Nutr Res Pract.  2008 Mar;2(1):46-49.

Identification and quantification of anthocyanin pigments in colored rice

• Kim MK
• Kim HA
• Koh K
• Kim HS
• Lee YS
• Kim YH

Abstract

Anthocyanin pigments from varieties of black, red and wild rice were identified and quantified to evaluate their potential as nutritional function, natural colorants or functional food ingredients. Anthocyanin extraction was conducted with acidified methanol with 0.1M HCl (85:15, v/v) and identification of anthocyanin, aglycone and sugar moieties was conducted by comparison with purified standards by HPLC, Ultraviolet-Visible absorption spectrophotometer and paper chromatography. Black and wild rice showed three different types of pigments by HPLC whereas red rice variety did not show any anthocyanins. Out of three pigments detected, one (peak 2) was characterized as cyanidin-3-glucoside (C3G) by comparison of spectroscopic and chromatographic properties with an authentic standard, and another (peak 3) was tentatively identified as cyanidin-fructoside on the basis of spectroscopic properties with lambdamax of aglycone in 1% HCl methanol at 537 nm, electrospray ionization mass spectra with major ions at 449 and 287 m/z and chromatographic properties. But another pigment (peak 1) has not been characterized. The most abundant anthocyanin in black and wild rice was C3G.

Keyword

Anthocyanin; black rice; wild rice; natural colorant; LC-MS

MeSH Terms

Absorption
Anthocyanins
Chromatography, High Pressure Liquid
Chromatography, Paper
Functional Food
Glucosides
Ions
Methanol
Anthocyanins
Glucosides
Ions
Methanol

Figure

• 

  Fig. 1 HPLC chromatograms of anthocyanin pigments separated from black rice (A) and wild rice (B) by Xterra MS C18 column. Peak 1, unknown; peak 2, cyanidin-glucoside (confirmed); peak 3, cyanidin-fructoside (presumed).

• 

  Fig. 2 LC-MS spectrum of unknown peak 3 shown in Fig. 1. (Cyanidin: major ions m/z 449 and 287).

Reference

1. Abdel-Aal E-SM, Hucl P. A rapid method for quantifying total anthocyanins in blue aleurone and purple pericarp wheats. Cereal Chemistry. 1999. 76:350–354.

2. Abdel-Aal E-SM, Hucl P. Composition and stability of anthocyanins in blue-grained wheat. J Agric Food Chem. 2003. 51:2174–2180.

3. Abdel-Aal E-SM, Young JC, Rabalski I. Anthocyanin composition in black, blue, pink, purple, and red cereal grains. J Agric Food Chem. 2006. 54:4696–4704.

4. Choi SW, Kang WW, Osawa T. Isolation and identification of anthocyanin pigments in black rice. Foods and Biotechnology. 1994. 3:131–136.

5. Francis FJ. Anthocyanins and betalains composition: composition and applications. Cereal Foods World. 2000. 45:208–213.

6. Fuleki T, Francis FJ. Quantitative methods for anthocyanins. 1. Extraction and determination of total anthocyanin in cranberries. J Food Sci. 1968. 33:72–77.

7. Harborne JB. Phytochemical methods. A guide to modern techniques of plant analysis. 1984. New York. USA: Chapman and Hall;33–99.

8. Hyun JW, Chung HS. Cyanidin and malvidin from Oryza sativa cv. Heugjinjubyeo mediate cytotoxicity against human monocytic leukemia cells by arrest of G₂/M phase and induction of apoptosis. J Agric Food Chem. 2004. 52:2213–2217.

9. Kamei H, Kojima T, Hasegawa M, Koide T, Umeda T, Yukawa T, Terabe K. Suppression of tumor cell growth by anthocyanins in vitro. Cancer Invest. 1995. 13:590–594.

10. Kim KI, Nam JH, Kwon TW. On the proximate composition, organic acids and anthocyanins of Omija, Schizandra chinensis Baillon. Korean Journal of Food Science and Technology. 1973. 5:178–182.

11. Mazza G, Gao L. Abdel-Aal E-SM, Peter PJ, editors. Blue and purple grains. Specialty Grains for Food and Feed. 2005. St. Paul. USA: American Association of Cereal Chemists;45–67.

12. Moreno YS, Sánchez GS, Hernández DR, Lobato NR. Characterization of antocyanin extracts from maize kernels. J Chromatogr Sci. 2005. 43:483–487.

13. Nam SH, Choi SP, Kang MY, Koh HJ, Kozukue N, Friedman M. Antioxidative activities of bran from twenty one pigmented rice cultivars. Food Chemistry. 2006. 94:613–620.

14. Philpott M, Gould KS, Lim C, Ferguson LR. In situ and in vitro antioxidant activity of sweetpotato anthocyanins. J Agric Food Chem. 2006. 54:1710–1715.

15. Ryu SN, Park SZ, Ho CT. High performance liquid chromatographic determination of anthocyanin pigments in some varieties of black rice. Journal of Food and Drug Analysis. 1998. 6:729–736.

16. Sakamura S, Francis FJ. The anthocyanins of the American cranberry. J Food Sci. 1961. 26:318–321.

17. Satue-Gracia M, Heinonen IM, Frankel EN. Anthocyanins as antioxidants on human low-density lipoprotein and lecithin-liposome system. J Agric Food Chem. 1997. 45:3362–3367.

18. Schou SA. Light absorption of several anthocyanins. Helv Chim Acta. 1927. 10:907–915.

19. Spaeth EC, Rosenblatt DH. Partition chromatography of synthetic anthocyanidin mixtures. Anal Chem. 1950. 22:1321–1326.

20. Strack D, Wray V. Methods in Plant Biochemistry. 1989. London. England: Academic Press;325–356.

21. Tsuda T, Horio F, Osawa T. Cyanidin 3-О-β-glucoside suppresses nitric oxide production during a zymosan treatment in rats. J Nutr Sci Vitaminol. 2002. 48:305–310.

22. Tsuda T, Horio F, Uchida K, Aoki H, Osawa T. Dietary cyanidin 3-О-β-D-glucoside-rich purple corn color prevents obesity and ameliorates hyperglycemia. J Nutr. 2003. 133:2125–2130.

23. Yoon HH, Paik YS, Kim JB, Hahn TR. Identification of anthocyanidins from Korean pigmented rice. Agricultural Chemistry and Biotechnology. 1995. 38:581–583.

24. Yoshinaga K. Liquor with pigments of red rice. Journal of Brewing Society of Japan. 1986. 81:337–342.

25. Zhao C, Giusti MM, Malik M, Moyer MP, Magnuson BA. Effects of commercial anthocyanin-rich extracts on colonic cancer and nontumorigenic colonic cell growth. J Agric Food Chem. 2004. 52:6122–6128.