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Method for determining cholesterol by using flow injection chemiluminescence with nano-copper oxide as catalyst

A nano-copper oxide and chemiluminescence technology, which is applied in the field of analytical chemistry and nanometers, can solve problems such as poor specificity and selectivity, unstable reagents, and difficult standardization, and achieve good reproducibility, less sample demand, and high sensitivity. Effect

Inactive Publication Date: 2015-07-15
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods often have disadvantages such as poor specificity and selectivity, unstable reagents, high cost and difficult standardization

Method used

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  • Method for determining cholesterol by using flow injection chemiluminescence with nano-copper oxide as catalyst
  • Method for determining cholesterol by using flow injection chemiluminescence with nano-copper oxide as catalyst
  • Method for determining cholesterol by using flow injection chemiluminescence with nano-copper oxide as catalyst

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Cholesterol was dissolved in isopropanol and Triton X-100 (18:7, V / V) to prepare a 5 mmol / L solution.

[0050] Add 62.5 μl of 5 mmol / L cholesterol solution, 25 μl of 27.7 U / ml cholesterol oxidase and 162.5 μl of 10 mmol / L phosphate buffer solution (pH 6.5) into the EP tube respectively, and incubate the mixture at 37°C for 10 minutes, and put the reaction The liquid was diluted 100 times, transported into the mixer by a peristaltic pump, mixed with a 1.0 mg / L nano-copper oxide colloid solution, and then mixed with a 0.2 mmol / L luminol solution (pH 11.5), and the three were mixed into the flow cell for reaction, resulting in The intensity of chemiluminescence was detected by a photomultiplier tube. In the blank control experiment, distilled water was used instead of the nano copper oxide colloidal solution. Such as figure 2 As shown, the nano-copper oxide can significantly enhance the chemiluminescence signal, and the enhancement factor is about 600.

Embodiment 2

[0052] Cholesterol was dissolved in isopropanol and Triton X-100 (18:7, V / V) to prepare a 5 mmol / L solution.

[0053] Add 62.5 μl of 5 mmol / L cholesterol solution, 25 μl of 27.7 U / ml cholesterol oxidase and 162.5 μl of 10 mmol / L phosphate buffer solution (pH 6.5) into the EP tube respectively, and incubate the mixture at 37°C for 10 minutes, and put the reaction The solution was diluted 100 times, transported by a peristaltic pump into a mixer and mixed with a 1.0 mg / L nano-copper oxide colloid solution, and then mixed with a 0.2 mmol / L luminol solution with different pH values ​​(take values ​​between 10.5 and 13). Mixed into the flow cell reaction, the intensity of chemiluminescence is detected by photomultiplier tube. Such as image 3 As shown, the ratio of chemiluminescence signal increases with the increase of pH and reaches the peak at pH 11.5, and the ratio of luminescence signal decreases when the pH value continues to increase.

Embodiment 3

[0055] Cholesterol was dissolved in isopropanol and Triton X-100 (18:7, V / V) to prepare a 5 mmol / L solution.

[0056] Add 62.5 μl of 5 mmol / L cholesterol solution, 25 μl of 27.7 U / ml cholesterol oxidase and 162.5 μl of 10 mmol / L phosphate buffer solution (pH 6.5) into the EP tube respectively, and incubate the mixture at 37°C for 10 minutes, and put the reaction The solution was diluted 100 times, transported by a peristaltic pump into the mixer and mixed with 1.0 mg / L nano copper oxide colloidal solution, and then mixed with different concentrations (take values ​​between 0.01 and 1.0 mmol / L) of luminol solution (pH 11.5), The three are mixed into the flow cell for reaction, and the intensity of the generated chemiluminescence is detected by a photomultiplier tube. Such as Figure 4 As shown, the chemiluminescence signal increased with the increase of luminol concentration and reached the peak value at 0.2 mmol / L, and the luminescence signal decreased with the increase of lu...

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Abstract

The invention discloses a method for determining cholesterol by using flow injection chemiluminescence with nano-copper oxide as a catalyst. The method comprises the following steps: mixing cholesterol, cholesterol oxidase and phosphatic buffer solution; warm bathing the mixture solution to obtain a reaction solution; after diluting the reaction solution, inputting the diluted reaction solution into a mixer through a peristaltic pump to mix with a nano-copper oxide colloidal solution; then mixing with a luminol solution; entering into a flow cell for reaction; and detecting the generated chemiluminescence intensity by a photomultiplier to determine the cholesterol. By using cholesterol oxidase for catalyzing the oxidation of cholesterol to generate hydrogen peroxide, combining the flow injection technique, the chemiluminescence signal in the determination of cholesterol is enhanced about 600 times. By using cholesterol-cholesterol oxidase-luminol-nano-copper oxide flow injection chemiluminescence to determine the cholesterol content, the linear range is 0.625-12.5 [mu]mol / L. the method can be used for determination of food cholesterol content and determination of the free cholesterol and total cholesterol in serum.

Description

technical field [0001] The invention relates to a method for measuring cholesterol by flow injection chemiluminescence using nanometer copper oxide as a catalyst, and belongs to the field of analytical chemistry and nanotechnology. Background technique [0002] Abnormal cholesterol levels can occur in anemia, sepsis, malnutrition, hypertension, coronary heart disease, arteriosclerosis, cerebral embolism, lipid metabolism disorders, myocardial infarction and other diseases. Therefore, the determination of cholesterol levels in blood and food has received extensive attention in the fields of clinical diagnosis and food quality control. Common cholesterol analysis methods include non-enzymatic methods, colorimetric methods, high performance liquid chromatography, and electrochemical methods. However, these methods often have disadvantages such as poor specificity and selectivity, unstable reagents, high cost and difficult standardization. Therefore, it is of great significanc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76
Inventor 陈伟洪磊刘爱林李光文林新华
Owner FUJIAN MEDICAL UNIV
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