Quantitative determination of CGP 53,437, a new HIV protease inhibitor, in plasma by high-performance liquid chromatography and fluorescence detection.
A specific and sensitive liquid chromatographic assay for CGP 53,437 (I), a potent HIV protease inhibitor, is described. The method is based on a deproteinization step, followed by a liquid-liquid extraction with diisopropyl ether. Then a deprotection step of the primary amine and derivatization using fluorescamine is performed. Chromatography is achieved by isocratic elution with a mobile phase of 63 mM borax buffer (pH 9)-acetonitrile (58:42, v/v). The flow-rate of the mobile phase is 1 ml/min. The derivatives of compound I and its internal standard CGP 54,451, II, fluoresce at 480 nm on excitation at 395 nm. The limit of quantitation which is the lowest concentration of the analyte that can be measured with a coefficient of variation and a deviation from theory of less than 20%, was 5 nmol/l plasma. The analyte is stable for at least seven months in spiked human plasma samples. It is also stable after freezing and thawing cycles. Different human plasma sources and plasma samples from three different species (dog, marmoset, and rat) were tested and no interferences from plasma constituents was observed.
Ciba-Geigy Ltd, Basle, Switzerland.
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Flesch
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Mann
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Boss
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Lang
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Degen
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Dieterle
D000818:Animals; D000998:Antiviral Agents / Q000097:blood*; D002144:Callithrix; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data; D004285:Dogs; D004355:Drug Stability; D004987:Ethers; D005260:Female; D005450:Fluorescamine; D017320:HIV Protease Inhibitors / Q000097:blood*; D006801:Humans; D015394:Molecular Structure; D009025:Morpholines / Q000097:blood* / Q000493:pharmacokinetics; D009842:Oligopeptides / Q000097:blood* / Q000493:pharmacokinetics; D011786:Quality Control; D051381:Rats; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013696:Temperature
D000998:Antiviral Agents; D004987:Ethers; D017320:HIV Protease Inhibitors; D009025:Morpholines; D009842:Oligopeptides; C084754:Boc-Phe-psi(CH(OH)CH2)-Phe-Val-Phe-morpholine; D005450:Fluorescamine; C011779:diisopropyl ether