Survey of Beet Red Contents in Foods using TLC, HPLC

TLC, HPLC를 이용한 식품 중 비트레드 함량조사

  • Jang, Yaung-Mi (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Lee, Tal-Soo (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Hong, Ki-Hyoung (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Park, Sung-Kwan (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Park, Sung-Kug (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Kwon, Yong-Kwan (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Park, Jae-Seok (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Chang, Sun-Young (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Hwang, Hye-Shin (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Kim, Eun-Jeong (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Han, Yeun-Jeong (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Kim, Byung-Sub (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration) ;
  • Won, Hye-Jin (Food Additives Team Nutrition & Functional Food Headquarters, Korea Food and Drug, Administration)
  • 장영미 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 이달수 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 홍기형 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 박성관 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 박성국 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 권용관 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 박재석 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 장선영 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 황혜신 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 김은정 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 한윤정 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 김병섭 (식품의약품안전청 영양기능식품본부 식품첨가물팀) ;
  • 원혜진 (식품의약품안전청 영양기능식품본부 식품첨가물팀)
  • Published : 2005.12.01

Abstract

This study was performed far development of new analytical method of beet red in foods. In this study, analysis of beet red in foods has been carried out by detection of betanine and isobetanine, the main color component of beet red as indicator compounds. The qualitative analysis technique consisted of clean-up of the colors with a $C_{18}$ cartridge, separation of the colors by cellulose TLC plate using acetone:3-methyl-1-butanol:distilled water (7:7:6) as a solvent system. Also, the quantitative analysis was performed using X-terra RP at wavelength 538 nm and $0.1\%$ phosphoric acid : methanol (90:10) as a solvent. The quantitative results of beet .ed were as follows:900.22$\∼$27701.60 $\mu$g/g for 60 item in nutrient supplement food, $21.95\∼713.40{\mu}g/g$ for 30 items and N.D. for 18 items in cindy, and $155.85{\∼}505.37{\mu}g/g$ for 12 items in ice creams, $43.52\∼64.75{\mu}g/g$ for 18 items and N.D. for 54 item in sauce, N.D. for 12 items in retort food.

비트레드는 '천연식품, 다류, 고춧가루 또는 실고추, 김치류, 고추장 및 식초'에 사용 금지토록 현행 식품첨가물공전에 사용기준이 규정되어 있으나, 식품 중 비트레드의 분석방법이 확립되어 있지 않은 실정이다. 따라서 국내유통 중인 식품 중 비트레드의 함유량을 조사하여 추후 식품 중 비트레드의 사용기준 준수여부 확인시 시험방법으로 사용하고자 식품 중 비트레드 분석법을 확립하였다. 본 연구는 LC-Mass로 비트레드의 주성분임을 확인한 베타닌(Betanine)과 이소베타닌 (Isobetanine)을 사용하여 HPTLC (High Performance Thin Layer Chromatography)와HPLC (High Performance Liquid Chromatography)로 신속하고 재현성이 높은 비트레드의 정성$\cdot$정량시험방법을 확립하였다. 이 때 정성분석조건은 셀룰로오스 박층판에, 아세톤:3-메틸-1-부탄올:물 (7:7:6) 전개용매를 이용하고, 정량분석은 역상계인 X-terra RP ($4.6\;mm{\times}250\;mm,\;5{\mu}m$)칼럼에 이동상, 온도 및 파장조건으로 각각 $0.1\%$ 인산:메탄올 (90:10), $40^{\circ}C$ 및 538 nm 조건이었을 때 가장 양호한 분석결과를 얻었다. 확립된 분석방법을 적용하여 시중에 유통되고 있는 식품 중 비트레드 사용실태를 확인하거나 사용가능성이 있는 영양보충용식품, 아이스크림, 소스류, 캔디류, 레토르트식품 등 총 5종 204품목에 함유된 비트레드의 함유량을 조사하였다. 그 결과, 영양 보충용식품은 60품목에서 $900.22\∼27,701.60{\mu}g/g$이 검출되었으며, 캔디류는 48품목 중 30품목에서 $21.95\∼713.40{\mu}g/g$이 검출, 나머지 18품목에서는 검출되지 않았다. 아이스크림에서는 12품목에서 $155.85{\∼}505.37{\mu}g/g$이 검출되었고, 소스류는 72품목 중 18품목에서 $43.52{\∼}64.75{\mu}g/g$이 검출, 나머지 54품목에서 검출되지 않았고 레토르트식품은 12품목에서 모두 검출되지 않았다.

Keywords

References

  1. Saito, K. and Miyakawa, K.I.: Sono-oscillatory mobilization of bound precarthamine from flower florets of dyer's saffron. Food Chem., 54, 269-271 (1995) https://doi.org/10.1016/0308-8146(95)00045-K
  2. Kim, S.H., Kim, S.H., Lee, J.N., An, S.W., Kim, K.S., Hwang B. and Lee, H.Y: Optimization of betacyanin production by red beet (Beta vulgaris L.) hairy root cultures. Kor. J Appl. microbiol. Biotechnol., 26(5), 435-441 (1998)
  3. Park, M.S. and Hong, I.K.: Analysis of color difference by mixed solvent composition in natural dyes extraction process. J Korean Ind, Eng. Chem. 13(8), 844-851 (2002)
  4. Macdougall, D.B.: Colour in food. CRC Press, New York, pp. 155-315 (2002)
  5. Gabriel, J.L.: Natural food colorants. Marcel Dekker Inc., New York, pp. 11-30 (2000)
  6. Methods of Analysis in Health Science, pp. 348-365 (2000)
  7. 藤井正義, 清水孝重, 中村幹雄, 新版天然色素, pp. 157-161, 坪城13年
  8. Florian, C.S. and Reinhold, C.: Structural Investigations on betacyanin pigments by LC NMR and 2D NMR spectroscopy. Trends in Food Sci. & Technol., 15, 19-38 (2004) https://doi.org/10.1016/j.tifs.2003.07.004
  9. Wybraniec, S. and Mizrahi, Y.: Influence of perfluorinated carboxylic acid on ion-pair reversed-phase high-performance liquid chromatographic separation of betacyanins and 17decarboxy- betacyanins. J Chromatogr. A, 1029, 97-101 (2004) https://doi.org/10.1016/j.chroma.2003.12.055
  10. Stuppner, H. and Egger, R.L.: Application of capillary zone electrophoresis to the analysis of betalains from Beta vulgaris. J Chromatogr. A, 735, 409-413 (1996) https://doi.org/10.1016/0021-9673(95)00885-3
  11. Fernandez-Lopez, J.A. and Almela, L.: Application of highperformance liquid chromatography to the characterization of the betalain pigment in prickly pear fruits. J Chromatogr. A, 913, 415-420 (2001) https://doi.org/10.1016/S0021-9673(00)01224-3
  12. Naoko, K., Jurgen, S., Manfred, N., Victor, W. and Willibald S.: Betalains from Chistmas cactus. Phytochem., 54, 419-426 (2000) https://doi.org/10.1016/S0031-9422(00)00129-1
  13. Kent, R.V. and Rovert, GS.: Separation and quantification of red beet betacyanins and betaxanthins by high-performance liquid chromatography. J Agric. Food Chem., 26(4),812-816 (1978) https://doi.org/10.1021/jf60218a026
  14. Florian, K., Florian, C.S. and Reinhold C.: Identification of betalains from petioles of differently coloreds swiss chard by high-performance liquid chromatography-electrospray ionization mass spectrometry. J Agric. Food Chem. 52,2975-2981 (2004) https://doi.org/10.1021/jf035491w
  15. Florian, C.S., Andreas, S. and Renihold, C.: Identification of betalains from yellow beet (Beta vulgaris L.) and cactus pear [Opuntia ficus-indica (L.) Mill.] by high-performance liquid chromatography-electrospray ionization mass spectrometry. J Agric. Food Chem., 50, 2302-2307 (2002) https://doi.org/10.1021/jf011305f
  16. Trezzini, GF. and Zryd, J.P.: Charaterization of some natural and semi-synthetic betaxanthins. Phytochem., 30(6), 1901-1903 (1991) https://doi.org/10.1016/0031-9422(91)85036-Y
  17. Escribano, J. and Pedreno, M.A.: Charaterization of the antiradical activity of betalains from beta vulgaris L. roots, Phytochem. Anal., 9, 124-127 (1998) https://doi.org/10.1002/(SICI)1099-1565(199805/06)9:3<124::AID-PCA401>3.0.CO;2-0
  18. Kuyala, T.S. and Loponen, J.M.: Phenolic and betacyanins in red beetroot(Beta vulgaris)root: distribution and effect of cold storage on the content of total phenolics and three individual compounds. J. Agaric. Food Chem., 48, 5338-5342 (2000) https://doi.org/10.1021/jf000523q
  19. Cai, Y., Sun, M., Schliemann, W. and Coke, H.: Chemical stability and colorant properties of betaxanthin pigment from Celosia argentea. J. Agric. Food Chem., 49, 4429-4435 (2001) https://doi.org/10.1021/jf0104735