Analysis of chitin oligosaccharides by capillary electrophoresis with laser-induced fluorescence.
A method, using capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection for analyzing chitin oligosaccharides is described. Chitin oligosaccharides were derivatized with 9-aminopyrene-1,4,6-trisulfonate (APTS) via reductive amination at 37 degrees C for 16 h (optimized conditions). The APTS-chitin oligosaccharides were analyzed using either an acidic citric acid-phosphate buffer or an alkaline borate buffer. The effects of buffer types, buffer pH values, and buffer concentrations on the separation were examined. The analytes were successfully separated by using a pH 4.6 citric acid-phosphate within 19 min. The APTS-derivatized chitin monosaccharide (D-glucosamine) migrated first. The analytes were also completely separated by using a pH 9.0 borate buffer within 24 min. Moreover, the specificity of enzyme digestion on chitin polysaccharides using the optimized APTS labeling procedure and the CE-LIF method was demonstrated.
Wang
Chin-Yu
CY
Department of Applied Chemistry, National Chiao Tung University, 1011 Ta-Hsueh Road, Hsinchu 300, Taiwan.
Hsieh
You-Zung
YZ
D002686:Chitin / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D007834:Lasers; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*
D009844:Oligosaccharides; D002686:Chitin