Dgu 20as degasser

MCs analyses were performed on a Shimadzu HPLC System with a DGU-20As Degasser, LC-20AD Pump, CTO-20AC Oven, and a SPD-M20A Diode Array Detector (Shimadzu do Brasil, Barueri, SP, Brazil). Chromatographic separation was achieved at 40 °C using a Luna C18 column (Phenomenex; 4.6 × 250 mm × 5 μm). The mobile phase consisted of an isocratic mixture of 20 mmol.L⁻¹ ammonium acetate and Acetonitrile (70:30; v:v). The flow rate was 1.0 mL.min⁻¹ and 20 μL of samples were injected into the HPLC System. MCs were detected at their maximum absorbance of 238 nm. The full chromatographic run was 10 min and the peak corresponding to MCs was observed at 6.36 min. All samples were analyzed in duplicate.

HPLC was conducted on a Shimadzu system consisting of a DGU-20AS degasser, SIL-20A HT autosampler, and a SPD-M20A UV–vis detector set at a wavelength of 230 nm. The system was fitted with an Eclipse XDB C18 column (4.6 × 150 mm, 3.5 μm particle size) and the oven temperature was set to 25 °C. The mobile phase consisted of ammonium acetate buffer (0.02 M, pH 5) and acetonitrile (43:57% v/v) at a flow rate of 1 mL/min, and the injection volume was 20 μL.

The OMW extracts were analysed using a Shimadzu UFLC XR system consisted of an electrospray ionization source (ESI) equipped with two LC-20ADXR solvent delivery units, a SIL-20AXR autosampler, an SCL-10A system controller, a CTO-20 AC column oven, a DGU-20AS degasser (Shimadzu, Kyoto, Japan). A volume of 5 μL each extract was injected at 0.5 mL/min to a Discover BIO Wide Pore C18 column (150 mm × 3 mm, 3 μm) at 40 °C and separated with two mobile phases: A (0.1% formic acid in water v/v) and B (0.1% formic acid in methanol v/v) following the programmed linear gradient elution : 0–14 min, from 10 to 20% B; 14–27 min, from 20 to 55% B; 27–37 min, from 55 to 100% B; 37–45 min, 100% B; and 45–50 min 10% B^{48 (link),49 (link)}. The ionization mode was negative and The ESI conditions were set as follows: capillary voltage of − 3.5 v, a nebulizing gas flow of 1.5 L/min, a dry gas flow rate of 15 L/min, a DL (dissolving line) temperature of 280 °C, a block source temperature of 400 °C, and a voltage detector of 1.35 V. Compounds were identified by comparing their retention time and mass spectra with those of reference standards. The validation of the method was determined as detailed in^{50 (link)}.