Bank filtration (BF) has been used for many years as an economical technique for providing high-quality drinking water. However, under anaerobic conditions, the aquifer release of undesirable metal(loid)s, such as iron manganese, and arsenic, reduces the bank filtrate quality and thus restricts the application of this technique. This study investigates the impact of the organic-matter composition of source water on the mobilisation of Fe, Mn, and As during the anaerobic BF process. A laboratory-scale column study was conducted at a controlled-temperature (30 ± 2 °C) using different feed water sources. The organic matter characteristics of the feed water were elucidated using excitation-emission spectroscopy techniques integrated with parallel factor framework clustering analysis (PFFCA) model. The experiment was performed at redox conditions between 66 mv and -185 mv. Moreover, batch studies were implemented to study the effect of natural organic matter type (humic, fulvic and tyrosine) and concentration on the mobilisation of the selected metal(loids). The laboratory experiments demonstrated that the mobilisation of Fe, Mn and As during the BF are varied with the organic water concentration and composition of the source water. The fluorescence results revealed that terrestrial and condensed structure humic compounds are more capable to release Fe into the filtrate water. In contrast, Mn exhibited an equal tendency of mobilisation towards all the humic compounds regardless of its origin and structure. However, at a humic concentration higher than 5 mg-C/L, Mn showed more affinity towards lower molecular weight humic compounds. Arsenic was found to be the least impacted by the alteration in the source water organic matter composition; its mobilisation was highly correlated with iron releasing process. On the other hand, the biodegradable organic matter at high concentration (>10 mg-C/L) was found to be highly effective to turn the infiltration area into Fe-reducing environment and thereby elevating Fe and As concentrations in the pumped water. In conclusion, this study revealed that the DOM composition and concentration of the raw water could play an important role in the mobilisation of metal(loids) during the BF processes.