A method for fluorescence detection of ferric ions

A technology of ferric iron and ions, applied in fluorescence/phosphorescence, biological testing, material inspection products, etc., can solve the problems of reduced fluorescence intensity and no fluorescence response, and achieve the effect of reduced fluorescence intensity

Inactive Publication Date: 2015-10-21
SHANXI UNIV
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  • Claims
  • Application Information

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Problems solved by technology

[0003] In recent years, people have begun to pay attention to the design and synthesis of Fe 3+ Fluorescent probes with specific responses, and synthesized several types of organic fluorescent probes, the representative ones are: coumarins, rhodamines, fluoresceins, fluorescent dyes BODIPY, and naphthylhydrazone derivatives. needle compound and Fe 3+ After the action, the fluorescence intensity is significantly reduced, and some other common metal cations have almost no significant fluorescence response

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  • A method for fluorescence detection of ferric ions
  • A method for fluorescence detection of ferric ions
  • A method for fluorescence detection of ferric ions

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Experimental program
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Embodiment 1

[0024] The preparation of embodiment 1 coumarin derivative 7-diethylaminocoumarin-4-formaldehyde

[0025] 7-Hydroxycoumarin-4-carbaldehyde was synthesized according to the literature, and its synthetic route:

[0026]

[0027] In xylene solution, add SeO 2 and 7-diethylamino-4-methylcoumarin (2.06 g, 8.9 mmol). Stirring was started, and the reaction mixture was heated to reflux for 3h. After the brown mixture cooled to room temperature, another portion of SeO was added 2 , the system continued to reflux for 3h. The dark brown residual oil was dissolved with acetone and the solid was filtered off. Concentration and purification by column chromatography in a hexane:acetone (4:1) solution system gave a dark yellow solid. Separation and purification yielded 620 mg of 7-diethylaminocoumarin-4-carbaldehyde with a yield of 28%.

[0028] 7-Diethylaminocoumarin-4-carbaldehyde 1 H NMR, 13 C NMR characterization, the results are as follows:

[0029] 1 H NMR (300MHz, CDCl 3 ...

Embodiment 2

[0031] Example 2Fe 3+ Fluorescence titration of 7-diethylaminocoumarin-4-carbaldehyde compound

[0032] In a fluorescent cuvette, add 1 μL of Fe every 10 s to 2 mL of HEPES buffer (pH=7.0) containing 7-diethylaminocoumarin-4-formaldehyde with a pipette gun 3+ For fluorescence titration experiments, a total of 18 μL was added. Detected on a fluorescence spectrophotometer, with the addition of the sample to be tested, the fluorescence intensity at 494nm is gradually weakened. Instrument parameters: the slit widths of the excitation wavelength and emission wavelength are 10.0nm and 10.0nm respectively, and in aqueous solution, the maximum excitation wavelength of the fluorescent probe solution is: λ ex 396nm and maximum fluorescence emission wavelength: λ em 494nm.

Embodiment 37- 2

[0033] Embodiment 37-Diethylaminocoumarin-4-formaldehyde compound to the fluorescence histogram of various metal cations

[0034] Add 2 mL of HEPES buffer solution with pH=7.0 and 21 μL of fluorescent stock solution of 7-diethylaminocoumarin-4-formaldehyde in the cuvette, and then add other metal ions (Cu 2+ 、Cu + , Fe 2+ , Mn 2+ , Mg 2+ , Hg 2+ , Pb 2+ , Zn 2+ 、Cr 3+ 、 Bi 3+ 、Ag + 、Co 2+ 、Ni 2+ 、K + 、Na + 、Cd 2+ 、La 3+ ), so that the final concentration is 1.8mM, then add Fe 3+ , so that the final concentration is 90 μM, measure the fluorescence spectrum respectively, and draw the histogram of the 494nm fluorescence intensity corresponding to different metal cations, see figure 2 . It has been proved by experiments that other metal cations do not interfere with the reaction of the system on Fe 3+ detection.

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Abstract

The invention provides a fluorescent detection method for quantitatively detecting ferric ions on the basis of a coumarin derivative (molecular formula: C14H5NO3), and the coumarin derivative 7-lignocaine coumarin-4-formaldehyde is synthesized according to documents. The method comprises the steps: coordinating Fe<3+> and two carbonyls in the 7-lignocaine coumarin-4-formaldehyde structure in a HEPES buffer solution in a pH value of 7.0, and detecting the ferric ions through the fluorescence quenching. The detection method is high in sensitivity and selectivity for the display of the ferric ions, the detection reagent is cheap, the detection process is simple, sensitive and rapid, and the detection result is accurate. In addition, a confocal fluorescence image further proves that a probe can be used for recognizing the Fe<3+> in a living cell.

Description

technical field [0001] The invention relates to a detection technology for ferric ions, and specifically belongs to a method for rapid and quantitative fluorescent detection of ferric ions by using 7-diethylaminocoumarin-4-formaldehyde as a fluorescent reagent. Background technique [0002] Iron is a very important trace element with important biological significance. For example, iron is the most abundant transition metal element in the brain. Brain tissue uses him as an important component of enzymes to participate in the transfer and metabolism of oxygen; iron is also an indispensable component of many metalloproteins; iron is also a catalyst for many enzymes Aerobic metabolism and electron transfer and more. In addition, the iron element in the human body is the raw material for hemoglobin and myoglobin, and it is also a component of the most important enzymes to maintain the normal life activities of the human body, and is closely related to energy metabolism. At the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/52G01N21/64
CPCG01N33/52
Inventor 钞建宾刘瑜红张永斌张艳
Owner SHANXI UNIV
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