Unfavorable controls, i.e., hepatocytes in sWME without the medicine and every drug separately in sWME with out hepatocytes, ended up incubated for 0 and 3 h in the same situations. Diclofenac also was incubated in the exact same conditions to be sure right metabolic action all through the experiments. Reactions were quenched with ice-cold acetonitrile and centrifugation. The incubates ended up saved at − eighty °C till Investigation.
All 4 compounds ended up detected only in constructive-ionization mode underneath the utilized analytical disorders.
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The controls have been saved while in the autosampler until the next validation run was well prepared. The analytes have been viewed as secure whenever they handed all quantitative conditions and/or didn't alter drastically (±twenty%) in response.