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. Author manuscript; available in PMC: 2017 Feb 2.

Published in final edited form as: J Am Chem Soc. 2016 Sep 12;138(37):12124–12141. doi: 10.1021/jacs.6b04437

P450 11A1/yeast alcohol dehydrogenase incubation of 20R,22R-dihydroxycholesterol (3) in the presence of ¹⁸O₂ followed by derivatization. (A) Scheme showing incubation followed by derivatization with picolinoyl chloride (14). (B) Selected ion chromatograms monitoring for picolinoyl esters containing one ¹⁸O atom (15-1) and no ¹⁸O atom (15-0). (C) Mass spectrum showing the absence of ¹⁸O-incorporated picolinoyl ester isotopologue (15-1). The peak of interest is at t_R 4.10 minutes.

P450 11A1/yeast alcohol dehydrogenase incubation of 20R,22R-dihydroxycholesterol (3) in the presence of ¹⁸O₂ followed by derivatization. (A) Scheme showing incubation followed by derivatization with picolinoyl chloride (14). (B) Selected ion chromatograms monitoring for picolinoyl esters containing one ¹⁸O atom (15-1) and no ¹⁸O atom (15-0). (C) Mass spectrum showing the absence of ¹⁸O-incorporated picolinoyl ester isotopologue (15-1). The peak of interest is at t_R 4.10 minutes.