title: Metabolite and matrix interference in phenytoin immunoassays.
abstract: The major phenytoin metabolite, 5-(p-hydroxyphenyl)-5-phenylhydantoin glucuronide (HPPG), was primarily responsible for the positive bias noted when uremic specimens were assayed with the Abbott TDx Free Phenytoin fluorescence polarization immunoassay. The amount of bias depended on both HPPG and phenytoin concentration, increasing with increases in either concentration. The new Abbott TDx II assays for phenytoin and free phenytoin exhibited no significant cross-reactivity with HPPG and no bias in clinical specimens from uremic patients. Both assays correlated well with Emit-based assays (r >0.98), had CVs of <3.5%, and had minimum detection limits of <0.1 mg/L. Calibration curves were stable for at least 6 weeks. All of the TDx assays cross-reacted with another metabolite, 5-(p-hydroxyphenyl)-5-phenylhydantoin (HPPH), but expected HPPH concentrations are too low to cause a clinically significant bias. The Emit-based phenytoin assay exhibited a significant matrix effect when calibrators were prepared in defibrinated plasma processed to resemble serum.
authors.affiliation: Department of Laboratory Medicine, Yale University School of Medicine, New Haven, CT 06504, USA. petrie.rainey@yale.edu
authors.forename: P M
authors.initials: PM
authors.lastname: Rainey
authors.forename: K E
authors.initials: KE
authors.lastname: Rogers
authors.forename: W L
authors.initials: WL
authors.lastname: Roberts
mesh_terms: D002138:Calibration; D017248:Enzyme Multiplied Immunoassay Technique / Q000592:standards / Q000706:statistics & numerical data; D005454:Fluorescence Polarization; D006801:Humans; D007118:Immunoassay / Q000592:standards / Q000706:statistics & numerical data*; D010672:Phenytoin / Q000031:analogs & derivatives* / Q000097:blood*; D012044:Regression Analysis; D012680:Sensitivity and Specificity; D014511:Uremia / Q000097:blood*
chemical_list: C030590:5-phenyl-5-(4-hydroxyphenyl)hydantoin glucuronide; D010672:Phenytoin