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Method for sensing and analyzing concentration of ascorbic acid online based on optical microscope and auxiliary color developing agent

A technology of optical microscopy and ascorbic acid, which is applied in the field of nanomaterials and analytical chemistry, can solve problems such as more environmental light requirements, and achieve the effects of easy-to-obtain raw materials, simple preparation, and real-time continuous analysis

Active Publication Date: 2018-04-06
CAPITAL NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this work can sensitively detect physiologically active substances, it is based on the principle of fluorescence luminescence, which requires more ambient light in the experiment, and there is no report on a simpler optical online sensing system based on visible light absorption.

Method used

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  • Method for sensing and analyzing concentration of ascorbic acid online based on optical microscope and auxiliary color developing agent
  • Method for sensing and analyzing concentration of ascorbic acid online based on optical microscope and auxiliary color developing agent
  • Method for sensing and analyzing concentration of ascorbic acid online based on optical microscope and auxiliary color developing agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1: the preparation of developer

[0039] (1) Preparation of cobalt oxyhydroxide suspension: first, 10mM CoCl 2 Mix 1mL of the solution with 250μL of 1.0M NaOH solution, and then sonicate for 1 minute; then add 50μL of 0.9M NaClO solution, and then sonicate for 10 minutes, and finally dilute to 40ml to obtain a 40 μg / mL CoOOH suspension for use.

[0040] (2) Preparation of 3,3',5,5'-tetramethylbenzidine solution: Disperse 10 mg of TMB in 1 mL of DMSO, add 1 mL of 0.1 mM CA solution, 20 μL of 0.5 M EDTA solution, and then constant volume to 40mL to obtain a 2mM TMB solution.

[0041] (3) Mix the above TMB solution (2mM) and CoOOH suspension (40μg / mL) in an equal volume ratio, and after mixing thoroughly, the TMB-CoOOH system can be used as a color developer for the detection of ascorbic acid, and placed in the refrigerator spare.

Embodiment 2

[0043] Using the color developer provided in Example 1, ascorbic acid was monitored in an optical online system. Specific steps are as follows:

[0044] (1) The chromogenic agent provided in Example 1 and deionized water are continuously fed into the transparent capillary with an inner diameter of 250nm fixed under the optical microscope at a speed of 3 μL / min, passed through after 15min with an optical microscope (objective lens 40X, Manual magnification 1.6X, black-and-white mode) within 150s, 30 pictures were continuously taken at equal intervals of the transparent capillary, and the light intensity signals of the 30 pictures obtained by continuous pictures were converted into 30 digital signals respectively with the image analysis software ImageJ. Average value, as light intensity value I 0 ;

[0045] (2) After deionized water was passed through for 20 minutes, 5 parts of ascorbic acid standards with concentrations of 1 μM, 5 μM, 10 μM, 20 μM, and 50 μM were sequentially...

experiment example 1

[0051] Experimental Example 1: Effects of Different Shooting Modes on the Sensitivity of Ascorbic Acid Detection

[0052] The color and black and white shooting modes in the microscope will affect the subsequent image recognition effect. Therefore designed the recognition experiment of ascorbic acid under different shooting modes to select optimal shooting mode; Described four kinds of shooting modes are specifically: the color shooting mode under the 20X objective lens, the color shooting mode under the 40X objective lens and the overall manual magnification 1.6X Shooting mode, black and white shooting mode under 20X objective lens, black and white shooting mode under 40X objective lens and overall manual magnification 1.6X. After mixing the chromogenic agent and ascorbic acid of different concentrations in a volume ratio of 10:2 in the centrifuge tube, use a syringe to manually pass the reaction system into the capillary tube, and take pictures under a microscope, wherein th...

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Abstract

The invention provides a method for sensing and analyzing the concentration of ascorbic acid online based on an optical microscope and an auxiliary color developing agent and relates to a color developing agent for assisting the online sensing and analysis of the concentration of ascorbic acid through the optical microscope. The color developing agent is a liquid color developing agent and is prepared by mixing hydroxy cobalt oxide and 3,3',5,5'-tetramethyl benzidine. The invention simultaneously provides the method for sensing and analyzing the concentration of ascorbic acid online based on the optical microscope. According to the method, a picture of a mixture of the color developing agent in a transparent capillary tube and ascorbic acids of different concentrations is taken by virtue of the microscope, image signals are converted into digital signals which are in a certain linear relation with the concentrations of ascorbic acids by virtue of software, and then the concentration detection of ascorbic acids is realized. The color developing agent is easy and convenient to prepare, can be used for rapidly and continuously analyzing the change condition of ascorbic acid in real time, is capable of monitoring the change condition of ascorbic acid in the physiological process of cranial nerves in a living body and has certain guiding significance to the establishment of other optical sensing system methods, and the raw materials are easily available.

Description

technical field [0001] The invention belongs to the field of nanometer materials and analytical chemistry, and relates to an optical detection method of ascorbic acid. Background technique [0002] Ascorbic acid is a water-soluble six-carbon sugar acid and an important neuromodulator with two dissociated protons (pKa=4.04, 11.34) under physiological conditions. Its electron-donating properties make it a famous small molecule antioxidant and free radical scavenger in cells, and some diseases are closely related to the level of ascorbic acid. Therefore, it is of practical significance to develop a sensing method for real-time monitoring of ascorbic acid changes in physiological and pathological processes. [0003] At present, there are many methods for the determination of the content of ascorbic acid using electrochemical detection of ascorbic acid, such as potentiostatic analysis, differential pulse voltammetry, and cyclic voltammetry. In addition, there are mainly high-pe...

Claims

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Application Information

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IPC IPC(8): G01N21/78G01N21/84
CPCG01N21/78G01N21/8483G01N2021/775
Inventor 林雨青王超丁永奇
Owner CAPITAL NORMAL UNIVERSITY
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