On In-capillary Processes of Capillary Electrophoresis and the Applications
Date Issued
2006
Date
2006
Author(s)
chen, Jong-Jyh
DOI
zh-TW
Abstract
Dynamic micro-scaled biochemical processes were designed and conducted within the separation capillary of capillary electrophoresis (CE) system, and several possible applications were demonstrated. To characterize the bio-affinity properties of chitosan, ACE (affinity capillary electrophoresis) techniques were applied to evaluate the binding constant. From the shift in migration times of hippurate / chitooligosacharide complex, the binding constant of hippurate with chitooligosacharides (around 5〜9 mer.) was calculated to be in the range of 0.56〜1.0174 M-1. Since the binding process occurred simultaneously with the separation process, the method is rapid and provides valuable information about the dynamic molecular interactions.
Secondly, in-capillary chromogenic reaction was used to enhance the separation and sensitivity of CE system. Two labeling procedures for glucosamine using o-phthalaldehyde (OPA) and borate were developed and compared. The OPA method is more sensitive (0.1~50mM, A340) and specific, but the reaction is too quick to reproducibly operate in an off-line manner. OPA-labeling reaction was successfully performed on-line within the capillary, and the glucosamine contents in nutraceuticals were accurately measured.
Thirdly, in-capillary micro-extraction process served as an efficient sample pretreatment procedure. Benzoate derivatives are common preservatives in soy sauce; however, the CEgrams of untreated samples were too complicated and can not be effectively resolved / analyzed. The complicated samples were acidified and then extracted with ethyl acetate. The organic extract was directly injected and analyzed in capillary zone electrophoresis mode. The organic sample plug (benzoates in ethyl acetate) was extracted and then separated in the alkaline running buffer, which simplified the electropherogram for quantification.
Subjects
毛細管電泳
葡萄糖胺
保健食品
微萃取
Capillary electrophoresis
Glucosamine
Nutraceutical
Micro-extraction
Type
thesis