Waters 2487 dual absorbance diode array detector

The total SA was analyzed by high-performance liquid chromatography (HPLC) [85 (link)]. The plant tissue (seedling) (0.2–0.3 g) was extracted using 20 mL of dH₂O (90–100 °C) and incubated at 100 °C for 30 min with subsequent cooling. Membrane filters (0.45 µm) (Chromafil Xtra PTFE–45/13, Macherey-Nagel GmbH Co, Duren, Germany) were used to filter the extracts. The analysis was caried out by a Waters Breeze chromatograph (Waters Corporation, Milford, MA, USA) with a Waters 2487 Dual & Absorbance diode array detector at 305 nm. A 250 × 4.6 mm Pursuit C18 5 µm column (Agilent Technologies, Santa Clara, CA, USA) was used. As a mobile phase, 0.5% solution of H₃PO₄: acetonitrile = 65:35 (1.0 mL min⁻¹) was used. A total of 20 µL of the extract was introduced into the chromatography system using an automated sampler Waters 2707 (Waters Corporation, Milford, MA, USA). The software calibration curve was used in calculating the total SA content.

The total endogenous SA was analyzed by high-performance liquid chromatography (HPLC) [66 (link)]. The seedlings (0.2–0.3 g) were extracted using 20 mL of dH₂O (90–100 °C) and incubated at 100 °C for 30 min with subsequent cooling. Membrane filters (0.45 µm) (Chromafil Xtra PTFE–45/13, Macherey-Nagel GmbH Co, Duren, Germany) were used to filter the extracts. The analysis was caried out using a Waters Breeze chromatograph (Waters Corporation, Milford, MA, USA) with a Waters 2487 Dual & Absorbance diode array detector at 305 nm. A Pursuit C18 column (250 × 4.6 mm, 5 µm) (Agilent Technologies, Santa Clara, CA, USA) was used. As a mobile phase, 0.5% H₃PO₄: acetonitrile = 65:35 (1.0 mL min⁻¹) was used. A total of 20 µL of the extract was injected into the chromatography system using a Waters 2707 automated sampler (Waters Corporation, Milford, MA, USA). The software calibration curve was used to calculate the total SA content.