Α novel method for the efficient simultaneous quantification of 67 phytoestrogens in plants and foodstuffs

Abstract

The significant beneficial effects of phytoestrogens on human and animal health have emerged the necessity to develop a sensitive, accurate and rapid method for their efficient quantitation in human and animal foods. Study herein concerns the development of a novel analytical method using high-performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) that allows the simultaneous quantitation of 67 natural phytoestrogens. The novel method was fully validated through the determination of the following parameters: selectivity, sensibility, recovery, precision, accuracy of detection and quantification limits for each analyte. Specifically, the novel method proved accurate displaying linear response with R2 values ≥ 0.9991, quantitation limits from 5.5 to 331.8 ng/mL for 98.5% of analytes, recovery values ranging between 64.8% and 150.3%, while the precision and accuracy scores remained below 20% for most analytes. The method was successfully applied on the following plant samples used in human and/or animal diet belonging to Fabaceae plant family: Trifolium repens, Vicia sativa, Pisum sativum and Glycine max (soybean meal). Furthermore, the novel method was applied for the determination of the phytoestrogenic content of two animal feeds (for livestock and fishes) and two foodstuffs (chicken meat and fish fillet).