[Direct UV detection of glucose and its derivatives in capillary zone electrophoresis].
UV labeling detection has been commonly used to determine the association constants between lectins and saccharides, but the interaction is always between the labeled carbohydrates, rather than the truly underivatized carbohydrates, and lectins. In order to directly detect saccharides during the study on the interaction of glucose and its derivatives with lectins (e.g., concanavalin A), a capillary zone electrophoretic method with detection at a wavelength of 195 nm has been developed. The influences of various separation conditions including buffer concentration, pH and voltage were investigated. By using an uncoated silica capillary (50 microns i.d., 375 microns o.d., 48.5 cm of total length, and 44.0 cm to the detector) and 50 mmol/L Na2HPO(4)-50 mmol/L NaH2PO4 solution (near to the physiological pH of 7.4) as buffer, the underivatized sugars, including glucosamine, N-acetylglucosamine, glucose, and sodium gluconate, were sufficiently separated within 11 min at an applied voltage of 10 kV. On-column UV monitoring allowed the detection of these compounds at less than 4 mmol/L level, and quantification by the peak area method allowed reproducible determination of them at least at their respective concentration ranges. The method is characterized by its simplicity, rapidity, and reproducibility, and should be useful for the analysis of the interaction of glucose and its derivatives with lectins.
Xu
Feng
F
Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. fengxu@dicp.ac.cn
Zhu
Wen-Jun
WJ
Mao
Xiu-Li
XL
Dai
Zhong-Peng
ZP
Zhang
Qing
Q
Du
Yu-Guang
YG
Liang
Xin-Miao
XM
Lin
Bing-Cheng
BC
D003208:Concanavalin A / Q000032:analysis / Q000302:isolation & purification; D019075:Electrophoresis, Capillary / Q000379:methods*; D005942:Gluconates / Q000032:analysis / Q000302:isolation & purification; D005947:Glucose / Q000031:analogs & derivatives / Q000032:analysis* / Q000302:isolation & purification; D037102:Lectins / Q000032:analysis* / Q000302:isolation & purification; D013056:Spectrophotometry, Ultraviolet
D005942:Gluconates; D037102:Lectins; D003208:Concanavalin A; D005947:Glucose; C030691:gluconic acid