Health, Dietary, Nutraceuticals, & Supplements
Single Laboratory Validation of UPLC-MS/MS Assay of Red Clover
With their origins in traditional medicine, botanical dietary supplements remain in widespread use. For example, preparations of red clover (Trifolium pratense L.) are currently used to treat respiratory problems (asthma, whooping cough and bronchitis), skin disorders (eczema and psoriasis), and menopausal issues in women, especially vasomotor symptoms. The active isoflavones are concentrated in the aerial parts of red clover. Daidzein and genistein are estrogenic, whereas biochanin A and formononetin can be metabolized to genistein and daidzein, respectively, and are considered proestrogens. In contrast, the isoflavones irilone and prunetin in red clover have progesterone activity.
For safe use and to ensure reproducible effects, red clover dietary supplements should be chemically standardized to the active isoflavones. In this study, a fast UHPLC-MS/MS based method was developed and validated for the quantitative of all 6 isoflavones in red clover dietary supplements using deuterium-labelled internal standards to correct for variations during sample preparation and analysis. Additionally, the UHPLC-MS/MS assay was validated using human serum matrix with the goal of supporting clinical research where serum samples will be measured for the various red clover isoflavones.