title: HPLC determination of ketoprofen enantiomers in human serum using a nonporous octadecylsilane 1.5 microns column with hydroxypropyl beta-cyclodextrin as mobile phase additive.
abstract: A sensitive and stereospecific high-performance liquid chromatography (HPLC) method for the quantitation of ketoprofen enantiomers in human serum was developed. The assay involves the use of an octadecylsilane solid-phase extraction for serum sample clean-up prior to HPLC analysis. Chromatographic resolution of the ketoprofen enantiomers was performed on a nonporous octyldecylsilane column with hydroxypropyl beta-cyclodextrin as the mobile phase additive. The composition of the mobile phase was 98:2 v/v aqueous 0.1% trifluoroacetic acid (TFA), pH 4.00 (adjusted with triethylamine (TEA))/acetonitrile containing 10 mM hydroxypropyl beta-cyclodextrin (beta-CD) at a flow rate of 0.8 ml min-1. Recoveries of R(-)-ketoprofen was 95.4 +/- 2.16% and for S(+)-ketoprofen 96.2 +/- 1.31%. Linear calibration curves were obtained in the range 0.025-15 micrograms ml-1 range for each enantiomer in serum. The detection limit based on a S/N = 3 ratio was 10 ng ml-1 for each enantiomer in serum with ultraviolet detection at 220 nm. The limit of quantitation for each enantiomer was 25 ng ml-1. Precision calculated as % relative standard deviation (%R.S.D.) and accuracy calculated as % error were in the range 0.2-5.2% and 0.3-2.2%, respectively, for the R enantiomer and 0.3-6.2 and 0.2-3.2%, respectively, for the S enantiomer.
authors.affiliation: Department of Medicinal Chemistry, College of Pharmacy, University of Georgia, Athens 30602-2352, USA.
authors.forename: E
authors.initials: E
authors.lastname: Ameyibor
authors.forename: J T
authors.initials: JT
authors.lastname: Stewart
mesh_terms: D000073738:2-Hydroxypropyl-beta-cyclodextrin; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000097:blood* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003505:Cyclodextrins / Q000737:chemistry; D006801:Humans; D007202:Indicators and Reagents; D007660:Ketoprofen / Q000097:blood* / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012821:Silanes / Q000737:chemistry; D013237:Stereoisomerism; D047392:beta-Cyclodextrins
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D003505:Cyclodextrins; D007202:Indicators and Reagents; D012821:Silanes; D047392:beta-Cyclodextrins; C024779:octadecylsilane; D000073738:2-Hydroxypropyl-beta-cyclodextrin; D007660:Ketoprofen