In this work, the Regions of Interest-Multivariate Curve Resolution (ROIMCR) method is proposed for the analysis of non-target metabolomics data. Samples from marine mussels (Mytilus galloprovincialis) exposed to a sublethal concentration (10 μg/L) of sulfamethoxazole (SMX) during 4 days in different seasonal conditions (summer and winter) were analyzed by High-Performance Liquid Chromatography – High-Resolution Mass Spectrometry (HPLC-HRMS) to study the effect of their exposure to SMX and the different seasonal conditions. The Regions of Interest (ROI) procedure has been applied for data filtering, compression, preprocessing and storage steps. Multivariate Curve Resolution-Alternating Least Squares (MCR-ALS) is then applied to the previously MS ROI preprocessed data sets to resolve the elution profiles and spectral fingerprints of the chemical constituents of the analyzed samples. The peak areas of the elution profiles of the chemical constituents resolved by the combined ROIMCR procedure were analyzed by Principal Component Analysis (PCA) and samples were clustered according to their experimental seasonal and SMX exposure. The effects of the two investigated factors and of their interaction on the concentrations of the metabolites were statistically assessed by ANOVA simultaneous component analysis (ASCA). Both types of analyses, PCA clustering and ASCA, confirmed that the seasonal conditions (summer versus winter) produced larger effects than those produced by the exposure to SMX and by the interaction of these two factors. The concentration changes of 16 identified endogenous metabolites were validated individually using a Wilcoxon statistical test, which confirmed the presence of significant disturbances in the levels of some of these metabolites (free fatty acids, amino acids and nucleic acids), and indicated the possible alteration of six different biological pathways, affected by the investigated seasonal and SMX exposure factors.

在这项工作中，提出了用于分析非目标代谢组学数据的感兴趣区域-多变量曲线分辨率 (ROIMCR) 方法。海洋贻贝(Mytilus galloprovincialis)样本通过高效液相色谱-高分辨率质谱 (HPLC-HRMS) 分析在不同季节条件下（夏季和冬季）4 天内暴露于亚致死浓度（10 μg/L）的磺胺甲恶唑 (SMX) 以研究他们接触 SMX 和不同季节条件的影响。感兴趣区域 (ROI) 程序已应用于数据过滤、压缩、预处理和存储步骤。然后将多元曲线分辨率-交替最小二乘法 (MCR-ALS) 应用于先前 MS ROI 预处理数据集，以解析所分析样品的化学成分的洗脱曲线和光谱指纹。通过组合 ROIMCR 程序解析的化学成分洗脱曲线的峰面积通过主成分分析 (PCA) 进行分析，并根据其实验季节和 SMX 暴露对样品进行聚类。通过方差分析同步成分分析 (ASCA) 统计评估了两个研究因素及其相互作用对代谢物浓度的影响。PCA 聚类和 ASCA 两种类型的分析都证实，季节性条件（夏季与冬季）产生的影响比暴露于 SMX 和这两个因素的相互作用产生的影响更大。使用 Wilcoxon 统计检验分别验证了 16 种已鉴定内源性代谢物的浓度变化，