GNPS - Supplemental Data For: Lipid Annotator: Towards Accurate Annotation in Non-Targeted LC-HRMS/MS Lipidomics Using A Rapid and User-Friendly Software

This data was used in the manuscript:
"Lipid Annotator: Towards Accurate Annotation in Non-Targeted LC-HRMS/MS Lipidomics Using A Rapid and User-Friendly Software"

Aliquots (40 uL for (+ mode) and 120 uL for (- mode)) of thawed plasma (NIST SRM 1950 Metabolites in Frozen Plasma, Sigma, St. Louis USA) were each extracted using a modified Folch extraction procedure and reconstituted in 100 uL of a methanol/chloroform mixture (9:1, v/v). LC separation was performed on an Agilent 1290 Infinity II LC System, with a 19 min gradient time on a reverse phase C18 column (Agilent InfinityLab Poroshell 120 EC-C18, 3.0 x 100 mm, 2.7 um). Mobile phase consisted of 10 mM ammonium acetate and 0.2 mM ammonium fluoride in 9:1 water/methanol, while mobile phase B consisted of 10 mM ammonium acetate and 0.2 mM ammonium fluoride in 2:3:5 acetonitrile/methanol/isopropanol. Negative and positive polarity data was acquired on the Agilent 6546 LC/Q-TOF using iterative MS/MS acquisition mode on 6 injections of extracted plasma for each polarity. Detailed experimental methods for chromatography and mass spectrometry can be found in in the Agilent application note 5994-0775en. Two methods were used, a high-load and a low-load method, to determine the effect of high injection volumes / concentration on the number of annotations using the Agilent 6546 LC/Q-TOF.