Dual-output sensor for lysozyme detection and preparation method of lysozyme
A dual-output, lysozyme technology, applied in the field of analytical chemistry, can solve the problems of complicated operation, limited application, a lot of time, etc., and achieve the effect of simple operation, obvious characteristics and low cost.
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Embodiment 1
[0021] According to references (Sivakumar, K.; Xie, F.; Cash, B. M.; Long, S.; Barnhill, H. N.; Wang, Q. A Fluorogenic 1,3-Dipolar Cyclodition Reaction of 3-Azidocoumarins and Acetylenes. Org. Lett ., 2004, 6, 4603-4606.) Preparation of 3-azido-7-hydroxycoumarin compound. The specific steps are as follows: Weigh 1.38 g of 2,4-dihydroxybenzaldehyde and 1.12 g of N-acetylglycine into 50 mL of anhydrous sodium acetate, and ultrasonically dissolve, then reflux and stir for 4 hours. After the reaction is completed, the resulting mixture is poured into ice water to obtain a yellow precipitate, filter the yellow precipitate, and wash the precipitate with ice water, and then reflux in 30 mL of a mixture of hydrochloric acid and ethanol (2:1) for 1 hour. After the reaction is complete, use an ice bath to dissolve the above Cool the solution, and dilute the above solution with 20 mL of ice water; add 20 mmol of sodium nitrite, stir for 10 minutes, then add 30 mmol of sodium azide, and s...
Embodiment 2
[0023] The preparation of the gold nanoparticle solution functionalized by triazole coumarin compound, the specific steps are as follows:
[0024] (1) According to the reference (Liu, D.; Chen, W.; Wei, J.; Li, X.; Wang, Z.; Jiang, X. A Highly Sensitive, Dual-Readout Assay Based on Gold Nanoparticles for Organophosphorus and Carbamate Pesticides. Anal. . 2012 , 84 , 4185-4191.) To prepare a gold nanoparticle solution with a diameter of 13 nm, the specific steps are as follows: first, clean the glassware such as the round bottom flask and reflux tube used in the preparation process with aqua regia, and then add chloroauric acid (0.206 mg / mL) into 25 mL water, stirred, heated to reflux, after boiling, added 1.0 mL of 1.3% sodium citrate solution to the above-mentioned hot water solution, continued to heat and reflux, and stirred vigorously for 15 minutes. During this process, the color of the solution changed from light red to Turned into wine red. Finally, stop the solutio...
Embodiment 3
[0028] Preparation method of dual-output sensor for detection of lysozyme: adding different concentrations of lysozyme to the solution of gold nanoparticles functionalized with triazole coumarin compound, shaking on a shaker for 30 minutes, and observing the gold nanoparticles with the naked eye The color of the solution gradually changed from wine red to blue-purple, and the process was accompanied by the recovery of fluorescence, which were detected by fluorescence spectrum and ultraviolet spectrum respectively. It was found that the fluorescence intensity of the solution increased gradually with the increase of the concentration of lysozyme, and the absorbance at 526 nm gradually decreased, and the absorption wavelength gradually shifted to the long-wave direction.
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