Retention Times of 5-oxo-12-HETE and 8-trans-5-oxo-12-HETE Under Different HPLC Conditions
| Stationary Phase* . | Mobile Phase† . | Retention Time (min) . | |
|---|---|---|---|
| 8t-5o-12-HETE . | 5o-12-HETE . | ||
| Silica | 3% iPrOH | 18.5 | 8.2 |
| ODS-silica | 40% MeCN | 33.6 | 41.2 |
| ODS-silica | 65% MeOH | 24.9 | 32.5 |
| Stationary Phase* . | Mobile Phase† . | Retention Time (min) . | |
|---|---|---|---|
| 8t-5o-12-HETE . | 5o-12-HETE . | ||
| Silica | 3% iPrOH | 18.5 | 8.2 |
| ODS-silica | 40% MeCN | 33.6 | 41.2 |
| ODS-silica | 65% MeOH | 24.9 | 32.5 |
5-oxo-12-HETE and 8-trans-5-oxo-12-HETE were prepared either chemically28 or biologically, by incubating 5-oxo-ETE with platelets in the presence of A23187. The chemically and biologically prepared compounds had identical retention times and cochromatographed with one another.
The stationary phases were a 5-μm particle size Econosphere column (250 × 4.6 mm; Alltech Associates, Deerfield, IL), a 5-μm particle size Spherisorb ODS-2 (250 × 3.2 mm; Phenomenex, Torrance, CA).
The mobile phases consisted of hexane/isopropanol/acetic acid (97:3:0.1) at 2 mL/min, water/acetonitrile/acetic acid (60:40:0.02) at 0.75 mL/min, and water/methanol/acetic acid (35:65:0.02) at 0.75 mL/min.