QOPNA, Mass Spectrometry Center, Department of Chemistry, University of Aveiro, Aveiro, Portugal.
Talanta. 2012 May 30;94:209-15. doi: 10.1016/j.talanta.2012.03.023. Epub 2012 Mar 30.
Sample preparation is a critical step for all kind of biological fluids analysis with serious implications in data retrieved. In this sense, efforts have been made to standardize biofluids' management procedures for diagnosis and research purposes. However, no agreement exists regarding saliva preparation. Aiming the delineation of an ideal preparation procedure for salivary peptidome analysis, the commonly used extraction methods such as selective precipitation with organic solvents, acid and addition of chaotropic agents in combination with filtration, were evaluated in the present study. Data concerning protein/peptide content, Tricine-gel electrophoresis and MALDI-TOF/TOF identification suggest that centrifugation, a generally used sample cleanup step, should be critically reconsidered based on the hydrophobic peptides that can be loss by aggregation with high molecular weight (MW) components. Although no individual method per se resulted in the identification of all MS identified peptides, the extraction method with bicarbonate/acetonitrile (ACN) followed by filtration resulted in the higher number of identified peptides.
样品制备是所有类型生物体液分析的关键步骤，对所获取的数据有重要影响。有鉴于此，人们已经努力使生物体液的管理程序标准化，以用于诊断和研究目的。然而，在唾液制备方面还没有达成共识。本研究旨在为唾液肽组分析制定理想的制备程序，评估了常用的提取方法，如选择性沉淀有机溶剂、酸和添加变性剂与过滤相结合。有关蛋白质/肽含量、Tricine-凝胶电泳和 MALDI-TOF/TOF 鉴定的数据表明，离心是一种常用的样品净化步骤，应根据疏水性肽与高分子量 (MW) 成分聚集而丢失的情况进行批判性重新考虑。尽管没有一种单一的方法本身能够鉴定所有 MS 鉴定的肽，但碳酸氢盐/乙腈 (ACN) 提取方法后过滤可鉴定出更多的肽。
