Urea Fluorescence Determination Method Based on Ammonia Etching Reaction
A technology for fluorescence measurement and urea, which is applied in the fields of analytical chemistry and nanometers, can solve the problems of scarcity, low luminous intensity, and poor stability, and achieve the effects of low processing requirements, good anti-interference, and low background signal
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example 1
[0031] The preparation process of copper nanoparticles was as follows: 1 mL of copper nitrate solution with a concentration of 0.1 mol / L was added dropwise to 10 mL of ascorbic acid solution with a concentration of 0.1 mol / L under stirring conditions, stirred at room temperature for 1 hour, and then added to the Continue to incubate at room temperature for 12 hours to obtain a copper nanoparticle solution.
example 2
[0033] 0.2 ml of the copper nanoparticle solution prepared in Example 1 was uniformly mixed with 0.8 ml of ammonia water with a concentration of 30 mmol / L, and reacted at room temperature for 15 minutes. Set up a group of blank controls without ammonia water. After the reaction, measure the fluorescence emission spectrum of the solution (excitation wavelength is 385 nm). The results show that after adding ammonia water, the etching reaction of copper nanoparticles occurs immediately to generate copper nanocluster fluorescent materials, and the fluorescence of the solution is obviously enhanced after the reaction (see figure 1 ), figure 1 Middle: A—copper nanoparticle solution control group; B—copper nanoparticle solution + ammonia water.
example 3
[0035] Add 0.3 mL of urease solution with a concentration of 2.5 U / mL into 0.75 mL of phosphate buffer containing 10 mmol / L urea (10 mmol / L, pH=7.40), shake well and incubate at 37°C for 30 minutes. After the reaction, 0.15 mL of 30 mmol / L sodium hydroxide and 0.3 mL of the copper nanoparticle solution obtained in Example 1 were added, mixed evenly and incubated at room temperature for 15 minutes. Finally, with 385 nm as the excitation wavelength, the emission light intensity value of the reaction solution at 498 nm (F 498), through the standard curve for the determination of urea. Set up the control group: A—copper nanoparticles solution; B—copper nanoparticles solution + urea; C—copper nanoparticles solution + urease; D—copper nanoparticles solution + urease + urea. After the reaction, observed under ultraviolet light, the control group had no obvious fluorescence ( figure 2 A, B and C in the experiment group), while the reaction solution of the experimental group produce...
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