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Analytical Method Validation and Determination of Genistein, Daidzein and Glycitein in Soybean Milk by RP-HPLC Method
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| Author:
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FLORENTINUS DIKA OCTA RISWANTO, ABDUL ROHMAN, SUWIDJIYO PRAMONO, SUDIBYO MARTONO
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| Abstract:
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Soybean milk or soya milk is a popular soy beverage widely consumed in Asia. The growing interest for studying content of isoflavone aglycones in soybean products was related to the biological activities of soy products towards human health. Hence, it was important to develop a validated analytical method to quantify content of aglycone isoflavones from soybean milk sample. In this study, a reversed-phase HPLC (RP-HPLC) method was validated and applied for determining content of genistein, daidzein, and glycitein in soybean milk. The developed RP-HPLC method was met the requirement of selectivity and sensitivity. Calibration curve formulas of genistein, daidzein, and glycitein were y = 99996x-9713.6 (r=0.9996), y = 71095x+8039.1 (r=0.9994), and y = 37116x –45855 (r=0.9994) in the range of 4.9136-29.4816, 5.0405-30.2430, and 4.2812-25.6872 µg/mL, respectively. Accuracy and intermediate precision were assessed on both standard solution and soybean milk samples spiked with standard solution of genistein, daidzein, and glycitein. It was found that both standard solution and the spiked samples solution were met the requirement of recovery of 85-110% and RSD of less than 4% for intraday and interday analysis. Content of genistein, daidzein, and glycitein in soybean milk sample (n=6) were 4.542?0.124, 6.813?0.224, and 2.038?0.070 µg/mL, respectively. It can be concluded that the RP-HPLC method was successfully validated and applied in the determination of genistein, daidzein, and glycitein in soybean milk.
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| Keyword:
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aglycones, isoflavones, RP-HPLC, soybean milk, validation
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| EOI:
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| DOI:
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https://doi.org/10.31838/ijpr/2021.13.01.343
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| Download:
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