Kato M, Dulay M T, Bennett B, Chen J, Zare R N
Department of Chemistry, Stanford University, CA 94305-5080, USA.
Electrophoresis. 2000 Sep;21(15):3145-51. doi: 10.1002/1522-2683(20000901)21:15<3145::AID-ELPS3145>3.0.CO;2-5.
A solution is prepared of 5 microm silica particles modified with (S)-N-3,5-dinitrobenzoyl-1-naphthylglycine (particle 1) or (S)-N-3,5-dinitrophenylaminocarbonyl-valine (particle 2) suspended in liquid tetraethylorthosilicate, ethanol, and aqueous hydrochloric acid. This solution is injected under pressure into a 30 cm long, 75 microm inner diameter capillary column and heated for 1 h at 120 degrees C after which the modified particles are embedded in a monolithic column of sol gel. The packed column measures approximately 15 cm from the inlet to the window used to view the laser-induced fluorescence. Thirteen different amino acids and three nonprotein amino acids are derivatized with the fluorogenic reagent 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) before injection onto the column for capillary electrochromatographic separation. The enantiomeric separation of the monolithic column packed with particle 1 results in a resolution ranging from 1.14 to 4.45, whereas that packed with particle 2 results in a resolution ranging from 0.79 to 1.17. On the basis of resolution and amount of chiral packing material the enantiomeric separation obtained by capillary electrochromatography is judged to be superior to that obtained previously with high performance liquid chromatography (HPLC).
制备一种溶液，其中含有用（S）-N-3,5-二硝基苯甲酰基-1-萘基甘氨酸（颗粒1）或（S）-N-3,5-二硝基苯基氨基羰基-缬氨酸（颗粒2）修饰的5微米二氧化硅颗粒，悬浮于液体原硅酸四乙酯、乙醇和盐酸水溶液中。将该溶液在压力下注入一根30厘米长、内径75微米的毛细管柱中，并在120℃下加热1小时，之后将修饰后的颗粒包埋在溶胶凝胶整体柱中。填充柱从进样口到用于观察激光诱导荧光的窗口的长度约为15厘米。在注入柱中进行毛细管电色谱分离之前，用荧光试剂4-氟-7-硝基-2,1,3-苯并恶二唑（NBD-F）对13种不同的氨基酸和3种非蛋白质氨基酸进行衍生化。填充颗粒1的整体柱对映体分离的分辨率范围为1.14至4.45，而填充颗粒2的整体柱对映体分离的分辨率范围为0.79至1.17。基于分辨率和手性填充材料的用量，判断毛细管电色谱法获得的对映体分离效果优于先前高效液相色谱法（HPLC）获得的效果。
