L-cysteine mediated rapid separation of Hg and MeHg by anion-exchange HPLC.

Owing to the on-going emission of Hg into the global environment, new insight into their bioinorganic chemistry in mammals is urgently required to better understand their adverse health effects and analytical methods to quantify Hg and MeHg in environmental samples are needed. Analytical separations can help to address both of these needs. While Hg and MeHg have been most frequently separated by cation and reversed-phase (RP) HPLC, we here report on using anion-exchange (AEX) HPLC in conjunction with a flame atomic absorption spectrometer (FAAS) to observe the retention behavior of these mercury species in the pH range 5.0-8.0 using mobile phases comprised of 10 mM l-cysteine (Cys) in 100 mM phosphate buffer. The results obtained for pH 5.0 served as a starting point to develop a rapid HPLC separation for these mercurials. The addition of 5-20 % methanol (MeOH) to this mobile phase revealed that MeOH did not appreciably change the retention of Hg, but significantly reduced the retention of MeHg. A 15 % MeOH-containing mobile phase offered the best compromise between achieving a rapid baseline separation in <400 s at affordable costs. To assess the suitability and robustness of the developed AEX-HPLC separation method for the analysis of environmental samples an inductively coupled plasma atomic emission spectrometer (ICP-AES) was employed as the mercury-specific detector. The developed AEX-HPLC-ICP-AES method allowed to achieve detection limits of 1.5 ppm for Hg and 2.9 ppm for MeHg and was successfully applied to analyze wastewater that had been spiked with Hg and MeHg.