CAS Key Laboratory of Separation Science for Analytical Chemistry, National Chromatographic R & A Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences (CAS), Dalian 116023, China.
J Chromatogr A. 2012 Mar 9;1228:263-9. doi: 10.1016/j.chroma.2011.07.048. Epub 2011 Jul 22.
A phenyl-silica hybrid monolithic column for capillary liquid chromatography (cLC) was prepared through "one-pot" process by con-currently using benzyl methacrylate and alkoxysilanes. The effects of the molar ratio of tetramethoxysilane/vinyltrimethoxysilane (TMOS/VTMS), polycondensation temperature, content of supramolecule template (cetyltrimethylammonium bromide, CTAB), ratio of N,N'-dimethylformamide/methanol (v/v), the volume of benzyl methacrylate on the morphologies of the prepared phenyl-silica hybrid monolithic columns were investigated in detail. The permeability of the hybrid monolithic column was calculated as 3.23 × 10⁻¹³ m², and the minimum plate height was determined as 8.38 μm which corresponding to 119,000 theoretical plates per meter. Separation of various neutral, polar and basic analytes as well as small peptides on the hybrid monolithic column was achieved by cLC and showed high efficiency and satisfactory reproducibility. Moreover, the prepared hybrid monolithic column was also applied for the analysis of tryptic digests of bovine serum albumin (BSA), ovalbumin, α-casein, cytochrome C and myoglobin by cLC tandem mass spectrometry (cLC-MS/MS), and the results showed that the separation performance was close to that of the octadecylsilane (C18) packed capillary column which demonstrating its potential in proteome analysis. Moreover, since the prepolymerization system was mainly consisted of organic solvents (methanol and N,N'-dimethylformamide), various hydrophobic monomers could be potentially used to prepare organic-silica hybrid monolithic columns through "one-pot" approach.
一种苯基-硅胶杂化整体柱通过“一锅法”制备，同时使用苯甲基丙烯酸酯和烷氧基硅烷。详细考察了四甲氧基硅烷/乙烯基三甲氧基硅烷（TMOS/VTMS）摩尔比、缩合温度、超分子模板（十六烷基三甲基溴化铵，CTAB）含量、N，N'-二甲基甲酰胺/甲醇（v/v）的比例、苯甲基丙烯酸酯的体积对制备的苯基-硅胶杂化整体柱形貌的影响。杂化整体柱的渗透性计算为 3.23×10⁻¹³m²，最小板高确定为 8.38μm，对应每米 119000 理论板。通过 cLC 对各种中性、极性和碱性分析物以及小肽进行分离，显示出高效和令人满意的重现性。此外，还通过 cLC 串联质谱（cLC-MS/MS）将制备的杂化整体柱应用于牛血清白蛋白（BSA）、卵清蛋白、α-酪蛋白、细胞色素 C 和肌红蛋白的胰蛋白酶消化物的分析，结果表明分离性能接近十八烷基硅烷（C18）填充毛细管柱，表明其在蛋白质组分析中的潜力。此外，由于预聚合体系主要由有机溶剂（甲醇和 N，N'-二甲基甲酰胺）组成，因此各种疏水性单体可以通过“一锅法”潜在地用于制备有机-硅胶杂化整体柱。
