Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
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Determination of β-Carotene in Infant Formulas by High-Performance Liquid Chromatography

HPLC를 이용한 조제유류 중 베타카로틴 함량 분석 연구

  • Hwang, Kyung Mi (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation) ;
  • Bae, Ji Won (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation) ;
  • Hu, Soo Jung (Nutrition and Functional Food Research Team, National Institute of Food and Drug Safety Evaluation) ;
  • Oh, Keum Soon (Food Additives Standard division, Ministry of Food and Drug Safety)
  • 황경미 (식품의약품안전평가원 영양기능연구팀) ;
  • 배지원 (식품의약품안전평가원 영양기능연구팀) ;
  • 허수정 (식품의약품안전평가원 영양기능연구팀) ;
  • 오금순 (식품의약품안전처 첨가물 기준과)
  • Received : 2019.06.25
  • Accepted : 2019.07.18
  • Published : 2019.08.30
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Abstract

A procedure based on high-performance liquid chromatography (HPLC) is described to determine ${\beta}-carotene$ in infant formulas. The method for ${\beta}-carotene$ analysis was performed on a C18 reversed-phase column using acetonitrile/methanol/dichloromethane (6:1:3, v/v/v) as a mobile phase. ${\beta}-Carotene$ was determined in HPLC with photo diode array (PDA) detector. The parameters of validation were specificity, linearity, LOD, LOQ, accuracy, precision and repeatability. The specificity was confirmed by the retention time and the linearity ($R^2$), which was over 0.999 in the range of 0.125~2 mg/L. The detection and quantification limits were 0.064 and 0.193 mg/L, respectively. The accuracy and precision of this method using an STD spiked sample were 80~119% and 1.02~2.05% respectively. The method was applied to the analysis of various infant formula and follow-up formulas products containing ${\beta}-carotene$, and all the products contained acceptable levels of ${\beta}-carotene$ for nutrition labeling.

본 연구는 조제유류 중 "식품의 기준 및 성분규격"에 기준 규격이 설정되어 있지 않으나 제품에 함유되어 있는 베타카로틴에 대해 분석법을 마련하고자 수행하였다. 조제유류에 함유된 베타카로틴 함량 분석을 위해 HPLC를 이용한 분석법을 확립하고 시중에 유통 중인 제품을 대상으로 적용성을 검토하였다. 베타카로틴 표준품을 이용하여 HPLC를 이용한 기기분석조건을 확립하고 시료 중의 베타카로틴을 추출하여 분석하였다. 분석법 검증은 특이성, 직선성, 검출한계 및 정량한계, 정확성, 정밀성에 대해 수행되었다. 0.125~2 mg/L의 농도범위에서 $R^2=0.999$ 이상의 우수한 직선성을 확인할 수 있었으며, LOD와 LOQ는 각각 0.1, 0.2 mg/L였다. 표준물질 첨가법을 이용하여 정확성을 검토하였으며, 81~120%의 회수율을 확인할 수 있었다. 정밀성을 검토한 결과 시료 채취량에 따른 반복성은 RSD값이 2.1~4.9%, 실험실간 교차검증을 통한 실험일자에 따른 재현성은 4.0 RSD(%)로 확인되었다. 본 연구에서 확립된 분석법을 적용하여 조제유류 13건, 성장기용조제식 7건 등 국내 유통 중인 제품 20건에 대해 적용성 검토를 실시한 결과 전체 시료에서 분석이 용이하였으며, 모두 표시기준에 적합함을 확인하였다.

Keywords

References

  1. Holick, C.N., Michaud, D.S., Stolzenberg-Solomon, R., Mayne S.T., Pietinen, P., Taylor, P.R., Virtamo, J., Albanes, D.: Dietary carotenoids, serum $\beta$-carotene, aand retinol and risk of lung cancer in the alpha-tocopherol, beta-carotene cohort study. Am. J. Epidemiol. 156(6), 536-547 (2002). https://doi.org/10.1093/aje/kwf072
  2. Noel W.S.Olson J.A. 2012. Chapter 11 Vitamin A. Present knowledge in nutrition. Tenth ed. Wiley Press, Washington D.C.
  3. Chytil, F.: Retinoids in lung development. FASEB J., 9, 986-992 (1996). https://doi.org/10.1096/fasebj.10.9.8801181
  4. Thompkinson, D.K., Kharb, S.: Aspects of infant food formulation. Comprehensive Rev. Food Sci. Food Safety, 6, 79-102 (2007). https://doi.org/10.1111/j.1541-4337.2007.00020.x
  5. Ok, K.H., Kim, K.S., Seo, J.S., Choi, Y.S., Shin, S.M.: A study on the nutrient intakes and supplemental food of infants in relation to the method of feeding practice. Kor. J. Nutr. 29(2), 143-152 (1996).
  6. CODEX ALIMENTARIUS: Standard for infant formula and formulas for special medical purposes intented for infants CODEX Stan 72-1981, FAO, WHO (2011).
  7. Food Code of Korea. Ministry of Food and Drug Safety, Cheongju, Korea (2018).
  8. AOAC official method 2005.07., Official methods of analysis of AOAC INTERNATIONAL 20th Ed. AOAC International, Gaithersburg, MD (2016).
  9. Thomas, J.B., Kline, M.C., Gill, L.M. Yen, J.H., Duewer, D.L., Sniegoski, L.T., Sharpless, K.E.: Preparation and value assignment of standard reference material 968c fat-soluble vitamins, carotenoids, and cholesterol in human serum. Clin. Chim. Acta, 305, 141-155 (2001). https://doi.org/10.1016/S0009-8981(00)00429-0
  10. David, C.W., Anja, B., Harvey, I., Adrienne, M.: Determination of vitamin A and vitamin E esters in infant formulae and fortified milk powders by HPLC : Use of internal standardisation. Food Chem. Acta, 197, 457-465 (2016). https://doi.org/10.1016/j.foodchem.2015.10.077
  11. AOAC official method 2016.13., Official methods of analysis of AOAC INTERNATIONAL 20th Ed. AOAC International, Gaithersburg, MD (2016).
  12. Standards for health functional food of Korea. Ministry of Food and Drug Safety, Cheongju, Korea (2018).
  13. Rodeney, J.B. : Determination of $\alpha$-and $\beta$-carotene in some raw fruits and vegetables by high-performance liquid chromatography. J. Agric. Food Chem. 34(3), 409-412 (1986). https://doi.org/10.1021/jf00069a006
  14. Official methods of analysis of AOAC INTERNATIONAL 19th Ed., Appendix L. AOAC INTERNATIONAL, Dr. George WL Jr, Recommended Guidelines for SPIFAN Single Laboratory Validation (SLV) (2012).