6540 agilent ultra high definition accurate mass q tof ms

An Agilent 1200-LC system (Agilent Technologies, Palo Alto, CA, USA) equipped with a vacuum degasser, autosampler, and binary pump was used for the chromatographic determination. The column used consisted of a Gemini^® NX-C18 (3 µM, 150 × 2 mm ID) (Phenomenex). The mobile phases were made of water (A) and acetonitrile (B), both with 0.1% of formic acid. The gradient program was as follows: 0–6 min, 50% B; 7–12 min, 100% B; 13–20 min, 50% B. The injection volume was set at 5 µL and the flow rate at 0.2 mL/min. Mass spectrometry (MS) analysis was performed employing a 6540 Agilent Ultra-High-Definition Accurate-Mass q-TOF-MS coupled to the HPLC, equipped with an Agilent Dual Jet Stream electrospray ionization (Dual AJS ESI) interface in positive and negative ionization modes under the following conditions: interface in positive and negative ionization modes; drying gas flow (N2) 12.0 L min⁻¹; nebulizer pressure, 50 psi; gas drying temperature, 370 °C; capillary voltage, 3500 V; fragmentor voltage, 160 V. Analysis was carried out in MS mode and MS spectra were collected within the scan range of 50–1500 m/z. Integration and data acquisition were performed employing the Mass Hunter Workstation software.

Power extracts from the extraction of 0.5 g of samples were redissolved in 3 mL of methanol/water (80:20 v/v). After their filtration, extracts were analyzed by HPLC-DAD-q-TOF-MS (Agilent 1200 series coupled to 6540 Agilent Ultra-High-Definition Accurate-Mass q-TOF-MS, Agilent Technologies, Palo Alto, CA, USA) according to the method proposed by Gómez-Caravaca et al. [13 (link)].
Identification was performed based on relative retention times, UV-Vis spectra, and mass spectra obtained by q-TOF-MS and from the literature. Quantification was carried out using calibration standards curves of gallic acid, coumaric acid, ferulic acid, vanillic acid, catechin, quercetin, ellagic acid, and mangiferin (Sigma Aldrich, St. Louis, MO, USA). Spectral and chromatographic data are compliled in Supplementary Table S1. Integration and data processing were performed using Mass Hunter Workstation Software, Qualitative Analysis, version B.07.00 (Agilent Technologies, Inc. 2014).