Hplc lc msd ultra trap system xct 6330

Analysis of extracts was performed on an HPLC-LC/MSD Ultra Trap System XCT 6330 (Agilent Technologies) equipped with a Luna Omega Polar C18 (5 µm, 150 × 2.1 mm; Phenomenex) column. Elution was performed with 0.1% formic acid (solvent A) and acetonitrile containing 0.06% formic acid (solvent B) at a flow rate of 0.4 ml/min using the following gradient: 0 to 100% solvent B over 20 min followed by an isocratic step of 100% B for 3 min. UV spectra were recorded between 230 and 600 nm by diode array detector. Mass spectrometry was performed using positive electrospray ionization (electrospray voltage 3.5 kV, heated capillary temperature 350° C, acquired mass range from 100 to 2200 m/z).

The methanolic fractions obtained from the size exclusion chromatography were analyzed by a HPLC-LC/MSD Ultra Trap System XCT 6330 (Agilent Technologies). For the HPLC measurements, a C18 Nucleosil 100 column (100 x 2 mm ID, 3 μm) and a pre-column (100 x 2 mm ID) (Dr. Maisch) were used. Solvent A (0.1% formic acid) and solvent B (0.06% formic acid in acetonitrile) were used as mobile phase applying the following gradient (t₀ = 10% B, t₁₅ = t₁₇ = 100% B, post-time 5 min. 10% B; flow rate 400 μl/min; injection volume 2.5 μl). UV signals were detected at: 220 nm (10 nm band width), 260 nm (20 nm), 280 nm (20 nm), 360 nm (20 nm), and 435 nm (40 nm). Data analysis was accomplished with the Agilent LC/MSD software ChemStation Rev. B.01.03, Agilent. The following parameters were used for the MS detection: ionisation: ESI (alternating positive and negative ionization) in ultra-scan mode; capillary voltage: 3.5 kV target mass: m/z 1500. Data analyses were performed with the software 6300 Series Trap Control Version 6.1.