The qualitative analysis of the PEO by LC-DAD-ESI-MS was performed using an Agilent 1100 series HPLC systems equipped with a vacuum degasser, diode array detector and a triple quadrupole mass spectrometer type MicromassAutospecUltimaPt (Kelso, UK) operating in negative mode. Separation was performed on a reverse-phase Uptisphere C18 (Interchim, Montfluçon, France) (2 mm × 100 mm, 5 μm particle size) using a mobile phase consisting of A (0.1% acetic acid in water) and B (acetonitrile) at a flow rate of 0.25 mL min−1. The gradient system was 0 min (5% B), 65 min (100% B), 67 min (100% B), and 70 min (5% B). The flow rate of the mobile phase was fixed at 0.25 mL min−1. The column temperature was 40 °C and the injection volume was 20 μL. UV spectra were recorded in the range 190–800 nm. The MS conditions were as follow: mass spectra were collected in negative ion mode from 135 and 700 m/z, capillary voltage, 3.2 kV; cone voltage, 40 V; probe temperature, 300 °C; and ion source temperature, 130 °C.
Uptisphere c18
Manufactured by Interchim
Sourced in France
Uptisphere C18 is a reversed-phase high-performance liquid chromatography (HPLC) column. The column features a silica-based stationary phase with chemically bonded C18 alkyl chains, providing a hydrophobic environment for the separation of a wide range of organic compounds.
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2 protocols using uptisphere c18
1
Qualitative Analysis of PEO by LC-DAD-ESI-MS
2
Spectroscopic Analysis of Organic Compounds
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Optical rotations were measured using an MCP-300 polarimeter (Anton Paar, Les Ulis, France). 1D and 2D NMR spectra were recorded on Avance 500 and 600 MHz spectrometers (Bruker, Wissembourg, France). The chemical shifts are relative to the residual signal solvent (CD3OD: δH 3.31; δC 49.20). High-resolution mass spectra were obtained on an LCT Premier XE spectrometer (Waters, Guyancourt, France) in electrospray ionization mode by direct infusion of the purified compounds. Chromatography on silica gel columns, was carried out on silica (Macherey-Nagel 60, 40–63 µm, Hoerdt, France), using a gradient of cyclohexane, ethyl acetate, methylene chloride, and methanol (Sigma-Aldrich, Saint-Quentin Fallavier, France). High performance liquid chromatography was performed on a 1260 Infinity system (Agilent, Les Ulis, France) equipped with a photodiode array detector (G1315C), an evaporative light scattering detector (G4260B), using an analytical reversed phase column Uptisphere C18 (4.6 mm × 250 mm, 5 µm, Interchim, Montluçon, France), and a semi-preparative Uptishere C18 (7.8 × 250 mm, 5 µm).
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