Analysis of Cyclic Adenosine Monophosphate (cAMP) Separation via RP-HPLC (reversed-phase high-performance liquid chromatography) by the Moment Method and the van Deemter Equation

Abstract

요 약고성능액체크로마토그래피(high-performance liquid chromatography, HPLC)에서 C18(octadecyl silica, ODS) 칼럼에서 고리형 아데노신 일인산(cyclic adenosine monophosphate, cAMP)의 크로마토그램을 얻은 후, 모멘트 분석을 수행하였 다. 일반속도 모델(general rate model, GR model)을 기반으로 first absolute moment와 second central moment를 계산 하였다. 모멘트 분석의 중요한 세 가지 계수인 분자확산계수(molecular diffusivity, D m ), 외부물질전달계수(external mass transfer coefficient, k f ), 입자내부확산계수(intra-particle diffusivity, D e )는 각각 Wilke-Chang 식, Wilson-Geankoplis 식을 이용하고 이론단수(theoretical plate number) 식과 van Deemter 식을 비교하여 계산하였다. 실험은 각각 세 가지의 이동상 조성, 용질 농도, 유량 조건에서 수행하였다. Van Deemter 그래프를 그려 모멘트 분석결과를 정성적으로 정리 했으며, 이론단 상당높이(height equivalent to a theoretical plate, HETP,Abstract − The moment analysis of cyclic adenosine monophosphate (cAMP) was performed using chromatograms that were obtained with the pulse input method from an octadecyl silica (ODS) high-performance liquid chromatography (HPLC) column. The general rate (GR) model was employed to calculate the first absolute moment and the second central moment. Three important coefficients for moment analysis, which are molecular diffusivity (D m ), external mass transfer coefficient (k f ), and intra-particle diffusivity (D e ), were estimated by the Wilke-Chang equation, Wilson-Geankoplis equation, and comparing van Deemter equation to theoretical plate number equation, respectively. Experiments were conducted by various conditions of flow rates, methanol volume ratio of the mobile phase, and solute concentration. After the moment analysis, results were organized by van Deemter plots. Also van Deemter coefficients were compared each other to effect H ax , H f , and H d on height equivalent to a theoretical plate (HETP, H total ). The value of intraparticle diffusion (H d ) was the primary factor which makes for HETP whereas external mass transfer (H f ) was disregardable factor.