ALKYLATION OF IONIC MERCURY TO METHYLMERCURY AND DIMETHYLMERCURY BY METHYLCOBALAMIN - SIMULTANEOUS DETERMINATION BY PURGE-AND-TRAP GC IN-LINE WITH FTIR

A new approach was used to determine the reaction products of methylcobalamin and ionic mercury: purge-and-trap gas chromatography in line with Fourier transform infrared spectroscopy (PT GC/FTIR). This technique simultaneously and specifically determines the spectrum of dimethylmercury (DMeHg) and methylmercury produced by the reaction. No interference from other known organic mercury species could be detected. The method is different from others because ft does not require solvent extraction of the organomercurials from aqueous solution, but relies on immediate volatilization from the reaction vessel by addition of 100 mul of 10 mM NaBH4. The sample was purged with nitrogen for 10 min. The volatile species of mercury were trapped in a column at -120-degrees-C, injected into the gas chromatograph and detected by FTIR. The efficiency of DMeHg and MeHg formation depended on different parameters: pH, temperature, reaction time, and the methylcobalamin/ionic mercury ratio. The initial reaction product was MeHg which was further transformed to DMeHg. The first methylation rate was two times faster than the second. MeHg formed first, reaching a maximum at higher temperatures (28-degrees-C and 37-degrees-C) and later decreasing as DMeHg formed. At lower temperatures (20-degrees-C) the rate of MeHg formation was slower, being similar to the formation rate of DMeHg. Different species of inorganic mercury such as HgSO4, Hg(NO3)2, Hg(SCN)2, HgCl2 and HgI2 were used to study differences in methylation by methylcobalamin under standard conditions of acidity, temperature and cofactor Hg(II) ratio.