Ropero M J Patiño, Fariñas N Rodríguez, Krupp E, Mateo R, Nevado J J Berzas, Martín-Doimeadios R C Rodríguez
Universidad de Castilla-La Mancha, 45.071 Toledo, Spain.
University of Aberdeen, Chemistry Department, AB 24 3UE Aberdeen, United Kingdom.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Jun 1;1022:159-166. doi: 10.1016/j.jchromb.2016.04.003. Epub 2016 Apr 3.
Mercury (Hg) is likely bound to large biomolecules (e.g. proteins) in living organisms, and in order to assess Hg metabolic pathways and possible toxicological effects, it is essential to study these Hg containing biomolecules. However, the exact nature of most metal binding biomolecules is unknown. Such studies are still in their infancy and information on this topic is scarce because the analysis is challenging, mainly due to their lability upon digestion or extraction from the tissue. New analytical methods that allow complex Hg-biomolecules to be analysed intact are needed and only few very recent studies deal with this approach. Therefore, as an initial step towards the characterization of Hg containing biomolecules, an analytical procedure has been optimised using size-exclusion chromatography (SEC) with inductively coupled plasma mass spectrometry (ICP-MS) detection. We applied this technique to elucidate the distribution and elution profile of Hg and Se, and some physiological important elements such as Fe, Ni, Zn and Cu, to assess metal binding profiles in liver and kidney samples of red deer (Cervus elaphus) and wild boar (Sus scrofa) who roam freely within the largest Hg mining district on Earth, Almadén in Spain. Elemental fractionation profiles of the extracts from different tissues were obtained using two different SEC columns (BioSep-SEC-S2000 GL 300-1kDa and Superdex 75 10/300 GL 70-3kDa). Similar profiles of Hg were observed in red deer and wild boar; however, significant differences were evident for liver and kidney. Moreover, the profiles of Se showed a single peak at high-medium molecular weight in all investigated tissues, while co-elution of Hg with Fe, Ni, Zn and Cu was observed.
汞（Hg）可能与生物体内的大分子（如蛋白质）结合，为了评估汞的代谢途径和可能的毒理学效应，研究这些含汞生物分子至关重要。然而，大多数金属结合生物分子的确切性质尚不清楚。此类研究仍处于起步阶段，关于这一主题的信息稀缺，因为分析具有挑战性，主要是由于它们在从组织中消化或提取时不稳定。需要新的分析方法来完整分析复杂的汞生物分子，而近期只有少数研究涉及这种方法。因此，作为表征含汞生物分子的第一步，已优化了一种分析程序，该程序使用尺寸排阻色谱（SEC）结合电感耦合等离子体质谱（ICP-MS）检测。我们应用该技术来阐明汞和硒以及一些生理重要元素（如铁、镍、锌和铜）的分布和洗脱曲线，以评估在西班牙阿尔马登（地球上最大的汞矿区）自由活动的马鹿（Cervus elaphus）和野猪（Sus scrofa）肝脏和肾脏样本中的金属结合情况。使用两种不同的SEC柱（BioSep-SEC-S2000 GL 300-1kDa和Superdex 75 10/300 GL 70-3kDa）获得了不同组织提取物的元素分级曲线。在马鹿和野猪中观察到了相似的汞曲线；然而，肝脏和肾脏存在明显差异。此外，在所有研究组织中，硒的曲线在中高分子量处显示出一个单峰，同时观察到汞与铁、镍、锌和铜的共洗脱。
