Method for quantitatively detecting alanine transaminase in solution based on copper nanocluster fluorescent probe
An alanine aminotransferase and copper nano-cluster technology, which is applied in fluorescence/phosphorescence, measuring devices, material analysis by optical means, etc., can solve the problems of low detection sensitivity, inaccuracy and high detection limit of detection purity, and achieves a synthesis method. Simple, good fluorescence performance, low detection limit effect
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Embodiment 1
[0025] The preparation of copper nanoclusters with glutathione as a stabilizer is carried out according to the following steps at room temperature:
[0026] (1) Preparation of 0.1 M copper chloride solution: weigh 1.7048 g CuCl 2 ∙2H 2 O was dissolved in 100 mL of high-purity water, fully dissolved for later use;
[0027] (2) Preparation of copper nanoclusters: at room temperature, 0.28 g glutathione was weighed and dissolved in 15 mL H 2 O, to which was added 450 mL CuCl 2 (0.1M), after fully reacting, add 0.1 g ascorbic acid (AA), then add 1 mL NaOH (1M), react for 1 h, until the white suspension is completely dissolved and turns into a light yellow clear solution, which proves that the copper nanoclusters form. By transmission electron microscopy (TEM) ( figure 1 ) It can be seen that the copper nanoclusters are uniformly dispersed and the particle size is small.
Embodiment 2
[0029] Copper nano-clusters are used as a method for fluorescent probe specific detection of alanine aminotransferase, which is characterized in that it is carried out according to the following steps:
[0030] (1) Preparation of tiopronin mother liquor: weigh 0.1000 g tiopronin and dissolve it in 5 mL of high-purity water; dilute the mother liquor to a low concentration of 1 mg / mL, and store it at low temperature for later use;
[0031] (2) Preparation of a series of different concentration solutions of alanine aminotransferase:
[0032] Prepare alanine aminotransferase solutions with concentrations of 1, 5, 50, 100, 500, and 1000 U / L respectively, and store them at low temperature for later use;
[0033] (3) Evenly disperse the prepared copper nanoclusters based on glutathione as a stabilizer in high-purity water, prepare a detection system with a concentration of 0.675 mM and a volume of 4 mL, and measure the concentration at this time using a fluorescence spectrophotometer...
Embodiment 3
[0039] 1. Refer to Example 1 for the preparation of copper nanoclusters using glutathione as a protective agent;
[0040] 2. Determination of excitation and emission spectra of copper nanoclusters stabilized with glutathione:
[0041] Disperse copper nanoclusters in high-purity water to measure the fluorescence excitation spectrum and fluorescence emission spectrum of the material, such as figure 2 As shown, the maximum excitation wavelength of copper nanoclusters is 354 nm, and under the excitation of the maximum excitation wavelength, the fluorescence emission wavelength is 632 nm.
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