New simplified microassay for the quantitation of theophylline and its major metabolites in serum by high-performance liquid chromatography

Journal of Chromatography
M B KesterH C Mansmann

Abstract

A high-performance liquid chromatographic technique is presented for simultaneous determination of theophylline, 3-methylxanthine, 1-methyluric acid, 1,3-dimethyluric acid and caffeine in serum using beta-hydroxyethyltheophylline (BHET) as an internal standard. An aliquot of a serum sample (100 microliters) was added to 40 microliters of an internal standard solution (BHET; 100 micrograms/ml) and vortexed. A 20% trichloroacetic acid solution (60 microliters) was then added; the mixture was vortexed, centrifuged and 100 microliters were injected onto a C8 column maintained at 45 degrees C. Inter- and intra-day variability of the assay for all compounds was less than 4.3%. Sensitivity ranged from 10 ng/ml for 1-methyluric acid to 25 ng/ml for theophylline. Drugs commonly coadministered with theophylline did not interfere with the assay.

References

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Citations

Jun 1, 1990·Antimicrobial Agents and Chemotherapy·M ParentM LeBel
Jun 1, 1990·Antimicrobial Agents and Chemotherapy·M LeBelM St-Laurent
Jul 22, 2015·Journal of Chromatographic Science·Fahad I Al-JenoobiAbdullah M Al-Mohizea

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