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Method using light on/off mode probe to measure solution pH

An optical switch and probe technology, applied in the field of analytical chemistry, can solve problems such as fluorescence intensity interference, and achieve the effect of expanding the application range and various measurement methods.

Inactive Publication Date: 2016-05-04
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that the fluorescence intensity of some types of fluorescent probes will be interfered to varying degrees by biologically relevant ions and small biological molecules such as amino acids and sugars that coexist in the system.

Method used

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  • Method using light on/off mode probe to measure solution pH
  • Method using light on/off mode probe to measure solution pH
  • Method using light on/off mode probe to measure solution pH

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1: the preparation method of each reagent in the method of the present invention.

[0044] (1) Probe solution: weigh 2.8 mg of probe (molecular formula: C 17 h 13 NO 3 Molecular weight: 279.09), dissolved in acetonitrile to prepare a 100mL solution with a concentration of 100μM.

[0045] (2) Tris-HCl buffer solution: prepared with a concentration of 50mM trishydroxymethylaminomethane (Tris) and 50mM HCl, and adjusted the pH to the desired value with a pH meter;

[0046] (3) HEPES-NaOH buffer solution: prepared with a concentration of 50mM 4-hydroxyethylpiperazineethanesulfonic acid (HEPES) and 50mM NaOH, and adjusted the pH to the desired value with a pH meter;

[0047] (4) Other coexisting ions and molecular solutions: Take analytically pure nitrates or perchlorates of various metals, dissolve them in double-distilled water, and prepare a double-distilled aqueous solution with a concentration of 20mM; take analytically pure various Amino acids, bovine se...

Embodiment 2

[0049] Example 2: Preparation of probe compounds.

[0050] Using 8-hydroxyquinaldine and 2,4-dihydroxybenzaldehyde as raw materials, using acetic anhydride and pyridine / water as solvents, firstly synthesize the intermediate, and then hydrolyze the intermediate in a mixed solvent of pyridine / water. The synthetic route is as follows:

[0051]

[0052] In the there-necked flask, in the acetic anhydride solution that is dissolved with 8-hydroxyquinaldine, add 2,4-dihydroxybenzaldehyde, by molar ratio 8-hydroxyquinaldine:2,4-dihydroxybenzaldehyde equals 1: 2. Under the protection of nitrogen, reflux, the reaction is completed, concentrated to remove the solvent acetic anhydride, and eluted by silica gel column chromatography to obtain an intermediate. Reaction temperature: 139°C (reflux), reaction time: 5h, reaction solvent: acetic anhydride, eluent: volume ratio chloroform: ethyl acetate (3:1).

[0053] N 2 Under protection, add the intermediate in the three-necked flask, py...

Embodiment 3

[0054] Example 3: Spectral measurement of pH probes.

[0055] Add the acetonitrile stock solution (0.1mM, 1mL) of the probe into a 10.0mL volumetric flask, and add Tris-HCl or HEPES-NaOH buffer solutions with different pH values, so that the solvent ratio of the test solution is acetonitrile / buffer solution (v / v, 3 / 2), dilute to the mark, shake well, and transfer to a 1cm quartz cuvette for fluorescence spectrum and UV-visible absorption spectrum determination. The maximum excitation wavelength measured by fluorescence spectroscopy is 440nm, and the maximum emission wavelength is 550nm or 580nm, respectively.

[0056] (1) Detection of pH by fluorescence spectroscopy

[0057] In a 1cm cuvette, add the probe solution with a concentration of 10μM, and then add the probe solution with a pH of 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, 3.4, 3.6, 3.8, 4.0, 4.2, 4.4, 4.6, 4.8, 5.0, 5.2, 5.4 Tris-HCl buffer solution, so that the solvent ratio of the test solution is (acetonitrile...

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Abstract

The invention discloses a method using a light on / off mode probe to measure the solution pH, and belongs to the field of analytical chemistry. A compound (E)-2-(2,4-dihydroxylphenyl)vinyl-8-hydroxyl quinoline is taken as the pH fluorescence probe, (probe for short). The method comprises the following steps: quantitatively measuring the pH of a solution in pH ranges of 2-5 and 10-12.4 through a fluorescence method; quantitatively measuring the pH of the solution in pH ranges of 2-5 and 5-10 through an ultraviolet-visible absorption spectroscopy method; qualitatively and semi-quantitatively measuring the pH of the solution in pH ranges of 2-5 and 10-13 through a visual method under the UV rays and sunlight; and using the probe to qualitatively and semi-quantitatively measure the pH of the solution in pH ranges of 2-5 and 10-13 under the UV rays and sunlight; wherein the pH florescence probe has a multi-stage light on / off effect: the fluorescence is on when pH is in a range of 2 to 4; the fluorescence is off when pH is in a range of 5 to 9; the fluorescence is on when pH is in a range of 10 to 12; and the fluorescence is off when pH is greater than 13.

Description

technical field [0001] The invention belongs to the field of analytical chemistry, in particular to an optical switch type probe method for measuring pH in solution. Background technique [0002] Accurate measurement of pH in solution is of great importance in research in the fields of chemistry, biology, medicine, agriculture, and environmental science. The conduct, completion and process control of chemical reactions, the measurement of intracellular pH in cell biology, cell proliferation and apoptosis, endocytosis, ion transport and homeostasis, multidrug resistance, bioanalytical chemistry and medical health All blood studies in the study are closely related to pH. Fluorescence spectroscopy is a detection method based on optical signals, which has the advantages of high sensitivity, high selectivity, fast reactivity, and reversible performance. The use of fluorescent probe method to measure the pH value of the solution has the characteristics of high sensitivity, no in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/80G01N21/64G01N21/31
CPCG01N21/314G01N21/643G01N21/80G01N2021/3155
Inventor 曾晞朱勤牟兰吴玉田曾莉冉琴
Owner GUIZHOU UNIV
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