The experimental design approach to the optimization of a simple quick easy cheap effective rugged and safe method for the analysis of phytoestrogens in complex soy‐based food

Abstract

The present paper deals with the multivariate optimization of an extractionpurification strategy for the determination of phytoestrogens (daidzein, genistein, coumestrol, formononetin, and biochanin A) in soy-based meat substitutes by high performance liquid chromatography with tandem mass spectrometry. For a reliable quantitation of these new concerning compounds in such a complex matrix, recovery and matrix effect must be carefully evaluated. Therefore, two sequential experimental designs were used to optimize the sample-pretreatment of soy-based burgers:the chosen technique was the quick, easy, cheap, effective, rugged and safe methodology, which does not require any particular facility or instrumentation. Thanks to the first screening design (Plackett-Burman), the significant factors influencing the studied responses were identified and further investigated through a response surface design (Box-Behnken). The optimal values of the variables (volume of extraction solvent mix/sample mass ratio and two clean-up sorbents) led to quantitative recoveries (97-104%) and low ion suppression (matrix effect 60-93%) for all analytes. This optimized method was characterized by low detection limits (0.2-1.5 ng/g) and excellent intraday precision (RSD 2-4%). It was applied to the determination of the considered compounds in several soy-burgers from the Italian market, detecting low ng/g levels (up to 40 ng/g) of coumestrol, formononetin, and biochanin A, and high concentrations (7.9-78 µg/g) of genistein and daidzein.

Homoisoflavonoids profiling of Ophiopogon japonicus by off‐line coupling high‐speed countercurrent chromatography with high‐performance liquid chromatography–diode array detector‒quadrupole time‐of‐flight tandem mass spectrometry

“…It was reported that about 50 homoisoflavoniods were purified and identified by repeated column fractionations [1]. HPLC with diode array detector (DAD) and MS/MS has been developed for rapid isolation and characterization of bioactive components from complex natural products [8–11]. However, information about comprehensive analysis of homoisoflavonoids, especially for minors, in O. japonicus by HPLC–DAD–MS/MS is restricted because of the existence of multicomponents, restriction of detection sensitivity, and coelution of minor and abundant compounds [12,13].…”