Editor's Note
The authors of this study compared the chemical dynamics between red clover traditional preparations of infusions, decoctions, and tinctures, as well as to identify the chemical variability over time in a traditional tincture. For this purpose, eight isoflavone aglycones as well as two  glucosides , ononin and sissotrin, were used as marker compounds. Infusions, decoctions and tinctures were shown to produce different chemical profiles and dynamic chemical variability of the red clover tincture was observed over time (one-month). The outcomes show that tinctures, taken over a period of time, are dynamic medicinal formulations that allow for time-controlled release of bioactive compounds.
Abstract

Ethnopharmacological relevance
The flowering tops of  Trifolium pratense  L., popularly known as red clover, are used in ethnic Western and  Traditional Chinese medicine , in a variety of preparations, including infusions, decoctions and  tinctures . Red clover has been reported to be helpful for treatment of menopausal symptoms,  premenstrual syndrome mastalgia , high cholesterol, and other conditions.

Aims of the study
The aims were to compare the chemical dynamics between traditional preparations of infusions, decoctions, and tinctures, as well as to identify the chemical variability over time in a traditional red clover tincture. For this purpose, eight  isoflavone   aglycones  as well as two  glucosides , ononin and sissotrin, were used as marker compounds.

Materials and methods
Quantitative NMR (qHNMR), LC-MS-MS, and UHPLC-UV methods were used to identify and quantitate the major  phenolic compounds  found within each extract.

Results
Infusions, decoctions and tinctures were shown to produce different chemical profiles.  Biochanin A  and  formononetin  were identified and quantified in infusion, decoction, and tinctures of red clover. Both infusion and decoction showed higher concentrations of  isoflavonoid  glucosides, such as ononin and sissotrin, than 45% ethanolic tinctures. Dynamic chemical variability (“dynamic residual complexity”) of the red clover tincture was observed over time (one-month), with biochanin A and formononetin reaching peak concentrations at around six days.

Conclusions
Insight was gained into why different formulation methods (infusions, decoctions, and tinctures) are traditionally used to treat different health conditions. Moreover, the outcomes show that tinctures, taken over a period of time, are dynamic medicinal formulations that allow for time-controlled release of bioactive compounds.

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