HPLC-UV Enrichment and LC-MS/MS Analysis

HPLC-UV clean-up and enrichment of the enzyme free samples were performed on a 4.6 mm × 150 mm Atlantis^® dC18 100 Å column (5 μm particle size, Waters, Milford, MA, USA), loaded with a 4.6 mm × 20 mm Guard Column 2 pK (Atlantis^® dC18 5 μm) on a Waters Alliance^® HPLC System (Waters e2695Separations Module, including a Waters 2998 Photodiode Array (PDA) detector as previously described [22 (link)]. The gradient program used an eluent composed by 2 mM ammonium formate, acetonitrile and methanol, while the fractions containing the lesions were collected, freeze-dried, pooled, freeze-dried again, and redissolved in Milli-Q water before been injected for LC-MS/MS analysis [21 (link),67 (link),68 (link)].

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Krokidis M.G., Louka M., Efthimiadou E.K., Zervou S.K., Papadopoulos K., Hiskia A., Ferreri C, & Chatgilialoglu C. (2019). Membrane Lipidome Reorganization and Accumulation of Tissue DNA Lesions in Tumor-Bearing Mice: An Exploratory Study. Cancers, 11(4), 480.

Publication 2019

2998 Photodiode Array (PDA) detector

Acetonitrile Ammonium formate Enzyme Freeze Hplc Methanol Ms ms

Corresponding Organization : Institute of Organic Synthesis and Photoreactivity

Other organizations : National and Kapodistrian University of Athens

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Variable analysis

independent variables

Gradient program used an eluent composed by 2 mM ammonium formate, acetonitrile and methanol

dependent variables

Fractions containing the lesions

control variables

4.6 mm × 150 mm Atlantis® dC18 100 Å column (5 μm particle size, Waters, Milford, MA, USA)
4.6 mm × 20 mm Guard Column 2 pK (Atlantis® dC18 5 μm)
Waters Alliance® HPLC System (Waters e2695Separations Module, including a Waters 2998 Photodiode Array (PDA) detector)

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