Method for screening aldose reductase inhibitor
A reductase inhibitor and aldehyde identification technology, which is applied in the field of analytical chemistry, can solve the problems of low sensitivity of spectrometry and easy to be subject to external interference, and achieve the effect of accurate sample detection.
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Embodiment 1
[0107] combine Figure 1-8 Illustrative Example 1
[0108] A method for identifying an aldose reductase inhibitor, the steps are as follows:
[0109] (1) Preparation of enzymatic reaction buffer
[0110] The buffer solution is ammonium acetate solution, the pH value is adjusted to 6.2 with acetic acid, and the buffer solution concentration is 100 mmol / L;
[0111] (2) Preparation of standard products
[0112]Sorbitol was made into concentrations of 0.1 micromol / L, 0.2 micromol / L, 0.5 micromol / L, 1 micromol / L, 2 micromol / L, 5 micromol / L, 10 Standard solutions of micromol / liter, 20 micromol / liter, and 50 micromol / liter, and each concentration of standard solution contains 1 micromol / liter of xylitol as an internal standard;
[0113] (3) Enzymatic reaction of blank sample and sample
[0114] Blank sample: a total reaction volume of 200 μl, containing aldose reductase in a volume of 30 μl, 2-mercaptoethanol at a final concentration of 5 mmol / L, NADPH at a final concentration o...
Embodiment 2
[0155] combine Figure 9 , Figure 10 Illustrative Example 2
[0156] A method for identifying an aldose reductase inhibitor, the steps are as follows:
[0157] (1) Preparation of enzymatic reaction buffer
[0158] The buffer solution is ammonium acetate solution, the pH value is adjusted to 6.2 with acetic acid, and the concentration is 100 mmol / L;
[0159] (2) Preparation of standard products
[0160] Use buffer solution and acetonitrile to prepare the standard products with concentrations of 0.1 micromol / L, 0.2 micromol / L, 0.5 micromol / L, 1 micromol / L, 2 micromol / L, 5 micromol / L, 10 Standard solutions of micromol / liter, 20 micromol / liter, and 50 micromol / liter, and each concentration of standard solution contains 1 micromol / liter of xylitol as an internal standard;
[0161] (3) Enzymatic reaction of blank sample and sample
[0162] Blank sample: a total reaction volume of 200 μl, containing aldose reductase in a volume of 30 μl, 2-mercaptoethanol at a final concentrat...
Embodiment 3
[0170] combine Figure 11 , Figure 12 Illustrative Example 3
[0171] A method for identifying an aldose reductase inhibitor, the steps are as follows:
[0172] (1) Preparation of enzymatic reaction buffer
[0173] The buffer solution is ammonium acetate solution, the pH value is adjusted to 6.2 with acetic acid, and the concentration is 100 mmol / L;
[0174] (2) Preparation of standard products
[0175]Use buffer solution and acetonitrile to prepare the standard products with concentrations of 0.1 micromol / L, 0.2 micromol / L, 0.5 micromol / L, 1 micromol / L, 2 micromol / L, 5 micromol / L, 10 Standard solutions of micromol / liter, 20 micromol / liter, and 50 micromol / liter, and each concentration of standard solution contains 1 micromol / liter of xylitol as an internal standard;
[0176] (3) Enzymatic reaction of blank sample and sample
[0177] Blank sample: a total reaction volume of 200 μl, containing aldose reductase in a volume of 30 μl, 2-mercaptoethanol at a final concentrat...
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