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COMPARISON OF FLUORESCENCE POLARIZATION IMMUNOASSAY AND HPLC FOR THE DETERMINATION OF THEOPHYLLINE IN SERUM
Author(s) -
ElSayed Y. M.,
Islam S. I.
Publication year - 1989
Publication title -
journal of clinical pharmacy and therapeutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.622
H-Index - 73
eISSN - 1365-2710
pISSN - 0269-4727
DOI - 10.1111/j.1365-2710.1989.tb00231.x
Subject(s) - theophylline , fluorescence polarization immunoassay , high performance liquid chromatography , chromatography , chemistry , caffeine , immunoassay , quantitative analysis (chemistry) , coefficient of variation , pharmacology , medicine , antibody , immunology
Summary Theophylline in serum was measured by fluorescence polarization immunoassay (FPIA) and by high‐performance liquid chromatography (HPLC). Within‐run precision studies using control samples in the subtherapeutic, therapeutic and toxic concentrations, resulted in coefficients of variation in the range of 2·86–3·12% (FPIA) and 2·1–3·66% (HPLC), respectively. Between‐run precision ranged from 2·76‐6·2% for FPIA and from 2·51–6·0% for HPLC. The mean recovery for three spiked controls was 98·9% for FPIA and 98·8% for HPLC. Comparison of 60 patients' samples, assayed with both methods, indicated an extremely good analytical correlation ( r = 0·990). The FPIA method displayed a slight but consistent positive bias in relation to the concentration of theophylline present in patients sera. Caffeine was found to exhibit a positive bias to 13%, over a caffeine concentration range of 10–40 μg/ml. The HPLC method offers an advantage for measurements of both caffeine and theophylline simultaneously. The FPIA offers significant advantages in speed of analysis and turnover‐time, while maintaining accuracy and precision compared with those of established HPLC procedures.