CN110186858A - A method of detection cysteine - Google Patents
A method of detection cysteine Download PDFInfo
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- CN110186858A CN110186858A CN201910537235.7A CN201910537235A CN110186858A CN 110186858 A CN110186858 A CN 110186858A CN 201910537235 A CN201910537235 A CN 201910537235A CN 110186858 A CN110186858 A CN 110186858A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
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Abstract
The present invention relates to a kind of methods for detecting cysteine, gold nano triangular plate solution is mixed with a series of cysteine solution of various concentrations, then NaAc_HAc buffer solution, TMB solution and the hydrogen peroxide of pH3.5-5.5 is added, after 10-50 DEG C of cultivation 5-30min, the absorption value at 652nm is detected, semicystinol concentration-absorption value standard curve is established；Then gold nano triangular plate solution is mixed with to test sample, NaAc_HAc buffer solution, TMB solution and the hydrogen peroxide of pH3.5-5.5 is added, after 10-50 DEG C of cultivation 5-30min, detect the absorption value at 652nm, and compared with semicystinol concentration-absorption value standard curve, it obtains to the semicystinol concentration in test sample.This method is without modifying sample, and detection process is easy, quick, and selectivity is high, the detection of semicystinol concentration suitable for the biological samples such as serum.
Description
Technical field
The present invention relates to a kind of methods of quickly detection cysteine, belong to technical field of chemical detection.
Background technique
Cysteine be it is a kind of important containing mercaptoamino acid in human body, play ten in the physiology of people and pathologic process
Divide important role, it can be adjusted plays in the activity of biological vivo protein and in the cell antioxidant defense system
Vital effect.The height of cysteine content can provide the foundation of science in life entity for the diagnosis of some diseases.
For example, the reduction of cysteine content in vivo will will lead to infant physical growth slow, pulmonary dysfunction etc.；And higher cysteine
Content is then closely related with the generation of certain cancers and cardiovascular disease, and abnormal cysteine content is also possible to cause AIDS
Disease, senile dementia, Parkinson's disease and renal failure etc..Therefore, the quantitative detection of cysteine is very significant, and also one
It has been exactly the hot spot of people's research since straight.
It mainly include enzyme parameters, high performance liquid chromatography, fluorescence in the prior art about the detection method of cysteine
Method etc., but these methods more or less come with some shortcomings, and for example, expensive equipment, cumbersome, time length needed etc. makes
It is restricted in practical applications, although electrochemical analysis method accuracy is high, measurement range is wide, and instrument and equipment is simpler,
It is at low cost, but poor selectivity, it cannot effectively distinguish the interference to cysteine such as ascorbic acid (AA), uric acid (UA).Therefore,
It is highly desirable to develop a kind of simple, quickly detection cysteine method.
Summary of the invention
It is an object of the invention to solve deficiency existing for cysteine detecting method in the prior art, a kind of detection is provided
The method of cysteine, this method is without modifying sample, and detection process is easy, quick, and speed is fast, and selectivity is high.
Technical solution
A method of detection cysteine includes the following steps:
(1) gold nano triangular plate solution is mixed with a series of cysteine solution of various concentrations, pH, which is then added, is
NaAc_HAc buffer solution, the 3,3' of 3.5-5.5,5,5'- tetramethyl benzidine (TMB) solution and hydrogen peroxide, gained is mixed
After zoarium ties up to 10-50 DEG C of cultivation 5-30min, the absorption value at 652nm is detected with ultraviolet spectrometry spectrometer, establishes half Guang ammonia
Acid concentration-absorption value standard curve；
(2) gold nano triangular plate solution is mixed with sample to be tested, the Acetic acid-sodium acetate that pH is 3.5-5.5 is then added
Buffer solution, 3,3', 5,5'- tetramethyl benzidine (TMB) solution and hydrogen peroxide cultivate gained mixed system at 10-50 DEG C
After 5-30min, detect absorption value at 652nm with ultraviolet spectrometry spectrometer, and with the semicystinol concentration-in step (1)
Absorption value standard curve compares, to obtain the semicystinol concentration in sample to be tested.
Further, it in step (1) and (2), the preparation method of the gold nano triangular plate solution: is sequentially added into test tube
Hexadecyltrimethylammonium chloride (CTAC) solution, the 4mLH of 800 μ L 0.1M2O, the HAuCl of 40 μ L 25.4mM4Solution, 37 μ L
The NaOH solution of the KI solution of 0.01M, 10 μ L 0.1M, stands 10min after shaking up, obtains growth-promoting media, and 50 μ L are added into growth-promoting media
The NaOH solution of the ascorbic acid solution of 64mM and 2.5 μ L 0.1M, stands 15min after shaking up, it is molten to obtain gold nano triangular plate
Liquid.
Further, in step (1) and (2), the concentration of the NaAc_HAc buffer solution is 0.05M, pH 4.2.
Further, in step (1) and (2), described 3,3', 5,5'- tetramethyl benzidine solution concentrations are 10-15mM.
Further, in step (1) and (2), the hydrogen peroxide concentration is 1-1.5M.
Further, in step (1) and step (2), cultivating temperature is 40 DEG C, and the cultivation time is 10min.
Beneficial effects of the present invention: the present invention provides a kind of method for detecting cysteine, this method is not necessarily to sample
It is modified, detection process is easy, quick, and speed is fast, and selectivity is high, and cysteine is dense suitable for the biological samples such as serum
The detection of degree.
Detailed description of the invention
Fig. 1 is the pH of NaAc_HAc buffer solution to AuNPs+TMB+H2O2The influence of system ultravioletvisible absorption value is bent
Line；
Fig. 2 is to cultivate temperature to AuNPs+TMB+H2O2The influence curve of system ultravioletvisible absorption value；
Fig. 3 is semicystinol concentration-absorption value canonical plotting in the embodiment of the present invention 3；
Fig. 4 is variety classes amino acid on the active influence of gold nano triangular plate.
Specific embodiment
The present invention will be further explained below with reference to the attached drawings and specific examples.
In following embodiments, the preparation method of gold nano triangular plate solution: sequentially add 800 μ L 0.1M's into test tube
Hexadecyltrimethylammonium chloride (CTAC) solution, 4mL H2O, the HAuCl of 40 μ L 25.4mM4The KI of solution, 37 μ L 0.01M
The NaOH solution of solution, 10 μ L 0.1M, stands 10min after shaking up, obtains growth-promoting media, is added 50 μ L 64mM's into growth-promoting media
The NaOH solution of ascorbic acid solution and 2.5 μ L 0.1M, stands 15min after shaking up, and obtains gold nano triangular plate solution.
Embodiment 1
The NaAc_HAc buffer solution of different pH is studied on the active influence of gold nano triangular plate:
To 100uL gold nano triangular plate solution, 300uL concentration is 3,3', 5, the 5'- tetramethyl biphenyl amine aqueous solutions of 12mM
(TMB) and 200uL concentration be 1.4M hydrogen peroxide mixed solution in, be separately added into a sequence difference pH (pH 3.5-5.5 it
Between) NaAc_HAc buffer solution, by gained mixed system after 40 DEG C of cultivation 10min, with ultraviolet spectrometry spectrometer detect
Absorption value at 652nm, the pH of NaAc_HAc buffer solution is to AuNPs+TMB+H2O2System ultravioletvisible absorption value
Influence curve is shown in Fig. 1.
As seen from Figure 1, when pH is 4.2, gold nano triangular plate has highest activity, and high activity is conducive to mention
The sensitivity of high detection.
Embodiment 2
Different cultivation temperature are studied on the active influence of gold nano triangular plate:
To 100uL gold nano triangular plate solution, 3,3', 5,5'- tetramethyl biphenyl amine aqueous solutions that 300uL concentration is 12mM,
The NaAc_HAc buffer solution that the hydrogen peroxide and pH4.2 concentration that 200 uL concentration are 1.4M are 0.05M, by gained mixture
System after 10 DEG C, 20 DEG C, 30 DEG C, 40 DEG C, 50 DEG C, 60 DEG C, 70 DEG C of cultivation 10min, is existed respectively with the detection of ultraviolet spectrometry spectrometer
Absorption value at 652nm cultivates temperature to AuNPs+TMB+H2O2The influence curve of system ultravioletvisible absorption value is shown in Fig. 2.
As seen from Figure 2, when cultivation temperature is 40 DEG C, gold nano triangular plate has highest activity, and high activity has
Conducive to the sensitivity for improving detection.
Embodiment 3
A method of detection cysteine includes the following steps:
(1) 100uL gold nano triangular plate solution is mixed with a series of cysteine solution of various concentrations of 100uL, is put
After setting 5min, the NaAc_HAc buffer solution for the pH 4.2 that 330uL concentration is 0.05M is added, 300uL concentration is 12 mM
3,3', 5,5'- tetramethyl biphenyl amine aqueous solutions and 200uL concentration be 1.4M hydrogen peroxide, by gained mixed system 40 DEG C train
After educating 10min, the absorption value at 652nm is detected with ultraviolet spectrometry spectrometer, it is bent to establish semicystinol concentration-absorption value standard
Line, as shown in Figure 3；
(2) gold nano triangular plate solution is directly mixed with sample to be tested (such as serum, drug), 330uL is then added
The 3,3' that the NaAc_HAc buffer solution for the pH 4.2 that concentration is 0.05M, 300uL concentration are 12mM, 5,5'- tetramethyl connection
The hydrogen peroxide that aniline (TMB) solution and 200uL concentration are 1.4M, by gained mixed system after 40 DEG C of cultivation 10min, use is ultraviolet
Sub-ray spectrometer detects absorption value at 652nm, and with the semicystinol concentration in step (1)-absorption value standard curve ratio
It is right, to obtain the semicystinol concentration in sample to be tested.
Test is tested using the selectivity of gold nano triangular plate detection cysteine:
100uL gold nano triangular plate solution is mixed with the different types of amino acid solution of 100uL 1.2mM, places 5min
Afterwards, the NaAc_HAc buffer solution for the pH 4.2 that 330uL concentration is 0.05M, 3,3' that 300uL concentration is 12mM is added,
The hydrogen peroxide that 5,5'- tetramethyl biphenyl amine aqueous solutions and 200uL concentration are 1.4M, by gained mixed system in 40 DEG C of cultivation 10min
Afterwards, the absorption value at 652nm is detected with ultraviolet spectrometry spectrometer.Figure is done to amino acid name with absorption value, obtains Fig. 4, Fig. 4
Be variety classes amino acid on the active influence of gold nano triangular plate, wherein 1-20 successively indicate water, cysteine, tyrosine,
Leucine, alanine, lysine, methionine, glutamine, aspartic acid, tryptophan, isoleucine, arginine, threonine,
Arginine monohydrochloride, phenylalanine, proline, glycine, histidine, serine, asparagine.
As seen from Figure 4, the only addition of cysteine, it is suppressed that the activity of gold nanoparticle makes the absorption of 652nm
It reduces, and other amino acid are added, and will not inhibit the activity of gold nanoparticle substantially, absorption value will not reduce or reduce amplitude
Far smaller than cysteine.
Claims (6)
1. a kind of method for detecting cysteine, which comprises the steps of:
(1) gold nano triangular plate solution is mixed with a series of cysteine solution of various concentrations, it is 3.5- that pH, which is then added,
5.5 NaAc_HAc buffer solution, 3,3', 5,5'- tetramethyl biphenyl amine aqueous solution and hydrogen peroxide, gained mixed system is existed
After 10-50 DEG C of cultivation 5-30min, the absorption value at 652nm is detected with ultraviolet spectrometry spectrometer, establishes semicystinol concentration-
Absorption value standard curve；
(2) gold nano triangular plate solution is mixed with sample to be tested, the Acetic acid-sodium acetate that pH is 3.5-5.5 is then added and buffers
Solution, 3,3', 5,5'- tetramethyl biphenyl amine aqueous solutions and hydrogen peroxide, by gained mixed system after 10-50 DEG C of cultivation 5-30min,
Detect absorption value at 652nm with ultraviolet spectrometry spectrometer, and with semicystinol concentration-absorption value standard in step (1)
Curve comparison, to obtain the semicystinol concentration in sample to be tested.
2. the method for detection cysteine as described in claim 1, which is characterized in that in step (1) and (2), the gold nano
The preparation method of triangular plate solution: hexadecyltrimethylammonium chloride solution, the 4mL of 800 μ L 0.1M are sequentially added into test tube
H2O, the HAuCl of 40 μ L 25.4mM4The NaOH solution of solution, the KI solution of 37 μ L 0.01M, 10 μ L 0.1M, stands after shaking up
10min obtains growth-promoting media, and the ascorbic acid solution of 50 μ L 64mM and the NaOH solution of 2.5 μ L 0.1M are added into growth-promoting media, shakes
15min is stood after even, obtains gold nano triangular plate solution.
3. the method for detection cysteine as described in claim 1, which is characterized in that in step (1) and (2), the acetic acid-vinegar
The concentration of sour sodium buffer solution is 0.05M, pH 4.2.
4. the method for detection cysteine as described in claim 1, which is characterized in that in step (1) and (2), described 3,3', 5,
5'- tetramethyl benzidine solution concentration is 10-15mM.
5. the method for detection cysteine as described in claim 1, which is characterized in that in step (1) and (2), the hydrogen peroxide
Concentration is 1-1.5M.
6. detecting the method for cysteine as described in any one of claim 1 to 5, which is characterized in that in step (1) and (2), training
Educating temperature is 40 DEG C, and the cultivation time is 10min.
Priority Applications (1)
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| CN201910537235.7A CN110186858A (en) | 2019-06-20 | 2019-06-20 | A method of detection cysteine |
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| CN201910537235.7A CN110186858A (en) | 2019-06-20 | 2019-06-20 | A method of detection cysteine |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104308179A (en) * | 2014-10-16 | 2015-01-28 | 苏州大学 | Method for quickly preparing high-yield gold triangular nanoprisms |
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- 2019-06-20 CN CN201910537235.7A patent/CN110186858A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104308179A (en) * | 2014-10-16 | 2015-01-28 | 苏州大学 | Method for quickly preparing high-yield gold triangular nanoprisms |
Non-Patent Citations (1)
| Title |
|---|
| RONG SHENG LI ET AL.: "Stable gold nanoparticles as a novel peroxidase mimic for colorimetric detection of cysteine", 《THE ROYAL SOCIETY OF CHEMISTRY》 * |
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Application publication date: 20190830 |