Bradlaw J A, Casterline J L
J Assoc Off Anal Chem. 1979 Jul;62(4):904-16.
Induction of aryl hydrocarbon hydroxylase (AHH) activity in rat hepatoma cell line serves as a simple and rapid method to detect minute (pg) amounts of certain classes of compounds, e.g., dibenzo-p-dioxins, dibenzofurans, and biphenyls. This method may provide a quick screen for such substances in extracts from foods prior to chemical identification. AHH activity is measured by conversion of benzo[a]pyrene (BP) to 3-hydroxy BP in homogenized cell extracts from control and treated cultures and is reported as pmol product formed/mg protein/min. Substances screened by this method include polyhalogenated analogs of dibenzo-p-dioxin (24 compounds), dibenzofuran (11 compounds), biphenyl (7 compounds), and extracts from several food sources. Response of the most reactive compound, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) was used to prepare a standard curve, and the AHH activity induced by mole doses of test substance is reported as an ED50 response (the estimated dose needed to produce 50% maximum enzyme induction). The AHH activity induced by food extracts is equated to the standard curve and reported as TCDD equivalents. A potent ED50 response in cell culture appears to correlate well with known toxic responses in other mammalian and avian systems for certain test substances. This correlation suggests that the cell culture enzyme induction method is a useful model for screening food extracts that are suspected to be contaminated with polychlorinated planar substances. A collaborative study would demonstrate the reproducibility of the method.
大鼠肝癌细胞系中芳烃羟化酶（AHH）活性的诱导，是检测微量（皮克）特定种类化合物（如二苯并 - p - 二恶英、二苯并呋喃和联苯）的一种简单快速的方法。该方法可在进行化学鉴定之前，对食品提取物中的此类物质进行快速筛选。通过将对照培养物和处理过的培养物的匀浆细胞提取物中的苯并[a]芘（BP）转化为3 - 羟基BP来测量AHH活性，结果以每毫克蛋白质每分钟形成的产物皮摩尔数表示。用该方法筛选的物质包括二苯并 - p - 二恶英的多卤代类似物（24种化合物）、二苯并呋喃（11种化合物）、联苯（7种化合物）以及几种食物来源的提取物。使用反应性最强的化合物2,3,7,8 - 四氯二苯并 - p - 二恶英（TCDD）的响应来绘制标准曲线，测试物质摩尔剂量诱导的AHH活性以ED50响应表示（产生最大酶诱导50%所需的估计剂量）。食物提取物诱导的AHH活性与标准曲线进行比对，并以TCDD当量表示。对于某些测试物质，细胞培养中的有效ED50响应似乎与其他哺乳动物和鸟类系统中已知的毒性响应密切相关。这种相关性表明，细胞培养酶诱导方法是筛选疑似被多氯平面物质污染的食物提取物的有用模型。一项合作研究将证明该方法的可重复性。
