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Enantioseparation of 5‐chloro‐2‐{2‐[3,4‐dihydroisoquinoline‐2(1 H )‐yl]ethyl}‐2‐methyl‐2,3‐dihydro‐1 H ‐inden‐1‐one (SYA 40247), a high‐affinity 5‐HT 7 receptor ligand, by HPLC–PDA using amylose tris‐(3, 5‐ dimethylphenylcarbamate) as a chiral stationary phase
Author(s) -
Onyameh Edem K.,
Bricker Barbara A.,
Ofori Edward,
Ablordeppey Seth Y.
Publication year - 2019
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4565
Subject(s) - chemistry , enantiomer , racemization , chromatography , diethylamine , high performance liquid chromatography , optical rotation , ethanol , hexane , stereochemistry , organic chemistry
In previous structure–activity relationship studies to identify new and selective 5‐HT 7 receptor (5‐HT 7 R) ligands, we identified the chiral compound, 5‐chloro‐2‐{2‐[3,4‐dihydroisoquinoline‐2(1 H )‐yl]ethyl}‐2‐methyl‐2,3‐dihydro‐1 H ‐inden‐1‐one (SYA 40247), with high‐affinity binding to the 5‐HT 7 R. Thus, it was of interest to separate the enantiomers in order to evaluate their affinity at the 5‐HT 7 R. To achieve this separation, a normal‐phase analytical method using HPLC–PDA and a 4.6 × 250 mm Chiralpak AD‐H column was developed. Optimized isocratic conditions of 1.00 mL/min 95:5:0.1 v/v/v hexane–ethanol–diethylamine and a 254 nm analysis wavelength yielded a 6.07 min baseline separation. The method was scaled up to a 10 × 250 mm Chiralpak AD‐H column, allowing 3 mg of racemate to be separated with a single injection, and 6 mg for an overlapping double injection in the same run. The separated enantiomers were reinjected into the analytical HPLC system, peak identities confirmed by retention time and PDA UV spectra, and the enantiomeric purities determined to be 100% for peak 1 and 100% for peak 2. A Jasco P‐1020 polarimeter was used to determine the specific rotation [ α ] of the enantiomers of peaks 1 and 2, which were −86.2 and +93.3 (deg mL)/(g dm) respectively. No racemization was observed, and the enantiomeric purity remained at 100% for each peak.