Advantages of achiral h.p.l.c. as a preparative step for chiral analysis in biological samples and its use in toxicokinetic studies.
1. Achiral reverse-phase h.p.l.c. with semi-automated post-column fraction collection and solid-phase sample reconcentration, has been applied as the purification procedure during the enantiomeric quantification of two widely differing experimental drugs; an HMG-CoA reductase inhibitor (I) and an alpha 2-adrenoceptor antagonist (II). 2. The robust and specific achiral methodologies were available prior to the need for chiral analyses and recovery of drug from the fractions provided clean samples from a variety of biological matrices, without the need to develop compatible achiral/chiral mobile phases. 3. Compared with direct chiral chromatography of plasma extracts, this approach decreased the potential for metabolites and endogenous components to interfere or impair the performance of the chiral stationary phase. 4. The availability of quantitative data from achiral analysis of samples negated the need for internal standardization of the chiral analyses, helped confirm assay specificity and provided potential to determine enantiomeric ratios where only one isomer could be accurately measured. 5. Routine enantiomeric analyses were successfully carried out on samples taken from animals dosed orally with the racemic drugs, providing important data on the possible levels of exposure to individual enantiomers during toxicity testing.
Drug Metabolism Department, Glaxo Group Research Limited, Greenford, Middlesex, UK.
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Beresford
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Caswell
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Chambers
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Kirk
D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004285:Dogs; D005260:Female; D006904:Hydroxymethylglutaryl-CoA Synthase / Q000037:antagonists & inhibitors; D007093:Imidazoles / Q000302:isolation & purification* / Q000493:pharmacokinetics / Q000633:toxicity; D008297:Male; D010883:Piperoxan / Q000031:analogs & derivatives* / Q000302:isolation & purification / Q000493:pharmacokinetics / Q000633:toxicity; D011758:Pyrroles / Q000302:isolation & purification* / Q000493:pharmacokinetics / Q000633:toxicity; D051381:Rats; D013237:Stereoisomerism
D007093:Imidazoles; D011758:Pyrroles; C078258:GR 92549; D010883:Piperoxan; D006904:Hydroxymethylglutaryl-CoA Synthase; C065161:fluparoxan