HPLC에서 이동상 변화와 온도에 따른 난백의 분리 = Separation of Egg White Using HPLC with Change of Mobile Phase and Temperature

다국어 초록 (Multilingual Abstract)

Lysozyme in egg white functions as bacteriolysis agent and ovalbumin plays a role as antigen in immune system. Egg white analysis methods usually include electrophoresis, gel permeation chromatography and reversed-phase HPLC(RP-HPLC). Among them, RP-HPLC was selected for rapid analysis and C18 column(Agilent, USA) was used as HPLC column. Optimum conditions were searched by changing mobile phase and temperature. Capacity factor and resolution were calculated and compared for various elution conditions. In the isocratic elution, mobile phase volume ratio was changed from 30/70/0.1 to 60/40/0.1(Acetonitrile(ACN)/Distilled water(DW)/Trifluoroacetic acid(TFA)). ACN composition was increased by 10% and temperature was set as 20℃ . In the gradient elution, ACN/DW ratio was changed from 10/90 to 60/40 during 20 minute and temperature was varied as 20, 30 and 40 ℃. In the isocratic elution, three peaks were separated at 50/50/0.1. Lysozyme and ovalbumin were confirmed as first and third peak in three peaks respectively. In the gradient elution, four peaks were separated at 30 ℃. Lysozyme and ovalbumin were confirmed as first peak and third peak in four peaks respectively.

참고문헌 (Reference)

1 김영준, "천식 마우스 모델에서 BCG백신의 투여시기에 따른 알레르기 염증반응 억제효과" 대한천식알레르기학회 30 (30): 307-313, 2010

2 김인호, "달걀 단백질 분석을 위한 HPLC 연구" 한국생물공학회 22 (22): 119-122, 2007

3 허윤석, "다중 이온교환크로마토그래피를 이용한 계란난백에서 리소짐의 분리" 한국생물공학회 18 (18): 122-126, 2003

4 Byun, M. W., "The Changes of Allergenic and Antigenic Properties of Egg White Albumin (Gal d 1) by Gamma Irradiation" 30 (30): 500-504, 2001

5 Nau, F., "Reversed-Phase Liquid Chromatography of Egg White Proteins. Optimization of Ovalbumin Elution" 22 (22): 1129-1147, 1999

6 Yang, J. S., "Feature of Egg White" 32 (32): 42-55, 1999

7 Byun, M. W., "Effects of Heat Treatment on the Antigenicity of Gamma-Irradiated Egg White Albumin" 30 (30): 848-853, 2001

8 Yu, I. J., "Effects of Egg White and Ion Exchange Resin Pretreatment on Separation of Egg White Lysozyme" 16 (16): 157-167, 1989

9 Abalo, C. A., "Comparison of Three Liquid Chromatographic Methods for Egg White Protein Analysis" 723 : 69-74, 1999

10 Lee, S. H., "Characteristics of the Egg White Lysozymes from Ogol Fowl and Fowl for the Lysis of an Alkalophilic and Thermophilic Bacillus sp. TA-11" 9 (9): 447-451, 1996

11 Jung, E. J., "A Study on the Allergenicity of Egg Protein" 11 (11): 228-236, 1998