Kuligowski Julia, Pérez-Guaita David, Escobar Javier, Lliso Isabel, de la Guardia Miguel, Lendl Bernhard, Vento Máximo, Quintás Guillermo
Neonatal Research Group, Health Research Institute La Fe, 46026 Valencia, Spain.
Department of Analytical Chemistry, University of Valencia, 46100 Burjassot, Spain.
Talanta. 2014 Sep;127:181-90. doi: 10.1016/j.talanta.2014.04.009. Epub 2014 Apr 15.
Liquid chromatography-mass spectrometry (LC-MS) has been increasingly used in biomedicine to study the dynamic metabolomic responses of biological systems under different physiological or pathological conditions. To obtain an integrated snapshot of the system, metabolomic methods in biomedicine typically analyze biofluids (e.g. plasma) that require clean-up before being injected into LC-MS systems. However, high resolution LC-MS is costly in terms of resources required for sample and data analysis and care must be taken to prevent chemical (e.g. ion suppression) or statistical artifacts. Because of that, the effect of sample preparation on the metabolomic profile during metabolomic method development is often overlooked. This work combines an Attenuated Total Reflectance-Fourier transform infrared (ATR-FTIR) and a multivariate exploratory data analysis for a cost-effective qualitative evaluation of major changes in sample composition during sample preparation. ATR-FTIR and LC-time of flight mass spectrometry (TOFMS) data from the analysis of a set of plasma samples precipitated using acetonitrile, methanol and acetone performed in parallel were used as a model example. Biochemical information obtained from the analysis of the ATR-FTIR and LC-TOFMS data was thoroughly compared to evaluate the strengths and shortcomings of FTIR biospectroscopy for assessing sample preparation in metabolomics studies. Results obtained show the feasibility of ATR-FTIR for the evaluation of major trends in the plasma composition changes among different sample pretreatments, providing information in terms of e.g., amino acids, proteins, lipids and carbohydrates overall contents comparable to those found by LC-TOFMS.
液相色谱 - 质谱联用（LC-MS）在生物医学中越来越多地用于研究生物系统在不同生理或病理条件下的动态代谢组学响应。为了获得系统的综合快照，生物医学中的代谢组学方法通常分析生物流体（如血浆），这些生物流体在注入LC-MS系统之前需要进行净化处理。然而，高分辨率LC-MS在样品和数据分析所需资源方面成本高昂，并且必须注意防止化学（如离子抑制）或统计假象。因此，在代谢组学方法开发过程中，样品制备对代谢组学谱图的影响常常被忽视。这项工作结合了衰减全反射傅里叶变换红外光谱（ATR-FTIR）和多变量探索性数据分析，以对样品制备过程中样品组成的主要变化进行经济高效的定性评估。使用一组平行进行的分别用乙腈、甲醇和丙酮沉淀的血浆样品分析得到的ATR-FTIR和液相飞行时间质谱（TOFMS）数据作为模型示例。对从ATR-FTIR和LC-TOFMS数据分析中获得的生化信息进行了全面比较，以评估FTIR生物光谱学在代谢组学研究中评估样品制备的优缺点。所得结果表明，ATR-FTIR可用于评估不同样品预处理之间血浆成分变化的主要趋势，提供的氨基酸、蛋白质、脂质和碳水化合物总含量等信息与LC-TOFMS所发现的信息相当。
