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The following steps were applied in data processing with MZmine 2.53: (i) Mass detection with a noise level of 1000, (ii) Chromatogram builder with a minimum time span of 0.08 min, minimum height of 1000 and a m/z tolerance of 0.006 m/z or 10.0 ppm, (iii) Chromatogram deconvolution using the local minimum search algorithm with a 70% chromatographic threshold, 0.05 min minimum RT range, 5% minimum relative height, 1200 minimum absolute height, a minimum ration of peak top/edge of 1.2 and a peak duration range of 0.085.0, (iv), Isotopic peak grouper with a m/z tolerance of 5.0 ppm, RT tolerance of 0.05 min, maximum charge of 2 and with the most intense isotope set as the representative isotope, (v) Join aligner with a m/z tolerance of 0.009 or 10.0 ppm and a weight for of 2, a RT tolerance of 0.15 min and a weight of 1 and with no requirement of charge state or ID and no comparison of isotope pattern, (vi) Peak list row filter with a minimum of 10% of the samples (vii) Gap filling using the same RT and m/z range gap filler algorithm with an m/z tolerance of 0.009 m/z or 11.0 ppm, (vii) Identification of lipids using a custom database search with an m/z tolerance of 0.008 m/z or 8.0 ppm and a RT tolerance of 0.25 min
doi: 10.1073/pnas.0711558105 297 YuL.WenG.ZhuS.HuX.HuangC.YangY
Once the platform was located the animal was allowed a 30 second rest period before the next trial started
