Detection of five mycotoxins in cocoa bean and its products with QuEChERS method using ultra high-performance liquid chromatography quadrupole-time-of-flight mass spectrometry

Cocoa beans are a vital raw material for chocolate and various food products; however, they are highly susceptible to contamination by ochratoxin A (OTA) and aflatoxins (B1, B2, G1, and G2), which are toxic mycotoxins of significant food safety concern. OTA is nephrotoxic, immunotoxic, and classified as a possible human carcinogen (Group 2B), while aflatoxin B1 is a Group 1 carcinogen strongly associated with liver cancer. Due to these severe health risks, international authorities such as the European Union and Codex Alimentarius have established strict regulatory limits, and non-compliance often results in trade restrictions. This study developed and validated an analytical method for simultaneous detection of OTA and aflatoxins in cocoa using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) technique coupled with ultra-performance liquid chromatography and quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) in positive MRM mode. The method demonstrated excellent linearity (R² > 0.999), with detection limits between 0.11 and 0.99 µg/kg and quantification limits from 0.34 to 2.99 µg/kg. Recovery rates ranged from 73.4% to 103.5%, with RSDs of 8.2–31.5%. The validated method is sensitive, accurate, and reliable, providing a robust tool for monitoring mycotoxins in cocoa to ensure consumer protection and compliance with international trade standards.