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HPLC-ELSD를 이용한 발효유 제품 중의 Isomaltooligosaccharides 분석법 개발

Determination of Isomaltooligosaccharides in Yoghurts by Using HPLC-ELSD

  • 고진혁 (경북대학교 식품공학부) ;
  • 이문석 (남양유업 중앙연구소 식품안전센터) ;
  • 곽병만 (남양유업 중앙연구소 식품안전센터) ;
  • 안장혁 (남양유업 중앙연구소 식품안전센터) ;
  • 박종수 (남양유업 중앙연구소 식품안전센터) ;
  • 권중호 (경북대학교 식품공학부)
  • Ko, Jinhyouk (School of Food Science and Biotechnology, Kyungpook National University) ;
  • Lee, Moon-Seok (Food Safety Center, Research and Development Institute, Namyang Dariy Co. Ltd.) ;
  • Kwak, Byung-Man (Food Safety Center, Research and Development Institute, Namyang Dariy Co. Ltd.) ;
  • Ahn, Jang-Hyuk (Food Safety Center, Research and Development Institute, Namyang Dariy Co. Ltd.) ;
  • Park, Jong-Su (Food Safety Center, Research and Development Institute, Namyang Dariy Co. Ltd.) ;
  • Kwon, Joong-Ho (School of Food Science and Biotechnology, Kyungpook National University)
  • 투고 : 2013.03.18
  • 심사 : 2013.06.11
  • 발행 : 2013.06.30

초록

본 연구를 통해 현행 이소말토올리고당 분석법의 한계를 극복하고 원료 시럽뿐만 아니라 발효유 중의 미량의 이소말토올리고당 함량을 신속, 정확하게 분석하기 위한 새로운 분석법을 개발하였다. IDF method와 dSPE 기술을 적용하여 전처리 방법을 개선하였고, 당 전용 컬럼과 ELSD를 이용하여 기기분석조건을 최적화하였다. 새롭게 개발된 분석법은 유효성 검증 절차에 의해 선택성, 직선성, 검출한계 및 정량한계, 회수율, 정확성 및 정밀성이 유효함을 확인하였다. 또한 시장에서 유통 중인 시럽 및 발효유 제품을 분석한 결과 이전에 발표된 연구결과와 일치하는 결론을 얻었으며(Goffin et al., 2011), 이소말토올리고당을 구성하는 성분 중 panose, isomaltose 및 isomaltotriose가 가장 많은 비율을 차지하는 것을 확인하였다. 본 연구 결과는 지방 및 단백질이 많고 유화의 특성을 가진 발효유 중에서의 이소말토올리고당 함량을 신속, 정확하게 분석할 수 있는 기술이 될 것으로 기대된다. 이러한 분석 기술은 향후 식품산업현장에는 물론 발효유를 소비하는 소비자들에게 발효유의 기능성에 대한 정확한 평가를 가능하게 하고, 이소말토올리고당 뿐만 아니라 당 분석을 위한 기초 연구자료로 활용될 것으로 기대된다.

A rapid and simple analytical method for the determination of 9 isomaltooligosaccharides (IMO) species in yoghurts was developed using dispersive solid phase extraction (dSPE) clean-up technic and high performance liquid chromatography with evaporative light-scattering detector (HPLC-ELSD). In this study, 9 IMO were extracted from samples simply with chemical reagent using ISO22662 IDF198 method and additional dSPE clean-up. The optimum instrument conditions for the determination were used carbohydrate ES $5{\mu}$ column with gradient elution of water and acetonitrile and ELS detector. The linearity of this method was expressed as the correlation coefficient ($r^2$), the results of IMO 9 species were shown in 0.9999. LOD and LOQ were respectively 7.9-22.1 mg/kg, 25.9-72.8 mg/kg. The accuracy of intra- and inter-day measurements were in the range from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$, and the preceision of the intra- and inter-day measurements were in the range from 0.8 to 7.7%. The recoveries were from $84.3{\pm}4.5$ to $104.9{\pm}6.5%$. The determination results of IMO 9 species for the 9 yoghurts circulated in the market were in the range from $0.317{\pm}0.007$ to $1.624{\pm}0.050$ g/100 g. The newly developed method is appropriate for the determination of IMO in yoghurts, is a rapid and simple method with excellent resolution in compared with previous method.

키워드

참고문헌

  1. AOAC Committee report. (1989) Appendix D: Guidelines for collaborative study procedures to validate characteristics of a method of analysis. J. Assoc. Off. Anal. Chem. 72, 694-704.
  2. AOAC international. (2007) Pesticide residues in foods by acetonitrile extraction and partitioning with magnesium sulfate. Maryland, USA.
  3. Crittenden, R. G. and Playne, M. J. (1996) Production, properties and applications of food-grade oligosaccharides. Trends Food Sci. Technol. 7, 353-361. https://doi.org/10.1016/S0924-2244(96)10038-8
  4. Goffin, D., Delzenne, N., Blecker, C., Hanon, E., Deroanne, C., and Paquot, M. (2011) Will isomalto-oligosaccharides, a well-established functional food in Asia, break through the European and American market? The status of knowledge on these prebiotics. Crit. Rev. Food Sci. Nutr. 5, 394-409.
  5. Goffin, D., Robert, C., Wathelet, B., Blecker, C., Malmendier, Y., and Paquot, M. (2009) A Step-forward method of quantitative analysis of enzymatically produced isomaltooligosaccharide preparations by AEC-PAD. Chromatogr. 69, 287-293. https://doi.org/10.1365/s10337-008-0875-0
  6. ICH Expert Working Group. (2005) Validation of analytical procedures: Text and methodology Q2(R1). ICH Harmonised tripartite guideline. Step 4, 1-18.
  7. ISO and IDF. (2007) ISO22662 IDF198 first edition. Milk and milk products-determination of lactose content by high-performance liquid chromatography (Reference method). Switzerland.
  8. KFDA (Korea Food and Drug Administration). (2008) The handbook for the method validation guidelines about drug products. Administrative publications 11-1470000-001693-01. Seoul, Korea.
  9. KFDA (Korea Food and Drug Administration). (2012a) Food Standards Codes. Korean Foods Industry Association, Seoul, Korea. pp. 65-71.
  10. KFDA (Korea Food and Drug Administration). (2012b) Food Standards Codes II. Korean Foods Industry Association, Seoul, Korea. pp. 12-13.
  11. KFDA (Korea Food and Drug Administration). (2012c) Food Standards Codes II. Korean Foods Industry Association, Seoul, Korea. pp. 35-36.
  12. Mussatto, S. I. and Mancilha, I. M. (2007) Non-digestible oligosaccharides: A review. Carbohydr. Polym. 68, 587-597. https://doi.org/10.1016/j.carbpol.2006.12.011
  13. Peng, C. A., Ferreira, J. F. S., and Wood, A. J. (2006) Direct analysis of artemisinin from artemisia annua L. using highperformance liquid chromatography with evaporative light scattering detector, and gas chromatography with flame ionization detector. J. Chromatogr. A. 1133, 254-258. https://doi.org/10.1016/j.chroma.2006.08.043
  14. Rivero-Urgell, M. and Santamaria-Orleans, A. (2001) Oligosaccharides: Application in infant food. Early Human Development. 65, S43-S52. https://doi.org/10.1016/S0378-3782(01)00202-X
  15. Robyt, J. F. and Mukerjea, R. (1994) Separation and quantitative determination of nanogram quantities of maltodextrins and isomaltodextrins by thin-layerchromatography. Carbohydr. Res. 251, 187-202. https://doi.org/10.1016/0008-6215(94)84285-X
  16. Sako, T., Matsumoto, K., and Tanaka, R. (1999) Recent progress on research and applications of non-digestible galactooligosaccharides. Int. Dairy J. 9, 69-80. https://doi.org/10.1016/S0958-6946(99)00046-1
  17. Sangeetha, P. T., Ramesh, M. N., and Prapulla, S. G. (2005) Recent trends in the microbial production, analysis and application of fructooligosaccharides. Trends Food Sci. Technol. 16, 442-457. https://doi.org/10.1016/j.tifs.2005.05.003
  18. Thompson, M., Ellison S. L. R, and Wood, R. (2002) Harmonized guidelines for single-laboratory validation of methods of analysis (IUPAC technical report). Pure Appl. Chem. 74, 835-855. https://doi.org/10.1351/pac200274050835
  19. Voragen, A. G. J. (1998) Technological aspects of functional food-related carbohydrates. Trends Food Sci. Technol. 9, 328-335. https://doi.org/10.1016/S0924-2244(98)00059-4

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