Determination of DuP 128, an ACAT inhibitor and its sulphoxide and sulphone metabolites in human plasma by liquid chromatography.
A sensitive and specific high-performance liquid chromatographic (HPLC) method with fluorescence detection was developed for the simultaneous determination of DuP 128 (N'-(2,4-difluorophenyl)-N-[5-(4,5-diphenyl-1H-imidazol-2-ylthio)p entyl]-N- hepthylurea), an ACAT inhibitor, its sulphone metabolite (XB277), and the separate determination of sulphoxide metabolite (XC164) in human plasma. After deproteinizing plasma samples with acetonitrile, the organic layer, created by adding approximately 0.25 g of NaCl, was removed, evaporated to dryness, and the residue then reconstituted with 400 microliters of acetonitrile. The acetonitrile layer was washed with 5 ml of hexane and then 50 microliters was injected into the HPLC. DuP 128 and XB277 were simultaneously quantified using a YMC basic column and fluorescence detection (lambda Ex = 270 nm and lambda Em = 385 nm). XC164 was quantified using a Waters microBondpack C18 reversed-phase column and fluorescence detection (lambda Ex = 270 nm and lambda Em = 365 nm). The relationship between the peak height and plasma concentrations best fit a power curve and showed an average correlation coefficient of > 0.99 over a concentration range of 1-200 ng ml-1 for DuP 128 and XC164 and 2.5-200 ng ml-1 for XB277. Good intraday and interday assay precisions (RSD < 10%) and accuracy (< 14%) for all three compounds were observed. The methods were sufficiently sensitive and selective to quantify plasma concentrations of DuP 128 and its sulphoxide and sulphone metabolites after oral administration of single or multiple dose(s) of > 350 mg of DuP 128 to healthy subjects.
Lai
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Drug Metabolism and Pharmacokinetics Section, DuPont Merck Pharmaceutical Company, Stine-Haskell Research Center, Newark, DE 19714.
Brogdon
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Quon
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Pieniaszek
H J
HJ
D000284:Administration, Oral; D000924:Anticholesteremic Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005453:Fluorescence; D006801:Humans; D007093:Imidazoles / Q000097:blood*; D008297:Male; D002785:Sterol O-Acyltransferase / Q000037:antagonists & inhibitors*; D013450:Sulfones / Q000097:blood; D013454:Sulfoxides / Q000097:blood; D014508:Urea / Q000031:analogs & derivatives* / Q000097:blood
D000924:Anticholesteremic Agents; D007093:Imidazoles; D013450:Sulfones; D013454:Sulfoxides; D014508:Urea; C084073:lecimibide; D002785:Sterol O-Acyltransferase