Vrije Universiteit Brussel, Department of Analytical and Environmental Chemistry (ANCH), Pleinlaan 2, 1050 Brussels, Belgium.
Talanta. 2011 Oct 15;85(5):2484-91. doi: 10.1016/j.talanta.2011.07.103. Epub 2011 Aug 5.
Since the CALUX (Chemically Activated LUciferase gene eXpression) bioassay is a fast and inexpensive tool for the throughput analysis of dioxin-like compounds in a large number of samples and requires only small sample volumes, the use of this technique in human biomonitoring programs provides a good alternative to GC-HRMS. In this study, a method for the separate analysis of PCDD/Fs and dioxin-like PCBs (dl-PCBs) in human serum with the new sensitive H1L7.5c1 mouse hepatoma cell line was optimized. Sample dilution factors of 5 and 2.4 were selected for routine analysis of respectively the PCDD/Fs and dl-PCBs. The validation studies showed that repeatability and within-lab reproducibility for the quality control (QC) standard were within the in-house criteria. A long-term within-lab reproducibility of 25% for the PCDD/F fraction and 41% for the dl-PCB fraction for the analysis of pooled serum samples, expressed as pg BEQ/g fat, was determined. CALUX recoveries of the spiked procedural blanks were within the acceptable in-house limits of 80-120% for both fractions and the LOQ was 30.3 pg BEQ/g fat for the PCDD/Fs and 14.5 pg BEQ/g fat for the dl-PCBs. The GC-HRMS recovery of a C13-spiked pooled serum sample was between 60 and 90% for all PCDD/F congeners and between 67 and 82% for the non-ortho PCBs. An adequate separation between both fractions was found. The CALUX/GC-HRMS ratio for a pooled serum sample was respectively 2.0 and 1.4 for the PCDD/Fs and the dl-PCBs, indicating the presence of additional AhR active compounds. As expected, a correlation was found between human serum samples analyzed with both the new H1L7.5c1 cell line and the more established H1L6.1c3 cell line. The geometric mean CALUX-BEQ values, reported for the adolescents of the second Flemish Environment and Health Study (FLEHS II) recruited in 2009-2010, were 108 (95% CI: 101-114) pg CALUX-BEQ/g fat for the PCDD/Fs and 32.1 (30.1-34.2) pg CALUX-BEQ/g fat for the dioxin-like PCBs.
由于 CALUX（化学激活的荧光素酶基因表达）生物测定法是一种快速且廉价的工具，可用于高通量分析大量样品中的类二恶英化合物，并且仅需要小体积的样品，因此该技术在人体生物监测计划中提供了一种替代 GC-HRMS 的良好选择。在这项研究中，优化了使用新型敏感的 H1L7.5c1 小鼠肝癌细胞系分别分析人血清中 PCDD/Fs 和类二恶英 PCB（dl-PCBs）的方法。选择了 5 和 2.4 的样品稀释因子，分别用于常规分析 PCDD/Fs 和 dl-PCBs。验证研究表明，QC 标准的重复性和实验室内部再现性在内部标准范围内。对于分析混合血清样本，PCDD/F 部分的长期实验室内部再现性为 25%，dl-PCB 部分为 41%，以 BEQ/g 脂肪表示。两个部分的加标程序空白的 CALUX 回收率均在可接受的内部 80-120%范围内，LOQ 分别为 PCDD/Fs 为 30.3 pg BEQ/g 脂肪和 dl-PCBs 为 14.5 pg BEQ/g 脂肪。C13 加标混合血清样品的 GC-HRMS 回收率对于所有 PCDD/F 同系物均在 60-90%之间，对于非邻位 PCB 则在 67-82%之间。发现两个部分之间有足够的分离。对于混合血清样品，CALUX/GC-HRMS 比分别为 PCDD/Fs 的 2.0 和 dl-PCBs 的 1.4，表明存在其他 AhR 活性化合物。如预期的那样，在使用新型 H1L7.5c1 细胞系和更成熟的 H1L6.1c3 细胞系分析的人血清样本之间发现了相关性。2009-2010 年招募的第二个佛兰芒环境与健康研究（FLEHS II）的青少年的几何平均 CALUX-BEQ 值，报告为 PCDD/Fs 的 108（95%CI：101-114）pg CALUX-BEQ/g 脂肪和 32.1（30.1-34.2）pg CALUX-BEQ/g 脂肪用于二恶英类 PCBs。
