The aim of the study was to develop a new method for mercury determination from plants grown on mercury-polluted soils as well as to evaluate the mobility and bioavailability of mercury in the soil and its transfer to different plant organs (root, stem, leaves) from three different plants, such as mustard (Sinapis alba), thyme (Thymus serpyllum) and wild thyme. A cold vapor atomic absorption spectrometry (CV-AAS) method for the detection of mercury in plants was developed. The developed method was validated in the laboratory and was verified using two different certified reference materials (CRMs) type matrices, namely: NIST 1515 Apple Leaves and BCR-482 Lichen.