Toxicological evaluation of mycotoxins in tetra-pack milk samples using advanced analytical techniques

Milk serves as a vital nutritional source globally, yet remains vulnerable to contamination by hazardous mycotoxins—particularly heat-stable aflatoxin M1 (AFM1), ochratoxin A (OTA), and emerging penicillic acid (PA). This risk is exacerbated in developing nations like Pakistan where climate-sensitive dairy supply chains face increasing contamination pressures from rising temperatures and humidity. This study investigates mycotoxin prevalence in Pakistani tetra-pack milk through integrated analytical and digital approaches, aiming to: (1) quantify contamination levels using advanced chromatography, (2) develop climate-sensitive predictive models, and (3) prototype an IoT–blockchain traceability system for real-time risk monitoring. We employed LC–MS/MS (LOD: 0.05 μ g/L) with confirmatory TLC analysis on 126 milk samples, coupled with IoT sensors tracking temperature ( ± 0.5 °C), relative humidity ( ± 3%), and pH ( ± 0.2) at critical control points. A machine learning model incorporating Arrhenius kinetics was developed to predict contamination patterns. Results revealed 57.1% co-occurrence of AFM1 (0.115 ± 0.026 μ g/L) and OTA (7.327 ± 0.351 μ g/L; 144% above EU limits), with PA in 14.3% of samples. Our predictive model achieved R 2 = 0 . 94 accuracy, demonstrating temperature-dependent accumulation ( E a = 45 . 2 kJ/mol for AFM1) and projecting 31% increased contamination under RCP8.5 scenarios. Alarmingly, children’s exposure reached 1736% above safety thresholds (91.8 ng/kg/day). These findings challenge assumptions about pasteurization efficacy and necessitate: (1) ≥ 2% nanoencapsulated antimicrobials in packaging, (2) IoT-enabled cold chain monitoring, and (3) blockchain-based traceability. The study provides both a methodological framework for tropical dairy systems and quantifiable evidence for digital transformation in food safety governance, particularly for climate-vulnerable regions balancing food security and safety imperatives.