Separation Study of Cytosine and Guanine by HPLC and Aspen Chromatography

Aspen Chromatography 전산모사와 HPLC를 이용한 구아닌 시토신의 분리특성연구

  • Park, Moon Bae (Department of Chemical Engineering, Chungnam National University) ;
  • Kim, In Ho (Department of Chemical Engineering, Chungnam National University)
  • 박문배 (충남대학교 화학공학과) ;
  • 김인호 (충남대학교 화학공학과)
  • Received : 2009.10.17
  • Accepted : 2009.12.19
  • Published : 2010.02.28

Abstract

DNA structure studies attract many interests in pharmaceutical, biochemical and medical disciplines. Among them, base pairs play a vital role in biological information transfer. Therefore, they need to be analyzed in various ways and the pair of guaninine and cytosine is the present analytical object. Separation of guanine and cytosine was researched by Aspen chromatography simulator and HPLC(High Performance Liquid Chromatography) experiments. Aspen chromatography simulation resulted in various chromatograms with changes of sample concentration, eluent flow rate and number of plate. The resolutions and yields of guanine and cytosine were calculated to obtain a best separation condition. $C_{18}$ HPLC column and water/methanol/acetic acid mixture(90/10/0.2) were used for separation of guanine and cytosine. HPLC parameters(resolution and number of theoretical plate) were calculated under different flow rates and sample concentrations. Aspen chromatography simulation and HPLC experimental results were compared with fair agreement.

DNA 구조를 밝히기 위해 의학, 약학 그리고 생명과학분야 등에서 활발한연구가 이루어지고 있다. 그 중 DNA의 염기쌍은 생명체의 정보 전달에 매우 중요한 역할을 하므로 염기쌍의 집중적인 분석이 필요하다. 그래서 DNA의 염기쌍 중 하나인 구아닌과 시토신을 선택하여 분석 실험을 하였다. 구아닌과 시토신의 분석은 Aspen chromatography 전산모사와 HPLC(High Performance Liquid Chromatography) 실험을 통하여 이루어졌다. Aspen Chromatography(ver. 2006 Aspen Tech. U.S.A)로 시료농도, 이동상 유속 그리고 이론단수를 변화시켜 전산모사하였다. HPLC 실험은 $C_{18}$ HPLC column 칼럼과 이동상 water/methanol/acetic acid 혼합액(90/10/0.1)을 이용하여 시료의 주입 농도와 이동상 속도를 변화시켰고 구아닌과 시토신의 크로마토그램의 분리도와 이론단수를 비교하였다. 실험과 전산모사 크로마토그래피 결과가 비교적 일치하였다.

Keywords

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