The method used to study the inhibitory effect of pqq on ages
A technology of inhibition and CEL, applied in the field of biological analysis, which can solve the problems of time-consuming, complicated operation, residual mass spectrometer, etc.
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Embodiment 1
[0039] Example 1 The research method of the inhibitory effect of PQQ on AGEs, the steps are:
[0040] (1) Preparation of standard solution and internal standard solution: Accurately weigh appropriate amounts of CML and CEL standard products, dissolve them in 70% methanol to prepare CML and CEL standard stock solutions with a concentration of 200 μg / mL, and weigh appropriate amounts of [ 2 h 2 ] - CML and [ 2 h 4 ]–CEL standard product, dissolved in 70% methanol solution to prepare a 50μg / mL internal standard stock solution, and stored in a refrigerator at 4°C; pipette an appropriate amount of CML and CEL standard solution, accurately add the internal standard solution, and dilute with water to determine The concentration of the series standard solution is about 1-800 ng / mL, and the concentration of the internal standard solution is 100 ng / mL.
[0041] (2) Plasma sample pretreatment: Take 50 µl plasma sample, add 400 µL borax buffer solution (pH 9.2, 0.2M), add 100 µL 100mM ...
Embodiment 2
[0055] Embodiment 2 The impact of different solid-phase extraction cartridges on the recovery rate of CML and CEL
[0056] Plasma samples containing high, medium and low concentrations of CML and CEL were added to the blank plasma respectively. Different solid-phase extraction cartridges were used to process plasma samples, and the recoveries were determined. The results are shown in Table 1.
[0057] Table 1 Effect of different SPE cartridges on the recovery of CML and CEL
[0058]
[0059] The experimental results show that the recovery rate of ordinary ODS-C18 solid-phase extraction cartridges is low, and Styre Screen® H2P is polymerized from hydrophilic N-vinylpyrrolidone and lipophilic divinylbenzene, and has strong adsorption for polar compounds ability. Compared with ODS-C18 solid phase extraction, the recovery rate is slightly improved. The Sepax CUALD solid phase extraction column uses silica gel as a filler, which is bonded with different covalent groups to fo...
Embodiment 3
[0060] Example 3 Effect of PQQ on plasma CML, CEL, protein-bound CML and protein-bound CEL
[0061] The plasma samples of each group were processed and determined according to the above method. For the measurement results, see Figure 5. After 6 weeks of D-gal-induced modeling, plasma free CML, CEL, protein-bound CML and protein-bound CEL increased in varying degrees compared with the control group, and the plasma CML content decreased significantly after administration of donepezil and PQQ; at the same time, PQQ can also Reduced plasma CEL and protein-bound CML; donepezil had no effect on both; however neither PQQ nor donepezil had any effect on plasma protein-bound CEL. It provides a new basis for the study of the mechanism of PQQ in aging diseases.
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