Isotopic labeling of isoflavones from plant cell and organ culture for metabolic tracking in animal models

Kudzu and red clover isoflavones are phytoestrogens that have been investigated as alternatives to estrogen replacement for treatment of osteoporosis, cardiovascular disease, and neurological disorders. However, the pharmacokinetics of these compounds are not clearly understood. In the present studies, we utilized 14C-radiolabeled crude isoflavone extracts to track the absorption, distribution, and elimination of kudzu and red clover isoflavones in rats. A dose of approx. 0.148 MBq (4µCi) of radiolabeled extract was found to be sufficient to be metabolically tracked in rats using a scintillation counter. Analysis showed radiolabel accumulated in all major tissues, including bones, and that kudzu isoflavone extracts underwent enterohepatic circulation. We then produced a 14C-radiolabeled kudzu isoflavone-enriched fraction from a high isoflavone yielding kudzu hairy root genotype in vitro, which was used to track the absorption, distribution, and elimination of kudzu isoflavones in rats. Pharmacokinetics showed that kudzu isoflavones were rapidly absorbed, distributed into all tissues including bones, underwent enterohepatic circulation, and was largely eliminated in the urine and feces. Femur bone extracts were analyzed by HPLC-ESI-MS and found to contain puerarin, daidzein, and puerarin glucuronide, which suggests that kudzu isoflavones may be able to have a role in promoting bone health. To improve kudzu isoflavone yields in vitro for further studies, kudzu hairy root cultures were elicited with acetate and salicylic acid in varied concentrations and treatment durations. However, acetate and salicylic acid failed to significantly improve isoflavone synthesis in kudzu hairy roots.

Name thesis:degree_name

M.S.

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Nutritional Sciences

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2010

Author dc:creator

• Mun, Jonathan G.

Contributors dc:contributor

• Lila, Mary Ann

• puerarin
• isoflavone
• kudzu
• hairy roots
• radiolabeling
• metabolic tracking
• pharmacokinetics
• elicitation

Statement dc:rights

• Copyright 2010 Jonathan G. Mun

Language dc:language

en

Handle dc:identifier

http://hdl.handle.net/2142/16006

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/16006