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K. D. Setchell, N. M. Brown and E. Lydeking-Olsen, “The Clinical Importance of the Metabolite Equol-A Clue to the Effectiveness of Soy and Its Isoflavones,” Journal of Nutrition, Vol. 132, No. 12, 2002, pp. 3577-3584.

has been cited by the following article:

  • TITLE: The Development of S-Equol Diastereoisomer Specific ELISA

    AUTHORS: Takayuki Minekawa, Akira Kambegawa, Kumiko Shindome, Shizuka Takehara, Hidetoshi Arakawa

    KEYWORDS: S-Equol; Diastereoisomer; Immunoassay; ELISA; Isoflavone; Deconjugation

    JOURNAL NAME: American Journal of Analytical Chemistry, Vol.3 No.6, June 21, 2012

    ABSTRACT: Equol is a metabolite of soybean isoflavone, daidzein, and many health benefits are expected. Endogenous equol in urine is S-equol and mostly exists as glucuronate or sulfate conjugate. In this study we preliminary established the simple enzyme-linked immunosorbent assay (ELISA) without deconjugation, then developed the S-equol specific ELISA involves deconjugation showing high stereospecificity to S-equol without using stereospecific antibody. For the simple ELISA, we used a polyclonal antibody that targets the regions not influenced by inhibition by conjugation of glucuronate and sulfate and achieved the correlation coefficient; r = 0.975, but the value was 30 % lower than high performance liquid chromatography (HPLC). Developing upon this we invented the specific ELISA established from S-equol homogeneous combination for the standard and enzyme-labeled antigen to enhance stereospecificity. The correlation with HPLC was favorable: r = 0.986, y = 0.996x – 6. Compared to the previous method using (R,S)-equol combination, cross-reactivity with R-equol was reduced from 65 to 13 %, and that with daidzein from 0.31% to 0.08%, markedly increased in the specificity. This study is expected to be applied for both simple clinical researches, and stereospecific immunoassays in which specific antibody preparation is difficult.