Acquity arc hplc

The racemic mixture of 7 was separated by normal phase HPLC. The semi-preparative instrument used was equipped with a 333 pump (Gilson, Villiers-le-Bel, France) and a UV 2487 detector (Waters, USA). PDR-Chiral Inc. software (PDR-Chiral Inc., West Palm Beach, FL, USA) was used for chromatographic data collection. The preparative column used was Chiralcel^® OD, with dimensions of 250 × 20 mm and a particle size of 5µm (Chiral Technologies, Illkirch-Graffenstaden, France). A mixture of heptane/ethanol/triethylamine (80/20/0.1 v/v/v) was used as an eluent at a flowrate of 18 mL/min. Each cycle, 25 mg of 7 was injected. The chromatographic peaks were detected at 250 nm.
Collected fraction were analyzed on an Acquity Arc HPLC equipped with a 2998 PDA detector, a Quaternary Solvent Manager—R, and a Column Manager -30S, all from Waters (Waters, USA), using the software Empower^® 3 (Waters, USA) for chromatographic data evaluation. The analytical column used was a CelluCoat^® 150 × 4.6 mm 3 µm column (Nouryon, Alby, Sweden), with a mixture of heptane/ethanol/trimethylamine (80/20/0.1 v/v/v) as the eluent at a flowrate of 0.8 mL/min. The column temperature was 25 °C, and the chromatographic peaks were detected at 260 nm.

HPLC analysis was performed on a Waters Acquity ARC HPLC (Waters, Ireland). Dried samples were reconstituted in 100 µL of 50% methanol and injected into the HPLC. Metabolites were separated on CORTECS C18 reverse-phase column 4.6 × 50 mm, 2.7 μM (Waters, Ireland), using a methanol/water gradient at 40 °C. The column effluent was analyzed using β-RAM model 3 in-line radioactivity detector (LABLOGIC, USA). Results represented the mean and standard deviation (s.d.) value from three repeated experiments. All HPLC studies were run in duplicate and repeated at least three times independently.