Biogenic amines are undesirable compounds formed through the decarboxylation of free amino acids by pathogenic microbes, They pose serious risks to public health and food safety. These amines serve as critical biomarkers for evaluating food quality and safety. Their physiological implications for human health necessitate vigilant monitoring, particularly in fish, where elevated concentrations can cause histamine poisoning and toxicity. This study employed liquid chromatography–electrospray ionization–tandem mass spectrometry (LC-ESI-MS/MS) to quantify spermidine and tyramine levels in Mackerel fish. Twenty random samples, comprising 10 frozen and 10 grilled, were collected from various markets and restaurants in the New Valley Governorate, Egypt. The analysis revealed that mean values of spermidine concentrations (mg/100g) were 233.2 ± 25.5 in raw frozen fish and 186.80 ±10.7 in grilled fish. The average concentrations of tyramine were 1.22 ± 0.09 in raw frozen fish and 3.54 ± 0.3 in grilled fish. Based on the established permissible limits recommended by EOS, all samples of frozen mackerel fish 100% and 90% of grilled fish were exceed the permissible spermidine limits. However, all frozen and grilled fish samples were within acceptable tyramine levels.