HPLC-MS/MS was used to quantify the AHLs produced by Pb1692 in the presence or absence of phloretin. The extraction of AHLs was performed as described previously, with minor modification (Ravn et al., 2001). Pb1692 was adjusted to approximately 5 × 106 CFU/mL in LB supplemented with 0.2 or 0.4 mM of phloretin and cultured for 24 h, 28°C, at 150 rpm. After incubation, each culture was centrifuged at 8,000 g for 15 min, 4°C and the supernatant was transferred to a sterile tube. AHLs were extracted from the supernatant with ethyl acetate and 0.1% formic acid (v/v). The extraction process was repeated twice. Extracts were dried using SpeedVac vacuum concentrator (Savant SPD 111V, Thermo Scientific, MA, United States). The extract was resuspended in 20 μL of 99.5% acetonitrile (Alfa Aesar, United States) and analyzed by LC-MS/MS.

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