Citrinin (CIT) is a mycotoxin contaminant in food commodities and can co-occur with ochratoxin A (OTA), another nephrotoxic contaminant in food and feed. Presence of OTA in maize from Bangladesh has been reported, but no data exist on CIT occurrence in food or feed in Bangladesh. Since biomonitoring provides the best approach to assess human exposure to contaminants from various sources and by all routes, a validated method for biomarker analysis has been used to investigate the presence of CIT and its metabolite dihydrocitrinone (HO-CIT) in urines from two Bangladeshi cohorts: Both analytes were determined in urine samples collected from inhabitants of a rural (n=32) and an urban (n=37) area in the Rajshahi district of Bangladesh. After cleanup by immunoaffinity columns, extracts were analyzed by LC-MS/MS; the limits of detection for CIT and HO-CIT in urine were 0.02 and 0.05 ng/mL, respectively. CIT and HO-CIT were detectable in 94 and 71% of all urine samples. Urinary biomarker levels did not show significant correlations with age, gender, and body mass index of the donors. However, excretion of CIT together with its metabolite HO-CIT was significantly higher (p<0.01) in the rural cohort (mean 1.1±1.9 ng/mL) than in the urban cohort (mean 0.14±0.14 ng/mL). This clearly indicates differences in mycotoxin exposure. As food habits differ between rural and urban people and also their main areas of occupation, further research is needed with regard to the major contributors of CIT exposure in the two cohorts. In conclusion, this first biomarker analysis indicates widespread and variable exposure to CIT in Bangladeshi adults.