In the past decade, various lipidomics methodologies have been developed using mass spectrometry based analytical technologies, enabling wide coverage lipid detection in a quantitative manner. Hence, lipidomics has become a widely-accepted approach for biomarker discovery and mechanism elucidation in both medical and biology research fields; however, there are still technical challenges. In this study, focusing on the sample preparation procedure, a single step deproteinization by a water-soluble organic solvent, such as methanol (MeOH), ethanol (EtOH), isopropanol (IPA) or acetonitrile (ACN), was evaluated and proved to be satisfactory for lipidomics analysis. Moreover, during this investigation ACN deproteinization was revealed to not be an effective method for lipid extraction because lipid decomposition was observed during the protein precipitation process through lipase activation, potentially due to the insufficient protein denaturation. Therefore, excluding ACN, protein precipitation by alcohol was evaluated as the lipid extraction reagent. Moreover, adding the MTBE-MeOH (mMM) method, one of the major liquid-liquid extraction methods for shotgun lipidomics, these four approaches were compared.

Lipids were extracted from mouse plasma by these four methods and used for exhaustive lipid profiling by liquid chromatography mass spectrometry (LC/MS) analysis. Comparison of these four methods revealed that alcohol based protein precipitation was a useful sample preparation procedure for LC/MS based lipidomics analysis. Whereas MeOH extraction was appropriate for hydrophilic lipid species, IPA was effective for hydrophobic lipids such as triacylglycerols (TG). In practice, EtOH extraction is thought to be the best approach to cover wide range of lipid species using a simple preparation procedure.

在过去的十年中，已经使用基于质谱的分析技术开发了多种脂质组学方法，从而能够以定量方式进行广泛的脂质检测。因此，脂质组学已成为医学和生物学研究领域中生物标志物发现和机理阐明的广泛接受的方法。但是，仍然存在技术挑战。在这项研究中，着重于样品制备程序，评估并证明了通过水溶性有机溶剂（例如甲醇（MeOH），乙醇（EtOH），异丙醇（IPA）或乙腈（ACN））进行的一步脱蛋白，并证明了对于脂质组学分析是令人满意的。而且，在这项研究中，ACN脱蛋白被认为不是一种有效的脂质提取方法，因为在蛋白质沉淀过程中，通过脂肪酶的活化可以观察到脂质的分解，这可能是由于蛋白质变性不足所致。因此，除了ACN以外，还评价了通过醇沉淀的蛋白质作为脂质提取试剂。此外，还添加了MTBE-MeOH（mMM）方法（这是shot弹枪脂质组学的主要液-液萃取方法之一），对这四种方法进行了比较。

通过这四种方法从小鼠血浆中提取脂质，并通过液相色谱质谱（LC / MS）分析将其用于详尽的脂质谱分析。这四种方法的比较表明，基于醇的蛋白质沉淀是基于LC / MS的脂质组学分析的有用样品前处理程序。甲醇萃取适合亲水性脂质，而IPA对于疏水性脂质（例如三酰基甘油（TG））有效。在实践中，EtOH提取被认为是使用简单的制备程序即可覆盖多种脂质种类的最佳方法。