Schirmer Kristin, Dayeh Vivian R, Bopp Stephanie, Russold Sandra, Bols Niels C
Department of Cell Toxicology, UFZ Centre for Environmental Research Leipzig-Halle, Permoserstr. 15, Leipzig 04318, Germany.
Toxicology. 2004 Dec 15;205(3):211-21. doi: 10.1016/j.tox.2004.06.053.
Methodology was developed in order to rapidly and cost-efficiently screen whole water samples without extraction for the presence of dioxin-like compounds using a cell bioassay approach. Presence of dioxin-like compounds was indicated by the induction in the rainbow trout (Oncorhynchus mykiss) liver cell line, RTL-W1, of cytochrome CYP1A, which was measured as 7-ethoxyresorufin-O-deethylase (EROD) activity. Two simple culture media, L-15/ex and Earle's-G, prepared in tissue culture water and supplemented with 5% serum, proved suitable for supporting RTL-W1 cell viability and induction of EROD activity by the model inducers, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and benzo[a]pyrene (BaP). Preparation of the same simplified media using whole surface and ground water instead of tissue culture water again allowed EROD induction by spiked TCDD and BaP to be detected but higher concentrations of inducers were necessary. Despite this reduced sensitivity, RTL-W1 cells responded to 4 out of 40 ground water samples from a former oil and lignite processing site with significant EROD induction. In the future, the value of the bioassay is as an inexpensive means of quickly screening ground and surface water samples to identify high contaminant levels particularly at industrial sites, where detailed site-investigations and long-term monitoring programs are required.
已开发出一种方法,以便使用细胞生物测定法快速且经济高效地筛选未经萃取的整个水样中是否存在二噁英类化合物。通过虹鳟(Oncorhynchus mykiss)肝细胞系RTL-W1中细胞色素CYP1A的诱导来表明二噁英类化合物的存在,细胞色素CYP1A的诱导通过7-乙氧基异吩噁唑酮-O-脱乙基酶(EROD)活性来测定。在组织培养用水中制备并添加5%血清的两种简单培养基L-15/ex和Earle's-G,被证明适合维持RTL-W1细胞活力以及由模型诱导剂2,3,7,8-四氯二苯并对二噁英(TCDD)和苯并[a]芘(BaP)诱导EROD活性。使用地表水和地下水而非组织培养用水制备相同的简化培养基,再次能够检测到加标TCDD和BaP诱导的EROD,但需要更高浓度的诱导剂。尽管灵敏度有所降低,但RTL-W1细胞对来自一个 former oil and lignite processing site的40个地下水样中的4个有显著的EROD诱导反应。未来,生物测定法的价值在于作为一种廉价手段,快速筛选地表水和地下水样,以识别高污染水平,特别是在需要详细场地调查和长期监测计划的工业场地。