São Paulo State University (UNESP), School of Agricultural Sciences, Av. Universitária 3780, Botucatu, SP, 18610-034, Brazil.
São Paulo State University (UNESP), School of Sciences, Av. Eng. Luiz Edmundo Carrijo Coube 14-01, Bauru, SP, 17033-360, Brazil.
J Chromatogr A. 2020 Dec 20;1634:461693. doi: 10.1016/j.chroma.2020.461693. Epub 2020 Nov 10.
From an environmental perspective, searching for useful compounds in agri-food by-products by employing inefficient and polluting analytical procedures is paradoxical. This work aimed to develop a green, simplified, and highly efficient experimental setup for extracting and tentatively identifying the broadest range of metabolites in sugarcane solid by-products collected directly within the industrial mills. Nine different extraction approaches were investigated side-by-side, including three reference methods. Based on the extraction and environmental performances assessed by two complementary metrics called Analytical-Eco Scale and the Analytical Greenness Calculator, it was possible to reach two highly efficient two liquid-phase extractions while avoiding harmful solvents and traditional time, energy, and solvent consuming sample preparation steps, such as solvent evaporation, metabolite concentration, re-suspension, and derivatization. The simultaneously produced hydroethanolic and n-heptane extracts were directly analyzed by ultra-high-performance liquid chromatography and gas chromatography, both coupled to mass spectrometry, respectively, leading to the annotation of a large dynamic range of compounds from information rich spectral data. Up to 111 metabolites were identified in a single matrix, from highly polar sucrose to nonpolar wax ester C53 in a single extraction. Orientin, apigenin-6-C-glucosylrhamnoside, 1-octacosanol, octacosanal, and other bioactive compounds were identified in these abundantly available by-products, which are currently just burned to produce energy. The best two methods developed here (Two-Liquid-Phase Ultrasound-Assisted Extraction with Probe and Two-Liquid-Phase Dynamic Maceration) appeared as a green, simplified, and highly efficient procedures to qualitatively profile metabolites in complex solid matrices.
从环境角度来看，通过采用低效和污染性的分析程序在农业食品副产物中寻找有用的化合物是矛盾的。本工作旨在开发一种绿色、简化和高效的实验装置，用于提取和初步鉴定直接在工业工厂内收集的甘蔗固体副产物中最广泛的代谢物。并排研究了九种不同的提取方法，包括三种参考方法。基于通过称为分析生态规模和分析绿色计算器的两个互补指标评估的提取和环境性能，可以达到两种高效的两相提取，同时避免使用有害溶剂和传统的耗时、耗能和消耗溶剂的样品制备步骤，例如溶剂蒸发、代谢物浓缩、再悬浮和衍生化。同时产生的水-乙醇和正庚烷提取物直接通过超高效液相色谱和气相色谱进行分析，分别与质谱联用，从而从富含信息的光谱数据中注释到大范围的化合物。在单个基质中鉴定出多达 111 种代谢物，从高度极性的蔗糖到单一提取中的非极性蜡酯 C53。在这些丰富的副产物中鉴定出了木犀草素、芹菜素-6-C-葡萄糖基鼠李糖苷、1-二十八烷醇、二十八醛和其他生物活性化合物，目前这些副产物只是燃烧来产生能量。这里开发的两种最佳方法（探头两相超声辅助提取和两相动态浸提）似乎是一种绿色、简化和高效的方法，可用于对复杂固体基质中的代谢物进行定性分析。
