title: Design of experiment assisted concurrent enantioseparation of bupropion and hydroxybupropion by high-performance thin-layer chromatography.
abstract: A simple and efficient high-performance thin-layer chromatographic method was developed for chiral separation of rac-bupropion (BUP) and its active metabolite rac-hydroxybupropion (HBUP). Design of experiment (DoE)-based optimization was adopted instead of a conventional trial-and-error approach. The Box-Behnken design surface response model was used and the operating variables were optimized based on 17 trials design. The optimized method involved impregnation of chiral reagent, L(+)-tartaric acid, in the stationary phase with simultaneous addition in the mobile phase, which consisted of acetonitrile : methanol : dichloromethane : 0.50% L-tartaric acid (6.75:1.0:1.0:0.25, v/v/v/v). Under the optimized conditions, the resolution factor between the enantiomers of BUP and HBUP was 6.30 and 9.26, respectively. The limit of detection and limit of quantitation for (R)-BUP, (S)-BUP, (R,R)-HBUP, and (S,S)-HBUP were 9.23 and 30.78 ng spot-1, 10.32 and 34.40 ng spot-1, 12.19 and 40.65 ng spot-1, and 14.26 and 47.53 ng spot-1, respectively. The interaction of L-tartaric acid with analytes and their retention behavior was thermodynamically investigated using van't Hoff's plots. The developed method was validated as per the International Conference on Harmonization guidelines. Finally, the method was successfully applied to resolve and quantify the enantiomeric content from marketed tablets as well as spiked plasma samples.
authors.affiliation: Chemistry Department, School of Sciences, Gujarat University, Ahmedabad, Gujarat, India.
authors.forename: Nejal M
authors.initials: NM
authors.lastname: Bhatt
authors.affiliation: Chemistry Department, School of Sciences, Gujarat University, Ahmedabad, Gujarat, India.
authors.forename: Vijay D
authors.initials: VD
authors.lastname: Chavada
authors.affiliation: Chemistry Department, St. Xavier's Science College, Ahmedabad, Gujarat, India.
authors.forename: Mallika
authors.initials: M
authors.lastname: Sanyal
authors.affiliation: Chemistry Department, School of Sciences, Gujarat University, Ahmedabad, Gujarat, India.
authors.forename: Pranav S
authors.identifier: http://orcid.org/0000-0002-1284-1558
authors.initials: PS
authors.lastname: Shrivastav
mesh_terms: D016642:Bupropion / Q000031:analogs & derivatives* / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002855:Chromatography, Thin Layer; D012107:Research Design; D013237:Stereoisomerism; D013644:Tartrates / Q000737:chemistry*
chemical_list: D013644:Tartrates; C580748:hydroxybupropion; D016642:Bupropion; C029768:tartaric acid
keywords: Box Behnken model; bupropion; hydroxybupropion; chiral separation; design of experiment; high-performance thin-layer chromatography