Moment Analysis (MA) of Lysozyme in Cation Exchange High Performance Liquid Chromatography (HPLC)

Abstract

요 약양이온교환 고성능액체크로마토그래피에서 라이소자임을 분석하고, 실험결과인 크로마토그램을 통해 모멘트 분석을 수행하였다. 용리 인산완충용액은 1.0, 0.75, 0.5 M의 소금을 포함하였다. 실험변수는 유량, 용리 완충용액중 소금 농 도, 시료의 농도로 하였다. General rate (GR) model을 도입하여 1차와 2차 모멘트를 해석하였다.Abstract − The moment analysis of lysozyme was implemented using chromatograms that were obtained from weak cation exchange column in high performance liquid chromatography system. Three elution sodium phosphate buffers containing 1.0, 0.75, 0.5 M sodium chloride were used. Experiments were conducted by varying flow rate, elution sodium chloride concentration, and lysozyme solute concentration. The general rate (GR) model was employed to calculate the first moment and the second moment. By plotting L/u 0 vs. (μ 1 -t 0 )/(1-ε e )(1-ε i )] equilibrium constants (K) were obtained from first moment analysis. Intra-particle diffusivity was obtained from theoretical plate number data. Based on the results of moment analysis, van Deemter plots were drawn in order to investigate the contributions of H ax , H f , and H d to total Height Equivalent to a Theoretical Plate (HETP, H total ). The effect of intra-particle diffusion (H d ) was the most dominant factor contributing to HETP while external mass transfer (H f ) was negligible factor.