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The toxic effects of mercury in earthworms and the potential alleviation effect of hydrogen-rich water (HRW) using ATR-FTIR and LC-MS analysis methods were investigated. Different concentrations of mercury chloride (H1 5 µg/mL, H2 10 µg/mL, H3 20 µg/mL, H4 40 µg/mL, and C1 control) and mercury chloride prepared in hydrogen-rich water (H5 5 µg/mL, H6 10 µg/mL, H7 20 µg/mL, H8 40 µg/mL, and C2 control) were injected into earthworms. The changes and reductions in some bands representing proteins, lipids, and polysaccharides (3280 cm-1,