Abstract: In this study,an efficient high performance liquid chromatography-mass spectrometry (LC-MS) method was established to quantitatively determine the isoflavone and anthocyanin concentrations in black soybean seed.Each sample was injected into Waters Xselect HSS T3 C₁₈ column for isoflavone analysis,the mobile phase consisted of acetonitrile and 0.1% acetic acid in HPLC water.For anthocyanin,X-Terra MS C₁₈ column was used,the mobile phase consisted of 10% formic acid and methanol/acetonitrile/water/formic acid (22.5/22.5/40/10,V/V).Selected ion monitoring (SIM) mode was used in the mass spectral acquisition.Identification of the isoflavones and anthocyanins were achieved by comparing the sample retention times and mass spectra with those of standard compounds.In this study,twelve isoflavones and eight anthocyanins were quantified in absolute terms via linear regression of their corresponding standards.The established analysis method was simple,stable and rapid.It can be used for the identification and quantitative determination of isoflavone and anthocyanin of black soybean seed.

Key words: black soybean, isoflavone, anthocyanin, LC-MS