Securityguard ultra cartridges

The UHPLC-HR-MS system was composed of a Vanquish Flex Binary pump LC and a Q Exactive Plus MS equipped with an electrospray ionization (ESI) source, the Orbitrap-based FT-MS system (Thermo Fischer Scientific Inc., Dreieich, Germany). The chromatography was performed on a Kinetex Biphenyl column (100 × 2.1 mm, 2.6 µm particle size) composed of SecurityGuardTM Ultra Cartridges (Phenomenex, Bologna, Italy). Elution was carried out with formic acid in H₂O 0.1% v/v (solvent A) and formic acid in acetonitrile 0.1% v/v (solvent B) with a linear solvent gradient (5 to 60% B within 16 min). Samples (5 µL) were injected into the LC system at a flow rate of 0.5 mL/min, maintaining autosampler and column oven temperatures at 4 and 35 °C, respectively. MS parameters were applied as previously reported [8 (link)].

Aromatic waters were diluted in MeOH (1:1, v/v), centrifuged (4000 rpm) and injected (5 μL injection volume) into the LC system composed by a Vanquish Flex Binary UHPLC coupled with a Vanquish DAD and a Q Exactive Plus mass spectrometer, Orbitrap-based FT-MS system, equipped by an ESI source (Thermo Fischer Scientific Inc., Bremem, Germany). Elution was performed on a Kinetex^® Biphenyl column (100 × 2.1 mm, 2.6 μm) provided of a SecurityGuard^TM Ultra Cartridges (Phenomenex, Bologna, Italy), using formic acid in MeOH 0.1% v/v (solvent A) and formic acid in H₂O 0.1% v/v (solvent B) as eluent and developing a linear solvent gradient of increasing 5 to 55% A within 15 min, at a flow rate 0.3 mL/min. During analysis, autosampler and column oven temperatures were maintained at 4 and 35 °C, respectively. UV data were registered using 254, 280 and 325 nm as preferential channels. A positive ion mode was used for ESI interface in a scan range of m/z 150–1200 and spectra were acquired both in full (70,000 resolution, 220 ms maximum injection time) and data dependent-MS/MS scan (17,500 resolution, 60 ms maximum injection time). The following ionization parameters were used: spray voltage 3500 V, capillary temperature 300 °C, sheath gas (N₂) 20 arbitrary unit, auxiliary gas (N₂) 3 arbitrary unit, collisionally activated dissociation (HCD) 18 eV. Data were elaborated with Xcalibur software.

A magnetic stirrer with a temperature controller from Selecta (Barcelona, Spain) was used for some separation phase steps. Centrifugation of the extracts was carried out by a Coulter Avanti J-25 centrifuge with a temperature controller (Beckman, Fullerton, USA). A rotatory evaporator Mod. LABOROTA 4000 from Heidolph (Schwabach, Germany) was used for the organic solvent evaporation. The 20 × 20 cm silica gel TLC plates with inorganic fluorescent indicator F254 from Merck Millipore (Billerica, MA, USA) was used for separation steps. Nylon filters with a pore size of 20 μm and an inner diameter of 13 mm from Millipore (Billerica, MA, USA) were used to remove solid particles from the extracts before the LC analysis. A 15 Gold LC System from Beckman Coulter (Fullerton, USA) equipped with a 26 Gold DAD detector (wavelength range 190–600 nm) was used for individual separation and UV detection. The instrumental setup was controlled by the Karat 3.0.7 software, which also enabled data acquisition and processing. Chromatographic separation was carried out using a Kinetex® EVO C18 column (150 mm, 4.6 mm id, 2.6 μm particle size) from Phenomenex Inc. (Torrance, CA, USA), furnished with a 4.6 mm SecurityGuard™ ULTRA cartridges.