CN107014787A - A kind of application of the stable gold nanoclusters of glutathione in detection cysteine and lysine content - Google Patents
A kind of application of the stable gold nanoclusters of glutathione in detection cysteine and lysine content Download PDFInfo
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Abstract
The invention discloses a kind of application of the stable gold nanoclusters of glutathione in detection cysteine and lysine content.By the freshly prepd glutathione aqueous solution and tetra chlorauric acid aqueous solution isoconcentration, mix with vigorous stirring in equal volume, then mixture is reacted 25 days under illumination condition, obtain pale yellow solution, centrifugation, supernatant is purified by dialyser, the gold nanoclusters of the glutathione stabilization prepared, it can separately or concurrently detect the application in cysteine and lysine as fluorescence probe and ratio fluorescent probe, with the characteristic such as high selectivity, high sensitivity, simplicity, easy, preparation method of the present invention is easy, green, gentle.
Description
Technical field
The invention belongs to amino acid detection field, and in particular to a kind of stable gold nanoclusters of glutathione detect half Guang ammonia
Application in acid and lysine content.
Background technology
Amino acid is that 20 kinds of amino acid have each not in the basic composition unit of biological function macro-molecular protein, human body
Same function, mutually promotes and influences each other.If human body lacks any essential amino acid, so that it may cause physiological function different
Often, what influence antibody was metabolized is normally carried out, and finally results in disease.Disease caused by amino acid starvation and dysbolism is related to
And the various diseases such as immune, nervous system, cardiovascular and cerebrovascular, kidney, metabolism, diabetes, influence growth in humans development, nutrition are good for
Each healthy link such as health, muscle skeleton growth, hormone secretion, function of detoxification.Cysteine is a kind of with physiological function
Amino acid, be constitutive protein matter 20 several amino acids in the only amino acid with reproducibility group sulfydryl (- SH), sulfydryl energy
With with Ag+、Hg2+、Cu2+、Pb2+Combined etc. heavy metal ion, be condensed with poisonous aromatic compound and play detoxication,
Also there is anti-oxidation function to remove interior free yl simultaneously.Lysine is one of essential amino acid, a kind of indispensable
Nutriment.Body development, enhancing immunologic function can be promoted, and be improved the effect of central nervous tissue function.It is wide at present
It is general to be applied to medicine, food and feed industry etc..Therefore, detection cysteine and lysine content are diagnosed to clinical medicine, eaten
Product examine is tested and industrial production etc. is all significant.
Measurement cysteine, the method for lysine content mainly have analytical photometry, High Performance Liquid Chromatography/Mass Spectrometry at present
Method, capillary electrophoresis, Flow Injection Analysis etc..Cysteine, the detection of lysine are generally by reaction time length, reaction condition
Such as pH value acid-base property, complex operation, sensitivity and the low influence of selectivity.Find new highly sensitive, high selectivity, it is simplicity, fast
Speed and can detect that the new method of cysteine and lysine content has important Research Significance simultaneously.
The content of the invention
Goal of the invention：In view of the problem of presently, there are with the detection method of cysteine, lysine content, the present invention is carried
For the application in a kind of glutathione stable gold nanoclusters detection cysteine and lysine content, the glutathione is prepared steady
The method of fixed gold nanoclusters is easy, green, gentle, and detection method has the spy such as high selectivity, high sensitivity, simplicity, easy
Property.
Technical scheme：
The stable gold nanoclusters of glutathione answering in detection cysteine content as fluorescence probe and ratio fluorescent probe
With.
The stable gold nanoclusters of glutathione are as fluorescence probe and ratio fluorescent probe in detection lysine content
Using.
The stable gold nanoclusters of glutathione as fluorescence probe and ratio fluorescent probe detect at the same time cysteine and
Application in lysine content.
The method that the stable gold nanoclusters of glutathione detect cysteine content, comprises the following steps：
Step (a1)：Under the exciting of 402nm wavelength lights, the stable gold nanoclusters of glutathione are determined at 570nm wavelength
Fluorescence intensity, is designated as I0；
Step (a2)：The Guang ammonia of various concentrations testing sample half is added into the stable gold nanoclusters of step (a1) glutathione
After acid, the stable gold nanoclusters of the glutathione containing various concentrations cysteine are determined in the case where 402nm wavelength lights are excited,
Fluorescence intensity at 570nm wavelength, is designated as I570, according to adding after various concentrations cysteine, the changing value of fluorescence intensity is counted
Calculate the changing value and the line of semicystinol concentration for adding the stable gold nanoclusters fluorescence intensity of glutathione before and after cysteine
Sexual intercourse；Step (a3)：Testing sample is added in the stable gold nanoclusters of glutathione, tested before and after it is added in 402nm
Under the exciting of wavelength light, the changing value of the fluorescence intensity at 570nm wavelength, the linear relationship obtained according to step (a2), meter
Calculate the quality or concentration for adding cysteine.
The specific continuous mode of step (a2) is as follows：The gold nanoclusters sample for taking the glutathione of 300 μ L preparations stable, plus
Enter into centrifuge tube, then add the cysteine working solution of appropriate volume, 3mL is diluted to distilled water, be well mixed, enter
Row is determined.Under the exciting of 402nm wavelength lights, its fluorescence spectrum is tested, the slit width of instrument is set as 5nm, experiment hair
There is maximum emission peak at present 570nm wavelength.Add after cysteine, test sample causes the change of fluorescence probe, record exists
The fluorescence intensity of the maximum emission peak position, is expressed as：Δ I=I570–I0, wherein, I0And I570Respectively glutathione is stablized
Gold nanoclusters itself and the fluorescence intensity added after cysteine.
The method that the stable gold nanoclusters of glutathione detect lysine content, comprises the following steps：
Step (b1)：Under the exciting of 402nm wavelength lights, the stable gold nanoclusters of glutathione are determined at 570nm wavelength
Fluorescence intensity, is designated as I0；
Step (b2)：Various concentrations testing sample lysine is added into the stable gold nanoclusters of step (b1) glutathione
Afterwards, the stable gold nanoclusters of the glutathione containing various concentrations lysine are determined in the case where 402nm wavelength lights are excited, in 473nm
Fluorescence intensity at wavelength, is designated as I473, the fluorescence intensity at 570nm wavelength is designated as I570, rely ammonia according to various concentrations are added
After acid, the changing value of fluorescence intensity, calculate add the stable gold nanoclusters of glutathione before and after lysine 473nm with
Fluorescence intensity ratio (is designated as I at 570nm473/I570) changing value and lysine concentration linear relationship；
Step (b3)：Testing sample is added in the stable gold nanoclusters of glutathione, tested before and after it is added in 402nm wavelength
Under the exciting of light, the changing value of the fluorescence intensity at 473nm and 570nm wavelength, the linear relationship obtained according to step (b2),
Calculate the quality or concentration for adding lysine.
The specific continuous mode of step (b2) is as follows：The gold nanoclusters sample for taking the glutathione of 300 μ L preparations stable, plus
Enter into centrifuge tube, then add the lysine working solution of appropriate volume, 3mL is diluted to distilled water, be well mixed, carry out
Determine.Under the exciting of 402nm wavelength lights, its fluorescence spectrum is tested, the slit width of instrument is set as 5nm, and experiment is found
There are two fluorescence emission peaks at 473nm and 570nm wavelength respectively.Add after lysine, test sample causes fluorescence probe
Change, records the fluorescence intensity ratio in 473nm and 570nm emitted at wavelengths peak position, is expressed as：I473/I570, wherein, I473
And I570Respectively the stable gold nanoclusters of glutathione add new emission peak fluorescence intensity and glutathione stabilization after lysine
The original emission peak fluorescence intensity of gold nanoclusters.
The method that the stable gold nanoclusters of glutathione detect cysteine and lysine content simultaneously, including following step
Suddenly：
Step (c1)：Under the exciting of 402nm wavelength lights, the stable gold nanoclusters of glutathione are determined at 570nm wavelength
Fluorescence intensity, is designated as I0；
Step (c2)：Two kinds of various concentrations, which are separately added into, in the stable gold nanoclusters of step (c1) glutathione treats test sample
After product cysteine and lysine, the stable gold nanoclusters of the glutathione containing various concentrations cysteine and lysine are determined
In the case where 402nm wavelength lights are excited, the fluorescence intensity at 570nm and 473nm wavelength respectively, the fluorescence at 570nm wavelength is strong
Degree, is designated as I570, the fluorescence intensity at 473nm wavelength is designated as I473.According to adding after various concentrations cysteine, fluorescence is strong
Degree fluorescence intensity at 570nm (is designated as I570) changing value, calculate and add the stable Jenner of glutathione before and after cysteine
The changing value and the linear relationship of semicystinol concentration of rice cluster fluorescence intensity；According to adding after various concentrations lysine, fluorescence is strong
The changing value of degree, calculates the gold nanoclusters of glutathione stably fluorescence intensity at 473nm before and after adding lysine and (is designated as
I473) changing value and lysine concentration linear relationship；
Step (c3)：Testing sample is added in the stable gold nanoclusters of glutathione, tested before and after it is added in 402nm wavelength
Under the exciting of light, the changing value of the fluorescence intensity at 473nm and 570nm wavelength, the linear relationship obtained according to step (c2),
The quality or concentration for adding cysteine and lysine are calculated respectively.
The specific continuous mode of step (c2) is as follows：
The gold nanoclusters sample for taking the glutathione of 300 μ L preparations stable, is added in centrifuge tube, then adds appropriate volume
Cysteine and lysine working solution, 3mL is diluted to distilled water, is well mixed, is measured.In 402nm wavelength lights
Excite down, test its fluorescence spectrum, the slit width of instrument is set as 5nm, and experiment is found respectively in 473nm and 570nm ripples
Strong point has two fluorescence emission peaks.The change for causing fluorescence probe after testing sample is added, is recorded in 473nm and 570nm wavelength
Locate the fluorescence intensity of emission peak positions, be expressed as：I473、I570, wherein, I473And I570The gold nano that respectively glutathione is stablized
Cluster adds new emission peak fluorescence intensity and the fluorescence intensity added after cysteine after lysine.
A kind of preparation method of the stable gold nanoclusters of glutathione, using glutathione as template, specifically includes following step
Suddenly：It is 1 that the freshly prepd glutathione aqueous solution and the tetra chlorauric acid aqueous solution are pressed into concentration ratio:1st, volume ratio is 1:1 ratio exists
Lower mixing is stirred vigorously, then mixture is reacted 2-5 days under illumination condition, pale yellow solution is obtained, is centrifuged, supernatant leads to
Dialyser purifying is crossed, the stable gold nanoclusters of glutathione are prepared.
The reaction temperature reacted under the illumination condition is 25-35 DEG C.
The gold nanoclusters for the glutathione stabilization that the preparation method is prepared, are presented spheric granules, are evenly distributed, have
There is good monodispersity, nano particle average diameter is about 2.2nm or so, size uniformity, with stable fluorescent characteristic.
Beneficial effect：
1st, the stable gold nanoclusters preparation method of glutathione of the invention is easy, green, gentle, the glutathione prepared
Detection of the stable gold nanoclusters to cysteine and lysine has the characteristic such as high selectivity, high sensitivity, simplicity, easy.
2nd, the detection method that the present invention is provided may apply to the measure of multiple fields, including food, medicine and animal are raised
In the detection for expecting cysteine and lysine content.
3rd, setting up for the detection method that the present invention is provided can carry for highly sensitive, the quick detection of cysteine and lysine
For new approaches.
Brief description of the drawings
Fig. 1 is ultraviolet-visible absorption spectroscopy, the fluorescent exciting of the stable gold nanoclusters of glutathione made from embodiment 1
Spectrum and fluorescence emission spectrum；
Fig. 2 is the transmission electron microscope picture of the stable gold nanoclusters of glutathione made from embodiment 1；
Fig. 3 is that the stable gold nanoclusters of glutathione add cysteine and the fluorescence emission spectrum added after lysine；
Fig. 4 a are the stable gold nanoclusters fluorescence intensity of glutathione with relation (the standard song for adding semicystinol concentration changing value
Line)；
Fig. 4 b are the stable gold nanoclusters fluorescence intensity of glutathione with relation (the standard song for adding lysine concentration changing value
Line).
Embodiment：
The embodiment of the present invention is described in detail below in conjunction with accompanying drawing.It is stable using glutathione in the present invention
Gold nanoclusters determine cysteine and lysine process be based on glutathione be template and reducing agent, in neutrallty condition
Under, the sulfydryl and Au of cysteine in glutathione3+Effect is reduced to zeroth order gold, while being used as paddy Guang of the template to formation
The gold nanoclusters of sweet stabilized peptide play stabilization.When adding cysteine, cysteine can pass through sulfydryl and gluathione
The gold nanoclusters of stabilized peptide, which are combined, reduces the Fluorescence Increasing that its surface defect causes the stable gold nanoclusters of glutathione；Work as addition
During lysine, lysine is assembled and being attached to the stable gold nanoclusters surface of glutathione makes it produce new fluorescence emission peak.
Material, reagent used in following embodiments etc. unless otherwise specified, are commercially obtained.
Quantitative determination experiment in following embodiments, is three repetition experiments, results averaged.
The preparation of the stable gold nanoclusters of the glutathione of embodiment 1 and sign
1st, by template of glutathione the preparation of gold nanoclusters (GSH-Au NCs)
There are the gold nanoclusters of fluorescent characteristic by templated synthesis of glutathione：The synthesis of fluorescence gold nanoclusters is according to document report
Road method [referring to：Chen W.B., Tu X.J., Guo X.Q.Chem.Commun., 2009,1736-1738.], do one
A little to improve, process is as follows：First, by the freshly prepd glutathione aqueous solution (5mM, 10mL) and the tetra chlorauric acid aqueous solution (5Mm,
10mL) mix, then react the mixture 2~5 days with vigorous stirring, obtain product for pale yellow solution.In this experiment,
Reactant feed concentration ratio is 1:1, reaction condition is：30 DEG C of reaction temperature, reaction environment is daylight illumination or fluorescent lamp illumination
Condition, after improvement, generated time is shorten to 2~5 days, the stable gold nanoclusters size uniformity of the glutathione prepared, and is had
Stable fluorescent characteristic.Product is centrifuged 20 minutes under rotating speed 15000rpm, supernatant passes through dialyser (molecular cut off
12000Da) it is further purified, the stable gold nanoclusters of the glutathione of preparation are kept in dark place standby under the conditions of 4 DEG C.
2nd, using glutathione as the sign of the gold nanoclusters of template
GSH-Au NCs fluorescence is characterized using XRF (Jasco FP-6500).By GSH-Au NCs with
Phosphate buffer solution (50mM PBS, pH 7.0) presses 1:1 dilution, takes 3mL in the cuvette of four sides printing opacity, sets luminoscope
Excitation wavelength 402nm, slit width 5nm, in wave-length coverage 480-680nm scanning record fluorescence spectrum.Take 10 μ L GSH-
Au NCs solution is diluted to 3mL in centrifuge tube, adds to be measured in cuvette, addition 3mL ultra-pure water conducts in another cuvette
Correcting sample, is scanned using ultraviolet-visible spectrometer (UV-2450) in 200-800nm wave-length coverages to sample.
Experiment measures GSH-Au NCs fluorescence spectrum and ultraviolet-visible spectrum, as shown in figure 1, GSH-Au NCs' swashs
Hair wavelength is 402nm, and launch wavelength is 570nm, and has larger Stokes shift (168nm).
This experiment prepare GSH-Au NCs present spheric granules, as shown in Fig. 2 and be evenly distributed, with good list
Dispersiveness.Nano particle average diameter is about 2.2nm or so.
The stable gold nanoclusters fluorescence probe detection cysteine of the glutathione of embodiment 2
The present embodiment will be apparent from how detecting half using the gold nanoclusters of the glutathione template prepared in embodiment 1
The process of cystine.
Utilize the gold nanoclusters probe in detecting cysteine specific steps that the fluorescence glutathione prepared is stable：Take
The gold nanoclusters sample of glutathione stabilization prepared by 300 μ L, is added in centrifuge tube, then adds a series of half Guang of concentration
Propylhomoserin working solution, detects the fluorescence signal for the gold nanoclusters probe that various concentrations cysteine is stablized to glutathione respectively
Influence (final concentration of 0,5,10,15,20,25,50,75,100,125,150,175,200,225,250,300 μM).Steamed with double
Water is diluted to final volume for 3mL, is well mixed, is measured.Under the exciting of 402nm wavelength lights, test glutathione is stable
Gold nanoclusters fluorescence spectrum, the slit width of instrument is set as the emission peak fluorescence intensity at 5nm, detection 570nm wavelength.
Test adds the fluorescence signal of sample after cysteine, records the fluorescence intensity in the maximum emission peak position, is expressed as：ΔI
=I570–I0, wherein, I0And I570The respectively stable gold nanoclusters itself of glutathione and the fluorescence added after cysteine are strong
Degree.Experiment is repeated 3 times, and as a result takes its average value.
As a result show, the gold nanoclusters fluorescence probe of the glutathione stabilization of the glutathione template prepared in embodiment 1,
Add after cysteine, Enhancement of Fluorescence occurs in the fluorescence signal of probe, refers to Fig. 3 and Fig. 4 a, and as cysteine is dense
The increase of degree, fluorescence probe signal intensity strengthens therewith, in 1-165 μM of concentration range, semicystinol concentration and probe it is glimmering
Luminous intensity shows good linear relationship, and its detection is limited to 50nM.
The stable gold nanoclusters ratio fluorescent probe in detecting lysine of the glutathione of embodiment 3
The present embodiment, which will be apparent from how detecting using the gold nanoclusters of the glutathione template prepared in embodiment 1, to be relied
The process of propylhomoserin.
Utilize the gold nanoclusters ratio fluorescent probe in detecting lysine specific steps that the glutathione prepared is stable：Take
The gold nanoclusters sample of glutathione stabilization prepared by 300 μ L, is added in centrifuge tube, then adds a series of bad ammonia of concentration
Sour working solution, detects influence of the various concentrations lysine to the fluorescence signal of the stable gold nanoclusters probe of glutathione respectively
(final concentration of 0,2.5,5,7.5,10,12.5,15,17.5,20,22.5,25 μM).It is 3mL that final volume is diluted to distilled water,
It is well mixed, it is measured.Under the exciting of 402nm wavelength lights, the fluorescence spectrum of the stable gold nanoclusters of test glutathione,
The slit width of instrument is set as 5nm, and experiment finds there are two fluorescence emission peaks at 473nm and 570nm wavelength respectively.Add
After lysine, test sample causes the change of fluorescence probe, records the fluorescence in 473nm and 570nm emitted at wavelengths peak position
Intensity rate, is expressed as：I473/I570, wherein, I473And I570The respectively stable gold nanoclusters of glutathione are added after lysine
New emission peak fluorescence intensity and the stable original emission peak fluorescence intensity of gold nanoclusters of glutathione.Experiment is repeated 3 times, knot
Fruit takes its average value.
As a result show, the gold nanoclusters ratio fluorescent probe of the glutathione template prepared in embodiment 1, add lysine
Afterwards, in the case where same excitation wavelength 402nm is excited, probe produces new emission peak at 473nm, and fluorescence occurs in fluorescence signal
Strengthen the emission peak at phenomenon, the stable gold nanoclusters 570nm of script glutathione and change little, only slight decrease, in detail
See Fig. 3 and Fig. 4 b, with the increase of lysine concentration, fluorescence signal intensity strengthens therewith at probe 473nm, according to 473nm and
Relation between the fluorescence intensity ratio of 570nm emitted at wavelengths peak position, in 0.02-5 μM of concentration range, lysine concentration with
The fluorescence ratio of ratio fluorescent probe shows good linear relationship, and its detection is limited to 2nM.It can thus be seen that according to reality
The gold nanoclusters ratio fluorescent probe for the glutathione stabilization that example 1 is obtained is applied, it is glimmering at two different wave lengths of probe by measuring
Light changes, using its ratio as signal parameter, is not influenceed by intensity of light source fluctuation and instrumental sensitivity.With traditional ratio fluorescent
Probe is compared, and extra fluorescent dye or other fluorescent nano materials without introducing, cost-effective, environmental protection, this experiment are carried
A kind of easy, high accuracy and the stable gold nanoclusters ratio fluorescent probe of highly sensitive new glutathione have been supplied, and has been carried
A kind of new method of the stable gold nanoclusters ratio fluorescent probe in detecting lysine of glutathione is supplied.
The stable gold nanoclusters fluorescence probe of the glutathione of embodiment 4 detects cysteine and lysine simultaneously
The present embodiment will be apparent from how examining simultaneously using the gold nanoclusters of the glutathione template prepared in embodiment 1
Survey the process of cysteine and lysine.
Using the stable gold nanoclusters ratio fluorescent probe of the glutathione prepared while detecting cysteine and relying
Propylhomoserin specific steps：The gold nanoclusters sample for taking the glutathione of 300 μ L preparations stable, is added in centrifuge tube, then adds
The cysteine and lysine working solution of a series of concentration, its step be the same as Example 2 and embodiment 3, detect different dense respectively
Spend the influence of cysteine and lysine to the fluorescence signal of the stable gold nanoclusters probe of glutathione.In 402nm wavelength lights
Exciting under, the fluorescence spectrum of the stable gold nanoclusters of test glutathione respectively at 473nm and 570nm wavelength.Add half
After cystine and lysine, test sample causes the change of fluorescence probe, records in 473nm and 570nm emitted at wavelengths peak positions
The fluorescence intensity put, is expressed as：I473、I570, wherein, I473And I570Respectively the stable gold nanoclusters of glutathione, which are added, relies ammonia
New emission peak fluorescence intensity and the fluorescence intensity added after cysteine after acid.Experiment is repeated 3 times, and as a result takes its average value.
As a result show that the stable gold nanoclusters fluorescence probe of glutathione is added after two kinds of amino acid, excited in same
Wavelength 402nm is excited down, probe produced at 473nm fluorescence signal at new emission peak, and 473nm and 570nm occur it is glimmering
Light strengthens phenomenon, with the increase of cysteine and lysine concentration, and fluorescence signal intensity is therewith at probe 473nm and 570nm
Enhancing.In this experiment, glimmering intensity light enhancing is attributed to the addition of lysine at 473nm, with the increase of lysine concentration, probe
Fluorescence signal intensity strengthens therewith at 473nm；Glimmering intensity light enhancing is attributed to the addition of cysteine at 570nm, with half Guang
Fluorescence signal intensity strengthens therewith at the increase of propylhomoserin concentration, probe 570nm.Change in fluorescence at two different wave lengths is not done mutually
Disturb, two kinds of amino acid can be detected simultaneously, it is stable that this experiment provides a kind of high selectivity, highly sensitive new glutathione
Gold nanoclusters fluorescence probe, and detect cysteine simultaneously there is provided a kind of stable gold nanoclusters fluorescence probe of glutathione
With the new method of lysine.Testing sample is added in the stable gold nanoclusters of glutathione, tested before and after it is added in 402nm
Under the exciting of wavelength light, the changing value of the fluorescence intensity at 473nm and 570nm wavelength, according to the linear relationship respectively obtained,
Calculate the concentration for adding cysteine and lysine.This method have high selectivity, high sensitivity, simplicity, it is easy the features such as.
The general principle and principal character and advantages of the present invention of the present invention has been shown and described above.The technology of the industry
Personnel are it will be appreciated that the present invention is not limited to the above embodiments, and described in above-described embodiment and specification is to illustrate this hair
Bright principle, without departing from the spirit and scope of the present invention, various changes and modifications of the present invention are possible, these changes
It all fall within the protetion scope of the claimed invention with improvement, its is equivalent by appended claims for the claimed scope of the invention
Thing is defined.
Claims (8)
1. the stable gold nanoclusters of glutathione are as fluorescence probe and ratio fluorescent probe in detection cysteine content
Using.
2. the stable gold nanoclusters of glutathione answering in detection lysine content as fluorescence probe and ratio fluorescent probe
With.
3. the stable gold nanoclusters of glutathione detect cysteine as fluorescence probe and ratio fluorescent probe and relied at the same time
Application in histidine content.
4. a kind of method that gold nanoclusters stable based on glutathione detect cysteine content, it is characterised in that including with
Lower step：
Step (a1)：Under the exciting of 402 nm wavelength lights, the stable gold nanoclusters of glutathione are determined at 570 nm wavelength
Fluorescence intensity, be designated as I0；
Step (a2)：The Guang ammonia of various concentrations testing sample half is added into the stable gold nanoclusters of step (a1) glutathione
After acid, the stable gold nanoclusters of the glutathione containing various concentrations cysteine are determined in the case where 402 nm wavelength lights are excited,
Fluorescence intensity at 570 nm wavelength, is designated as I570 ,According to adding after various concentrations cysteine, the changing value of fluorescence intensity is counted
Calculate the changing value and the line of semicystinol concentration for adding the stable gold nanoclusters fluorescence intensity of glutathione before and after cysteine
Sexual intercourse；
Step (a3)：Testing sample is added in the stable gold nanoclusters of glutathione, tested before and after it is added in 402 nm ripples
Under the exciting of long light, the changing value of the fluorescence intensity at 570 nm wavelength, the linear relationship obtained according to step (a2) is calculated
Go out to add the quality or concentration of cysteine.
5. a kind of method that gold nanoclusters stable based on glutathione detect lysine content, it is characterised in that including following
Step：
Step (b1)：Under the exciting of 402 nm wavelength lights, the stable gold nanoclusters of glutathione are determined at 570nm wavelength
Fluorescence intensity, is designated as I0；
Step (b2)：Various concentrations testing sample lysine is added into the stable gold nanoclusters of step (b1) glutathione
Afterwards, the stable gold nanoclusters of the glutathione containing various concentrations lysine are determined in the case where 402 nm wavelength lights are excited, 473
Fluorescence intensity at nm wavelength, is designated as I473, the fluorescence intensity at 570 nm wavelength is designated as I570, according to addition various concentrations
After lysine, the changing value of fluorescence intensity, the gold nanoclusters of glutathione stably are in 473 nm before and after calculating addition lysine
(I is designated as with fluorescence intensity ratio at 570 nm473/I570) changing value and lysine concentration linear relationship；
Step (b3)：Testing sample is added in the stable gold nanoclusters of glutathione, tested before and after it is added in 402 nm ripples
Under the exciting of long light, the changing value of the fluorescence intensity at 473 nm and 570 nm wavelength, according to step (b2) obtain it is linear
Relation, calculates the quality or concentration for adding lysine.
6. a kind of gold nanoclusters stable based on glutathione detect the method for cysteine and lysine content, its feature simultaneously
It is, comprises the following steps：
Step (c1)：Under the exciting of 402 nm wavelength lights, the stable gold nanoclusters of glutathione are determined at 570 nm wavelength
Fluorescence intensity, be designated as I0；
Step (c2)：Two kinds of various concentrations, which are separately added into, in the stable gold nanoclusters of step (c1) glutathione treats test sample
After product cysteine and lysine, the stable gold nanoclusters of the glutathione containing various concentrations cysteine and lysine are determined
In the case where 402 nm wavelength lights are excited, the fluorescence intensity at 570 nm and 473 nm wavelength, glimmering at 570 nm wavelength respectively
Luminous intensity, is designated as I570, the fluorescence intensity at 473 nm wavelength is designated as I473.It is glimmering according to adding after various concentrations cysteine
Luminous intensity fluorescence intensity at 570 nm (is designated as I570) changing value, glutathione is stable before and after calculating addition cysteine
Gold nanoclusters fluorescence intensity changing value and the linear relationship of semicystinol concentration；After addition various concentrations lysine,
The changing value of fluorescence intensity, calculates and adds the gold nanoclusters of glutathione stably fluorescence intensity at 473 nm before and after lysine
(it is designated as I473) changing value and lysine concentration linear relationship；
Step (c3)：Testing sample is added in the stable gold nanoclusters of glutathione, tested before and after it is added in 402 nm ripples
Under the exciting of long light, the changing value of the fluorescence intensity at 473 nm and 570 nm wavelength, according to step (c2) obtain it is linear
Relation, calculates the quality or concentration for adding cysteine and lysine respectively.
7. the preparation method of the stable gold nanoclusters of any one of claim 1 ~ 6 glutathione, it is characterised in that with paddy Guang
Sweet peptide is template, by the freshly prepd glutathione aqueous solution and tetra chlorauric acid aqueous solution isoconcentration, it is isometric with vigorous stirring
Mixing, then mixture is reacted 2-5 days, obtain pale yellow solution under illumination condition, is centrifuged, and supernatant is pure by dialyser
Change, prepare the stable gold nanoclusters of glutathione.
8. a kind of preparation method of the stable gold nanoclusters of glutathione according to claim 7, it is characterised in that the light
The reaction temperature reacted according under the conditions of is 25-35 DEG C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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