Electrophoretic separation with 2-mm inner diameter fused-silica microcolumn packed with quartz microncrystals and its application.
The feasibility of a microcolumn electrophoresis technique was investigated with a 100mm length, 2mm I.D. fused-silica microcolumn packed with uniform quartz microncrystals prepared by hydrothermal synthesis. To evaluate the separation technique, tryptophan, phenylalanine and tyrosine were primarily separated by the microcolumn electrophoresis and detected at 216 nm without derivatization by an ordinary spectrophotometer. The separation conditions of the amino acids were optimized. With 1.5 mmol/L disodium phosphate buffer solution (pH 11.5) containing 25% (v/v) methanol and 10% (v/v) acetonitrile, the three amino acids were separated and the separation efficiency of tryptophan was 4.5x10(4)plates/m. The limits of detection were 0.035, 0.22 and 0.20 micromol/L, respectively. The sample capacity of the electrophoretic microcolumn achieved 35 microL. The proposed method was used to determine these amino acids in compound amino acid injection samples without derivatization. For the simplicity and portability of the microcolumn electrophoresis, it is studied as one of the high-performance separation techniques for an in situ and real-time electrokinetic flow analysis system. For its high detection sensitivity and large sample capacity, it can be developed for preparative electrophoresis.
Li
Lian
L
Department of Chemistry, University of Science and Technology of China, Hefei, Anhui 230026, China.
He
You-Zhao
YZ
Gan
Wu-Er
WE
Wang
Xiao-Kui
XK
Xie
Hai-Yang
HY
Gao
Yong
Y
D024322:Amino Acids, Aromatic / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods / Q000592:standards; D008722:Methods; D011791:Quartz; D013056:Spectrophotometry, Ultraviolet
D024322:Amino Acids, Aromatic; D011791:Quartz