Copper-based nanopesticides are increasingly being used in the agriculture, especially in organic farming. This has triggered some concerns about their risk to environmental and human health. In this study, 24-day-old lettuce plants grown in soil were exposed via the leaves to different concentrations (0, 1050 and 1555 mg/L) of Cu(OH) 2 nanopesticides for one month. Results showed Cu was mainly localized in lettuce leaves (823-1111 and 1353-2008 mg/kg in vascular and photosynthetic tissues), which may potentially increase Cu intake and impact human health. In addition, foliar application of Cu(OH)2 nanopesticide significantly increased potassium concentration in lettuce leaves by 6-7% and 21-28%, in vascular and photosynthetic tissues. A Gas Chromatography-Time-of-Flight Mass Spectrometry (GC-TOF-MS) based metabolomics approach was applied to determine hundreds of organic compounds simultaneously. Using relative quantitation and Partial Least Squares-Discriminant Analysis (PLS-DA) clustering of all compounds, clear differences were observed in the metabolite profiles of control and Cu(OH) 2 nanopesticides treated leaves. Discriminating compounds include amino acids, organic acids, polyamines, vitamin C and polyphenols. Dehydroascorbic acid and cis-caffeic acid, which are important antioxidants, were significantly decreased (19-33% and 5-8%) due to foliar exposure to the nanopesticide. Total antioxidant capacity was significantly decreased 20-23% after exposure to Cu(OH) 2 nanopesticides. There was also up-and down-regulation of a number of amino acids, particularly 4-hydroxybutytic acid (GABA) which decreased 50% compared to the control, potentially affecting the overall nutritional value of lettuce leaves exposed to the Cu(OH) 2 nanopesticides. In future work, determining an appropriate level of nanopesticide will be important to obtain the antifungal benefits without resulting in a significant decrease in nutritional value.