This paper describes the optimization and validation of an acetonitrile based method for simultaneous extraction of multiple pesticides and mycotoxins from raw coffee beans followed by LC-ESI-MS/MS determination. Before extraction, the raw coffee samples were milled and then slurried with water. The slurried samples were spiked with two separate standard solutions, one containing 131 pesticides and a second with 35 mycotoxins, which were divided into 3 groups of different relative concentration levels. Optimization of the QuEChERS approach included performance tests with acetonitrile acidified with acetic acid or formic acid, with or without buffer and with or without clean-up of the extracts before LC-ESI-MS/MS analysis. For the clean-up step, seven d-SPE sorbents and their various mixtures were evaluated. After method optimization a complete validation study was carried out to ensure adequate performance of the extraction and chromatographic methods. The samples were spiked at 3 concentrations levels with both mycotoxins and pesticides (with 6 replicates at each level, n = 6) and then submitted to the extraction procedure. Before LC-ESI-MS/MS analysis, the acetonitrile extracts were diluted 2-fold with methanol, in order to improve the chromatographic performance of the early-eluting polar analytes. Calibration standard solutions were prepared in organic solvent and in blank coffee extract at 7 concentration levels and analyzed 6 times each. The method was assessed for accuracy (recovery %), precision (RSD%), selectivity, linearity (r2), limit of quantification (LOQ) and matrix effects (%).

中文翻译：

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通过 LC-ESI-MS/MS 分析同时测定生咖啡中的 117 种农药和 30 种真菌毒素，无需净化

本文介绍了一种基于乙腈的方法的优化和验证，该方法用于从生咖啡豆中同时提取多种农药和真菌毒素，然后进行 LC-ESI-MS/MS 测定。在提取之前，将生咖啡样品研磨，然后用水调成浆状。浆状样品中加入两种不同的标准溶液，一种含有 131 种农药，另一种含有 35 种霉菌毒素，分为 3 组不同的相对浓度水平。QuEChERS 方法的优化包括使用乙酸或甲酸酸化的乙腈进行性能测试，使用或不使用缓冲液，以及在 LC-ESI-MS/MS 分析之前对提取物进行净化或不净化。对于净化步骤，评估了七种 d-SPE 吸附剂及其各种混合物。方法优化后，进行了完整的验证研究，以确保提取和色谱方法具有足够的性能。样品中添加了 3 个浓度水平的真菌毒素和农药（每个水平 6 次重复，n = 6），然后进行提取程序。在 LC-ESI-MS/MS 分析之前，乙腈提取物用甲醇稀释 2 倍，以提高早期洗脱极性分析物的色谱性能。在有机溶剂和咖啡空白提取物中制备 7 个浓度水平的校准标准溶液，各分析 6 次。对该方法的准确度（回收率 %）、精密度 (RSD%)、选择性、线性 (r2)、定量限 (LOQ) 和基质效应 (%) 进行了评估。