Nichkova Mikaela, Park Eun-Kee, Koivunen Marja E, Kamita Shizuo G, Gee Shirley J, Chuang Jane, Emon Jeanette M Van, Hammock Bruce D
Department of Entomology, Cancer Research Center, University of California, 303 Briggs Hall, One Shields Avenue, Davis, CA 95616, USA.
Talanta. 2004 Aug 8;63(5):1213-23. doi: 10.1016/j.talanta.2004.05.030.
Polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) are considered highly toxic contaminants and the environmental and biological monitoring of these compounds is of great concern. Immunoassays may be used as screening methods to satisfy the growing demand for rapid and low cost analysis. In this work, we describe the application of an immunoassay that uses 2,3,7-trichloro-8-methyldibenzo-p-dioxin (TMDD) as a surrogate standard for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) to sediment and human serum samples. Sample extraction and preparation methods were developed with the aim to establish the simplest, cost-effective and efficient removal of the matrix interferences in the enzyme-linked immunosorbent assay (ELISA). The overall method for sediments is based on a hexane extraction; clean up by a multilayered silica gel column and an activated carbon column; an organic solvent exchange with DMSO-Triton X-100 and ELISA measurement. The gas chromatography-high resolution mass spectrometry (GC-HRMS) validation studies (n = 13) revealed that the method is suitable for the toxic equivalents (TEQ) screening of dioxin in sediments with a method detection limit of about 100pgg(-1) dry sediment with a precision of 13-33% R.S.D. The analysis of a large number of samples originating from different sources would be required to establish more precisely the screening level, as well as the number of false positives and negatives of dioxin TEQ by the immunoassay for sediments. The immunoassay method for sediment analysis offers improvement in speed, sample throughput, and cost in comparison to GC-HRMS. Dioxins were determined in serum samples after a simple liquid-liquid extraction and solvent exchange into DMSO-Triton X-100 without further dilution. The current method (approximate method LOQ of 200pgml(-1) serum) is not sufficiently sensitive for the determination of dioxins in serum to measure acceptable exposure limit.
多氯代二苯并 - 对 - 二噁英(PCDDs)和多氯代二苯并呋喃(PCDFs)被视为剧毒污染物,对这些化合物进行环境和生物监测备受关注。免疫测定法可用作筛选方法,以满足对快速低成本分析日益增长的需求。在本研究中,我们描述了一种免疫测定法的应用,该方法使用2,3,7 - 三氯 - 8 - 甲基二苯并 - 对 - 二噁英(TMDD)作为2,3,7,8 - 四氯二苯并 - 对 - 二噁英(TCDD)的替代标准物,用于沉积物和人血清样本。开发了样品提取和制备方法,目的是在酶联免疫吸附测定(ELISA)中建立最简单、经济高效且能有效去除基质干扰的方法。沉积物整体方法基于己烷萃取;通过多层硅胶柱和活性炭柱净化;与二甲基亚砜 - 聚乙二醇辛基苯基醚(DMSO - Triton X - 100)进行有机溶剂交换并进行ELISA测定。气相色谱 - 高分辨率质谱(GC - HRMS)验证研究(n = 13)表明,该方法适用于沉积物中二噁英的毒性当量(TEQ)筛选,方法检测限约为100 pg g⁻¹干沉积物,精密度为相对标准偏差(R.S.D.)的13 - 33%。需要分析大量来自不同来源的样本,以更精确地确定筛选水平,以及沉积物免疫测定法中二噁英TEQ的假阳性和假阴性数量。与GC - HRMS相比,沉积物分析的免疫测定法在速度、样品通量和成本方面有改进。血清样本经简单液 - 液萃取并将溶剂交换为DMSO - Triton X - 100后,无需进一步稀释即可测定二噁英。当前方法(血清的方法定量下限约为200 pg ml⁻¹)对血清中二噁英的测定不够灵敏,无法测量可接受的暴露限值。