Sidki A M, Pourfarzaneh M, Rowell F J, Smith D S
Ther Drug Monit. 1982;4(4):397-403. doi: 10.1097/00007691-198212000-00011.
A polarization fluoroimmunoassay for the direct determination of phenobarbital in serum or plasma was developed, optimized, and validated. The drug was derivatized by the introduction of a p-amino function in the phenyl group to enable coupling to carrier protein (used to raise antiserum in sheep) and to fluorescein label. The assay required simply the mixing of prediluted sample, labeled drug, and antiserum, followed by a short incubation and measurement of fluorescence polarization. Normal reliability criteria were satisfied and there was good correlation with established chromatographic and radioimmunoassay methods. Nonseparation, nonisotopic immunoassays are inherently susceptible to sample interferences, but it was shown that the polarization fluoroimmunoassay was subject to inaccuracy only in the presence of markedly elevated levels of bilirubin (above 500 mumol/L), or with gross hemolysis or lipemia.
开发、优化并验证了一种用于直接测定血清或血浆中苯巴比妥的偏振荧光免疫分析法。通过在苯基中引入对氨基官能团，使药物衍生化，以便与载体蛋白（用于在绵羊体内产生抗血清）偶联并进行荧光素标记。该分析只需将预稀释的样品、标记药物和抗血清混合，然后进行短时间孵育并测量荧光偏振。满足了正常的可靠性标准，并且与既定的色谱法和放射免疫分析法有良好的相关性。非分离、非同位素免疫分析法本质上易受样品干扰，但结果表明，偏振荧光免疫分析法仅在胆红素水平显著升高（高于500 μmol/L）、或存在严重溶血或脂血的情况下会出现不准确。
