Spices Board Quality Evaluation Laboratory, SIPCOT, Gummidipoondi, Chennai, 601201, India.
Spices Board Quality Evaluation Laboratory, Suganda Bhavan, Palarivattom, Cochin, 682025, India.
Mycotoxin Res. 2024 Nov;40(4):559-568. doi: 10.1007/s12550-024-00546-1. Epub 2024 Jul 11.
UPLC-MS/MS analytical conditions for the analysis of aflatoxins in spices were optimized and validated in this study. Liquid-liquid partition-based protocols for the cleaning up of extracts using common organic solvents such as acetonitrile, hexane, and ethyl acetate were developed and validated. The developed liquid-liquid partition methods were compared with immuno-affinity column and QuEChERS clean-up methods for the UPLC-MS/MS analysis of aflatoxins in 8 spices. The reduction of lipophilic components using the partition with hexane is particularly useful in spices like red pepper that have higher levels of fatty acids, carotenoids, sterols, triterpenoids, etc. The subsequent partitioning with ethyl acetate considerably reduced the matrix interference from the polar components and increased the sensitivity. The cleaning up of spice extracts using liquid-liquid partition techniques resulted in limits of quantification (LOQ) of 2-5 µgL in UPLC-MS/MS analysis. Trueness, repeatability, and reproducibility of the methods were in acceptable ranges. The accuracy of the developed methods was further verified by analyzing aflatoxins in naturally incurred samples of spices and comparing the results with those obtained from the immuno-affinity column cleanup-HPLC-FD method.
本研究优化并验证了用于分析香料中黄曲霉毒素的 UPLC-MS/MS 分析条件。开发并验证了基于液液分配的提取净化方案，使用常见有机溶剂，如乙腈、正己烷和乙酸乙酯。将开发的液液分配方法与免疫亲和柱和 QuEChERS 净化方法进行了比较，用于 8 种香料中黄曲霉毒素的 UPLC-MS/MS 分析。使用正己烷进行分配以去除亲脂性成分对于红辣椒等含有较高水平脂肪酸、类胡萝卜素、甾醇、三萜等的香料特别有用。随后用乙酸乙酯进行分配可大大减少极性成分的基质干扰并提高灵敏度。使用液液分配技术净化香料提取物，在 UPLC-MS/MS 分析中的定量限（LOQ）为 2-5 µgL。方法的准确度、重复性和再现性均处于可接受范围内。通过分析受污染的香料样品中的黄曲霉毒素，并将结果与免疫亲和柱净化-HPLC-FD 方法的结果进行比较，进一步验证了所开发方法的准确性。
