Kynurenine pathway utilizes tryptophan to generate the biologically important metabolites (including kynurenine and kynurenic acid) associated with different pathologies. NAD+ is the final product of this pathway, but it can be also produced from nicotinic acid (niacin) in the salvage pathway. There is small number of literature available and great desire for analytical tools for quantification of these metabolites in foods. We developed and validated the chromatography/mass spectrometry-based method for the simultaneous determination of nicotinic acid, kynurenine, and kynurenic acid in raw poultry (chicken and turkey) meat. Using this method, the raw meat from different body parts (breast, drumstick, thigh, mid joint wing, back, neck) of chicken and turkey were analyzed. Nicotinic acid from 0.06 to 46.01 mg/kg was found in tested chicken and turkey meat samples. Most samples (∼84%) contained quantifiable amounts of kynurenine (0.013–0.274 mg/kg), but kynurenic acid (0.004–0.138 mg/kg) was determined in only ∼3% of poultry meat.