Optimization of a rapid microwave assisted extraction method for the liquid chromatography-electrospray-tandem mass spectrometry determination of isoflavonoid aglycones in soybeans

Dipartimento di Chimica Generale ed Inorganica, Chimica Analitica, Chimica Fisica, Università degli Studi di Parma, Parco Area delle Scienze 17/A, 43100 Parma, Italy.
Journal of Chromatography A (Impact Factor: 4.17). 06/2007; 1152(1-2):274-9. DOI: 10.1016/j.chroma.2007.03.112
Source: PubMed


A very fast chromatographic separation of isoflavonoids genistein, daidzein, formononetin and biochanin A was developed on a C18 high-speed column under isocratic conditions. The method was validated in terms of detection limits, quantitation limits (LOQs), linearity and precision. LOQs in 0.04-0.2 microg/g range were calculated, making feasible the determination of these compounds of nutritional concern at trace levels. Good linearity was demonstrated over three concentration orders of magnitude for each analyte (r2 0.990-1.000). The intra-day and inter-day repeatability was evaluated in terms of relative standard deviation (RSD%) at two concentration levels for each analyte (RSD% <9%). An optimization strategy was adopted to find the best conditions for the extraction of isoflavonoid aglycones from yellow soybeans using microwave-assisted extraction. The most relevant parameters resulted to be the microwave power, the extraction time and the acid concentration, optimal values being 600 W, 1 min and 12 M, respectively. When performing sample treatment on a fortified soybean sample, high recovery percentage was obtained for both compounds (94+/-8% for daidzein and 97+/-5% (n = 4) for genistein). The concentration level at which daidzein and genistein were found in the soybean sample were 1.21+/-0.15 mg/g and 2.38+/-0.09 mg/g (n=4), respectively.

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    • "), Daidzein 6 (Careri et al., 2007) Formononetin 7 (Kim et al., 2007) Maackiain 8 (Liu et al., 2005) and Medicarpin 9 (Tsiri, Chinou, Halabalaki, Haralampidis, & Ganis-Spyropoulos, 2009) and five compounds from peas such as Afrormosin 10 (Carlsen et al., 2008; Gong, Wang, Chen, Liu, & Yu, 2009), Anhydropisatin 11 (Mizuno, Tanaka, Katsuragawa, Saito, & Iinuma, 1990;), Pisatin 12 (Mizuno et al., 1990;), Pseudobaptigenin 13 (Wu, Wang, & Simon, 2003) have been identified by LC-MS/MS/MS analysis. These compounds belong to the classes of flavanones, Isoflavones, pterocarpans and their glyco conjugates (Figure 1). "
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