TABLE 1.
Conditions for the production of enriched cholesterol and applications
| Growth Medium |
Cholesterol |
Av.% of 13C Multiplicity |
|||||
|---|---|---|---|---|---|---|---|
| Dilution Lab./Nat. (g) | Enrichment in % (g) | (lJCC) |
(nJCC)a |
||||
| Labeled glucose | s | d | s | d | Applications and Remarks | ||
| (u-13C6, 99%) | 1:0 (6.75 g) | 94b (5.4 mg) | 0.5c | 20.6d | 0.0c | 1.1 | Experiments not affected by dense 13C-13C coupling networks |
| (u-13C6, 99%) | 1:0.5 | 65.9 | 3.0 | 25.1 | 0.7 | 5.2 | DQ experimentse |
| (u-13C6, 99%) | 1:2 | 32.2 | 4.5 | 18.3 | 2.1 | 8.7 | DQ experimentse |
| (u-13C6, 99%) | 1:8.5 | 10.09 | 2.5 | 6.4 | 2.2 | 5.3 | DQ experimentse |
| (u-13C6, 24.0%) | 2:3 (9.9:15 g) | 9.49 (46 mg) | 6.8 | 2.5 | 6.6 | 3.3 | For all C for SQ experimentsf |
| (1-C-13C6, 99%) | 1:0 | 27.25 | 45.0 | 2.7 | 35.5 | 8.9 | For C 1, 3, 5, 7, 9, 15, 19, 21, 22, 24, 26, 27 at low res. SQg |
| (1-C-13C6, 99%) | 1:2(1 g/2 g) | 8.94 (1.7 mg) | 15.1 | 0.3 | 13.9 | 1.8 | For C 1, 3, 5, 7, 9, 15, 19, 21, 22, 24, 26, 27 at low-res SQg |
| (2-C-13C6, 99%) | 1:0 | 22.11% | 47.6 | 1.3 | 25.8 | 15.0 | For C 2, 4, 6, 10, 16, 20, 23, 25 at low-res SQg |
| (2-C-13C6, 99%) | 1:2 | 7.34% | 13.2 | 0.9 | 10.3 | 14.7 | For C 2, 4, 6, 8, 10, 11, 13, 14, 16, 20, 23, 25 at low-res SQg |
a
The criterion for split is JCC > 1 Hz.
b
Based on MS data. The yeast extract decreased slightly the enrichment level.
c
High-multiplicity spectra, negligible amount of singlets.
d
This is due to the 5 methyls coupling with unit probabilities with their directly-bound carbons.
e
“DQ” stands for experiments based on 13C-13C double quantum coherences (ADEQUATES, INADEQUATES, etc.).
f
“SQ” stands for experiments based on isolated 13C (HSQC, 13C spectra, DEPT, etc.).
g
JCC > 25 Hz.