Assignments for signals from H-1 and C-13 in the NMR spectra of pregne
nolone (1), 16-dehydropregnenolone (2), and the 3-acetate of 1 (3) wer
e validated by two-dimensional correlated spectroscopy (2D COSY) and h
eteronuclear single quantum coherence (HSQC). The narrow band of overl
apping signals from H-7, H-2, and H-4 was resolved by exploiting three
-bond coupling in the 20 COSY spectra and heteronuclear correlation. A
ssignments were based on high intensity cross peaks from long-range co
upling by H-18 with H-17 and H-12 (axial). Similar cross peaks were ob
served for H-17 with H-21. Low intensity cross peaks were seen for H-4
coupled with H-6 and H-7, and also H-16 (quasi-arial) with H-14 of 1
and 3. Assignments bared on 20 COSY spectra were confirmed by correlat
ion peaks from HSQC. This now corrects the earlier conflicts among ass
ignments reported for C-13 signals of 1 and 3. Accurate assignments we
re similarly derived for signals from C-2, C-7, C-8, and C-21 of 1 and
3, and C-15 and C-16 of 1, 2, and 3. The complete sets of H-1 and C-1
3 NMR data for pregnenolone, pregnenolone 3-acetate, and 16-dehydropre
gnenolone serve as reference standards.