Goto-Inoue Naoko, Hayasaka Takahiro, Taki Takao, Gonzalez Tania Valdes, Setou Mitsutoshi
Department of Molecular Anatomy, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 431-3192, Japan.
J Chromatogr A. 2009 Oct 16;1216(42):7096-101. doi: 10.1016/j.chroma.2009.08.056. Epub 2009 Aug 27.
Thin-layer chromatography (TLC) is a highly established convenient technique for lipid separation and partial characterization of neutral and acidic glycosphingolipids (GSLs) and phospholipids, in mixtures. Meanwhile, imaging mass spectrometry (IMS) is a promising tool for lipidomics. However, some lipid classes are detected more sensitively than others, which can lead to suppression effects when complex mixtures are analyzed. Therefore to analyze complex lipid mixtures, a precise separation into the individual lipid classes is necessary. Here we present our highly sensitive and convenient analytical technology that combines TLC and IMS, namely the TLC-Blot-MALDI-IMS method, to visualize whole lipids and individual molecular species with high sensitivity compared with common staining methods. This method allows for easy visualization of all lipids with a linear range of approximately one order of magnitude and precision <16% RSD, making it useful for differential display analysis of lipids.
薄层色谱法（TLC）是一种成熟且便捷的技术，用于分离脂质混合物中的中性和酸性糖鞘脂（GSLs）及磷脂，并对其进行部分表征。同时，成像质谱（IMS）是脂质组学中一种很有前景的工具。然而，某些脂质类别比其他类别检测得更灵敏，在分析复杂混合物时可能会导致抑制效应。因此，为了分析复杂的脂质混合物，需要将其精确分离为各个脂质类别。在此，我们展示了一种高度灵敏且便捷的分析技术，它将TLC和IMS相结合，即TLC-印迹-基质辅助激光解吸电离-IMS方法，与常用染色方法相比，该方法能够以高灵敏度可视化整个脂质和单个分子种类。此方法能够轻松可视化所有脂质，线性范围约为一个数量级，精密度<16%相对标准偏差，使其可用于脂质的差异显示分析。
