P.O. Manjri Farm, National Reference Laboratory, ICAR-National Research Centre for Grapes, Pune, 412307, India.
Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, Belfast, UK.
Mycotoxin Res. 2022 Aug;38(3):193-203. doi: 10.1007/s12550-022-00460-4. Epub 2022 Jul 14.
Chilli powder, a popular spice, is predominantly contaminated with aflatoxins (AFs) and ochratoxin A (OTA), posing a menace to public health. As no validated method exists for the simultaneous and direct analysis of AFs and OTA in chilli powder, it was imperative to develop one to ensure their effective monitoring and promote trade. In this research, we developed and validated a multi-mycotoxin analysis method that allows the simultaneous determination of AFs (AFB, AFB, AFG and AFG) and OTA in chilli powder with high sensitivity, accuracy and precision. The optimised sample preparation workflow started with the extraction of chilli powder (25 g) with methanol-water (100 mL, 80:20). An aliquot (3 mL) was cleaned on a multi-mycotoxin, immunoaffinity column (AFLAOCHRA PREP) and analysed using ultrahigh performance liquid chromatography with fluorescence (UHPLC-FLD) and tandem mass spectrometric (LC-MS/MS) detection in a single chromatographic run. The method performance was evaluated through intra- and inter-laboratory validation (ILV) studies, and also by analysing a certified reference material. A direct analysis using UHPLC-FLD (without derivatisation) provided the limits of quantification (LOQ) of 0.25 and 1 ng/g for AFs and OTA, respectively, while the LOQ for all these mycotoxins in LC-MS/MS was 0.5 ng/g. These LOQs are much lower than the maximum levels (MLs) specified by the European Commission. The recoveries of these analytes at LOQ and higher levels were above 75% (RSD < 12%). The ILV study demonstrated satisfactory method-reproducibility (RSD < 25%). The analysis of the certified reference material provided accuracies of AFs and OTA in the range of 83-101%. The analysis by UHPLC-FLD and LC-MS/MS provided very similar results. The incurred levels of B1 in market samples were estimated with a precision-RSD of < 6%. Considering its efficiency and alignment with the regulatory requirements, this method can be implemented for the routine analysis of AFs and OTA in chilli powder.
辣椒粉是一种流行的香料，主要受到黄曲霉毒素（AFs）和赭曲霉毒素 A（OTA）的污染，对公众健康构成威胁。由于目前尚无用于同时直接分析辣椒粉中 AFs 和 OTA 的验证方法，因此必须开发一种方法来确保对其进行有效监测并促进贸易。在这项研究中，我们开发并验证了一种多真菌毒素分析方法，该方法能够高灵敏度、准确性和精密度同时测定辣椒粉中的 AFs（AFB、AFB、AFG 和 AFG）和 OTA。优化的样品制备工作流程从用甲醇-水（100mL，80:20）提取 25g 辣椒粉开始。取 3mL 等分试样，用多真菌毒素免疫亲和柱（AFLAOCHRA PREP）净化，然后在单个色谱运行中使用超高效液相色谱-荧光（UHPLC-FLD）和串联质谱（LC-MS/MS）检测进行分析。通过实验室间验证（ILV）研究和分析认证参考物质来评估方法性能。使用 UHPLC-FLD 进行直接分析（无需衍生化），可分别为 AFs 和 OTA 提供定量限（LOQ）为 0.25 和 1ng/g，而在 LC-MS/MS 中所有这些真菌毒素的 LOQ 均为 0.5ng/g。这些 LOQ 远低于欧盟委员会规定的最大限量（MLs）。在 LOQ 和更高水平下，这些分析物的回收率均高于 75%（RSD<12%）。ILV 研究表明方法重现性良好（RSD<25%）。认证参考物质的分析结果表明 AFs 和 OTA 的准确度在 83-101%范围内。UHPLC-FLD 和 LC-MS/MS 的分析结果非常相似。通过该方法可以精确估计市场样品中 B1 的含量，其精密度 RSD<6%。考虑到其效率和与监管要求的一致性，该方法可用于辣椒粉中 AFs 和 OTA 的常规分析。
