KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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HPLC Study for Egg White Analysis

달걀 단백질 분석을 위한 HPLC 연구

  • Jeon, Young-Ju (Department of Chemical engineering, Chungnam National University) ;
  • Lee, Eun (Department of Chemical engineering, Chungnam National University) ;
  • Kim, In-Ho (Department of Chemical engineering, Chungnam National University)
  • 전영주 (충남대학교 화학공학과) ;
  • 이은 (충남대학교 화학공학과) ;
  • 김인호 (충남대학교 화학공학과)
  • Published : 2007.04.30
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Abstract

Since egg white contains various protein, it is important to research the protein distribution of egg white. Specially, lysozyme and ovalbumin important proteins, are used in medicine and food industry. Reversed phase high performance liquid chromatography has been used for separation of egg white, and column of RP-HPLC is available in variety. We have used C4, C8 and C18 columns to obtain chromatograms by varying carbon chain length of stationary phase. Long carbon chain length of stationary phase has good separation of egg white. Also, we have changed the composition of mobile phase (acetonitrile, water, and trifluoroacetic acid) to find optimum chromatograms. Acetonitrile and water composition of 50 : 50 show many peaks from egg white. Isocartic and gradient elution in RP-HPLC were used to compare the chromatography of egg white.

RP-HPLC에서 주요 변수인 이동상의 조성과 컬럼의 종류를 변화시켜 난백 단백질을 실험하였다. C4, C8 C18 컬럼을 비교 실험하여 C18 컬럼에서 가장 많은 피크를 보여 C18 컬럼을 선택하였다. 등용매 용리에서 ACN : water의 조성이 50 : 50에서 가장 많은 피크를 보여 이 결과를 토대로 기울기 용리를 하여 실험하였다. 기울기 용리에서 얻어진 4가지 피크를 확인하기 위하여 단일성분인 lysozyme와 ovalbumin의 체류시간을 측정하여 확인하였고 논문을 통해 2가지 피크를 예측할 수 있었다.

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References

  1. Lu, J., Y. Wan, and Z. Cui (2006), Strategy to separate lysozyme and ovalbumin from CEW using UF, Desalination, 200, 477-479 https://doi.org/10.1016/j.desal.2006.03.402
  2. Kim, W. K. and B. H. Chung (1999), Optimization of chromatographic separation of lysozyme from homogenate of hen egg white by comparison of breakthough behavior, Korean J. Biotechnol. Bioeng. 14(3), 279-283
  3. Kim, H. W. and I. H. Kim (2003), Comparison lysozyme purification from egg white between ion exchange chromatography and precipitation, HWAK KONGHAK 41(3), 332-336
  4. Huntingston, J. A. and P. Stein (2001), Structure and properties of ovalbumin, J. Chromatoar. B 756, 189-198 https://doi.org/10.1016/S0378-4347(01)00108-6
  5. Nau, F., A. Mallard, J. Pages, and G. Brule (1999), Reversed-phase liquid chromatography of egg white proteins. Optimization of ovalbumin elution, J. Liq. Chrom.&ReI. Technol. 22(8), 1129-1147 https://doi.org/10.1081/JLC-100101722
  6. Guiochon, G., Golshan-Shirazi, A .M, S. Katti (1994), Fundamentals of preparative and nonlinear chromatography, P393-397, Academic Press, Boston
  7. Awade, A. C. and T. Efstathiou (1999), Comparison of three liquid chromatographic methods for egg-white protein analysis, J. Chromatoar. B 723, 69-74 https://doi.org/10.1016/S0378-4347(98)00538-6
  8. Itoh, H., N. Noriyuki, T. Kinosita, N. Nagae, and N. Mitsugu (1991), Fast protein separation by reversed-phase high-performance liquid chromatography on octadecylsilyl-bonded nonporous silica gel, Anal. Biochem. 199, 7-10 https://doi.org/10.1016/0003-2697(91)90261-Q