A rapid method for the determination of methylmercury and inorganic mercury species in whole blood by liquid chromatography with detection using vapor generation ICP-MS/MS.

Speciation methods provide a more detailed picture regarding human exposure to toxic metals/metalloids and their effects on human health. The toxicity of methylmercury (MeHg) differs considerably from inorganic mercury (iHg), such that their separation and quantification in whole blood is helpful in identifying sources and possible pathways of exposure. Liquid chromatography (LC) has several advantages over gas chromatography (GC) for the separation of iHg from MeHg due to the former's compatibility with uptake rates of common nebulizer systems used with ICP-MS and the latter's requirement for a derivatization step to produce gaseous Hg species for an effective separation. Here we report an improved method that was developed to separate and quantify MeHg and iHg species in whole blood using isocratic LC elution with determination by vapor generation (VG) coupled with ICP-MS/MS. Chromatographic separation of MeHg and iHg is achieved in ∼4 minutes on a C8 reversed phase column. In those rare cases where there may be human exposure to ethylmercury (EtHg), or where a certified reference material (CRM) is known to contain EtHg (e.g., NIST SRM 955c), all three Hg species can be separated by extending the LC elution time to 8 minutes. Adding VG post column boosts the signal-to-noise ratio, and lowers the LOD. With optimized sample preparation, the LC-VG-ICP-MS/MS method LOD for both iHg and MeHg is 0.2 μg L-1. Method validation was conducted using NIST SRM 955c Toxic Metals in Caprine Blood and NIST SRM 955d Toxic Elements and Metabolites in Frozen Human Blood. Additional validation data were generated using archived blood reference materials from multiple Proficiency Testing programs and External Quality Assessment schemes. Blood-based quality control materials, previously analyzed for Hg species using isotope dilution with GC coupled to ICP-MS, were provided by the US CDC.