11 deoxycortisol

The remainder of the methanol extracts from the ground hair were subjected to reversed-phase HPLC with UV detection (Beckman Coulter) using a Luna 5 μM Kinetex C18 column (Phenomenex, Torrance, CA). Isocratic flow of 30 % acetonitrile and 70% water was used for 20 minutes and fractions were collected in 1-minute increments using a Fractionator. The fractions were transferred to vials with scintillation fluid to determine the fraction(s) in which [³H] was detected. Cortisol and cortisol metabolite standards were run through the same chromatography and the elution profile of the standard was compared with the fractions containing [³H]. Cortisol and cortisone standards were purchased from Cerilliant (Round Rock, TX). All other cortisol metabolites (α-cortolone, β-cortolone, α-cortol, tetrahydrocortisol, tetrahydrocortisone, 5β-dihydrocortisol, 5-dihydrocortisone, 20-dihydrocortisol, 20-dihydrocortisol, 11-deoxycortisol, 6-hydroxycortisol) were purchased from Steraloids (Newport, RI).-dihydrocortisol, 5β-dihydrocortisone, 20α-dihydrocortisol, 20β-dihydrocortisol, 11-deoxycortisol, 6β-hydroxycortisol) were purchased from Steraloids (Newport, RI).

Internal standards testosterone-¹³C₃, Androstenedione-¹³C₃, 17-hydroxyprogesterone-¹³C₃, and cortisol-d₄ were purchased from Cerilliant (Round Rock, TX) and were determined to have 99.99% purity by the manufacturer. progesterone-¹³C₃ was purchased from Cambridge Isotopes (Tewksbury, MA) and was determined to have 98% purity by the manufacturer. Standards for calibration curves were purchased from various vendors. Androstenedione, testosterone, 17-hydroxyprogesterone, progesterone, corticosterone, cortisone, and cortisol were purchased from Sigma Aldrich (St. Louis, MO). 11-Deoxycorticosterone and 11-deoxycortisol were purchased from Steraloids (Newport, RI). The stated purity of all calibration standards was ≥98%.