A total of 105 different types of poultry feed samples from South Africa were simultaneously analysed for the presence of 16 mycotoxins using ultra-high-performance liquid chromatography coupled to a triple quadrupole mass spectrometer (UHPLC-MS/MS). The data revealed the presence of 16 mycotoxins in the various poultry feed samples. Fumonisin B1 (FB1) was the most dominant recovered from 100% of samples analysed at concentrations ranging between 38.7 and 7125.3 µg/kg. This was followed by zearalenone (ZEN) (range: 0.1–429 µg/kg) and deoxynivalenol (DON) (range: 2.5–154 µg/kg). Samples were also found to be contaminated with fumonisin B2 (FB2) (range: 0.7–125.1 µg/kg), fumonisin B3 (FB3) (range: 0.1–125.1 µg/kg), α-zearalenol (α-ZEL) (range: 0.6–20 µg/kg ), β-zearalenol (β-ZEL) (range: 0.2–22.1 µg/kg), 3-acetyldeoxynivalenol (3-ADON) (range: 0.1–12.9 µg/kg) and 15-acetyldeoxynivalenol (15-ADON) (range: 1.7–41.9 µg/kg). Alternaria mycotoxin, i.e., Alternariol monomethyl ether (AME) was recovered in 100% of samples at concentrations that ranged from 0.3–155.5 µg/kg. Aflatoxins (AFs) had an incidence rate of 92% with generally low concentration levels ranging from 0.1–3.7 µg/kg. Apart from these metabolites, 2 type A trichothecenes (THs), i.e., HT-2 toxin (HT-2) (range: 0.2–5.9 µg/kg) and T-2 toxin (T-2) (range: 0.1–15.3 µg/kg) were also detected. Mycotoxin contamination in South African poultry feed constitutes a concern as correspondingly high contamination levels, such as those observed herein are likely to affect birds, which can be accompanied by severe health implications, thus compromising animal productivity in the country. Such exposures, primarily to more than one mycotoxin concurrently, may elicit noticeable synergistic and or additive effects on poultry birds.