Sorption by layered double hydroxides (LDH) is gaining substantial interest for remediating emerging contaminants, including pharmaceuticals from wastewaters. Findings from a sorbent material performing successfully in lab-based studies using non-environmental (laboratory-sourced) water cannot be assumed to translate to equal performance under environmental downstream applications. However, studies evaluating sorbent material performance for removal of pollutants and understanding material interactions with environmental waters are limited. This study evaluates the removal of the antibiotic amoxicillin (AMX) using a Mg2Al–NO3-LDH sorbent material from laboratory-grade water and wastewater effluent (WWE). AMX is successfully removed (94.53 ± 4.30 % within 24 h) in laboratory-grade water (under batch sorption conditions: 100 μg/L AMX, 0.2 g/L LDH, 20 °C). The comparison of LDH removal performance in laboratory grade and WWE shows a decreased maximum removal of AMX in WWE (13.39 ± 5.53 %). A lower final AMX concentration is observed in the WWE without the presence of LDH, compared to the ‘removal’ experiments in WWE with the presence of LDH, indicating a contribution of non-sorption removal pathways of AMX. This is proposed to be due to the difference in metal concentrations in the WWE with and without LDH present. The presence of LDH is found to decrease concentrations of metal pollutants in WWE, such as Zn concentration decreasing by 85 % over 24 h, changing water characteristics. Overall, this paper reports that an LDH performs differently in laboratory-sourced water and a wastewater effluent. This provides evidence that sorbent material performance needs to be evaluated in complex water matrices to ensure that it is representative of how a sorbent material will perform in an environmental application, which is the end goal of developing such technologies. Finally, good practice recommendations are provided for future lab-scale sorption experiments evaluating the performance of any new sorbent materials for water treatment applications.