CN110437175A - Two excitation fluorescence probe EuШ- dtpa-bis (HBT) and its application in detection hydrazine - Google Patents
Two excitation fluorescence probe EuШ- dtpa-bis (HBT) and its application in detection hydrazine Download PDFInfo
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Abstract
The invention discloses two excitation fluorescence probe EuШ- dtpa-bis (HBT) and its application in detection hydrazine.Diethylenetriamine pentaacetic acid, acetic anhydride are taken, pyridine is stirred at reflux for 24 hours at 80 DEG C, and cooling, decompression filters, and washs, dry；The diethylenetriamine pentaacetic acid dianhydride and triethylamine that will be obtained, dimethylformamide (DMF), 2- (2- hydroxy phenyl) benzothiazole (HBT) are stirred at reflux for 24 hours in 100 DEG C, cooling, are rotated, and are washed, dry；Obtained diethylenetriamine pentaacetic acid-bis- (HBT) and Eu (NO3)3·6H2O obtains target product in 100 DEG C of heating stirring 3h.By EuIII- dtpa-bis (HBT) detects hydrazine (N as probe combination fluorescent method2H4).The method of the present invention is simple and novel, at low cost, high-efficient, and can be applicable in environmental water sample and biological cell.
Description
Technical field
The invention belongs to the synthesis of analytical chemistry field more particularly to a kind of two excitation fluorescence probe and in actual water sample and
Hydrazine (N is detected in biological cell2H4) in application.
Background technique
Hydrazine (N2H4) it is a kind of colourless, oily liquid, there is strong pungent taste.Due to hydrazine have it is stronger alkalinity and
Reproducibility is widely used in the fields such as catalyst, medicine intermediate, emulsifier, dyestuff and agricultural.In addition, due to the energy of hydrazine
Metric density is higher, so that it is had blast characteristics, is commonly used as the high-energy fuel propellant of guided missile, rocket and satellite.However, hydrazine
It is also a kind of potential carcinogen, when that can accelerate mutagenic effect with skin contact, direct oral cavity sucking can cause respiratory tract sense
Dye can damage kidney, liver, lung and central nervous system through lung's sucking, serious to generate acute forms poison
Property simultaneously has carcinogenesis.If hydrazine is dealt with improperly in manufacture, use and transportational process, the health of human body can be not only threatened,
Also environment water will be caused to seriously affect.Therefore, a kind of simple and effective detection method is established, to control environmental pollution,
Ensure that human health is of great significance.
The method of traditional detection hydrazine mainly has titration, chemoluminescence method, liquid chromatography, spectrophotometry, electrification
Learn analytic approach etc..Although these methods can detect the hydrazine in water body, sample preparation is complicated in detection process, time-consuming
The special precision instrument etc. with needs.Therefore, designing the method that one kind easily and fast, effectively detects hydrazine has important meaning
Justice.
Summary of the invention
An object of the present invention is that design synthesizes and a kind of can be used for effectively detecting hydrazine in actual water sample and biological cell
Two excitation fluorescence probe EuШ-dtpa-bis(HBT)。
It is a kind of easy to operate the second object of the present invention is to provide, it is at low cost, it is sensitive quick, and the detection hydrazine that selectivity is good
Method.
The technical solution adopted by the present invention is that: two excitation fluorescence probe Eu provided by the inventionШ- dtpa-bis (HBT),
Preparation method includes the following steps:
1) diethylenetriamine pentaacetic acid, acetic anhydride and pyridine are uniformly mixed, are stirred at reflux at 80 DEG C for 24 hours, are cooled to room
Temperature, decompression are filtered, are successively washed with acetic anhydride and anhydrous ether, dry at 60 DEG C, obtain diethylenetriamine pentaacetic acid dianhydride
(dtpaa).Preferably, in molar ratio, diethylenetriamine pentaacetic acid: acetic anhydride: pyridine=1:4:6.
2) by diethylenetriamine pentaacetic acid dianhydride (dtpaa), triethylamine, anhydrous DMF and 2- (2- hydroxy phenyl) benzothiazole
(HBT) it is uniformly mixed, is stirred at reflux for 24 hours, is cooled to room temperature at 100 DEG C, rotary evaporation is added ice water, is vigorously stirred, obtains light
Yellow solid is successively washed with ice water and anhydrous ether, and decompression filters, and in 60 DEG C of dryings, it is bis- to obtain diethylenetriamine pentaacetic acid-
(HBT)(dtpa-bis(HBT)).Preferably, in molar ratio, diethylenetriamine pentaacetic acid dianhydride: triethylamine: HBT=1:3:2.
3) by diethylenetriamine pentaacetic acid-bis- (HBT) and Eu (NO3)3·6H2O adds Tris-HCl buffer solution to dissolve respectively,
Then it mixes, it is cooling in 100 DEG C of agitating and heating 3h, obtain EuШ-dtpa-bis(HBT).Preferably, in molar ratio, Diethylenetriamine
Pentaacetic acid-is bis- (adenine): Eu (NO3)3·6H2O=1:1.
Above-mentioned fluorescence probe EuШ- dtpa-bis (HBT) hydrazine (N in qualitative and quantitative detection actual water sample2H4) in
Using.
The method of hydrazine in qualitative detection actual water sample: taking the actual water sample containing hydrazine, and above-mentioned two excitation fluorescence probe is added
EuШThe aqueous solution of-dtpa-bis (HBT), is sufficiently mixed, and carries out fluorescence inspection under the excitation of 270nm and 325nm wavelength light respectively
It surveys, observes the variation of fluorescence spectrum.It can be found that with the increase of hydrazine content, the fluorescence of two excitation fluorescence probe in actual water sample
Intensity is remarkably reinforced.
The method of hydrazine in quantitative detection actual water sample: being 5.0 × 10 in concentration-4The above-mentioned two excitation fluorescence probe of mol/L
EuШThe hydrazine (1.0 × 10 of various concentration is separately added into the aqueous solution of-dtpa-bis (HBT)-5mol/L-2.0×10-4mol/
L), fluorescence detection is carried out under the excitation of 270nm and 325nm wavelength light respectively, obtains the hydrazine and Eu of various concentrationIII-dtpa-
Bis (HBT) Standardization curve for fluorescence intensity.According to standard curve, EuIII- dtpa-bis (HBT) comes as two excitation fluorescence probe
Detect the concentration of hydrazine in actual water sample.
Above-mentioned fluorescence probe EuШ- dtpa-bis (HBT) hydrazine (N in qualitative detection biological cell2H4) in application.
The method of hydrazine in qualitative detection biological cell: taking the biological cell containing hydrazine, and above-mentioned two excitation fluorescence probe is added
EuШThe aqueous solution of-dtpa-bis (HBT), is sufficiently mixed, and carries out fluorescence imaging under bright field and under blue light respectively, observes cell
The variation of shape and cell imaging.
The beneficial effects of the present invention are:
1. the present invention is directed to the design feature of detected material hydrazine, dtpa is modified using esterification, is then passed through
Hydrazinolysis realizes that the detection to hydrazine, design have synthesized a kind of novel two excitation fluorescence probe.
2. by means of the present invention, fluorescence probe EuШ- dtpa-bis (HBT) can carry out hydrazine sensitive and specific
Detection.Compared with the method for other detection hydrazines, have simply, quickly, at low cost, selectivity is good, is not influenced by external interference object
The advantages that.
Detailed description of the invention
Fig. 1 is two excitation fluorescence probe EuШThe synthetic route chart of-dtpa-bis (HBT).
Fig. 2 a is Fourier transform infrared spectroscopy (FT-IR) figure of dtpa.
Fig. 2 b is Fourier transform infrared spectroscopy (FT-IR) figure of 2- (2- hydroxy phenyl) benzothiazole (HBT).
Fig. 2 c is Fourier transform infrared spectroscopy (FT-IR) figure of dtpa-bis (HBT).
Fig. 2 d is dtpa-bis (N2H4) Fourier transform infrared spectroscopy (FT-IR) figure.
Fig. 3 is HBT, EuШ- dtpa-bis (HBT) and EuШ- dtpa-bis (HBT)+hydrazine (N2H4) ultra-violet absorption spectrum
Figure.
Fig. 4 a is two excitation fluorescence probe EuШ-dtpa-bis(HBT)(λex=270nm) to hydrazine (N2H4) detection fluorescence
Spectrogram.
Fig. 4 b is two excitation fluorescence probe EuШ-dtpa-bis(HBT)(λex=325nm) to hydrazine (N2H4) detection fluorescence
Spectrogram.
Fig. 5 a-1 is two excitation fluorescence probe EuШ- dtpa-bis (HBT) is to hydrazine (N2H4) mixed respectively with different material
Interfere fluorescence spectrum (λex=270nm) comparison diagram.
Fig. 5 a-2 is the histogram of Fig. 5 a-1.
Fig. 5 b-1 is two excitation fluorescence probe EuШ- dtpa-bis (HBT) is to hydrazine (N2H4) mixed respectively with different material
Interfere fluorescence spectrum (λex=325nm) comparison diagram.
Fig. 5 b-2 is the histogram of Fig. 5 b-1.
Fig. 6 a-1 is two excitation fluorescence probe EuШ- dtpa-bis (HBT) is to hydrazine (N in actual water sample2H4) detection fluorescence
Spectrum (λex=270nm) comparison diagram.
Fig. 6 a-2 is the histogram of Fig. 6 a-1.
Fig. 6 b-1 is two excitation fluorescence probe EuШ- dtpa-bis (HBT) is to hydrazine (N in actual water sample2H4) detection fluorescence
Spectrum (λex=325nm) comparison diagram.
Fig. 6 b-2 is the histogram of Fig. 6 b-1.
Fig. 7 is two excitation fluorescence probe EuШToxotest of-the dtpa-bis (HBT) to BV2 cell.
Fig. 8 is two excitation fluorescence probe EuШ- dtpa-bis (HBT) is to hydrazine (N in biological cell2H4) detection bio-imaging
Figure；
Wherein, a-1: under bright field, BV2 cell+EuIII-dtpa-bis(HBT)；
A-2: under blue light, BV2 cell+EuIII-dtpa-bis(HBT)；
B-1: under bright field, BV2 cell+EuIII- dtpa-bis (HBT)+hydrazine；
B-2: under blue light, BV2 cell+EuIII- dtpa-bis (HBT)+hydrazine.
Specific embodiment
1 two excitation fluorescence probe Eu of embodimentШ-dtpa-bis(HBT)
Synthetic route is as shown in Figure 1.
(1) preparation method
1, the preparation of diethylenetriamine pentaacetic acid dianhydride (dtpaa)
Weigh 7.8670g (0.02mol) diethylenetriamine pentaacetic acid (dtpa), acetic anhydride 16.0mL (0.08mol), pyridine
10.0mL (0.12mol) is placed in three neck round bottom, and heating is slowly stirred at 80 DEG C, is condensed back for 24 hours.Stop heating and
Stirring, product at reduced pressure is filtered, successively wash (3 × 10mL) three times respectively with acetic anhydride and anhydrous ether after being cooled to room temperature,
And depressurize suction filtration, by product in drying box 60 DEG C of dryings to get diethylenetriamine pentaacetic acid dianhydride (dtpaa).
2, the preparation of diethylenetriamine pentaacetic acid-bis- (2- (2- hydroxy phenyl) benzothiazole) (dtpa-bis (HBT))
Take diethylenetriamine pentaacetic acid dianhydride (dtpaa) 1.9652g (5.5mmol), the triethylamine of 2.334mL
(16.5mmol), anhydrous DMF (50mL), 2- (2- hydroxy phenyl) benzothiazole (HBT) 2.4971g (11mmol), in three necks circle
In the flask of bottom.It under the conditions of 100 DEG C of constant temperature, quickly stirs, is condensed back for 24 hours.It is stood after fully reacting, after being cooled to room temperature, rotation
Turn evaporation of solvent DMF, 100mL ice water is added, is vigorously stirred, flaxen solid is precipitated.Decompression filters, and successively uses ice water
It is washed respectively with anhydrous ether three times, decompression filters.It dries under the conditions of 60 DEG C to get the bis- (2- (2- of diethylenetriamine pentaacetic acid-
Hydroxy phenyl) benzothiazole) (dtpa-bis (HBT)).
3, fluorescence probe EuШThe preparation of-dtpa-bis (HBT)
Weigh dtpa-bis (HBT) (0.1010g, 0.125mmol) be dissolved in 50mL Tris-HCl ([Tris-HCl]=
50mmol/L, pH=7.50) in buffer solution, heating stirring is to being completely dissolved.Eu (NO is weighed again3)3·6H2O(0.0558g,
0.125mmol) in Yu Shangshu mixed solution, 100 DEG C of heating stirrings flow back 3 hours.Solution after reaction is cooled to room temperature, so
After be transferred in 250mL volumetric flask, with Tris-HCl buffer solution constant volume to 250mL, obtain EuIII- dtpa-bis (HBT) storage
Standby liquid (5.00 × 10-4mol/L)。
(2) it detects
(1) .dtpa, 2- (2- hydroxy phenyl) benzothiazole (HBT), dtpa-bis (HBT) and dtpa-bis (N2H4)
FT-IR figure.Such as Fig. 2 a, 2b, 2c, shown in 2d.Comparison diagram 2a and Fig. 2 c are it can be found that the stretching vibration peak of C=O is located at
1752cm-1And 1749cm-1Place.Compared with Fig. 2 a, the absorption peak blue shift 3cm of C=O in Fig. 2 c-1.In Fig. 2 c, C-O-C's is not right
Stretching vibration and symmetrical stretching vibration is claimed to be located at 1254cm-1And 1151cm-1.It can be found that C-H's is flexible from Fig. 2 c
Vibration peak appears in 851cm-1Place.Compared with Fig. 2 b, the stretching vibration peak red shift 34cm of C-H in Fig. 2 c-1.These characteristic peaks
The mobile formation for illustrating ester bond, i.e. dtpa-bis (HBT) are successfully synthesized.In addition, from Fig. 2 d it can be found that the flexible vibration of N-H
Dynamic peak appears in 3419cm-1Place.The stretching vibration peak of C=O appears in 1660cm-1, compared with Fig. 2 a, C=O's stretches in Fig. 2 d
Contracting vibration peak blue shift 92cm-1.The movement of the two characteristic peaks illustrates dtpa-bis (N2H4) be generated by hydrazinolysis.
(2) .HBT, EuШ- dtpa-bis (HBT) and EuШ-dtpa-bis(HBT)+N2H4Uv absorption spectra such as Fig. 3
It is shown.From figure 3, it can be seen that EuШ- dtpa-bis (HBT) has a weak absorption peak at 220nm.However when hydrazine is added into
EuIIIAfter-dtpa-bis (HBT) solution, EuIII-dtpa-bis(HBT)+N2H4The absorption peak strength of mixed solution is remarkably reinforced.
It can predict EuШVery big change can occur after hydrazine is added for the fluorescence intensity of-dtpa-bis (HBT).
2 two excitation fluorescence probe Eu of embodimentШApplication of-the dtpa-bis (HBT) in detection hydrazine
(1) two excitation fluorescence probe EuШThe fluorescence spectrum that-dtpa-bis (HBT) detects hydrazine
Experiment condition: measuring 5mL concentration respectively is 5.00 × 10-4The Eu of mol/LIII- dtpa-bis (HBT) stock solution adds
Enter in 2 50mL volumetric flasks, then measuring 5mL concentration is 5.00 × 10-4The hydrazine solution of mol/L is added in above-mentioned volumetric flask, uses
Tris-HCl buffer solution constant volume is to 50mL.The variation of fluorescence spectrum is observed under the excitation of 270nm and 325nm wavelength light respectively
Carry out fluoremetry.
As a result such as Fig. 4 a, shown in 4b.Under the excitation of 270nm and 325nm wavelength light, two excitation fluorescence probe EuШ-
Dtpa-bis (HBT) shows two weak emission peaks at 470nm and 480nm respectively.It is added in probe solution when hydrazine
Afterwards, the fluorescence of probe is all significantly enhanced.
(2) different coexisting substances are mixed with hydrazine to two excitation fluorescence probe EuШThe influence of-dtpa-bis (HBT) detection
Experiment condition: measuring 5ml concentration respectively is 5.00 × 10-4The Eu of mol/LIII- dtpa-bis (HBT) stock solution adds
Enter in 18 50mL volumetric flasks.Measuring 5mL concentration respectively again is 5.00 × 10-4The hydrazine solution of mol/L, which is added to 18 50mL, to be held
In measuring bottle.Sequentially adding 5mL concentration in 17 50mL volumetric flasks later is 5.00 × 10-4The K of mol/L+, Na+, Zn2+, Mg2+,
Ba2+, Ni2+, Mn2+, Ca2+, Cr3+, Al3+, Fe3+, Cl-, ClO-, SO4 2-, glucose, H2O2And NH3·H2O stock solution is used
For Tris-HCl buffer solution constant volume to 50mL, obtaining each material concentration is 5.00 × 10-5mol/L.It additionally needs and is prepared separately
One 5.00 × 10-3The hydrazine solution of mol/L.The variation of fluorescence spectrum is observed under the excitation of 270nm and 325nm wavelength light respectively
Carry out fluoremetry.
As a result as shown in Fig. 5 a-1, Fig. 5 a-2, Fig. 5 b-1 and Fig. 5 b-2.As can be seen that In from Fig. 5 a-1 and Fig. 5 b-1
Under the excitation of 270nm and 325nm wavelength light, probe solution issues weak fluorescence at 470nm and 480nm respectively.When hydrazine is added
After into probe solution, the fluorescence of probe is all significantly enhanced.However work as K+, Na+, Zn2+, Mg2+, Ba2+, Ni2+, Mn2+, Ca2 +, Cr3+, Al3+, Fe3+, Cl-, ClO-, SO4 2-, glucose, H2O2And NH3·H2The coexisting substances such as O are separately added into probe and hydrazine
After in mixed solution, the fluorescence of mixed solution hardly happens change.This explanation, other substances coexisted with hydrazine will not interfere spy
For the detection of hydrazine.Fig. 5 a-2 and Fig. 5 b-2 can more intuitively see the variation of fluorescence intensity.
(3) two excitation fluorescence probe EuШ- dtpa-bis (HBT) detects hydrazine in actual water sample
Experiment condition: this experiment chooses water and reservoir in Reservoir in Shen nest reservoir and goes out saliva, there are also tap water and drinks
Four water samples of water, control group using deionized water solution.
Firstly, taking 2mL, 5mL and 10mL hydrazine concentration respectively is 5.00 × 10-4The stock solution of mol/L in 3 volumetric flasks,
Obtain hydrazine final concentration of 2.00 × 10-5Mol/L, 5.00 × 10-5Mol/L and 10.00 × 10-5mol/L.In addition 16 50ml are taken
Volumetric flask, be divided into three groups every group 5.The concentration that 5mL is added in each volumetric flask is 5.00 × 10-4The Eu of mol/LIII-
Dtpa-bis (HBT) stock solution.Then, it is 2.00 × 10 that 5mL concentration is all separately added into every group of 5 volumetric flasks-5Mol/L,
5.00×10-5Mol/L and 10.00 × 10-5The hydrazine solution of mol/L.Last every group 5 volumetric flask Fen Bie Yong Shen nest Reservoir in
Water, reservoir go out saliva, tap water, drinking water and deionized water and are settled to 50mL, respectively swashing in 270nm and 325nm wavelength light
The variation for giving observation fluorescence spectrum carries out fluoremetry.
As a result as shown in Fig. 6 a-1 and Fig. 6 b-1.Under the excitation of 270nm and 325nm wavelength light, EuIII-dtpa-bis
(HBT) there is weak emission peak at 470nm and 480nm respectively, after four kinds of different water samples are added, for same concentration
Hydrazine, EuIIIThe fluorescence intensity of-dtpa-bis (HBT) solution is basically unchanged；However, the concentration with hydrazine increases 2.00 × 10- 5mol/L-10.00×10-5Within the scope of mol/L, EuIII- dtpa-bis (HBT) fluorescence intensity shows good rising gradient.
Clearer comparison can be found out from Fig. 6 a-2 and Fig. 6 b-2, for each concentration of hydrazine, the fluorescence intensity of four kinds of water samples
Hydrazines essentially identical and close to addition same concentrations in control water sample.Therefore, under the excitation of 270nm and 325nm wavelength light,
Demonstrate EuIII- dtpa-bis (HBT) can specifically detect the hydrazine in actual water sample.
(4) two excitation fluorescence probe EuШToxotest of-the dtpa-bis (HBT) to BV2 cell
Experiment condition: firstly, BV2 cell is seeded on the culture plate in 96 holes, by cell incubator (37 DEG C, 5%
CO2With 95% air) Eu of middle various concentration (0-50 μm of ol/L)III- dtpa-bis (HBT) hydroponics 24 hours.So
Afterwards, 10 μ L 3- (4,5- dimethyl -2- sulphur azoles) -2.5- hexichol -2H- thiazolyl blue (MTT) (5mg/mL) is taken to be added to respectively each
In hole, continue culture 4 hours.The DMSO solution of 100mL is finally taken to be respectively added to dissolve crystal in each hole.Finally, will training
It supports plate to stir 10 minutes, analyzes the cell in each hole with microplate reader.
It can be seen from figure 7 that there is no fluorescence probe Eu in the mediumIIIIn the presence of-dtpa-bis (HBT),
BV2 cell survival rate is 100%.Even if working as EuIIIThe concentration of-dtpa-bis (HBT) solution reaches 5.00 × 10-5Mol/L, BV2
The survival rate of cell only reduces 11%.This shows fluorescence probe EuIII- dtpa-bis (HBT) has lower poison to BV2 cell
Property.Therefore, two excitation fluorescence probe Eu can be predictedIII- dtpa-bis (HBT) can be used for the detection of hydrazine in living cells.
(5) two excitation fluorescence probe EuШ- dtpa-bis (HBT) is to hydrazine (N in biological cell2H4) detection bio-imaging
Experiment condition: use high glucose medium (5% cow's serum and 1% double antibody), air jet flow case (37 DEG C, 5%CO2/
95%) BV2 cell is cultivated in.Again by these cell inoculations in 6 orifice plates and make its attach 12 hours.It, will for fluorescence imaging
One group of cell is in EuIIIIt is cultivated 1.0 hours in-dtpa-bis (HBT) (20 μM) complex solution, another group of cell is in EuIII-
dtpa-bis(HBT)+N2H4It is cultivated 1.0 hours in (20 μM) mixed solution.Then culture medium is removed, is rushed respectively with PBS buffer solution
It washes cell three times, removes extracellular compounds.Gained cell is imaged on Laser Scanning Confocal Microscope, blue channel record fluorescence at
The image of picture.
As a result as shown in Figure 8.Under bright field, the imaging of BV2 cell is as shown in a-1 in Fig. 8.In EuIII-dtpa-bis
(HBT) in the presence of, BV2 cell can normally be grown, and illustrate EuIII- dtpa-bis (HBT) is to BV2 cell without poison
Property.A-2 is the imaging for the BV2 cell cultivated under blue channel in Fig. 8.It can be seen from the figure that in EuIII-dtpa-bis
(HBT) in the presence of, for the BV2 cell of culture almost without fluorescence, this illustrates do not have hydrazine molecule in cell.When in the medium plus
After entering hydrazine, under bright field (b-1 in Fig. 8), the size and shape of BV2 cell all has apparent difference with a-1 in Fig. 8,
This illustrates that the presence of hydrazine affects the normal growth of BV2 cell.B-2 is the Eu under blue channel in Fig. 8III-dtpa-bis
(HBT) and N2H4When existing simultaneously, BV2 cell shows apparent blue-fluorescence.The experimental results showed that EuIII-dtpa-bis
(HBT) it can be used for the detection of hydrazine in living cells as two excitation fluorescence probe.
Claims (8)
1. two excitation fluorescence probe EuШ- dtpa-bis (HBT), which is characterized in that the two excitation fluorescence probe EuIII-dtpa-
The preparation method of bis (HBT) includes the following steps:
1) diethylenetriamine pentaacetic acid (dtpa), acetic anhydride and pyridine are uniformly mixed, are stirred at reflux for 24 hours, are cooled at 80 DEG C
Room temperature, decompression filter, are successively washed with acetic anhydride and anhydrous ether, in 60 DEG C of dryings, obtain diethylenetriamine pentaacetic acid dianhydride
(dtpaa)；
2) by diethylenetriamine pentaacetic acid dianhydride (dtpaa), triethylamine, anhydrous DMF and 2- (2- hydroxy phenyl) benzothiazole (HBT)
It is uniformly mixed, is stirred at reflux for 24 hours, is cooled to room temperature at 100 DEG C, rotary evaporation is added ice water, is vigorously stirred, obtains faint yellow
Solid is successively washed with ice water and anhydrous ether, and decompression filters, and in 60 DEG C of dryings, it is bis- (HBT) to obtain diethylenetriamine pentaacetic acid-
(dtpa-bis(HBT))；
3) by diethylenetriamine pentaacetic acid-bis- (HBT) (dtpa-bis (HBT)) and Eu (NO3)3·6H2O adds Tris-HCl slow respectively
Solution dissolution is rushed, is then mixed, it is cooling in 100 DEG C of agitating and heating 3h, obtain EuIII-dtpa-bis(HBT)。
2. two excitation fluorescence probe Eu as described in claim 1Ш- dtpa-bis (HBT), which is characterized in that in step 1), press
Molar ratio, diethylenetriamine pentaacetic acid: acetic anhydride: pyridine=1:4:6.
3. two excitation fluorescence probe Eu as described in claim 1Ш- dtpa-bis (HBT), which is characterized in that in step 2), press
Molar ratio, diethylenetriamine pentaacetic acid dianhydride: triethylamine: 2- (2- hydroxy phenyl) benzothiazole (HBT)=1:3:2.
4. two excitation fluorescence probe Eu as described in claim 1Ш- dtpa-bis (HBT), which is characterized in that in step 3), press
Molar ratio, diethylenetriamine pentaacetic acid-are bis- (HBT): Eu (NO3)3·6H2O=1:1.
5. the described in any item two excitation fluorescence probe Eu of claim 1-4Ш- dtpa-bis (HBT) is in qualitative and quantitative detection
Hydrazine (N in environmental water sample2H4) in application.
6. application as claimed in claim 5, which is characterized in that method is as follows: in the actual water sample of Yu Hanyou hydrazine, right is added
It is required that the described in any item two excitation fluorescence probe Eu of 1-4ШThe aqueous solution of-dtpa-bis (HBT) is uniformly mixed, respectively at
Fluorescence detection is carried out under the excitation of 270nm and 325nm wavelength light.
7. the described in any item two excitation fluorescence probe Eu of claim 1-4Ш- dtpa-bis (HBT) is thin in qualitative detection biology
Hydrazine (N in born of the same parents2H4) in application.
8. the use as claimed in claim 7, which is characterized in that method is as follows: in the biological cell of Yu Hanyou hydrazine, right is added
It is required that the described in any item two excitation fluorescence probe Eu of 1-4ШThe aqueous solution of-dtpa-bis (HBT) is uniformly mixed, respectively at bright
Off field with fluorescence imaging is carried out under blue light.
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| CN113884475A (en) * | 2021-10-12 | 2022-01-04 | 青岛农业大学 | Tetracycline detection method based on europium-doped carbon quantum dot ratio fluorescent probe |
| WO2022099659A1 (en) * | 2020-11-13 | 2022-05-19 | 大连理工大学 | Leukocyte classification reagent, erythrocyte analysis reagent, reagent kit, and analysis method |
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| WO2022099659A1 (en) * | 2020-11-13 | 2022-05-19 | 大连理工大学 | Leukocyte classification reagent, erythrocyte analysis reagent, reagent kit, and analysis method |
| CN113884475A (en) * | 2021-10-12 | 2022-01-04 | 青岛农业大学 | Tetracycline detection method based on europium-doped carbon quantum dot ratio fluorescent probe |
| CN113884475B (en) * | 2021-10-12 | 2024-02-13 | 青岛农业大学 | Tetracycline detection method based on europium-doped carbon quantum dot ratio fluorescent probe |
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