Acrolein and its precursor
3-HPA produced by S. Typhimurium and L. reuteri were quantified by ion-exclusion chromatography
with pulsed-amperometric detection (IC-PAD) as described.1 (link) Immediately after collection of the supernatant,
360 μL was transferred to a 0.7 mL PP Crimp/Snap LC vial (BGB
Analytik) containing 40 μL of freshly prepared 2% hydroquinone
in water for acrolein stabilization.29 (link) All
of the steps were performed over ice to avoid acrolein evaporation.
Analysis was performed on an ICS-5000+ system (Thermo Fisher Scientific).
Analytes were separated with a IonPac ICE-AS1 4 × 250 mm ion-exclusion
column (Thermo Fisher Scientific) with a guard column, operated at
30 °C. The injection volume was 10 μL, and methanesulfonic
acid (100 mM) at a flow rate of 0.2 mL min–1 was
used as eluent for 36 min in isocratic conditions. Solutions of acrolein
(Sigma-Aldrich, ≥95.0%) and chemically synthesized 3-HPA15 (link) were used as external standards. Detection limits
were 7.5 μM for 3-HPA and 4.4 μM for acrolein.
3-HPA produced by S. Typhimurium and L. reuteri were quantified by ion-exclusion chromatography
with pulsed-amperometric detection (IC-PAD) as described.1 (link) Immediately after collection of the supernatant,
360 μL was transferred to a 0.7 mL PP Crimp/Snap LC vial (BGB
Analytik) containing 40 μL of freshly prepared 2% hydroquinone
in water for acrolein stabilization.29 (link) All
of the steps were performed over ice to avoid acrolein evaporation.
Analysis was performed on an ICS-5000+ system (Thermo Fisher Scientific).
Analytes were separated with a IonPac ICE-AS1 4 × 250 mm ion-exclusion
column (Thermo Fisher Scientific) with a guard column, operated at
30 °C. The injection volume was 10 μL, and methanesulfonic
acid (100 mM) at a flow rate of 0.2 mL min–1 was
used as eluent for 36 min in isocratic conditions. Solutions of acrolein
(Sigma-Aldrich, ≥95.0%) and chemically synthesized 3-HPA15 (link) were used as external standards. Detection limits
were 7.5 μM for 3-HPA and 4.4 μM for acrolein.
