Gas-liquid chromatographic analysis of fluoropyrimidine nucleosides and fluorouracil in plasma and urine |
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| Authors: | W M Williams B S Warren F Lin |
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| Institution: | 1. Department of Anesthesiology, University of Tennessee Health Science Center, Memphis, TN, USA;2. School of Medicine, Royal College of Surgeons in Ireland, Dublin, Ireland;1. Department of Clinical Sciences, Section for Anaesthesiology and Intensive Care, Lund University, SE-22185 Lund, Sweden;2. Department of Intensive and Perioperative Care, Skåne University Hospital, SE-22185 Lund, Sweden;1. Paul C Lauterbur Research Centre for Biomedical Imaging, Institute of Biomedical and Health Engineering, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, China;2. Centers for Biomedical Engineering, College of Information Science and Technology, University of Science and Technology of China, Hefei, China;3. Department of Electrical and Computer Engineering, Texas A&M University, College Station, TX, USA;4. School of Information Science and Technology, Northwest University, Xian, China |
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| Abstract: | A gas-liquid chromatographic method employing on-column alkylation and a nitrogen-sensitive detector was developed for the analysis of 5-fluoro-2'-deoxyuridine, 5-fluorouridine, and 5-fluorouracil in plasma and urine. Samples (0.72 ml) containing the fluoropyrimidine and internal standard (5-chloro-2'-deoxyuridine for nucleoside analyses and 6-methyluracil for 5-fluorouracil analyses) were prepared for gas-liquid chromatography by sequential cation-exchange and anion-exchange column chromatography. Recoveries of fluoropyrimidines were 71-95% over the concentration ranges studied. The dried eluate from the anion-exchange column was dissolved in p-tolyltrimethylammonium hydroxide in methanol before gas-liquid chromatographic analysis. Columns packed with either 3% SP-2100 on Supelcoport or 3% OV-1 on Gas-Chrom Q were suitable for nucleoside analyses; a column packed with 0.75% Carbowax 20M-5% KOH on Chromsorb G was used for 5-fluorouracil analyses. The fluoropyrimidine nucleosides were well separated from each other and from the potentially interfering endogenous compounds 2'-deoxyuridine and uridine; 5-fluorouracil was well separated from uracil. Linear standard curves (peak area ratio method) were obtained for plasma containing 0.025 to 20 micrograms FdUrd (0.1 to 81 microM) or 0.05 to 1.0 microgram FUrd (0.2 to 3.8 microM), and for urine containing 0.2 to 1.0 microgram (0.8 to about 4 microM) of the nucleosides. Standard curves for 5-fluorouracil (1.5 to 7.9 microM) and 2'-deoxyuridine (0.9 to 4.4 microM) were also linear. A measurable amount of 5-fluorouracil, equivalent to 4 to 7% of the 5-fluoro-2'-deoxyuridine injected, was formed from the nucleoside on the gas-liquid chromatographic column, requiring correction of 5-fluorouracil concentrations measured in the presence of 5-fluoro-2'-deoxyuridine. |
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