A Rapid, Simple, and High Throughput UPLC-MS/MS Method for Determination of isoflavones in soybean.

• Main Authors: Yu-Hsun Chen, 陳昱勲
• Other Authors: Chien-Chen Lai
• Format: Others
• Language: zh-TW
• Published: 2019
• Online Access: http://ndltd.ncl.edu.tw/handle/6td2af

碩士 === 國立中興大學 === 分子生物學研究所 === 107 === Soybean reportedly contain high quantity of isoflavones. The goal of this study was to develop an effective, simple and rapid ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for the analysis of twelve isoflavones in soybean. A rapid and effective homogenization method is applied for the isoflavone isomers extraction. The entire extraction time is less than 8 minutes, and the extraction efficiency is 60% higher than the traditional method. A complete separation of all compounds was achieved within 5 min using a reverse-phase CORTECS® T3 column (50 x 2.1 mm, 1.6 μm). The quantitative analysis was performed with UHPLC-MS/MS via multiple reaction monitoring (MRM) in polarity switching mode. Method validation was performed in terms of linearity, the relative standard deviations (RSDs) of the intra-day and inter-day, and it was found that linearity was good (r2 ＞ 0.99), precisions were within 15 % and the accuracy ranged from 84.6% to 117.5 %. The recovery was more than 40 %. The limit of detection (LOD) and limit of quantification(LOQ) values of the twelve analytes varied between 0.375 to 1.5 ng/mL and 1.5 to 6.25 ng/mL, respectively. The matrix effect are ranged from -8.5% to +13.72%. The concentration of isoflavines of different varieties of soybean ranged from 1207.28 to 21654.36 mg/kg. Overall, this robust and high-throughput method has a short run time (extraction plus analysis ＜ 13 min).