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Method for measuring glucose by rhodamine S double-enzyme catalysis spectrophotometry

A spectrophotometry, glucose technology, applied in the field of analytical chemistry, to achieve the effect of easy operation, high sensitivity and low price of the instrument

Inactive Publication Date: 2008-12-10
GUILIN UNIVERSITY OF TECHNOLOGY
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Problems solved by technology

At present, there is no report on the method of spectrophotometric determination of trace glucose using potassium iodide-horseradish peroxidase-glucose oxidase-rhodamine S catalytic system

Method used

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  • Method for measuring glucose by rhodamine S double-enzyme catalysis spectrophotometry

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Embodiment

[0009] In a 5mL stoppered colorimetric tube, add 0.15mL pH4.6 HAc-NaAc buffer solution and 0.35mL 0.04mol / L KI solution respectively, then add 0.60mL 0.20μg / mL horseradish peroxidase, then add 0.01 , 0.05, 0.10, 0.20, 0.30, 0.35mL 1.01×10 -4 mol / L glucose solution, dilute to the scale of 2.5mL with twice distilled water, shake well, let stand for 30 minutes, then add 0.35mL 2.00×10 -4 mol / L rhodamine S solution, dilute to 4.0mL with twice distilled water, shake well, and let stand for 10 minutes. Measure the absorbance value A at a wavelength of 526nm on a UV-visible spectrophotometer 526nm ; Make reagent blank A without adding glucose 0 , calculate ΔA 526nm =A 0 -A 526nm value, its absorbance value A 526nm The linear regression equation with glucose concentration is ΔA 526nm =0.0152C-0.003. Take another appropriate amount of glucose injection, dilute it with water, take an appropriate amount, measure the absorbance value in the same way, and substitute it into the abo...

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Abstract

The invention discloses a glucose determining method using rhodamine S double enzymatic spectrophotometry. 0.05-0.50mL HAc-NaAc buffer solution of pH4.6 and 0.05-0.50mL KI solution of 0.04mol / L are respectively added in a color-comparison tube of 5mL. Then, 0.05-1.0mL horseradish peroxides of 0.20Mug / mL are added and then 0.01-2.5mL 1.01*10-4mol / L glucose solution is added. Secondary distilled water is used for diluting the solution to the scale of 2.5mL. The mixed solution is shaken up and is arranged to be standing for 2-60 minutes. Then, 0.10-0.60mL 2.00*10-4mol / L rhodamine S solution is added. Secondary distilled water is used for diluting to 4.0mL. The mixed solution is shaken up and is arranged to be standing for 10 minutes. Arranged on an ultraviolet visible spectrophotometer, the absorbance value A526nm is measured at the wavelength of 526nm. Solution which is not added with glucose is used as agent blank A0 for calculating the value of Delta A526nm which is equal to the difference between A0 and A526nm. In addition, proper amount of glucose injection liquid is prepared, and the absorbance value is measured through the same method. The content of the glucose in the injection liquid can be worked out. The glucose determining method using rhodamine S double enzymatic spectrophotometry has the advantages of high sensitivity, convenient operation and cheap instrument.

Description

technical field [0001] The invention relates to the field of analytical chemistry, in particular to a method for measuring glucose by using rhodamine S double-enzyme catalyzed spectrophotometry. Background technique [0002] Glucose has extremely important physiological significance in organisms. It is the source of energy in the human body, and it is also a weathervane for predicting certain diseases (such as diabetes and its complications). As we all know, the level of glucose in body fluid is an important indicator of the health status of the human body, especially in the monitoring of body fluid glucose in diabetic patients. The sudden increase and decrease of glucose concentration indicates the occurrence of abnormal conditions. Therefore, accurate determination of glucose is of great importance to clinical Treatment, and the prevention of disease is of great significance. Since proteins, nucleic acids and glucose are very important biological macromolecules, they carr...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N21/33
Inventor 蒋治良汤亚芳梁月园黄勇
Owner GUILIN UNIVERSITY OF TECHNOLOGY
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