HPLC Analysis of Isoflavones on Ascentis^® Phenyl

column

Ascentis Phenyl, 15 cm × 4.6 mm I.D., 5 μm particles (581616-U)

mobile phase

[A] 10 mM ammonium formate (pH 3.0 with concentrated formic acid), [B] methanol

flow rate

1 mL/min split to the MS

column temp.

35 °C

detector

MS, ESI(+) in selected ion recording (SIR) mode

injection

10 μL

sample

glycitein, genistein at 1 μg/mL, equol at 50 μg/mL and daidzin, glycitin, genistin at 10 μg/mL in 60:40 10 mM ammonium formate (pH 3.0 with concentrated formic acid):methanol