Method for determining content of primary amine groups in gelatin
A technology based on primary amino groups and gelatin, which is applied in the field of chemical detection, can solve problems such as inaccurate measurement results, and achieve the effects of sensitive and rapid measurement, high precision and accurate results
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Embodiment 1
[0039] A method for measuring primary amino group content in gelatin, comprising steps as follows:
[0040](1) Dissolve 0.022g of glycine in 25mL of a buffer solution with a pH of 10, dilute ten times with the same buffer solution, weigh 2.2mg of fluorescamine and dissolve it in 5mL of acetone to prepare a solution of fluorescamine in acetone, and pipette three equal portions of each Add 375 μL, 560 μL, and 750 μL of fluorescamine acetone solution to 250 μL of diluted glycine buffer solution, and react for 10 minutes in the dark. After the reaction is completed, each portion is diluted to 25 mL with buffer solution.
[0041] The fluorescence intensity of each solution was measured with a fluorescence spectrometer (HITACHI F-4600). When measuring, the excitation wavelength was set to 380nm, the voltage was set to 600v, and the maximum relative fluorescence intensity when the emission wavelength was 503nm was 1146, 1339, and 1367, respectively. Select the glycine fluorescamine s...
Embodiment 2
[0046] A method for measuring primary amino group content in gelatin, comprising steps as follows:
[0047] (1) Weigh 0.039g leucine and dissolve it in 25mL pH 10 buffer solution, dilute ten times with the same buffer solution, weigh 2.2mg fluorescamine and dissolve it in 5mL acetone to prepare fluorescamine acetone solution, pipette three equal parts Add 375 μL, 560 μL, and 750 μL of fluorescamine-acetone solution to 250 μL of diluted leucine buffer solution, and react for 10 minutes in the dark. After the reaction is completed, each portion is diluted to 25 mL with buffer solution.
[0048] The fluorescence intensity of each part was measured with a fluorescence spectrometer (HITACHI F-4600). When measuring, the excitation wavelength was set to 380nm, the voltage was set to 600v, and the maximum relative fluorescence intensity when the emission wavelength was 503nm were: 934,989,1099.
[0049] Select the leucine fluorescamine solution with the highest fluorescence intensity,...
Embodiment 3
[0054] A method for measuring primary amino group content in gelatin, comprising steps as follows:
[0055] (1) Weigh 0.0258g L-arginine and dissolve it in a buffer solution with a pH of 10, dilute ten times with the same buffer solution, weigh 2.2 mg of fluorescamine and dissolve it in 5 mL of acetone to prepare a fluorescamine-acetone solution, and pipette three equal parts For each 250 μL diluted arginine buffer solution, add 375 μL, 560 μL, and 750 μL of fluorescamine acetone solution, and react for 10 minutes in the dark. After the reaction is completed, add buffer solution and dilute to 25 mL.
[0056] The fluorescence intensity of each part was measured with a fluorescence spectrometer (HITACHI F-4600). When measuring, the excitation wavelength was set to 380nm, the voltage was set to 600v, and the maximum relative fluorescence intensity when the emission wavelength was 503nm were: 855, 887, and 910, respectively. Select the arginine fluorescamine solution with the high...
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