Advanced analytical method for determination of Aflatoxins in chilli powder using LC-MS/MS

Chilli powder, a widely used spice, is commonly contaminated with aflatoxins (AFs) posing significant health risks. AFs, particularly Aflatoxin B1, a highly carcinogenic mycotoxin produced by Aspergillus species and pose serious health risks, especially in agricultural commodities like chilli, where contamination must be closely monitored. Traditional detection methods such as direct chromatographical analysis using HPLC often face challenges due to the complex matrix of chilli powder. Due to the lack of a validated method for the direct detection AFs in chilli powder, it was essential to develop one for effective monitoring and to support trade. In this study, we established and validated a sensitive, accurate, and precise analytical method capable of detecting AFs (AFB1, AFB2, AFG1, AFG2) in chilli powder. The optimized sample preparation involved QuEChERS method followed by analysis through liquid chromatography with tandem mass spectrometry (LC–MS/MS). This direct analysis provided the quantification limits (LOQs) of 0.25 ng/g for each AF well below the maximum levels set by the European Commission. Recoveries at and above the LOQ levels les in range of 80-108%, with RSD below 8%. The QuEChERS extraction protocol compared with the Immuno affinity clean up. Market sample analysis showed AFB1 levels with precision RSDs below 6%. Given its high efficiency and compliance with regulatory standards, this method is suitable for routine AF and OTA testing in chilli powder.