Identification and quantification of the main isoflavones and other phytochemicals in soy based nutraceutical products by liquid chromatography–orbitrap high resolution mass spectrometry

• Use of Orbitrap analyzer during the determination of phytochemicals in soy products.
• Development of an in-house database containing 60 phytochemicals.
• Adequate separation of some isomeric compounds included in the database.
• Determination of the main isoflavones and other phytochemicals in soy nutraceuticals.
• Comparison of the obtained results with the labelled data.

The specific phytochemicals composition of soy nutritional supplements is usually not labelled. Hence, 12 dietary supplements were analyzed in order to detect and identify the main phytochemicals present in these samples, using a database containing 60 compounds. Ultra-high performance liquid chromatography coupled to single-stage Orbitrap high resolution mass spectrometry (UHPLC–Orbitrap–MS) has been used. Two consecutive extractions, using as extraction solvent a mixture of methanol:water (80:20, v/v), were employed, followed by two dilutions (10 or 100 times depending on the concentration of the components in the sample) with a mixture of an aqueous solution of ammonium acetate 30 mM:methanol (50:50, v/v). The method was validated, obtaining adequate recovery and precision values. Limits of detection (LODs) and quantification (LOQs) were calculated, ranging from 2 to 150 μg L−1. Isoflavones were the predominant components present in the analyzed supplements with values higher than 93% of the total amount of phytochemicals in all cases. The aglycones (genistein, daidzein, glycitein and biochanin A) as well as their three conjugated forms, β-glucosides (genistin, daizin and glycitin) were detected and quantified, being daidzein the isoflavone detected at higher concentration in 8 out of 12 samples reported, with values ranging from 684 to 35,970 mg kg−1, whereas biochanin A was detected at very low concentrations, ranging from 18 to 50 mg kg−1. Moreover, other phytochemicals as flavones, flavonols, flavanones and phenolic acids were also detected and quantified.