In this work, the impact of biological matrices, such as plasma and urine, was evaluated under SFCHRMS in the field of metabolomics. For this purpose, a representative set of 49 metabolites were selected. The assessment of the matrix effects (ME), the impact of biological fluids on the quality of MS/MS spectra and the robustness of the SFCHRMS method were each taken into consideration. The results have highlighted a limited presence of ME in both plasma and urine, with 30% of the metabolites suffering from ME in plasma and 25% in urine, demonstrating a limited sensitivity loss in the presence of matrices. Subsequently, the MS/MS spectra evaluation was performed for further peak annotation. Their analyses have highlighted three different scenarios: 63% of the tested metabolites did not suffer from any interference regardless of the matrix; 21% were negatively impacted in only one matrix and the remaining 16% showed the presence of matrix-belonging compounds interfering in both urine and plasma. Finally, the assessment of retention times stability in the biological samples, has brought into evidence a remarkable robustness of the SFCHRMS method. Average RSD (%) values of retention times for spiked metabolites were equal or below 0.5%, in the two biological fluids over a period of three weeks. In the second part of the work, the evaluation of the Sigma Mass Spectrometry Metabolite Library of Standards containing 597 metabolites, under SFCHRMS conditions was performed. A total detectability of the commercial library up to 66% was reached. Among the families of detected metabolites, large percentages were met for some of them. Highly polar metabolites such as amino acids (87%), nucleosides (85%) and carbohydrates (71%) have demonstrated important success rates, equally for hydrophobic analytes such as steroids (78%) and lipids (71%). On the negative side, very poor performance was found for phosphorylated metabolites, namely phosphate-containing compounds (14%) and nucleotides (31%).

中文翻译：

---

超临界流体色谱-质谱法在代谢组学中的适用性。II-代谢物综合库的评估和生物基质的评估。

在这项工作中，根据SFCHRMS在代谢组学领域评估了生物基质（例如血浆和尿液）的影响。为此，选择了代表性的49种代谢物。分别考虑了基质效应（ME）的评估，生物流体对MS / MS光谱质量的影响以及SFCHRMS方法的鲁棒性。结果表明血浆和尿液中都存在有限的ME，血浆中30％的代谢物患有ME，尿液中25％的代谢物表明在基质存在下灵敏度有限。随后，进行了MS / MS谱图评估，以进行进一步的峰注释。他们的分析突出了三种不同的情况：63％的被测代谢物不受基质干扰，21％的化合物仅在一种基质中受到不利影响，其余16％的化合物在尿液和血浆中均存在干扰。最后，对生物样品中保留时间稳定性的评估已证明SFCHRMS方法具有显着的鲁棒性。在两种生物液中，加标代谢物的保留时间的平均RSD（％）值在三周内等于或低于0.5％。在工作的第二部分中，在SFCHRMS条件下，对包含597种代谢物的西格玛质谱代谢物标准库进行了评估。商业库的总可检测性达到了66％。在检测到的代谢产物家族中，其中一些满足了很高的百分比。高极性代谢物，例如氨基酸（87％），核苷（85％）和碳水化合物（71％）已显示出重要的成功率，对于类固醇（78％）和脂质（71％）等疏水性分析物同样如此。不利的一面是，发现磷酸化代谢产物的性能很差，即含磷酸盐的化合物（14％）和核苷酸（31％）。