Laboratório de Toxicologia e Essencialidade de Metais, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
Talanta. 2010 Jan 15;80(3):1158-63. doi: 10.1016/j.talanta.2009.09.001.
Despite the necessity to differentiate chemical species of mercury in clinical specimens, there are a limited number of methods for this purpose. Then, this paper describes a simple method for the determination of methylmercury and inorganic mercury in blood by using liquid chromatography with inductively coupled mass spectrometry (LC-ICP-MS) and a fast sample preparation procedure. Prior to analysis, blood (250microL) is accurately weighed into 15-mL conical tubes. Then, an extractant solution containing mercaptoethanol, l-cysteine and HCl was added to the samples following sonication for 15min. Quantitative mercury extraction was achieved with the proposed procedure. Separation of mercury species was accomplished in less than 5min on a C18 reverse-phase column with a mobile phase containing 0.05% (v/v) mercaptoethanol, 0.4% (m/v) l-cysteine, 0.06molL(-1) ammonium acetate and 5% (v/v) methanol. The method detection limits were found to be 0.25microgL(-1) and 0.1microgL(-1) for inorganic mercury and methylmercury, respectively. Method accuracy is traceable to Standard Reference Material (SRM) 966 Toxic Metals in Bovine Blood from the National Institute of Standards and Technology (NIST). The proposed method was also applied to the speciation of mercury in blood samples collected from fish-eating communities and from rats exposed to thimerosal. With the proposed method there is a considerable reduction of the time of sample preparation prior to speciation of Hg by LC-ICP-MS. Finally, after the application of the proposed method, we demonstrated an interesting in vivo ethylmercury conversion to inorganic mercury.
尽管有必要在临床标本中区分汞的化学物质，但为此目的的方法数量有限。然后，本文描述了一种简单的方法，用于通过使用液相色谱 - 电感耦合等离子体质谱（LC-ICP-MS）和快速样品制备程序来测定血液中的甲基汞和无机汞。在分析之前，将准确称重的 250μL 血液加入 15mL 锥形管中。然后，在超声处理 15min 后，向样品中加入含有巯基乙醇、L-半胱氨酸和 HCl 的提取剂溶液。通过所提出的程序实现了定量的汞提取。在 C18 反相柱上，使用含有 0.05％（v/v）巯基乙醇、0.4％（m/v）L-半胱氨酸、0.06molL（-1）乙酸铵和 5％（v/v）甲醇的流动相，在不到 5min 的时间内完成了汞形态的分离。无机汞和甲基汞的方法检测限分别为 0.25μg L（-1）和 0.1μg L（-1）。方法准确性可追溯到国家标准与技术研究所（NIST）的牛血中有毒金属标准参考物质（SRM）966。该方法还应用于从吃鱼的社区和暴露于硫柳汞的大鼠中采集的血液样品中汞的形态分析。使用所提出的方法，在通过 LC-ICP-MS 对 Hg 进行形态分析之前，大大减少了样品制备的时间。最后，在应用所提出的方法之后，我们证明了体内乙基汞向无机汞的有趣转化。
