Stereospecific HPLC method for the quantitation of the enantiomers of MK-0571, a potent leukotriene D4 receptor antagonist, in biological specimens.
A stereospecific HPLC bioanalytical method was developed for quantitation of the enantiomers of MK-0571, a leukotriene D4 receptor antagonist. The procedure involves the addition of an internal standard analog to the biological matrix followed by extraction of the free acids into ethyl acetate. The acids are subsequently reacted with the homochiral reagent, (+)-(R)-alpha-(1-naphthyl)ethylamine (NEA) to form diastereomers. Following removal of excess reagent and side products by a dilute acid wash, the NEA-MK-0571 diastereomers are separated on a phenyl urea chiral column using a mobile phase containing hexane, isopropanol, and acetonitrile and are detected with a fluorescence detector. The sensitivity of the method is such that 50 ng of each enantiomer can be quantitated. In the 0.05 to 10 micrograms range the recoveries of the enantiomers of MK-0571 from plasma were 100.4 +/- 7.9% and 100.0 +/- 7.2%. NMR and mass spectral data confirmed the structure of the derivative. The method has been utilized in drug safety evaluation studies to demonstrate enantioselectivity in disposition of the enantiomers of MK-0571 in rats and monkeys but not in mice.
Lee
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Department of Safety Assessment, Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania 19486.
Zacchei
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D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005021:Ethylamines; D008253:Macaca mulatta; D009682:Magnetic Resonance Spectroscopy; D008297:Male; D013058:Mass Spectrometry; D008722:Methods; D009281:Naphthalenes; D011422:Propionates / Q000032:analysis* / Q000097:blood; D011804:Quinolines / Q000032:analysis* / Q000097:blood; D051381:Rats; D011971:Receptors, Immunologic / Q000037:antagonists & inhibitors; D018077:Receptors, Leukotriene; D015203:Reproducibility of Results; D013237:Stereoisomerism
D005021:Ethylamines; D009281:Naphthalenes; D011422:Propionates; D011804:Quinolines; D011971:Receptors, Immunologic; D018077:Receptors, Leukotriene; C055450:1-(1-naphthyl)ethylamine; C059141:verlukast