An On-Column Enzyme Mediated Fluorescence-Amplification Method for Plasma Total Cholesterol Measurement by Capillary Electrophoresis with LIF Detection

摘要

Total cholesterol monitoring in human plasma is one of the most important routine analysis performed in the clinical laboratory. Here we report a new capillary electrophoresis method for determination of plasma total cholesterol based on on-column enzymatic assay and laser-induced fluorescence detection. In this method, the capillary is used not only as a separation medium but also as the reaction chamber which minimizes the amount of samples and the consumption of the reagents. The whole procedure can be automated which minimizes the possible errors and saves experimental time. The analytical characteristics of the proposed method, such as linearity and repeatability of the peak area and the migration time, were evaluated. The RSD of the migration time and peak area were 1.0 and 3.5 % (intraday), and 2.2 and 2.9 % (interday), respectively. The obtained LOQ values were at the nanomolar concentration level, therefore the developed method is sufficient for determination and quantification of plasma total cholesterol.