A kind of carboxymethyl lysine remover and its application
A carboxymethyllysine and elimination agent technology, which is applied in the field of carboxymethyllysine elimination preparations, can solve the problems of inability to eliminate and completely inhibit AGEs, and achieve the reduction of CML levels, compensation for the ineffectiveness of AGEs, and good elimination The effect of action
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Embodiment 1
[0072] Identification of 4-methyl-phthaloquinone (4MBQ) capture CML ability and identification of the structure of the additive product, specifically including the following steps:
[0073] (1) Prepare 4MBQ solution with 0.2M phosphate buffer (pH 4.5) so that its concentration is 0.08mM;
[0074] (2) Prepare a solution of 1-10 mM CML with 0.2 M phosphate buffer, so that after mixing with the above (1) solution in equal volume, the pH is 5.0, 7.0 or 8.0;
[0075] (3) Use Stopped-flow to measure the reaction kinetics of 4MBQ and CML by monitoring the absorbance change of the reaction system in the ultraviolet-visible region (200-750nm) at 25°C;
[0076] (4) Prepare a solution in which CML is 10 mM and 4MBQ is 1 mM with 0.2 M phosphate buffer (pH 7.0);
[0077] (5) Stir the mixed liquid described in (4) at room temperature for 1 min, and process it through a 0.22 μm microfiltration membrane to obtain the test solution;
[0078] Finally, LC-ESI-MS / MS was used to detect the prepa...
Embodiment 2
[0097] The structural identification of epicatechin (EC) quinones capturing CML addition products specifically includes the following steps:
[0098] (1) Use 0.2M phosphate buffer (pH 8.0) to prepare solutions with 0.1M lysine (Lys) and 0.1M glucose (Glu) respectively to simulate the food system;
[0099] (2) Take 1mL of the simulated food system described in (1), and add 2mM EC quinone;
[0100] (3) Heat the sample in a water bath at 80°C for 1 hour;
[0101] (4) Take 1 mL of the heated sample, add distilled water to make up to 5 mL;
[0102] (5) Take 1 mL of the diluted solution and pass it through a 0.22 μm microfiltration membrane, and use UPLC-ESI-MS / MS to carry out structural identification of the adduct.
[0103] Described UPLC adopts Agilent SB1290 model, and detection condition is:
[0104] Column: Agilent SB-C 18 Chromatographic column 2.1×50mm, 1.8μm
[0105] Column temperature: 25°C
[0106] Mobile phase: A) formic acid: water = 1:1000 (v:v), B) acetonitrile ...
Embodiment 3
[0119] The structural identification of epicatechin (EC) quinones capturing CML addition products specifically includes the following steps:
[0120] (1) Use 0.2M phosphate buffer (pH 8.0) to prepare solutions with 0.1M lysine (Lys) and 0.1M glucose (Glu) respectively to simulate the food system;
[0121] (2) Take 1mL of the simulated food system described in (1), and add 2mM EC quinone;
[0122] (3) Heat the sample in a water bath at 100°C for 1 hour;
[0123] (4) Take 1 mL of the heated sample, add distilled water to make up to 5 mL;
[0124] (5) Take 1 mL of the diluted solution and pass it through a 0.22 μm microfiltration membrane, and use UPLC-ESI-MS / MS to identify the structure of the adduct.
[0125] Described UPLC adopts Agilent SB1290 model, and detection condition is:
[0126] Column: Agilent SB-C 18 Chromatographic column 2.1×50mm, 1.8μm
[0127] Column temperature: 25°C
[0128] Mobile phase: A) formic acid: water = 1:1000 (v:v), B) acetonitrile
[0129] Gr...
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