Rapid Screening and Identification of Daidzein Metabolites in Rats Based on UHPLC-LTQ-Orbitrap Mass Spectrometry Coupled with Data-Mining Technologies.

Wenjing Zhao,Jiayu Zhang,Moran Huang,Zhanpeng Shang,Wenbin He,Qinqing Li,Zhibin Wang

doi:10.3390/molecules23010151

Abstract

Daidzein, the main bioactive soy isoflavone in Nature, has been found to possess many biological functions. It has been investigated in particular as a phytoestrogen owing to the similarity of its structure with that of the human hormone estrogen. Due to the lack of comprehensive studies on daidzein metabolism, further research is still required to clarify its in vivo metabolic fate and intermediate processes. In this study, an efficient strategy was established using UHPLC-LTQ-Orbitrap mass spectrometry to profile the metabolism of daidzein in rats. Meanwhile, multiple data-mining methods including high-resolution extracted ion chromatogram (HREIC), multiple mass defect filtering (MMDF), neutral loss fragment (NLF), and diagnostic product ion (DPI) were utilized to investigate daidzein metabolites from the HR-ESI-MS1 to ESI-MSn stage in both positive and negative ion modes. Consequently, 59 metabolites, including prototype compounds, were positively or tentatively elucidated based on reference standards, accurate mass measurements, mass fragmentation behaviors, chromatographic retention times, and corresponding calculated ClogP values. As a result, dehydration, hydrogenation, methylation, dimethylation, glucuronidation, glucosylation, sulfonation, ring-cleavage, and their composite reactions were ascertained to interpret its in vivo biotransformation. Overall, our results not only revealed the potential pharmacodynamics forms of daidzein, but also aid in establishing a practical strategy for rapid screening and identifying metabolites of natural compounds.

Highlights

Daidzein (40,7-dihydroxyisoflavone), one of the most prominent soy isoflavones, is largely restricted to leguminous plants, such as Trifolium pretense L., Medicago sativa L. and Pueraria lobataOhwi [1,2]
The preponderance of liquid chromatography coupled with high resolution mass spectrometry (LC-HR-MS), such as LTQ-Orbitrap MS, FT-ICR-MS, etc., in the structural characterization of known and unknown metabolites due to the properties of high speed, efficiency, selectivity and detection sensitivity has been fully proved by disparate research teams [11,12,13]
A novel and integrated strategy (Figure 1) was established for profiling the in vivo daidzein metabolism based on UHPLC-LTQ-Orbitrap MS coupled with multiple post-acquisition data-mining methods

Summary

Introduction

Daidzein (40 ,7-dihydroxyisoflavone), one of the most prominent soy isoflavones, is largely restricted to leguminous plants, such as Trifolium pretense L., Medicago sativa L. and Pueraria lobataOhwi [1,2]. The preponderance of liquid chromatography coupled with high resolution mass spectrometry (LC-HR-MS), such as LTQ-Orbitrap MS, FT-ICR-MS, etc., in the structural characterization of known and unknown metabolites due to the properties of high speed, efficiency, selectivity and detection sensitivity has been fully proved by disparate research teams [11,12,13]. It can provide precise elemental composition from accurate mass measurement, which is tremendously helpful to identify of major-to-minor metabolites in vitro and in vivo [14]. Efficient data-mining methods including isotope pattern filtering (IPF) [19], diagnostic product ion (DPI) [15,20], neutral loss filtering (NLF) [21], extracted ion chromatogram (EIC) [22], mass defect filter (MDF), and multiple mass defect filters (MMDFs) [23] have been successful applied to systematically profile the in vivo drug metabolism of drugs

Methods

Results

Conclusion