基于亚微米二氧化硅颗粒薄膜的蛋白质 UTLC-MALDI-MS。
Protein UTLC-MALDI-MS using thin films of submicrometer silica particles.
机构信息
Department of Chemistry, Purdue University, 560 Oval Drive, West Lafayette, IN 47907, USA.
出版信息
J Chromatogr A. 2011 Oct 7;1218(40):7196-202. doi: 10.1016/j.chroma.2011.07.098. Epub 2011 Aug 6.
Abstract
Slides for ultra thin-layer chromatography (UTLC) were made by coating nonporous silica particles, chemically modified with polyacrylamide, as 15 μm films on glass or silicon. Three proteins, myoglobin, cytochrome c and lysozyme, are nearly baseline resolved by the mechanism of hydrophilic interaction chromatography. A plate height as low as 3 μm, with 3900 plates, is observed in 14 mm. Varying silica particle diameter among 900, 700 and 350 nm showed that decreasing particle diameter slightly improves resolution but slows the separation. Matrix-assisted laser desorption/ionization (MALDI)-MS of the proteins after separation is demonstrated by wicking sufficient sinapinic acid into the separation medium.
摘要
超薄层色谱（UTLC）的幻灯片是通过在玻璃或硅上涂覆 15μm 厚的聚酰胺改性无孔硅颗粒制成的。三种蛋白质（肌红蛋白、细胞色素 c 和溶菌酶）通过亲水相互作用色谱机制几乎实现了基线分离。在 14mm 处观察到板高低至 3μm，理论塔板数高达 3900 块。在 900、700 和 350nm 之间改变硅胶粒径表明，粒径的略微减小可略微提高分辨率，但会减缓分离速度。通过将足够量的 3,5-二甲氧基-4-羟基肉桂酸吸入分离介质中，展示了分离后蛋白质的基质辅助激光解吸/电离（MALDI）-MS。
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