Environmental exposure to persistent organic pollutants (POPs) has been suggested as a contributing factor for the increased rate of T2D and obesity. A complex mixture of 29 POPs (Total mixture), based on human blood concentrations was used to expose a glucagon-like peptide 1 (GLP-1) secreting enteroendocrine cell line (pGIP/neo: STC-1) in vitro, for 3 and 24 h. Significant increases of GLP-1 occurred when cells were exposed to Total Mixture at x 500 blood levels. Six sub-mixtures representing chlorinated (Cl), brominated (Br), and perfluorinated chemicals (PFAA), and their combinations (Cl + Br, Cl + PFAA, Br + PFAA) were also tested at x 500. Secretion levels seen for these, remained lower than for the Total mixture, and the Br mixture had no effect. After 24 h, increased secretion was seen with all mixtures at x 1 blood levels. Cytotoxicity was present for x 100 and x 500 blood levels. When tested in a GLP-1 receptor translocation assay (U2OS-GLP1R-EGFP), neither agonistic, nor antagonist effects on receptor internalisation were seen for any of the mixtures. We conclude individual classes of POP, alone or in combination, can affect GLP-1 secretion and might contribute as a molecular mechanism, linking environmental toxicants and diabetes.