In vitro production of radiolabeled red clover (Trifolium pratense) isoflavones

Nancy J. Engelmann, Adam Reppert, Gad Yousef, Randy B. Rogers, Mary Ann Lila

Research output: Contribution to journalArticlepeer-review


Red clover isoflavones are increasingly used in dietary supplements for their purported estrogenic effects. However, little is known about their metabolism in animals due to a lack of commercially available isotopically labeled tracers. The goal of this research was to establish red clover cell culturing methodology for 14C-biolabeling of isoflavones. When root, leaf, and petiole-derived suspension cultures were grown in darkness or light, dark-grown, petiole-derived solution cultures produced the highest concentrations of the two major red clover isoflavones, formononetin (0.67 mg/g FM inoculum) and biochanin A (0.13 mg/g FM inoculum). Varying levels and timing of copper chloride elicitor did not significantly affect isoflavone accumulation. Approximately 38% of the 14C-sucrose dose accumulated in the cells. Eighteen percent of the initial labeled dose was detected in the isoflavone-rich methanolic extract and of that, 22% accumulated in isoflavones.

Original languageEnglish (US)
Pages (from-to)147-156
Number of pages10
JournalPlant Cell, Tissue and Organ Culture
Issue number2
StatePublished - Aug 2009


  • Copper chloride
  • Isoflavones
  • Metabolic tracers
  • Plant cell culture
  • Radiolabeling
  • Red clover
  • Trifolium pratense

ASJC Scopus subject areas

  • Horticulture


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