Formalin can be added as preservative to fresh foods to prevent spoilage and extend shelf life. Formalin contains 37-40% formaldehyde, which is classified as carcinogenic to humans. To assess the public health risk associated with formaldehyde exposure in freshwater fish in Southern Bangladesh, formaldehyde concentrations (mg/kg) were determined in tilapia, Indian major carp rui, Chinese carp and a minor carp from local market and in laboratory simulations (0.5, 1.0, 2.0 and 4.0% formaldehyde solution for 5, 15, 30 and 60 min) with spectrophotometric and high performance liquid chromatography (HPLC) methods. A food frequency questionnaire was used to collect fish consumption (kg/kg BW. d) data from 400 respondents. A probabilistic exposure assessment was conducted using @Risk7.0 software. Fish treated with formalin at increasing concentrations and exposure time showed increased trends of formaldehyde acquisition irrespective of fish species and analytical methods used (p < .05). Compared to spectrophotometry, the HPLC method was shown to be more sensitive and is therefore the preferred method for formalin quantification. Maximum exposure to formaldehyde (0.28 mg/kg BW. d) was calculated for tilapia using HPLC analysis. Margin of exposure (MoE) provides high priority (<10,000) for tilapia and Indian major carp rui at P99 under spectrophotometric analysis whereas as determined using HPLC, tilapia had MoE values much lower than 10,000 at P99, P95 and P90 (both total population and consumers). Exposure to formaldehyde associated with freshwater fish consumption is a public health concern in Southern Bangladesh and needs further assessment and risk management strategies.