The impact of nanoparticles on the separation of DNA and amines by capillary electrophoresis
Date Issued
2005
Date
2005
Author(s)
Yu, Wan-Chi
DOI
zh-TW
Abstract
In this thesis, we focus on the impact of nanoparticles on the separation of biological molecules by capillary electrophoresis (CE) in conjunction with laser-induced fluorescence (LIF) detection. In the first part of this thesis, the impact of additives in 0.35% 2-hydroxyethyl cellulose (HEC, Mw 1,300 000 g/mole) on DNA separation by CE-LIF is studied. By using 0.35% HEC prepared in 40 mM glycine buffer (pH 9.9) containing 1% thiourea, 32-nm gold nanoparticles, and 1.27μM ethidium bromide, 9 DNA fragments with their sizes ranging from 394 to 653 bp were well resolved within 5 min. The separation is fast, highly efficient, and reproducible, mainly because of the change in HEC morphologies as a result of the interaction among thiourea, HEC, and gold nanoparticles. In the second part of this thesis, we have investigated the separation of biological amines and acids by CE-LIF using capillaries coated with poly-L-lysine (PLL) and silica nanoparticles (SiONPs). The capillaries coated with (PLL-SiONP)3 and (PLL-SiONPs)2-PLL are suitable for the separations of amines and acids, respectively. By using a (PLL-SiONP)3 capillary filled with a solution consisted of 10 mM formic acid (pH 3.7) and 3% acetonitrile (ACN), the limits of detection (LOD) at signal-to-noise ratio (S/N) = 3 are 1.2 nM and 35.0 nM for 5-hydroxyindole-3-acetic acid (5-HIAA) and 5-hydroxytryptamine (5-HT), respectively. The LODs are 0.09 nM and 19.0 nM for TA and 5-HIAA, respectively, when using a (PLL-SiONPs)2-PLL capillary filled with 10 mM formic acid ( pH 3.7). The practical application of the method has been validated by the analysis of urine samples, with great sensitivity, high speed, efficiency, and good reproducibility.
Subjects
奈米粒子
毛細管電泳
DNA分離
生物胺分離
nanoparticles
capillary electrophoresis
separation of DNA and biological amines
Type
thesis
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