R. Lovlin et al., RAPID, SENSITIVE AND DIRECT CHIRAL HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR KETOPROFEN ENANTIOMERS, Journal of chromatography B. Biomedical applications, 679(1-2), 1996, pp. 196-198
Citations number
6
Categorie Soggetti
Chemistry Analytical","Biochemical Research Methods
Journal title
Journal of chromatography B. Biomedical applications
The stereospecific HPLC assays reported for ketoprofen (KT) mainly uti
lize indirect approaches. These assays involve the formation of amide
diastereomeric derivatives, which are then separated by chromatography
. The advantages of indirect methods include versatility, good sensiti
vity and cost effectiveness; however, lengthy preparation time is ofte
n required. Therefore, we have developed a new direct stereospecific H
PLC assay for KT enantiomers to improve preparation time and sensitivi
ty. The KT enantiomers and indomethacin, internal standard (I.S.), wer
e resolved using a Chiralpac AD column attached to 5 cm Supelcosil LC-
SI at constant temperature (30 degrees C). The mobile phase consisted
of hexane-isopropanol-trifluoroacetic acid (90:10:0.1). Under chromato
graphic conditions employed R-KT, S-KT and I.S. were eluted at 12, 14
and 16 min, respectively. A linear concentration response relationship
was found (0.05-5.0 mu g/ml of enantiomers) which covered normally ob
served concentrations in plasma after conventional doses of KT. The mi
nimum quantifiable concentration of the assay was found to be 0.025 or
0.25 mu g/ml based on 1 mi of human or 0.1 mi of rat plasma samples,
respectively. This direct HPLC method is suitable for pharmacokinetic
studies of KT enantiomers and offers the advantages of shorter sample
preparation and run time. This method is at least as sensitive as assa
ys currently in use.