Rapid High Performance Liquid Chromatographic Quantification of Major Isoflavones in Soybeans and Soybean Pastes

  • Kim, Won-Chan (Division of Applied Biology & Chemistry, College of Agriculture & Life Sciences, Kyungpook National University) ;
  • Kwon, Soon-Ho (Division of Applied Biology & Chemistry, College of Agriculture & Life Sciences, Kyungpook National University) ;
  • Rhee, In-Koo (Division of Applied Biology & Chemistry, College of Agriculture & Life Sciences, Kyungpook National University) ;
  • Hur, Jong-Moon (Division of Applied Biology & Chemistry, College of Agriculture & Life Sciences, Kyungpook National University) ;
  • Jeong, Hyun-Hee (Division of Applied Biology & Chemistry, College of Agriculture & Life Sciences, Kyungpook National University) ;
  • Choi, Sun-Ha (Department of Biochemistry, School of Medicine, Konyang University) ;
  • Lee, Kyung-Bok (Department of Biochemistry, School of Medicine, Konyang University) ;
  • Kang, Young-Hwa (Division of Applied Biology & Chemistry, College of Agriculture & Life Sciences, Kyungpook National University) ;
  • Song, Kyung-Sik (Division of Applied Biology & Chemistry, College of Agriculture & Life Sciences, Kyungpook National University)
  • Published : 2006.02.28

Abstract

A simple HPLC quantification method was developed for genistein, genistin, daidzein, and daidzin in soybeans and soybean products. The procedure used a $4.6{\times}100\;mm$ $Chromolith^{(R)}$ RP-18e column with a mobile phase of 1% HOAc in 20% MeOH to 1% HOAc in 80% MeOH for 10 min. The injection volume was $2\;{\mu}L$ at a flow rate of 2 mL/min. Detection was carried out under UV at 254 nm. Under these conditions, the major isoflavones daidzein, daidzin, genistein, and genistin in soybean and soybean pastes were eluted within 7 min with baseline separation. Optimal extraction of the above four major isoflavones was achieved when 40 g of soybean or soybean paste was refluxed in 100 mL of 95% ethanol for 2 hr. Ten different soybean cultivars and nine commercial soybean pastes were analyzed by this method. The total isoflavone content was highest in the cultivar Somyung ($2,497\;{\mu}g/g$ dry weight). The isoflavone content in soybean pastes varied widely from manufacturer to manufacturer (an almost five-fold difference between the highest and lowest values). Such variations were presumably due to differences in fermentation conditions, type of soybeans used, and levels of such additives as starch and salt.

Keywords

References

  1. Tham DM, Gardner CD, Haskell WL. Clinical review 97: Potential health benefits of dietary phytoestrogens: a review of the clinical, epidemiological, and mechanistic evidence. J. Clin. Endocrinol. Metab. 83: 2223-2235 (1998) https://doi.org/10.1210/jc.83.7.2223
  2. Nam HY, Min SG, Shin HC, Kim HY, Fukushima M, Han KH, Park WJ, Choi, KD, Lee CH. The protective effect ofisoflavone extracted from soybean paste in free radical inhibitor treated rats. Food Sci. Biotechnol. 14: 586-592 (2005)
  3. Bingham SA, Atkinson C, Liggins J, Bluck L, Coward A. Phytoestrogens: where are we now? Br. J. Nutr. 79: 393-406 (1998) https://doi.org/10.1079/BJN19980068
  4. Bingham SA. Diet and colorectal cancer prevention. Biochem. Soc. Transact. 28: 12-16 (2000) https://doi.org/10.1042/bst0280012
  5. Jacobsen BK, Knutsen SF, Fraser GE. Does high soy milk intake reduce prostate cancer incidence? The Adventist Health Study (United States). Cancer Causes Control. 9: 553-557 (1998) https://doi.org/10.1023/A:1008819500080
  6. Bradlow HL, Sepkovic ow. Diet and breast cancer. An. New York Acad. Sci. 963: 247-267 (2002)
  7. Alkel DL, Sirtori CR, Muanza DN, Vasquez A, Cannell J, Grant WP, van der Schouw YT, Grobbee DE, Kok L, Kreijkamp-Kaspers S. Isoflavones and postmenopausal women. J. Am. Med. Assoc. 292: 2336-2337 (2004)
  8. van der Schouw YT, de Kleijn MJ, Peeters PH, Grobbee DE. Phytooestrogens and cardiovascular disease risk. Nutr. Metab. Cardiovasc. Dis. 10: 154-67 (2000)
  9. Anderson JJ, Garner SC. Phytoestrogens and bone. Bailliere's Clin. Endocrinol. Metab. 12: 543-557 (1998)
  10. Ghosh P, Fenner GP. Improved method for gas chromatographic analysis of genistein and daidzein from soybean (Glycine max) seeds. J. Agric. Food Chem. 47: 3455-3456 (1999) https://doi.org/10.1021/jf990157e
  11. Liggins J, Bluck LJC, Runswick S, Atkinson C, Coward WA, Bingham SA. Daidzein and genistein contents of vegetables. Br. J. Nutr. 84: 717-725 (2000)
  12. Nurmi T, Mazur W, Heinonen S, Kokkonen J, Adlercreutz H. Isoflavone content of the soy based supplements. J. Pharm. Biomed. Anal. 28: 1-11 (2002) https://doi.org/10.1016/S0731-7085(01)00612-4
  13. Satterfield M, Black DM, Brodbelt JS. Detection of the isoflavone aglycones genistein and daidzein in urine using solid-phase microextraction-high-performance liquid chromatography-electro spray ionization mass spectrometry. J. Chromatogr. B. 759: 33-41 (2001) https://doi.org/10.1016/S0378-4347(01)00200-6
  14. Bala S, Uniyal GC. High-performance liquid chromatographic analysis of genistein and other isoflavonoids from heartwood of Ougeinia dalbergioides. Phytochem. Anal. 13: 211-214 (2002) https://doi.org/10.1002/pca.646
  15. Mitani K, Narimatsu S. Kataoka H. Determination of daidzein and genistein in soybean foods by automated on-line in-tube solid-phase microextraction coupled to high-performance liquid chromatography. J. Chromatogr. A. 986: 169-177 (2003) https://doi.org/10.1016/S0021-9673(02)02014-9
  16. Williamson AP, Wahala K, Williamson G. Identification and quantification of polyphenol phytoestrogens in foods and human biological fluids. J. Chromatogr. B. 777: 93-109 (2002) https://doi.org/10.1016/S1570-0232(02)00095-8
  17. Kao TH, Chen BH. An improved method for determination of isoflavones in soybean powder by liquid chromatography. Chromatographia 56: 423-430 (2002) https://doi.org/10.1007/BF02492004
  18. Coward L, Smith M, Kirk M, Barnes S. Chemical modification of isoflavones in soy foods during cooking and processing. Am. J. Clin. Nutr. 68: 1486S-1491S (1998)
  19. Barnes S, Kirk M, Coward L. Isoflavones and their conjugates in soy foods: extraction conditions and analysis by HPLC mass spectrometry. J. Agric. Food Chem. 42: 2466-2474 (1994) https://doi.org/10.1021/jf00047a019
  20. Simmonne AH, Smith M, Weaver DB, Vail T, Barnes S, Wei CI. Retention and changes of soy isoflavones and carotenoids in immature soybean seeds (Edarnmame) during processing. J. Agric. Food Chem. 48: 6061-6069 (2000)
  21. Coward L, Smith M, Kirk M, Barnes S. Chemical modification of isoflavones in soy foods during cooking and processing. Am. J. Clin. Nutr. 68: 1486S-1491S (1998)
  22. Mahungu SM, Diaz-Mercado S, Li J, Schwenk M, Singletary K, Faller J. Stability of isoflavones during extrusion processing of com! soy mixture. J. Agric. Food. Chem. 47: 279-284 (1999) https://doi.org/10.1021/jf980441q
  23. Setchell KDR, Cassidy A. Dietary isoflavones: biological effects and relevance for human health. J. Nutr. 129: 758S-767S (1999) https://doi.org/10.1093/jn/129.3.758S
  24. Fang, NB, Yu SG, Badger TM. Characterization of isoflavones and their conjugates in female rat urine using LCIMSIMS. J. Agric. Food. Chem. 50: 2700-2707 (2002) https://doi.org/10.1021/jf011384v
  25. Murphy PA, Baruna K, Hank CC. Solvent extraction selection in the determination of isoflavones in soy foods. J. Chromatogr. B. 777: 129-138 (2002) https://doi.org/10.1016/S1570-0232(02)00342-2
  26. Wang HJ, Murphy PA. Isoflavone contents in commercial soybean foods. J. Agric. Food Chem. 42: 1666-1673 (1994) https://doi.org/10.1021/jf00044a016
  27. Wang HJ, Murphy PA. Isoflavone composition of American and Japanese soybeans in Iowa: Effects of variety, crop year, and location. J. Agric. Food Chem. 42: 1674-1677 (1994) https://doi.org/10.1021/jf00044a017
  28. WU Q, Wang M, Sciarappa WJ, Simon JE. LC/UV/ESI-MS analysis of isoflavones in Edarnarne and tofu soybeans. J. Agric. Food Chem. 52: 2763-2769 (2004) https://doi.org/10.1021/jf035053p
  29. Dixon RA, Ferreira D. Genistein. Phytochemistry 60: 205-211 (2002) https://doi.org/10.1016/S0031-9422(02)00116-4
  30. Krenn L, Unterrieder I, Ruprechter R. Quantification of isoflavones in red clover by high-performance liquid chromatography. J. Chromatogr. B. 777: 123-128 (2002) https://doi.org/10.1016/S1570-0232(02)00079-X