects Gu Bingzhuan as fluorescence probe
The feasibility of adnosine deaminase；
(6) the linear measurement of the content of glutamic-pyruvic transaminase in solution is detected
0.2 ~ 3.4 mL high purity water, 0.4 ~ 3.6 mL copper nano-cluster solution, 0.1 general sieve of mL sulphur are separately added into centrifuge tube
Peaceful solution is separately added into the glutamic-pyruvic transaminase solution of 0.1 mL various concentration, 1 ~ 1000 U/L into mixed solution, sufficiently reacts 1
~ 15 min detect the fluorescence intensity before and after glutamic-pyruvic transaminase is added with sepectrophotofluorometer respectively；The copper nano-cluster is
Refer to the copper nano-cluster based on glutathione for stabilizer.
2. the method based on glutamic-pyruvic transaminase in copper nanocluster fluorescence probe quantitative detection solution described in claim 1, detection
The range of linearity is wide, and detection is limited to 0.61 U/L.
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| CN111157505A (en) * | 2020-01-17 | 2020-05-15 | 天津师范大学 | Method for detecting sulfur-containing pollutant thioglycollic acid in solution |
| CN113189232A (en) * | 2021-04-28 | 2021-07-30 | 玉林市食品药品检验检测中心 | Method for determining glutathione, tiopronin and inosine in liver-protecting Chinese patent medicine |
| CN116514901A (en) * | 2023-07-04 | 2023-08-01 | 北京建工环境修复股份有限公司 | Double-response fluorescent iron nanocluster probe and preparation method and application thereof |
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| CN116514901A (en) * | 2023-07-04 | 2023-08-01 | 北京建工环境修复股份有限公司 | Double-response fluorescent iron nanocluster probe and preparation method and application thereof |
| CN116514901B (en) * | 2023-07-04 | 2023-09-29 | 北京建工环境修复股份有限公司 | Double-response fluorescent iron nanocluster probe and preparation method and application thereof |
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