Kitagawa Fumihiko, Inoue Keisuke, Hasegawa Takayuki, Kamiya Masato, Okamoto Yukihiro, Kawase Masaya, Otsuka Koji
Department of Material Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Kyoto 615-8510, Japan.
J Chromatogr A. 2006 Oct 20;1130(2):219-26. doi: 10.1016/j.chroma.2006.03.077. Epub 2006 Apr 4.
Avidin was immobilized covalently onto the inner surface of fused silica capillaries as a stationary phase in an open tubular capillary electrochromatography (OT-CEC) for chiral separations. The modification was attained by a combination of glutaraldehyde with both an amino-silylated fused silica surface and avidin using a Schiff base formation reaction. This method couples the advantage of high efficiency and small consumption of a chiral selector with the possibility of UV detection without limitations of protein absorption. In addition, the prepared capillary was stable for 50 days with over 100 runs. To evaluate the electrochromatographic performance of the prepared capillaries, the chiral separation of abscisic acid and arylpropionic acids were investigated. Effects of the modification condition of protein, pH of running buffer, and an organic modifier on the enantioseparation were also investigated. In addition, the avidin immobilized capillary was employed for the selective chiral analysis with an electrospray ionization mass spectrometry (ESI-MS) detection scheme.
在开管毛细管电色谱（OT-CEC）中，抗生物素蛋白通过共价键固定在熔融石英毛细管的内表面，作为手性分离的固定相。通过戊二醛与氨基硅烷化的熔融石英表面以及利用席夫碱形成反应的抗生物素蛋白相结合来实现修饰。该方法将手性选择剂的高效率和低消耗优势与无蛋白质吸收限制的紫外检测可能性相结合。此外，制备的毛细管在进行100多次运行后可稳定50天。为了评估制备的毛细管的电色谱性能，研究了脱落酸和芳基丙酸的手性分离。还研究了蛋白质修饰条件、运行缓冲液的pH值和有机改性剂对对映体分离的影响。此外，固定有抗生物素蛋白的毛细管被用于采用电喷雾电离质谱（ESI-MS）检测方案的选择性手性分析。
