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LC-MS/MS analysis of the recombinant CYP76B74 enzyme assays using GHQ as the substrate. The assay samples were incubated with shaking for 4 h at 28°C. A, LC-MS chromatograms of extracts from the reaction containing CYP76B74 microsomes with NADPH (pESC-Trp-CYP76B74), empty vector control (pESC-Trp), and microsome-free control (GHQ). B, Mass spectrum of the reaction product. C, Direct conversion of GHQ (1) to GHQ-3′′-OH (1a).
