title: Chiral separation of amides using supercritical fluid chromatography.
abstract: Nine amide derivatives bearing α-stereocenters as well as different substitutions on the amide nitrogen were synthesized via an n-propanephosphonic acid cyclic anhydride (T3P)-mediated coupling, and their enantiomeric pairs were separated using supercritical fluid chromatography (SFC). Five polysaccharide-based chiral stationary phases (CSPs), Chiralcel OD-H, and OJ-H, and Chiralpak AD-H, AS-H and IC columns were explored for the chiral separation of these compounds. None of the compounds could be resolved on all five columns, and no single column could separate all nine pairs of enantiomers. Comparatively, the IC and OD-H columns showed the best results for this group of amides, yielding baseline separations for eight of nine pairs. The type of polar functional group and aromatic substitution in the CSPs and the substitutions on the amide nitrogen had a significant impact on the enantiomeric resolution of the compounds in the interaction between the analyte and the stationary phases. The potential separation mechanism and the effect of substitutions in the CSPs and amide solutes on the separation are discussed. The effects of the organic modifiers, modifier composition, mobile phase additives, and temperature were investigated for the separation of these amides on the IC or the OD-H column. Baseline resolution was achieved under optimized chromatographic conditions using an IC or an OD-H column. Linearity, reproducibility, and limit of quantitation were also demonstrated for the compound 9. Approximately three-fold improvement in signal-to-noise was observed using a SFC system with better instrument design.
authors.affiliation: Analytical Research and Development, Pharmaceutical Sciences, Pfizer Worldwide Research and Development, Eastern Point Road, Groton, CT 06340, USA. yanqiao.xiang@mpi.com
authors.forename: Yanqiao
authors.initials: Y
authors.lastname: Xiang
authors.forename: Joshua R
authors.initials: JR
authors.lastname: Dunetz
authors.forename: Michael
authors.initials: M
authors.lastname: Lovdahl
mesh_terms: D000577:Amides / Q000737:chemistry / Q000302:isolation & purification*; D025924:Chromatography, Supercritical Fluid / Q000295:instrumentation / Q000379:methods*; D016014:Linear Models; D063065:Organophosphonates; D011407:Propane / Q000031:analogs & derivatives; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D059629:Signal-To-Noise Ratio; D013237:Stereoisomerism
chemical_list: D000577:Amides; D063065:Organophosphonates; C569209:n-propanephosphonic acid anhydride; D011407:Propane