Jereb Vesna, Horvat Milena, Drobne Damjana, Pihlar Boris
Department of Environmental Sciences, Jozef Stefan Institute, Jamova 39, SI-1000 Ljubljana, Slovenia.
Sci Total Environ. 2003 Mar 20;304(1-3):269-84. doi: 10.1016/S0048-9697(02)00574-0.
The biological cycle of mercury in the terrestrial isopod Porcellio scaber was investigated. Testing the possibility of in vivo Hg(2+) methylation was divided into two methodologically different parts. Firstly, concentrations of total mercury and MeHg in isopods P. scaber and their environment from a Hg-unpolluted area were measured by the use of validated methods (CV AAS, CV AFS). The data obtained show that the percentage of MeHg in leaves, soil and faeces was less than 1%. In contrast, the percentage of MeHg in gut and hepatopancreas was increased to 14 and 77%, respectively, indicating methylation of Hg(2+) in the gut and its further accumulation in glands. To confirm this assumption, the second methodology was applied-a radiotracer technique with 203Hg(2+) of high specific activity. There are few radiotracer techniques for Hg-methylation assays; for our work we chose the method of Czuba et al. which includes alkaline leaching of Hg species, their extraction into dithizone-toluene, followed by specific separation of Hg dithizonates by thin-layer chromatography and gamma counting. All steps of the analytical protocol were checked and optimised by the use of aqueous solutions of 203Hg(2+) and Me(203)Hg(+). The most important finding was that cleaning-up the extract through a florisil column is not appropriate, because the column retains different percentages of Hg(2+) and MeHg(+) and consequently affects the accuracy of the final result. This optimised protocol was then applied to Hg transformation studies in the terrestrial isopod P. scaber. Leaching Hg species from P. scaber fed with 203Hg(2+) or Me(203)Hg(+) dosed food was completely efficient only at elevated temperatures. Preliminary results of methylation/demethlytion studies are rather variable but they show that both processes (Hg(2+)<-->MeHg(+)) take place in the isopod P. scaber. Additionally, an assessment of the mass balance of Hg in isopods P. scaber exposed to 203Hg(2+) indicates that volatile Hg species are also formed.
对陆生等足动物鼠妇体内汞的生物循环进行了研究。检测体内汞(II)甲基化可能性的实验分为两个方法不同的部分。首先,使用经过验证的方法(冷蒸气原子吸收光谱法、冷蒸气原子荧光光谱法)测量了来自汞未污染地区的鼠妇及其环境中总汞和甲基汞的浓度。所得数据表明,叶片、土壤和粪便中甲基汞的百分比低于1%。相比之下,肠道和肝胰腺中甲基汞的百分比分别增至14%和77%,这表明汞(II)在肠道中发生了甲基化,并进一步在腺体中积累。为了证实这一假设,采用了第二种方法——使用高比活度的203汞(II)的放射性示踪技术。用于汞甲基化分析的放射性示踪技术很少;在我们的研究中,我们选择了祖巴等人的方法,该方法包括汞物种的碱性浸出、将其萃取到双硫腙 - 甲苯中,然后通过薄层色谱法和伽马计数对双硫腙汞进行特定分离。通过使用203汞(II)和甲基(203)汞(+)的水溶液对分析方案的所有步骤进行了检查和优化。最重要的发现是,通过弗罗里硅土柱净化提取物不合适,因为该柱对汞(II)和甲基汞(+)的保留率不同,从而影响最终结果的准确性。然后将这个优化后的方案应用于陆生等足动物鼠妇的汞转化研究。仅在高温下,从喂食了203汞(II)或甲基(203)汞(+)的鼠妇中浸出汞物种才完全有效。甲基化/去甲基化研究的初步结果差异较大,但它们表明这两个过程(汞(II)⇄甲基汞(+))在鼠妇体内均会发生。此外,对暴露于203汞(II)的鼠妇体内汞的质量平衡评估表明,还会形成挥发性汞物种。