Mole M L, Sanders L, Oglesby L A
Environmental Protection Agency, Research Triangle Park, North Carolina 27711.
Anal Biochem. 1988 Nov 15;175(1):74-84. doi: 10.1016/0003-2697(88)90363-6.
A high-performance liquid chromatographic method has been modified for the evaluation of both Phase I and II metabolism of biphenyl by hepatocytes maintained in an embryo/hepatocyte co-culture medium. Extracts of the media, before and after hydrolysis of conjugates, are directly injected onto the HPLC and the major hydroxylated metabolites plus unmetabolized biphenyl are detected by fluorescence after separation under gradient or isocratic conditions. The method is almost free of interferences and is relatively simple and rapid. In the case of the monohydroxylated derivatives, the minimum media concentrations which can be measured are 7 to 20 nM (0.07 to 0.2 pmol on-column). Recoveries from culture medium to which known amounts of biphenyl and metabolites had been added were quantitative (90-103%) and the reproducibility good (interassay CV less than 5%). The assay was applied to cultures of hepatocytes derived from rabbit and from phenobarbital induced and noninduced rat.
一种高效液相色谱法已被改进，用于评估在胚胎/肝细胞共培养基中培养的肝细胞对联苯的I相和II相代谢。在结合物水解前后的培养基提取物直接注入高效液相色谱仪，在梯度或等度条件下分离后，通过荧光检测主要的羟基化代谢物和未代谢的联苯。该方法几乎没有干扰，相对简单且快速。对于单羟基化衍生物，可测量的最低培养基浓度为7至20 nM（柱上0.07至0.2 pmol）。向已知添加了联苯和代谢物的培养基中的回收率是定量的（90 - 103%），重现性良好（批间变异系数小于5%）。该测定法应用于来自兔子以及苯巴比妥诱导和未诱导的大鼠的肝细胞培养物。
