Bayu Refindra Fitriadi, Mustofa Ahda, Addriani Mardika Dewi, Ayutia Ciptaningtyas Putri
Peanuts are a widely traded agricultural commodity, with Indonesia importing substantial quantities from countries in Africa and Asia. However, peanuts are highly susceptible to fungal growth, particularly when stored and transported under suboptimal conditions. These conditions can promote the production of aflatoxins by Aspergillus flavus and Aspergillus parasiticus, posing serious health risks. Effective monitoring of aflatoxin levels is therefore essential to ensure food safety. Indonesia has set maximum residue limits (MRLs) at 15 µg/kg for aflatoxin B1 and 20 µg/kg for total aflatoxins (sum of aflatoxins B1 + B2 + G1 + G2). In this study, a robust, sensitive, simple, and cost-effective method was developed and validated for the determination of total aflatoxins and aflatoxin B1 in peanuts. The method employs ultrasonic-assisted extraction and ultra-performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS), and its validation was conducted in accordance with SANTE 11312/2021 v2 guidelines and the Guidance document on identification of mycotoxins and plant toxins in food and feed. The extraction method used was dissolving the sample in acetonitrile, followed by sonication for 1 h and centrifugation at a speed of 3700 rpm for 4 min. This method has the advantages of being cheaper, easier, and faster in the sample preparation process compared to previous methods. The limit of quantification (LOQ) was determined to be 15 µg/kg for total aflatoxins and 6 µg/kg for aflatoxin B1. Calibration curves showed excellent linearity (R2 > 0.999), with residuals ranging from −5 to 19%. The method demonstrated high specificity, as reagent and matrix blanks remained below 30% of the LOQ. Accuracy ranged from 101 to 113% for total aflatoxins and 75 to 109% for aflatoxin B1, with precision values (%RSD) of 4.48% and 16.8%, respectively. Participation in an international proficiency test confirmed the method’s reliability, with all z-scores falling within the satisfactory range (|z|≤ 2). From 2023 to 2024, the method was applied to 40 peanut samples originating from various countries. The results revealed that only 25 samples complied with the legal aflatoxin limits. © The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2025.
Center for Plantation Seeds and Plant Protection Surabaya, Ministry of Agriculture, Surabaya, Indonesia; Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Ahmad Dahlan University, Yogyakarta, Indonesia; Ahmad Dahlan Halal Center, Research Center, Ahmad Dahlan University, Yogyakarta, Indonesia; Department of Chemistry, Faculty of Matematics and Science, Brawijaya University, Malang, Indonesia
