Automated high-performance liquid chromatographic analysis of (-)-2'-deoxy-3'-thiacytidine in biological fluids using the automated sequential trace enrichment of dialysate systems.
A fully automated HPLC procedure was developed for the analysis of a small volume of perfusion solutions from an isolated perfused rat kidney study. The method involved separation of (-)-2'-deoxy-3'-thiacytidine (3TC) from the matrix by dialysis with 10 mM potassium phosphate buffer pH 3.0. 3TC was subsequently separated from the dialysate as it flowed through a SCX cation-exchange cartridge. The trapped 3TC was then eluted with a mobile phase of 50 mM ammonium acetate buffer (pH 5.5)-methanol (90.5:9.5, v/v) at a flow-rate of 1.0 ml/min for 2 min. The eluent was directed to the HPLC system and chromatographed with a BDS C18 analytical column at a temperature of 45 degrees C. Detection of 3TC was carried out by UV absorption at 274 nm. The procedure was validated from 25 to 10,000 ng/ml. Coefficients of variance (C.V.) of 3TC quality control samples were less than 9%. C.V.s of the standard curve samples were also less than 10% except for the 25 ng/ml samples (11.5%). The mean interpolated concentrations were within 8% of the nominal concentrations for all samples. No interference from concurrent drugs was observed. Preliminary results suggested that this procedure may also be used for human serum and urine samples.
Hsyu
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PH
Department of Clinical Pharmacology, Glaxo Research Institute, Triangle Park, NC 27709.
Lloyd
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TL
D000818:Animals; D000998:Antiviral Agents / Q000032:analysis*; D001322:Autoanalysis; D001826:Body Fluids / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods* / Q000706:statistics & numerical data; D003956:Dialysis; D006801:Humans; D007668:Kidney / Q000378:metabolism; D019259:Lamivudine; D051381:Rats; D012680:Sensitivity and Specificity; D016047:Zalcitabine / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000652:urine
D000998:Antiviral Agents; D019259:Lamivudine; D016047:Zalcitabine