Comparative determination of phenytoin in plasma by fluorescence polarization immunoassay and high performance liquid chromatography.
The need for careful monitoring of plasma concentrations of phenytoin during use of the drug in the treatment of epilepsy is well recognized; there can be great intersubject variation in the absorption rate and clearance rate of the drug, and its therapeutic ratio is narrow. In this study, two methods for determining plasma phenytoin concentrations were compared. One, based on fluorescence polarisation immunoassay (FPIA), is utilised in a commercially-available kit. The other, our own modification of a published procedure, was based on high performance liquid chromatography (HPLC). The accuracy and precision of both methods were evaluated, and the coefficients of variation (C.V.) were calculated. The C.V.'s ranged from 0.71 to 1.86% for the FPIA method, and from 2.81 to 8.69% for the HPLC method. Corresponding bias values were 1.20 to 1.60%, and 2.81 to 8.69%, respectively. A good correlation coefficient (0.977) was obtained, but estimated phenytoin concentrations were significantly higher (95% confidence level) using the HPLC method. We conclude that both methods perform adequately for clinical purposes. The HPLC method is, however, less expensive than the FPIA method.
Othman
S
S
Faculty of Pharmacy, University of Jordan, Amman.
al-Turk
W A
WA
Awidi
A S
AS
Daradkeh
T K
TK
Shaheen
O
O
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004827:Epilepsy / Q000188:drug therapy; D005455:Fluorescent Antibody Technique; D006801:Humans; D010672:Phenytoin / Q000097:blood* / Q000627:therapeutic use; D011933:Reagent Kits, Diagnostic
D011933:Reagent Kits, Diagnostic; D010672:Phenytoin