Detection method of angiotensin converting enzyme (ACE) inhibitory activity of antihypertensive peptide
An angiotensin and inhibitory activity technology, applied in measuring devices, instruments, scientific instruments, etc., can solve problems such as inability to detect ACE inhibitory activity, and achieve the effect of saving detection time and improving accuracy
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example 1
[0027] 1. Dissolve a certain amount of fish scale antihypertensive peptide in pH 8.3, 0.1mol / L boric acid buffer, prepare a sample solution with a concentration of 1mg / ml, absorb 25μL of antihypertensive peptide, and add 75μL of 60mU·ml -1 The ACE was incubated at 37°C for 5min, and then 112μL of 5.0mmol / L HHL was added. In the blank control group, the volume of the antihypertensive peptide was replaced by the same volume of boric acid buffer solution, incubated in a constant temperature water bath at 37°C for 60min, and then 25μL of 0.1 %TFA terminated the reaction.
[0028] 2. LC-MS detection and analysis of reaction products, high performance liquid chromatography conditions: Ug120-C18 reversed-phase chromatographic column, mobile phase is acetonitrile: ultrapure water = 20:80 (containing 0.1% TFA, volume ratio), flow rate 1.0mL·min -1 , The detection wavelength is 228nm, and the injection volume is 10μL. The mass spectrometry conditions are: electrospray ionization posit...
example 2
[0031] 1. Dissolve a certain amount of peanut antihypertensive peptide in pH 8.3, 0.1mol / L boric acid buffer, prepare a sample solution with a concentration of 1mg / ml, absorb 25μL of antihypertensive peptide, and add 75μL of 60mU·ml -1 The ACE was incubated at 37°C for 5min, and then 112μL of 5.0mmol / L HHL was added. In the blank control group, the volume of the antihypertensive peptide was replaced by the same volume of boric acid buffer solution, incubated in a constant temperature water bath at 37°C for 60min, and then 25μL of 0.1 %TFA terminated the reaction.
[0032]2. LC-MS detection and analysis of reaction products, high performance liquid chromatography conditions: Ug120-C18 reversed-phase chromatographic column, mobile phase is acetonitrile: ultrapure water = 20:80 (containing 0.1% TFA) (volume ratio), flow rate 1.0mL·min -1 , The detection wavelength is 228nm, and the injection volume is 10μL. The mass spectrometry conditions are: electrospray ionization positive ...
example 3
[0035] 1、 Dissolve a certain amount of glycine-phenylalanine (Gly-Phe) in pH 8.3, 0.1mol / L boric acid buffer, prepare a sample solution with a concentration of 1mg / ml, absorb 25μL of antihypertensive peptide, add 75μL60mU·ml -1 The ACE was incubated at 37°C for 5min, and then 225μL of 5.0mmol / L HHL was added. In the blank control group, the same volume of boric acid buffer was used to replace the volume of the antihypertensive peptide, incubated in a constant temperature water bath at 37°C for 60min, and then 25μL of 0.1 %TFA terminated the reaction.
[0036] 2、 LC-MS detection and analysis of reaction products, high performance liquid chromatography conditions: Ug120-C18 reversed-phase chromatographic column, mobile phase is acetonitrile: ultrapure water = 20:80 (containing 0.1% TFA) (volume ratio), flow rate 1.0mL min -1 , The detection wavelength is 228nm, and the injection volume is 10μL. The mass spectrometry conditions are: electrospray ionization positive ion mode,...
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