Laboratório de Toxicologia e Essencialidade de Metais, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, SP, Brazil.
Anal Chim Acta. 2013 Jan 25;761:11-7. doi: 10.1016/j.aca.2012.11.038. Epub 2012 Nov 29.
A simple and sensitive method with a fast sample preparation procedure is proposed for the determination of mercury species in plasma/serum. The method combines online high-performance liquid chromatography separation, Hg cold-vapor formation and inductively coupled plasma mass spectrometry detection. Prior to analysis, plasma (250 μL) was accurately pipetted into 15 mL conical tubes. Then, an extractant solution containing mercaptoethanol, L-cysteine and HCl was added to the samples following sonication for 10 min. Quantitative mercury extraction was achieved with the proposed procedure. Separation of mercury species was accomplished in less than 8 min on a C8 reverse phase column with a mobile phase containing 3% v/v methanol + 97% v/v (0.5% v/v 2-mercaptoethanol + 0.05% v/v formic acid). The method detection limits were found to be 12 ng L(-1), 5 ng L(-1) and 4 ng L(-1) for inorganic mercury, ethylmercury and methylmercury, respectively. Method accuracy is traceable to Standard Reference Material (SRM) 966 Toxic Metals in Bovine Blood from NIST. Additional validation was provided by the analysis of a secondary reference serum sample from the INSQ-Canada. Finally, the method was successfully applied for the speciation of mercury in plasma samples collected from volunteers exposed to methylmercury through fish consumption. For the first time to our knowledge, levels of different species of Hg in plasma samples from riverside populations exposed to MeHg were determined.
一种简单灵敏的方法,具有快速的样品制备程序,用于测定血浆/血清中的汞形态。该方法结合了在线高效液相色谱分离、汞冷蒸气形成和电感耦合等离子体质谱检测。在分析之前,将 250 μL 的血浆准确吸取到 15 mL 锥形管中。然后,在样品中加入含有巯基乙醇、L-半胱氨酸和 HCl 的萃取剂溶液,超声 10 min。采用所提出的程序可实现定量提取汞形态。在 C8 反相柱上,流动相含有 3% v/v 甲醇+97% v/v(0.5% v/v 2-巯基乙醇+0.05% v/v 甲酸),不到 8 min 即可完成汞形态的分离。对于无机汞、乙基汞和甲基汞,方法检测限分别为 12 ng L(-1)、5 ng L(-1)和 4 ng L(-1)。方法的准确性可追溯到 NIST 的牛血中有毒金属标准参考物质(SRM)966。通过对来自 INSQ-Canada 的二级参考血清样本的分析,提供了额外的验证。最后,该方法成功应用于通过鱼类摄入暴露于甲基汞的志愿者血浆样品中的汞形态分析。据我们所知,这是首次测定接触 MeHg 的河边人群血浆样品中不同种类 Hg 的水平。