Abstract
A procedure for the extraction and determination of methyl mercury and mercury (II) in fish muscle tissues and sediment samples is presented. The procedure involves extraction with 5% (v/v) 2-mercaptoethanol, separation and determination of mercury species by HPLC-ICPMS using a Perkin-Elmer 3μm C8 (33mm×3mm) column and a mobile phase 3 containing 0.5% (v/v) 2-mercaptoethanol and 5% (v/v) CH3OH (pH 5.5) at a flow rate 1.5mlmin−1 and a temperature of 25°C. Calibration curves for methyl mercury (I) and mercury (II) standards were linear in the range of 0–100μgl−1 (r2=0.9990 and r2=0.9995 respectively). The lowest measurable mercury was 0.4μgl−1 which corresponds to 0.01μgg−1 in fish tissues and sediments. Methyl mercury concentrations measured in biological certified reference materials, NRCC DORM – 2 Dogfish muscle (4.4±0.8μgg−1), NRCC Dolt – 3 Dogfish liver (1.55±0.09μgg−1), NIST RM 50 Albacore Tuna (0.89±0.08μgg−1) and IRMM IMEP-20 Tuna fish (3.6±0.6μgg−1) were in agreement with the certified value (4.47±0.32μgg−1, 1.59±0.12μgg−1, 0.87±0.03μgg−1, 4.24±0.27μgg−1 respectively). For the sediment reference material ERM CC 580, a methyl mercury concentration of 0.070±0.002μgg−1 was measured which corresponds to an extraction efficiency of 92±3% of certified values (0.076±0.04μgg−1) but within the range of published values (0.040–0.084μgg−1; mean±s.d.: 0.073±0.05μgg−1, n=40) for this material. The extraction procedure for the fish tissues was also compared against an enzymatic extraction using Protease type XIV that has been previously published and similar results were obtained. The use of HPLC-HGAAS with a Phenomenox 5μm Luna C18 (250mm×4.6mm) column and a mobile phase containing 0.06moll−1 ammonium acetate (Merck Pty Limited, Australia) in 5% (v/v) methanol and 0.1% (w/v) l-cysteine at 25°C was evaluated as a complementary alternative to HPLC-ICPMS for the measurement of mercury species in fish tissues. The lowest measurable mercury concentration was 2μgl−1 and this corresponds to 0.1μgg−1 in fish tissues. Analysis of enzymatic extracts analysed by HPLC-HGAAS and HPLC-ICPMS gave equivalent results.
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