Saponin identification from Korean oat grain was performed using
the UPLC system coupled with quadrupole time-of-flight (QToF) mass
spectrometry (SCIEX X500R, SCIEX Co., MA, USA). Chromatographic conditions
were set up as follows: column, CORTECS UPLC T3, 2.1 × 150 mm,
1.6 μm (Waters Co., Milford, MA, USA); precolumn, CORTECS UPLC
Vanguard T3, 2.1 × 50 mm, 1.6 μm, (Waters Co.); column
temperature, 30 °C; sample injection volume, 1 μL; flow
rate, 0.35 mL/min; mobile phase, 0.1% formic acid in DW (A), 0.1% formic acid in ACN (B). Gradient conditions
used: 0–10 min, 15% B; 10 min, 25% B; 40–45 min, 50% B; 50–60
min, 15% B. Mass spectra were multiscanned in the
range of m/z 100–2000 of
the positive ionization mode through an electrospray ionization (+
ESI) source, with the the following parameters: ion source gas, 50
psi; curtain gas, 30 psi; ion source temperature, 450 °C; declustering
potential (DP), 80 V; collision energy (CE), 15 ± 10 V; spray
voltage, 5500 V. Avenacoside A was externally quantified based on
multiple-reaction monitoring (MRM) mode using UPLC-Triple Q-MS/MS
(SCIEX QTRAP 4500, SCIEX CO.), whereas other saponins were internally
quantified using UPLC-QToF-MS (SCIEX X500R, SCIEX CO.). Optimized
MRM conditions for avenacoside A: precursor to product ion pair, 1063.2
→ 1063.2; DP, 161 V; CE, 13 V.
the UPLC system coupled with quadrupole time-of-flight (QToF) mass
spectrometry (SCIEX X500R, SCIEX Co., MA, USA). Chromatographic conditions
were set up as follows: column, CORTECS UPLC T3, 2.1 × 150 mm,
1.6 μm (Waters Co., Milford, MA, USA); precolumn, CORTECS UPLC
Vanguard T3, 2.1 × 50 mm, 1.6 μm, (Waters Co.); column
temperature, 30 °C; sample injection volume, 1 μL; flow
rate, 0.35 mL/min; mobile phase, 0.1% formic acid in DW (A), 0.1% formic acid in ACN (B). Gradient conditions
used: 0–10 min, 15% B; 10 min, 25% B; 40–45 min, 50% B; 50–60
min, 15% B. Mass spectra were multiscanned in the
range of m/z 100–2000 of
the positive ionization mode through an electrospray ionization (+
ESI) source, with the the following parameters: ion source gas, 50
psi; curtain gas, 30 psi; ion source temperature, 450 °C; declustering
potential (DP), 80 V; collision energy (CE), 15 ± 10 V; spray
voltage, 5500 V. Avenacoside A was externally quantified based on
multiple-reaction monitoring (MRM) mode using UPLC-Triple Q-MS/MS
(SCIEX QTRAP 4500, SCIEX CO.), whereas other saponins were internally
quantified using UPLC-QToF-MS (SCIEX X500R, SCIEX CO.). Optimized
MRM conditions for avenacoside A: precursor to product ion pair, 1063.2
→ 1063.2; DP, 161 V; CE, 13 V.