Determination of cordycepin metabolites in liver microsomes by liquid chromatography-mass spectrometry
A technology of liver microsomes and LC-MS, which is applied in the field of medical testing, can solve the problems of in vitro metabolism research of liver microsomes that have not been seen, and achieve the effect of high detection sensitivity and good selectivity
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[0056] A liquid mass spectrometry method for determining cordycepin metabolites in liver microsomes, comprising the following steps
[0057] 1) Incubation of cordycepin liver microsomes
[0058] The transformation system is 2 mL, add 154 μL of liver microsome solution, 166 μL of NADP solution, 164 μL of NADH solution, 166 μL of G-6-P solution, 144 μL of G-6-P DH solution, MgCl 2 Add 840 μL tris(hydroxymethyl)aminomethane hydrochloride buffer solution to 166 μL solution, shake well, put it into a preheated 37°C full-temperature shaker and incubate for 5 minutes, then add 200 μL cordycepin solution to start the reaction. During the reaction process, oxygen was passed on the liquid surface for 30 seconds every 20 minutes. After 30 minutes, the culture bottle was taken out from the shaker, and 6 mL of cold methanol was quickly added to terminate the metabolic reaction.
[0059] 2) Sample processing
[0060] The incubation product was vortexed for 30s, centrifuged (rotating speed...
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