<Originals>High-performance liquid chromatographic determination of theophylline by direct serum injection

概要

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Isocratic high-performance liquid chromatography employing direct injection of microserum has been developed for theophylline quantitation in human serum. The packing material of the analytical column consists of a hybrid ligand containing hydrophobic and hydrophilic phases. Disodium hydrogenphosphate buffer (0.1M, pH7.0) is used as the mobile phase, and theophylline is directly detected at a wavelength of 280nm. Serum samples of 5μl were directly injected to the analytical column, and total analytical time was 5 minutes. Chromatograms showed high separation factors, and linearities of the calibration curve were good. Coefficients of variation for within-day reproducibility was 1.14〜2.52% for the concentration range of 16.6〜223μmol/l ; for between-day reproducibility it was 2.16% for the concentration of 48.5μmol/l, and the recovery rate was 100〜104%. The minimum limit of detection was 2.0μmol/l at a signal-to-noise ratio of 3 : 1. No interference from metabolites or many currently used drugs in serum was found. Correlation coefficient between the homogenous enzymemultiplied immunoassay and this method was 0.984. The method is simple, specific, fast and less expensive in routine clinical laboratory testing.
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