Journal of Food Hygiene and Safety (한국식품위생안전성학회지)

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

DOI QR Code

The Optimization and Verification of an Analytical Method for Sodium Iron Chlorophyllin in Foods Using HPLC and LC/MS

식품 중 철클로로필린나트륨의 HPLC 및 LC/MS 최적 분석법과 타당성 검증

  • Chong, Hee Sun (Research Center for Food and Bio Convergence, Department of Food Science, Sun Moon University) ;
  • Park, Yeong Ju (Research Center for Food and Bio Convergence, Department of Food Science, Sun Moon University) ;
  • Kim, Eun Gyeom (Research Center for Food and Bio Convergence, Department of Food Science, Sun Moon University) ;
  • Park, Yea Lim (Research Center for Food and Bio Convergence, Department of Food Science, Sun Moon University) ;
  • Kim, Jin Mi (Research Center for Food and Bio Convergence, Department of Food Science, Sun Moon University) ;
  • Yamaguchi, Tokutaro (Research Center for Food and Bio Convergence, Department of Food Science, Sun Moon University) ;
  • Lee, Chan (Advanced Food Safety Research Group, BrainKorea21 Plus, Department of Food Science and Technology, Chung-Ang University) ;
  • Suh, Hee-Jae (Research Center for Food and Bio Convergence, Department of Food Science, Sun Moon University)
  • 정희선 (선문대학교 식품바이오 융합연구소) ;
  • 박영주 (선문대학교 식품바이오 융합연구소) ;
  • 김은겸 (선문대학교 식품바이오 융합연구소) ;
  • 박예림 (선문대학교 식품바이오 융합연구소) ;
  • 김진미 (선문대학교 식품바이오 융합연구소) ;
  • 야마구치토쿠타로 (선문대학교 식품바이오 융합연구소) ;
  • 이찬 (중앙대학교 식품공학부) ;
  • 서희재 (선문대학교 식품바이오 융합연구소)
  • Received : 2019.03.11
  • Accepted : 2019.04.03
  • Published : 2019.04.30
Download PDF
⟨ Previous Next ⟩

Abstract

An optimized analytical method for sodium iron chloriphyllin in foods was established and verified by using high performance liquid chromatography with attached diode array detection. An Inertsil ODS-2 column and methanol-water (80:20 containing 1% acetate) as a mobile phase were employed. The limit of detection and quantitation of sodium iron chloriphyllin were 0.1 and 0.3 mg/kg, respectively, and the linearity of calibration curve was excellent ($R^2=0.9999$). The accuracy and precision were 93.9~104.95% and 2.0~7.7% in both inter-day and intra-day tests. Recoveries for candy and salad dressing were ranged between 93 and 104% (relative standard deviation, (RSD) 0.3~4.3%), and between 83 and 115% (RSD 1.2~2.0%), respectively. Liquid chromatography mass spectrometry was used to verify the main components of sodium iron chlorophyllin which were Fe-isochlorin e4 and Fe-chlorin e4.

본 연구는 현재 우리나라에서 착색료로 사용허가 되어 있지만 아직 분석법이 개발되지 않은 철클로로필린나트륨의 최적 분석법을 개발하기 위하여 진행되었다. 철클로로필린나트륨은 HPLC-PDA (390 nm), Inertsil ODS-2 column, 1% 초산을 함유한 메탄올-물(80:20, v/v)을 이동상으로 분석할 때 가장 우수한 분리능과 재현성을 나타내었다. UPLC/MS를 통하여 철클로로필린나트륨의 주유도체를 확인한 결과 철 클로린 e4(Fe-chlorin e4)와 철 이소클로린 e4(Fe-isochlorin e4)가 피크의 주요 구성성분임을 확인하였다. 최적 분석법의 타당성을 확보하기 위해 직선성, 검출한계, 정량한계, 정확도, 정밀도, 회수율 분석을 실시하였다. 검량선의 $R^2$은 0.9999, 검출한계는 0.1 mg/kg, 정량한계는 0.3 mg/kg로 나타나 직선성, 검출 한계, 정량한계가 모두 우수하였다. 정확도는 일내분석 93.9~100.8%, 일간분석 99.3~104.95%로 나타났고, 정밀도는 일내분석 2.0~5.4%, 일간분석 4.6~7.7%로 나타났다. 회수율은 수용성 식품(캔디)과 지용성 식품(샐러드드레싱)에서 각각 93.3~104.4%(RSD 0.3~4.3%)과 82.6~114.9%(RSD 1.2~2.0%)로 나타나 국제표준화 기구(ISO)의 기준에 적합하였다. 본 연구는 식품 중 함유된 철클로로필린나트륨을 분석하고 그 유도체를 확인한 결과로 향후 공인분석 방법으로 활용 가치가 클 것으로 기대된다.

Keywords

References

  1. MFDS: Standards and specifications for food additives. In Korea Food Additives Code. Ministry of Food and Drug Safety, Osong, Korea. No. 2017-100 (2017).
  2. Nonomura, Y., Yamaguchi, M., Hara, T., Furuya, K., Yoshioka, N., Inoue, H.: High-performance liquid chromatographic separation of iron (III) chlorophyllin. J. chromatography A, 721, 350-354 (1996). https://doi.org/10.1016/0021-9673(95)00775-X
  3. Chong, H.S., Sim, S., Yamaguchi, T., Park, J., Lee, C., Kim, M., Suh, H. J.: Simultaneous determination of sodium iron chlorophyllin and sodium copper chlorophyllin in food using high-performance liquid chromatography and ultra-performance liquid chromatography-mass spectrometry. Food chemistry, 276, 390-396 (2019). https://doi.org/10.1016/j.foodchem.2018.10.015
  4. KFDA: Standards and specifications for food additives. In Korea Food Additives Code. Korea Food and Drug Administration, Osong, Korea. No. 2013-2184 (2013).
  5. Standards for Use, according to Use Categories Japan. Available from: https://www.ffcr.or.jp/en/tenka/standards-for-use/standards-for-use-of-food-additives.html. Accessed 10 Jul 2017.
  6. Toyoda, T., Cho, Y.M., Mizuta, Y., Akagi, J.I., Nishikawa, A., Ogawa, K.: A 13-week subchronic toxicity study of sodium iron chlorophyllin in F344 rats. J. Toxicol. Sci., 39, 109-119 (2014). https://doi.org/10.2131/jts.39.109
  7. EFSA Panel on Food Additives and Nutrient Sources added to food (ANS): Scientific Opinion on re-evaluation of copper complexes of chlorophylls (E 141 (i)) and chlorophyllins (E 141 (ii)) as food additives. EFSA J., 13, 4151 (2015). https://doi.org/10.2903/j.efsa.2015.4151
  8. Zhan, R., Wu, J., Ouyang, J.: In vitro Antioxidant Activities of Sodium Zinc and Sodium Iron Chlorophyllins from Pine Needles. Food Technol. Biotechnol., 52, 505-510 (2014). https://doi.org/10.17113/ftb.52.04.14.3592
  9. MFDS: Korea Food Additives Code. Ministry of Food and Drug Safety, Osong, Korea. pp. 504 (2015).
  10. JECFA: Purity specifications for chlorophyllins, copper complexes sodium and potassium salts. (1987) Available from: http://www.fao.org/fileadmin/user_upload/jecfa_additives/docs/Monograph1/Additive-127.pdf. Accessed 10 Nov. 2017.
  11. Scotter, M.J., Castle, L., Roberts, D.: Method development and HPLC analysis of retail foods and beverages for copper chlorophyll (E141 [i]) and chlorophyllin (E141 [ii]) food colouring materials. Food Add. Contam., 22, 1163-1175 (2005). https://doi.org/10.1080/02652030500306885
  12. Yasuda, K., Tadano, K., Ushiyama, H., Ogawa, H., Kawai, Y., Nishima, T.: Investigation to find an indicator substance for the analysis of sodium copper chlorophyllin in foods. Food Hyg. Safety Sci., (Shokuhin Eiseigaku Zasshi), 36, 710-716_1 (1995). https://doi.org/10.3358/shokueishi.36.710
  13. Ann, J.H., Lee, J.W., Kim, J.S., Koo, H.M., Yoo, Y.J., Jung, K.S., So, B.H.: Developing of the direct analytical method and monitoring for copper chlorophyll in foods. (2015). Available from: http://www.ndsl.kr/ndsl/commons/util/ndsl-OriginalView.do?dbt=TRKO&cn=TRKO201600010309&rn=&url=&pageCode=PG18. Accessed 14 Jul 2017.
  14. Chong, H.S., Park, H.I., Cho, S.R., Lee, O.K., Park J.H., Lee, C., Suh, H.J.: Establishment and Validation of an Optimized Analytical Method for Sodium Copper Chlorophyllin in Food Using HPLC and LC/MS. J. Korean Soc. Food. Sci. Nutr., 47, 550-558 (2018). https://doi.org/10.3746/jjkfn.2018.47.5.550
  15. Chong, H.S.: Development of the analytical methods for Sodium Copper Chlorophyllin and Sodium Iron Chlorophyllin, food additives by using HPLC and LC/MS. (2018). Available from: http://sunmoon.dcollection.net/public_resource/pdf/200000117182_20190226040927.pdf. Accessed 26 Feb 2019.
  16. EMEA: Validation of analytical procedures. Text and methodology. European Medicines Agency, London, UK. pp. 8-15 (2006).
  17. Mortensen, A., Geppel, A.: HPLC-MS analysis of the green food colorant sodium copper chlorophyllin. Innovative food science & emerging technologies, 8, 419-425 (2007). https://doi.org/10.1016/j.ifset.2007.03.018
  18. ISO: Accuracy (Trueness and Precision) of Measurement Methods and Results-Part 1. Introduction and basic principles. (2011). Available from: https://www.iso.org/obp/ui/ #iso:std:iso:5725:-1:ed-1:v1:en:en. Accessed 10 Jul 2017.
  19. European Commission: Method validation and quality control procedures for pesticide residues analysis in food and feed (Document No. SANCO/2007/3131). (2007). Available from: http://www.crl-pesticides.eu/library/docs/allcrl/Aqc-Guidance_Sanco_2007_3131.pdf. Accessed 10 Mar 2018.