Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

The Korean Society of Food Hygiene and Safety (한국식품위생안전성학회)
권호 표지
DOI QR코드

DOI QR Code

Optimization of Analytical Method for Annatto Pigment in Foods

식품 중 안나토색소 분석법 최적화 연구

  • Lee, Jiyeon (Department of Food Science and Technology, Chung-Ang University) ;
  • Park, Juhee (Department of Food Science and Technology, Chung-Ang University) ;
  • Lee, Jihyun (Department of Food Science and Technology, Chung-Ang University) ;
  • Suh, Hee-Jae (Department of Food Science, Sun Moon University) ;
  • Lee, Chan (Department of Food Science and Technology, Chung-Ang University)
  • Received : 2021.07.05
  • Accepted : 2021.08.11
  • Published : 2021.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

In this study we sought to develop a simultaneous analysis method for cis-bixin and cis-norbixin, the main components, to detect annatto pigment in food. To establish the optimal test method, the HPLC analysis methods of the European Food Safety Authority (EFSA), Japan's Ministry of Health, Labor and Welfare (MHLW), and National Institute of Food and Drug Safety Evaluation (NIFDS) were compared and reviewed. In addition, a new pretreatment method applicable to various foods was developed after selecting conditions for simultaneous high-performance liquid chromatography (HPLC) analysis in consideration of linearity, limit of detection (LOD), limit of quantification (LOQ), and analysis time. The HPLC analysis method of NIFDS showed the best linearity (R2 ≥ 0.999), exhibiting low detection and quantification limits for cis-norbixin and cis-bixin as 0.03, 0.05 ㎍/mL, and 0.097, 0.16 ㎍/mL, respectively. All previously reported pretreatment methods had limitations in various food applications. However, the new pretreatment method showed a high recovery rate for all three main food groups of fish meat and meat products, processed cheese and beverages. This method showed an excellent simultaneous recovery rate of 98% or more for cis-bixin and cis-norbixin. The HPLC analysis method with a new pretreatment method showed high linearity with a coefficient of determination (R2) of 1 for both substances, and the accuracy (recovery rate) and precision (%RSD) were 98% and between 0.4-7.9, respectively. From this result, the optimized analytical method was considered to be very suitable for the simultaneous analysis of cis-bixin and cis-norbixin, two main components of annatto pigment in food.

본 연구의 목적은 식품 중 안나토색소를 검출하기 위하여 주성분인 cis-bixin과 cis-norbixin의 동시분석법을 개발하는 것이다. 최적 시험법 확립을 위해 국내외 기관 중 유럽식품안전청, 일본 후생노동성, 우리나라 식품의약품안전평가원의 HPLC분석법들을 비교 및 검토하였다. 그리고 직선성, 검출한계, 정량한계 및 분석시간을 고려하여 최적 HPLC 동시분석조건을 선택 후 여러 식품에 적용가능한 최적 전처리법을 개발하였다. 식품의약품안전평가원 HPLC 분석법이 가장 우수한 직선성(R2≥0.999)을 보였으며, cis-norbixin 및 cis-bixin에 대한 검출한계와 정령한계가 각각 0.03과 0.05 ㎍/mL 그리고 0.097과 0.16 ㎍/mL로 낮게 나타났다. 모든 보고된 전처리방법은 여러 식품적용에 한계가 있었으며, 어육 및 육가공품, 가공치즈, 음료의 주요 세 식품군 모두에 대해서 높은 회수율을 보이는 최적 전처리법이 새롭게 확립되었다. 이 전처리 방법은 cis-norbixin과 cis-bixin에 대해서 98% 이상의 우수한 동시 회수율을 나타냈었다. 새로운 전처리방법이 적용된 분석법은 두 성분에 대해 모두 결정계수(R2) 1로서 높은 직선성을 나타냈으며, 정확도(회수율)와 정밀도(%RSD)가 각각 평균 98% 및 0.4-7.9 이었다. 이러한 결과로부터 최적화된 분석시험법은 식품 중 안나토색소의 cis-norbixin과 cis-bixin 두 성분 동시분석에 매우 적합한 것으로 판단되었다.

Keywords

Acknowledgement

이 연구는 2020년도 식품의약품안전처의 연구개발비(18162식품위009)로 수행되었으며, 이 논문은 2020년도 중앙대학교 연구년 결과물로 제출됨.

References

  1. Scotter, M.J., Thorpe, S.A., Reynolds, S.L., Wilson, L.A., Strutt, P.R., Characterization of the principal colouring components of annatto using high performance liquid chromatography with photodiode-array detection. Food Addit. Contam., 11, 301-315 (1994). https://doi.org/10.1080/02652039409374229
  2. EFSA ANS Panel (EFSA Panel on Food Additives and Nutrient Sources added to Food), Safety of annatto E and the exposure to the annatto colouring principles bixin and norbixin (E 160b) when used as a food additive, EFSA J., 17, 5626, (2019).
  3. Sabuz, A.A., Khan, M.H.H., Rahman, M.T., Rana, R., Brahma, S., Stability of organic food colorant extracted from annatto seeds on food matrix, Int. J. Food Sci. Nutr., 5, 10-16 (2020).
  4. EFSA ANS Panel (EFSA Panel on Food Additives and Nutrient Sources added to Food), Safety of annatto extracts (E 160b) as a food additive, EFSA J., 14, 4544, (2016).
  5. Scotter, M.J., Castle, L., Honeybone, C.A., Nelson, C., Method development and analysis of retail foods for annatto food colouring material. Food Addit. Contam., 19, 205-222 (2002). https://doi.org/10.1080/02652030110085386
  6. Attokaran, M., 2011. Natural Food Flavors and Colorants, second ed. Jhon Wiley & Sons Ltd; Wiley-Blackwell, Chicago, IL, USA, pp. 63-67.
  7. FAO, (2021. August 23). Annatto Extracts (Chemical and Technical Assessment). Retrieved from http://www.fao.org/3/at973e/at973e.pdf
  8. Joint FAO/WHO Expert Committee on Food Additives. Meeting (67th: 2006: Rome, Italy), World Health Organization & Food and Agriculture Organization of the United Nations. (2007). Evaluation of certain food additives and contaminants: sixty-seventh report of the Joint FAO/WHO Expert Committee on Food Additives. World Health Organization. Retrieved from https://apps.who.int/iris/handle/10665/43592
  9. CODEX Alimentarius, (2021, May 18). GSFA Online. Retrieved from http://www.fao.org/gsfaonline/additives/index.html
  10. Joint FAO/WHO Expert Committee on Food Additives. Meeting (19th: 1983: Rome, Italy), World Health Organization, Evaluation of certain food additives and contaminants: Twenty-sixth report of the Joint FAO/WHO Expert Committee on Food Additives. World Health Organization. Retrieved from https://apps.who.int/iris/handle/10665/41546
  11. European Union (EU), Commission Regulation (EU) 2020/771 of 11 June 2020 amending Annexes II and III to Regulation (EC) No 1333/2008 of the European Parliament and of the Council and the Annex to Commission Regulation (EU) No 231/2012 as regards the use of Annatto, Bixin, Norbixin (E 160b) (Text with EEA relevance). OJEU, 184, 25-42 (2020).
  12. Food additives code. (2021, August 23). Annatto, Water-Soluble. Retrieved from http://www.foodsafetykorea.go.kr/foodcode/04_03.jsp?idx=222
  13. Food additives code. (2021, August 23). Annatto Extract. Retrieved from http://www.foodsafetykorea.go.kr/foodcode/04_03.jsp?idx=291
  14. Ministry of Health, Labour, Welfare. (2021, August 23). Japan's Specifications and Standards for Food Additives, D. Monographs, Annatto Extract, Bixin, Norbixin, Annatto, Water-soluble. Retrieved from http://www.nihs.go.jp/dfa/_src/541/D-210224.pdf
  15. Ministry of Health, Labour, Welfare. (2021, August 23). Food additives analysis methods. Retrieved from https://www.ffcr.or.jp/webupload/168bb6f7b3f31735b345b5e65defb1c98a675b07.pdf
  16. Food additives analytical methods in foods. (2021, August 23). Food additive analysis method in food. Retrieved from https://www.haccp.or.kr/site/haccp/boardView.do?post=59051&page=16&boardSeq=773&key=2216&category=&searchType=&searchKeyword=&subContents=
  17. Noppe, H., Abuin Martinez, S., Verheyden, K., Van Loco, J., Companyo Beltran, R., De Brabander, H.F, Determination of bixin and norbixin in meat using liquid chromatography and photodiode array detection. Food Addit. Contam., 26, 17-24 (2009). https://doi.org/10.1080/02652030802322564
  18. ICH Harmonised Tripartite Guideline. (2021, August 23). Validation of Analytical Procedures: Text and Methodology Q2(R1). Retrieved from https://database.ich.org/sites/default/files/Q2%28R1%29%20Guideline.pdf
  19. De Oliveira Rios, A., Mercadante, A.Z., Novel method for the determination of added annatto colour in extruded corn snack products. Food Addit. Contam., 21, 125-133 (2004). https://doi.org/10.1080/02652030310001642771
  20. Food and Drug Administration, 2019. Guidelines for the Validation of Chemical Methods in Food, Feed, Cosmetics, and Veterinary Products. Food and Drug Administration, Silver Spring. MD, USA, pp. 1-39.
  21. Bareth, A., Strohmar, W., Kitzelmann, E., HPLC and spectrophotometric determination of annatto in cheese. Eur. Food Res. Technol., 215, 359-364 (2002). https://doi.org/10.1007/s00217-002-0541-y
  22. van Scheppingen, W.B., Boogers, I.A.L.A., Duchateau, A.L.L., Study on decomposition products of norbixin during bleaching with hydrogen peroxide and a peroxidase by means of UPLC-UV and mass spectrometry. Food Chem., 132, 1354-1359 (2012). https://doi.org/10.1016/j.foodchem.2011.11.118
  23. Breithaupt, D.E., Simultaneous HPLC determination of carotenoids used as food coloring additives: applicability of accelerated solvent extraction, Food Addit. Contam., 86, 449-456 (2004).
  24. Lancaster, F.E., Lawrence, J.F., Determination of annatto in high?fat dairy products, margarine and hard candy by solvent extraction followed by high-performance liquid chromatography, Food Addit. Contam., 12, 9-19 (1995). https://doi.org/10.1080/02652039509374274