Analytical and semipreparative enantioseparation of 9-hydroxyrisperidone, the main metabolite of risperidone, using high-performance liquid chromatography and capillary electrophoresis. Validation and determination of enantiomeric purity.
The HPLC semipreparative enantioseparation of 9-hydroxyrisperidone (9-OHRisp) was studied by optimizing various experimental conditions: the nature of the chiral stationary phase (CSP), mobile phase composition, temperature and analyte loading. This semipreparative enantioseparation was successfully completed using the polysaccharide Chiralcel OJ chiral stationary phase and a n-hexane/ethanol/methanol (50/35/15, v/v/v) ternary mobile phase. To assess the enantiomeric purity of both isolated isomers, three analytical methods using UV detection were developed and validated: one CE method using dual cyclodextrin mode and two HPLC methods using either the Chiralcel OJ CSP in normal-phase mode or the alpha-acid glycoprotein (alpha-AGP) CSP in reversed-phase mode. The three methods make it possible to obtain excellent enantioseparations (R(s) >3) with analysis times lower than 15 min, and the calculated limits of detection allow for the determination of minor enantiomeric impurities (0.1%). Enantiomeric purity obtained for dextrorotatory and levorotatory enantiomers was superior to 99.9% and equal to 98.9%, respectively, which proved the success of the semipreparative enantioseparation. A brief comparison of the performances of the analytical methods completes this work.
Danel
Cécile
C
Laboratoire de Chimie Analytique EA 4034, Faculté des Sciences Pharmaceutiques et Biologiques, Université de Lille 2, BP 83, 3 rue du Pr. Laguesse, 59006 Lille Cedex, France.
Barthélémy
Christine
C
Azarzar
Dalila
D
Robert
Hugues
H
Bonte
Jean-Paul
JP
Odou
Pascal
P
Vaccher
Claude
C
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D019075:Electrophoresis, Capillary / Q000379:methods*; D007555:Isoxazoles / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D015394:Molecular Structure; D000068882:Paliperidone Palmitate; D011743:Pyrimidines / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D015203:Reproducibility of Results; D018967:Risperidone / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D013237:Stereoisomerism
D007555:Isoxazoles; D011743:Pyrimidines; D018967:Risperidone; D000068882:Paliperidone Palmitate