CN109232589A - A kind of fluorescence probe and its preparation and application - Google Patents
A kind of fluorescence probe and its preparation and application Download PDFInfo
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- CN109232589A CN109232589A CN201811222729.8A CN201811222729A CN109232589A CN 109232589 A CN109232589 A CN 109232589A CN 201811222729 A CN201811222729 A CN 201811222729A CN 109232589 A CN109232589 A CN 109232589A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
- C07D487/22—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains four or more hetero rings
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D219/00—Heterocyclic compounds containing acridine or hydrogenated acridine ring systems
- C07D219/02—Heterocyclic compounds containing acridine or hydrogenated acridine ring systems with only hydrogen, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the ring system
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- C09K11/00—Luminescent, e.g. electroluminescent, chemiluminescent materials
- C09K11/06—Luminescent, e.g. electroluminescent, chemiluminescent materials containing organic luminescent materials
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6402—Atomic fluorescence; Laser induced fluorescence
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- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1029—Heterocyclic compounds characterised by ligands containing one nitrogen atom as the heteroatom
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- C09K2211/00—Chemical nature of organic luminescent or tenebrescent compounds
- C09K2211/10—Non-macromolecular compounds
- C09K2211/1018—Heterocyclic compounds
- C09K2211/1025—Heterocyclic compounds characterised by ligands
- C09K2211/1044—Heterocyclic compounds characterised by ligands containing two nitrogen atoms as heteroatoms
Abstract
The invention discloses a kind of fluorescence probe and its preparations and application.Probe is made of eight yuan of melon rings (Q [8]) with acridine hydrochloride.Preparation method is to take Q [8], and Q [8] is dissolved in water, solution A is obtained；Acridine hydrochloride is taken, acridine hydrochloride is dissolved in water, solution B is obtained；Solution A and solution B are mixed, reaction, which adjusts its pH with concentrated hydrochloric acid, at normal temperature can be prepared by probe for 4.Its application is mainly used for detecting the basic L-amino acid (L-arginine, L-Histidine, L-lysine) in water, L-phenylalanine and L-Trp.Probe of the invention can be to water neutral and alkali l-amino acid (L-arginine, L-Histidine, L-lysine), L-phenylalanine and L-Trp are detected, and have the characteristics that higher sensitivity, lower testing cost, sample treatment are simple and convenient to operate, measure quick and real-time detection.
Description
Technical field
The present invention relates to a kind of fluorescence probe and its preparation and application, water neutral and alkali L- amino is applied in especially a kind of detection
Sour (L-arginine, L-Histidine, L-lysine), fluorescence probe of L-phenylalanine and L-Trp and preparation method thereof and
Using.
Background technique
Amino acid plays its extremely important effect during organism growth and development: 1. be the composition of tissue
Part；2. constituting the intracorporal various substances of people；3. supplying heat；4. immunological regulation；5. playing person's fortune as carrier important in vivo
Defeated effect；6. oxidative function.Amino acid is the basic component of protein, and intake amino acid is that human body obtains nitrogen source only
One mode, absorption of human body amino acid rear portion are used directly synthetic proteins matter, and a part is oxidized decomposition, wherein nitrogenous portion
Point be used to synthesize other must amino acid, a part be broken off as energy, excreted in the form of urea.
Therefore the detection of amino acid content in food and its type is particularly important, and fluorescence probe is a kind of novel
Detection reagent, due to it is simple and convenient to operate with higher sensitivity, lower testing cost, sample treatment, measure quickly with
And the advantages of real-time detection and the favor by people.Study the fluorescence probe and fluorescence of l-amino acid in novel detection water
Probe-detection methods are necessary.
Summary of the invention
The object of the present invention is to provide a kind of fluorescence probe and its preparations and application.Probe of the present invention can be to in water
Basic L-amino acid (L-arginine, L-Histidine, L-lysine), L-phenylalanine and L-Trp are detected, a spy
Needle detects a variety of amino acid, is able to achieve detection of the Single probe to plurality of target amino acid, and testing cost is low, and detection efficiency is high, and
Be conducive to the analysis to complicated microscopic system.When furthermore using probe in detecting of the present invention, also there is higher sensitivity, sample treatment
It is simple and convenient to operate, measures the characteristics of quick and real-time detection.
Technical solution of the present invention: a kind of fluorescence probe, the molecular formula of the probe are C48H48N32O16@C13H10NCl。
A kind of preparation method of fluorescence probe above-mentioned is made of eight yuan of melon rings (Q [8]) with acridine hydrochloride.
In the preparation method of fluorescence probe above-mentioned, the molar ratio of the Q [8] and acridine hydrochloride is 1:0.5-2.
In the preparation method of fluorescence probe above-mentioned, the molar ratio of the Q [8] and acridine hydrochloride is 1:1.
The preparation method of fluorescence probe above-mentioned, specifically includes the following steps:
(1) it takes Q [8], Q [8] is dissolved in water, and obtains solution A, concentration 1.0*10-4mol/L；
(2) acridine hydrochloride is taken, acridine hydrochloride is dissolved in water, obtains solution B, concentration 1.0*10-3mol/L；
(3) solution A and solution B are mixed in 10:1 ratio, i.e. molar ratio 1:1 reacts at normal temperature, and adjusting its pH is 4
It can be prepared by probe.
In the preparation method of fluorescence probe above-mentioned, the water being added in the step (1) and step (2) is；The two of pH=7
Secondary water adjusts its pH=4 with concentrated hydrochloric acid.
A kind of application of fluorescence probe above-mentioned, for detecting a variety of l-amino acids in water.
In the application of fluorescence probe above-mentioned, described is for detecting a variety of l-amino acids in water；For detecting in water
Basic L-amino acid, L-phenylalanine and L-Trp；The basic L-amino acid is L-arginine, L-Histidine and L-
Lysine.
In the application of fluorescence probe above-mentioned, for detecting the basic L-amino acid in water, L-phenylalanine and L- color ammonia
When sour, specific detection method is:
1) fluorescence probe is taken, it is 2.0*10 that the water for adding pH to be 4, which is diluted to concentration,-5It is molten to obtain probe standard for the solution of mol/L
Liquid；
2) water to be detected is added into probe standard solution made from step 1), places 10-20min, is then swashed with fixed
It sends out wavelength 346nm and carries out fluorescence emission spectrum measurement, and the variation for drawing the fluorescence intensity of the laser wave strong point inspired is bent
Line；
3) fluorescence being added in fluorescence probe solution under water correspondence 478nm to be measured is calculated according to the curve of step 2) to send out
Spectral intensity changing value Δ F is penetrated, it can be to the basic L-amino acid (L-arginine, L-Histidine, L-lysine) in water, L- benzene
Alanine and L-Trp are detected.
In the application of fluorescence probe above-mentioned, in the step 3), when the fluorescence being added under water correspondence 478nm to be measured
Intensity of emission spectra changes or occurs blue shift, then shows to contain basic L-amino acid (L-arginine, L- group in water to be detected
Propylhomoserin, L-lysine), L-phenylalanine and L-Trp；When the fluorescence emission spectrum being added under water correspondence 478nm to be measured
Qiang Budu changes or blue shift does not occur, then shows in water to be detected without containing basic L-amino acid (L-arginine, L- group ammonia
Acid, L-lysine), L-phenylalanine and L-Trp.
To verify beneficial effects of the present invention, inventor has carried out a large amount of experimental study, part Experiment process and result
It is as follows:
Experimental example 1 probes into the appropriate molar ratios of Q [8] Yu the formed probe of acridine hydrochloride
In order to probe into the appropriate molar ratios of Q [8] Yu the formed probe of acridine hydrochloride, composed using UV absorption light method and glimmering
Light spectroscopic methodology investigates the interaction between Subjective and Objective.
Mole ratio method measures ultra-violet absorption spectrum between each system and fluorescence Spectra and fluorescence data, specific method
Are as follows: by G and Q [8 } aqueous solution (pH=4) that is configured to 1.0mmol/L and 0.1mmol/L respectively is spare, fixed object concentration is
0.04mmol/L changes the concentration of Q [8], configures NQ[8]/NGIt is 0,0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6,1.8,
2.0,2.2,2.4,2.6,2.8,3.0 aqueous solution (pH=4), the uv-visible absorption spectra for measuring solution at room temperature are (real
Testing data is attached drawing 1A, 1B)；Fixed object concentration is 0.02mmol/L, changes the concentration of Q [8], configures NQ[8]/NGIt is 0,0.2,
0.4,0.6,0.8,1.0,1.2,1.4,1.6,1.8,2.0,2.2,2.4,2.6,2.8,3.0 aqueous solution is in excitation wavelength
346nm, exciting slit 5nm, transmite slit 5nm, voltage measures solution fluorescence emission spectrum under conditions of being 540V are (real
Testing data is attached drawing 2A, 2B) then uses ultra-violet absorption spectrum between asmus method (JOB method) measurement system
And fluorescence Spectra, the total concentration of fixed Subjective and Objective are that 4.0mmol/L is constant, by constantly change the substance between Subjective and Objective amount it
Than making a series of different mol ratio NQ[8]/(NQ[8]+NG)=0.1,0.20.8,0.9 solution to be measured,
And according to said method, ultra-violet absorption spectrum and fluorescence Spectra are measured (experimental data is attached drawing 1C, 2C).
2 quantitative analysis of experimental example
It is 2.0*10 to concentration produced by the present invention-5The fluorescence probe standard solution (being prepared by embodiment 1) of mol/L
The middle solution containing L-Phe that different volumes score is added is detected, and testing result is as shown in Figure 6, it can be seen that is added not
The concentration of L-Phe is not also identical in standard solution after androgynous fraction, and the L-Phe of various concentration can be such that fluorescence probe solution occurs
Different degrees of fluorescence sensitivity, and the range of linearity of L-Phe response is (1.0-30.0) * 10-5Mol/L, detection are limited to 2.108*
10-6mol/L.Then the nuclear-magnetism titration experiments (inspection when solution that various concentration contains L-Phe is added in probe standard solution has been carried out
It is as shown in Fig. 8 to survey result), and conclude therefrom that the binding mode figure of L-Phe and probe as shown in 8 illustration of attached drawing.
It is 2.0*10 to concentration produced by the present invention-5The fluorescence probe standard solution (being prepared by embodiment 1) of mol/L
The middle solution containing L-Trp that different volumes score is added is detected, and testing result is as shown in Figure 7, it can be seen that is added not
The concentration of L-Trp is not also identical in standard solution after androgynous fraction, and the L-Trp of various concentration can be such that fluorescence probe solution occurs
Different degrees of fluorescence sensitivity, and the range of linearity of L-Trp response is (1.0-30.0) * 10-5Mol/L, detection are limited to 4.750*
10-6mol/L.Then the nuclear-magnetism titration experiments (inspection when solution that various concentration contains L-Phe is added in probe standard solution has been carried out
It is as shown in Fig. 9 to survey result), and conclude therefrom that the binding mode figure of L-Trp and probe as shown in 9 illustration of attached drawing.
Compared with prior art, the invention has the following advantages:
1, fluorescence probe of the invention can to water neutral and alkali l-amino acid, (L-arginine, L-Histidine, L- rely ammonia simultaneously
Acid), L-phenylalanine and L-Trp are detected simultaneously, as long as wherein containing one or more amino acid, can detect that,
Have the advantages that detection range is wide and specific.That is a variety of amino acid of probe in detecting, are able to achieve Single probe to plurality of target
The detection of amino acid, testing cost is low, and detection efficiency is high, and is conducive to the analysis to complicated microscopic system.
2, the present invention is by forming super molecular complex using Q [8] and anti-acridine hydrochloride, when detecting above-mentioned amino
When sour, basic L-amino acid (L-arginine, L-Histidine, L-lysine) can destroy probes probes acid or alkali environment to make probe
Fluorescence blue shift occurs, phenylalanine and tryptophan destroy probe to form new compound, occur that the fluorescence of probe again
Sensitization, therefore, can it is simple, quickly, sensitively to the basic L-amino acid in water, (L-arginine, L-Histidine, L- rely
Propylhomoserin), L-phenylalanine and L-Trp are detected.
3, compared with traditional detection method, testing cost substantially reduces the present invention, and operates and also facilitate, Ke Yishi
When detect.
Therefore, probe of the present invention can be to water neutral and alkali l-amino acid (L-arginine, L-Histidine, L-lysine), L-
Phenylalanine and L-Trp are detected, and a variety of amino acid of probe in detecting are able to achieve Single probe to plurality of target amino
The detection of acid, testing cost is low, and detection efficiency is high, and is conducive to the analysis to complicated microscopic system.Furthermore it is visited using the present invention
When needle detects, also have the characteristics that higher sensitivity, sample treatment are simple and convenient to operate, measure quick and real-time detection.
Detailed description of the invention:
Fig. 1 is Q [8] and acridine hydrochloride mole ratio method (A), (B), and JOB method (C) is ultraviolet；
Fig. 2 is Q [8] and acridine hydrochloride mole ratio method (A), (B), JOB method (C) fluorescence；
Fig. 3 is Q [8] and acridine hydrochloride nuclear-magnetism titrates and inclusion ideograph；Wherein, (A) G；(B) [8]=1 G:Q:
0.17；(C) G:Q [8]=1:0.42；(D) G:Q [8]=1:0.69；(E) G:Q [8]=1:1.04；(F)Q[8]；
Fig. 4 is formed the mass spectrogram of probe by Q [8] and acridine hydrochloride inclusion；
Fig. 5 is the fluorescent spectrum curve when solution for containing different L-type amino acid is added in probe standard solution；
Fig. 6 is the fluorescent spectrum curve (A) when the solution that various concentration contains L-Phe is added in probe standard solution, glimmering
Luminous intensity (B) and detection limit simulation drawing (C)；
Fig. 7 is the fluorescent spectrum curve (A) when probe standard solution addition various concentration contains the solution of L-Trp, fluorescence
Intensity (B) and detection limit simulation drawing (C)；
Fig. 8 is nuclear-magnetism titration figure when probe standard solution addition various concentration contains the solution of L-Phe；Wherein, (A) G；
(B) G:Q [8]=1:1；(C) [8] G:Q: L-Phe=1:1:7.15；(D) [8] G:Q: L-Phe=1:1:14.33；(E)G:Q
[8]: L-Phe=1:1:21.26；(F) [8] G:Q: L-Phe=1:1:27.78；(G)L-Phe；
Fig. 9 is nuclear-magnetism titration figure when probe standard solution addition various concentration contains the solution of L-Trp；Wherein, (A) G；
(B) G:Q [8]=1:1；(C) [8] G:Q: L-Trp=1:1:0.7；(D) [8] G:Q: L-Trp=1:1:1.75 (E) G:Q [8]: L-
Trp=1:1:3.97；(F) [8] G:Q: L-Trp=1:1:5.88；(G) [8] G:Q: L-Trp=1:1:7.63；(H)G:Q[8]:
L-Trp=1:1:10.68；(I) [8] G:Q: L-Trp=1:1:17.01；(D)L-Trp.
Specific embodiment
Embodiment 1:
The fluorescence probe of a variety of amino acid in a kind of detection water, the molecular formula of the probe are C48H48N32O16@
C13H10NCl, the probe patterns figure of the probe such as 3 illustration of attached drawing, the probe is by Q [8] and acridine hydrochloride in molar ratio 1:
1 is made.
The preparation method of the fluorescence probe of a variety of l-amino acids in above-mentioned detection water, steps are as follows:
1) Q [8] are taken, the secondary water of Q [8] plus pH=7 are dissolved, solution A, concentration 1.0*10 are obtained-4mol/L；
2) acridine hydrochloride, the secondary water dissolution of acridine hydrochloride pH=7, obtains solution B, concentration 1.0*10-3mol/
L；
3) solution A and solution B are mixed into (i.e. molar ratio 1:1) in 10:1 ratio, this result is by Q [8] and acridine hydrochloride
Ultraviolet, the nuclear-magnetism titration of the molar ratio of fluorescence, JOB experiment, Mass spectrometry experiments and Q [8] and acridine hydrochloride show that (experiment is tied
Fruit such as attached drawing 1, attached drawing 2, attached drawing 3 and attached drawing 4), it reacts at normal temperature, adjusting its pH with concentrated hydrochloric acid can be prepared by probe for 4.
Above-mentioned fluorescence probe detects water neutral and alkali l-amino acid (L-arginine, L-Histidine, L-lysine), L- phenylpropyl alcohol ammonia
The method of acid and L-Trp is as follows:
1) probe is taken, the water for being 4 with pH is added to dilute, producing concentration is 2.0*10-5The probe standard solution of mol/L；
2) water to be detected is added into probe standard solution made from step 1), 15min is placed, then to fix excitation wave
Long 346nm carries out fluorescence emission spectrum measurement, and draws the change curve of the fluorescence intensity of the laser wave strong point inspired；
3) fluorescence being added in fluorescence probe solution under water correspondence 478nm to be measured is calculated according to the curve of step 2) to send out
Spectral intensity changing value Δ F is penetrated, when obvious blue shift occurs for the fluorescence emission spectral intensity being added under water correspondence 478nm to be measured
When, then show (L-arginine, L-Histidine, L-lysine) containing basic L-amino acid in water to be detected；Before water to be measured is added
When corresponding to the fluorescence emission spectral intensity under 478nm afterwards and being remarkably reinforced, then show to contain L-phenylalanine and L- in water to be detected
Tryptophan.When both the above situation not occurring to the fluorescence emission spectral intensity under 478nm before and after water to be measured is added, then show
Basic L-amino acid (L-arginine, L-Histidine, L-lysine) is free of in water to be detected, L-phenylalanine and L-Trp.
Testing result is as shown in Fig. 5.
Embodiment 2:
The fluorescence probe of a variety of amino acid in a kind of detection water, by Q [8] and acridine hydrochloride 1:0.5 system in molar ratio
At.
The preparation method of the fluorescence probe of a variety of l-amino acids in above-mentioned detection water, steps are as follows:
1) Q [8] are taken, the secondary water of Q [8] plus pH=7 are dissolved, solution A, concentration 1.0*10 are obtained-4mol/L；
2) acridine hydrochloride, the secondary water dissolution of acridine hydrochloride pH=7, obtains solution B, concentration 1.0*10-3mol/
L；
3) solution A and solution B are mixed into (i.e. molar ratio 1:1) in 10:1 ratio, this result is by Q [8] and acridine hydrochloride
Ultraviolet, the nuclear-magnetism titration of the molar ratio of fluorescence, JOB experiment, Mass spectrometry experiments and Q [8] and acridine hydrochloride show that (experiment is tied
Fruit such as attached drawing 1, attached drawing 2, attached drawing 3 and attached drawing 4), it reacts at normal temperature, adjusting its pH with concentrated hydrochloric acid can be prepared by probe for 4.
Above-mentioned fluorescence probe detects water neutral and alkali l-amino acid (L-arginine, L-Histidine, L-lysine), L- phenylpropyl alcohol ammonia
The method of acid and L-Trp is as follows:
1) probe is taken, the water for being 4 with pH is added to dilute, producing concentration is 2.0*10-5The probe standard solution of mol/L；
2) water to be detected is added into probe standard solution made from step 1), 15min is placed, then to fix excitation wave
Long 346nm carries out fluorescence emission spectrum measurement, and draws the change curve of the fluorescence intensity of the laser wave strong point inspired；
3) fluorescence being added in fluorescence probe solution under water correspondence 478nm to be measured is calculated according to the curve of step 2) to send out
Spectral intensity changing value Δ F is penetrated, when obvious blue shift occurs for the fluorescence emission spectral intensity being added under water correspondence 478nm to be measured
When, then show (L-arginine, L-Histidine, L-lysine) containing basic L-amino acid in water to be detected；Before water to be measured is added
When corresponding to the fluorescence emission spectral intensity under 478nm afterwards and being remarkably reinforced, then show to contain phenylalanine and color ammonia in water to be detected
Acid.When both the above situation does not occur for the fluorescence emission spectral intensity being added under water correspondence 478nm to be measured, then show to
It detects in water and is free of basic L-amino acid (L-arginine, L-Histidine, L-lysine), L-phenylalanine and L-Trp.
Embodiment 3:
The fluorescence probe of various metals ion in a kind of detection water, by Q [8] and acridine hydrochloride 1:2 system in molar ratio
At.
The preparation method of the fluorescence probe of a variety of l-amino acids in above-mentioned detection water, steps are as follows:
1) Q [8] are taken, the secondary water of Q [8] plus pH=7 are dissolved, solution A, concentration 1.0*10 are obtained-4mol/L；
2) acridine hydrochloride, the secondary water dissolution of acridine hydrochloride pH=7, obtains solution B, concentration 1.0*10-3mol/
L；
3) solution A and solution B are mixed into (i.e. molar ratio 1:1) in 10:1 ratio, this result is by Q [8] and acridine hydrochloride
Ultraviolet, the nuclear-magnetism titration of the molar ratio of fluorescence, JOB experiment, Mass spectrometry experiments and Q [8] and acridine hydrochloride show that (experiment is tied
Fruit such as attached drawing 1, attached drawing 2, attached drawing 3 and attached drawing 4), it reacts at normal temperature, adjusting its pH with concentrated hydrochloric acid can be prepared by probe for 4.
Above-mentioned fluorescence probe detects water neutral and alkali l-amino acid (L-arginine, L-Histidine, L-lysine), L- phenylpropyl alcohol ammonia
The method of acid and L-Trp is as follows:
1) probe is taken, the water for being 4 with pH is added to dilute, producing concentration is 2.0*10-5The probe standard solution of mol/L；
2) water to be detected is added into probe standard solution made from step 1), 15min is placed, then to fix excitation wave
Long 346nm carries out fluorescence emission spectrum measurement, and draws the change curve of the fluorescence intensity of the laser wave strong point inspired；
3) fluorescence being added in fluorescence probe solution under water correspondence 478nm to be measured is calculated according to the curve of step 2) to send out
Spectral intensity changing value Δ F is penetrated, when obvious blue shift occurs for the fluorescence emission spectral intensity being added under water correspondence 478nm to be measured
When, then show (L-arginine, L-Histidine, L-lysine) containing basic L-amino acid in water to be detected；Before water to be measured is added
When corresponding to the fluorescence emission spectral intensity under 478nm afterwards and being remarkably reinforced, then show to contain L-phenylalanine and L- in water to be detected
Tryptophan.When both the above situation does not occur for the fluorescence emission spectral intensity being added under water correspondence 478nm to be measured, then table
Basic L-amino acid (L-arginine, L-Histidine, L-lysine), L-phenylalanine and L- color ammonia are free of in bright water to be detected
Acid.
Claims (10)
1. a kind of fluorescence probe, it is characterised in that: the molecular formula of the probe is C48H48N32O16@C13H10NCl。
2. a kind of preparation method of fluorescence probe as described in claim 1, it is characterised in that: by eight yuan of melon rings (Q [8]) and a word used for translation
Thiamine hydrochloride is made.
3. the preparation method of fluorescence probe as claimed in claim 2, it is characterised in that: the Q [8] is rubbed with acridine hydrochloride
You are than being 1:0.5-2.
4. the preparation method of fluorescence probe as claimed in claim 3, it is characterised in that: the Q [8] is rubbed with acridine hydrochloride
You are than being 1:1.
5. the preparation method of fluorescence probe as claimed in claim 2, it is characterised in that: specifically includes the following steps:
(1) it takes Q [8], Q [8] is dissolved in water, and obtains solution A, concentration 1.0*10-4mol/L；
(2) acridine hydrochloride is taken, acridine hydrochloride is dissolved in water, obtains solution B, concentration 1.0*10-3mol/L；
(3) solution A and solution B are mixed in 10:1 ratio, i.e. molar ratio 1:1 reacts at normal temperature, and adjusting its pH is 4
Probe is made.
6. the preparation method of fluorescence probe as claimed in claim 5, it is characterised in that: add in the step (1) and step (2)
The water entered is；The secondary water of pH=7 adjusts its pH=4 with concentrated hydrochloric acid.
7. a kind of application of the fluorescence probe as described in claim 1-6, it is characterised in that: for detecting a variety of L- amino in water
Acid.
8. the application of fluorescence probe as claimed in claim 7, it is characterised in that: described is used to detect a variety of L- amino in water
Acid is；For detecting the basic L-amino acid in water, L-phenylalanine and L-Trp；The basic L-amino acid is L- essence
Propylhomoserin, L-Histidine and L-lysine.
9. the application of fluorescence probe as claimed in claim 8, it is characterised in that: for detect the basic L-amino acid in water,
When L-phenylalanine and L-Trp, specific detection method is:
1) fluorescence probe is taken, it is 2.0*10 that the water for adding pH to be 4, which is diluted to concentration,-5The solution of mol/L obtains probe standard solution；
2) water to be detected is added into probe standard solution made from step 1), 10-20min is placed, then to fix excitation wave
Long 346nm carries out fluorescence emission spectrum measurement, and draws the change curve of the fluorescence intensity of the laser wave strong point inspired；
3) it is calculated in fluorescence probe solution according to the curve of step 2) and the fluorescence emission under water correspondence 478nm to be measured is added
Spectral intensity changing value Δ F, can be to the basic L-amino acid (L-arginine, L-Histidine, L-lysine) in water, L- phenylpropyl alcohol ammonia
Acid and L-Trp are detected.
10. the application of fluorescence probe as claimed in claim 9, it is characterised in that: in the step 3), before water to be measured is added
The fluorescence emission spectral intensity under 478nm is corresponded to afterwards to change or occur blue shift, then shows to contain alkalinity L- in water to be detected
Amino acid (L-arginine, L-Histidine, L-lysine), L-phenylalanine and L-Trp；When addition water correspondence to be measured
Fluorescence emission spectrum under 478nm, which is not spent by force, to change or blue shift does not occur, then shows in water to be detected without containing alkalinity L-
Amino acid (L-arginine, L-Histidine, L-lysine), L-phenylalanine and L-Trp.
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