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MB Sample ID: SA237204

Local Sample ID:PCa-G12
Subject ID:SU002456
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606

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Combined analysis:

Analysis ID AN003862
Analysis type MS
Chromatography type None (Direct infusion)
Chromatography system none
Column none
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name ABI Sciex 5500 QTrap
Ion Mode POSITIVE
Units Peak Area (cps)

MS:

MS ID:MS003603
Analysis ID:AN003862
Instrument Name:ABI Sciex 5500 QTrap
Instrument Type:Triple quadrupole
MS Type:ESI
MS Comments:NanoESI mass spectrometry of lipid extracts was performed using a hybrid triple quadrupole linear ion trap mass spectrometer (QTRAP® 5500, SCIEX, Framingham, MA, USA) equipped with an automated chip-based nanoelectrospray source (TriVersa Nanomate®, Advion Biosciences, NY, USA). Spray parameters were set at a gas pressure of 0.4 psi and a voltage of 1.2 kV. PC and SM data were acquired in positive ion mode using a precursor ion scan of m/z 184 at a scan rate of 200 Da/s across a mass range of 640 – 850 m/z. Declustering potential was set at 100 V, entrance potential at 10 V, collision energy at 47 V, and collision cell exit potential at 8V (31). Aspiration of 10 μl of sample from each well generated a stable spray time of ≥30 minutes. Lipids were identified from acquired data using Lipidview™ software (v1.2b, SCIEX, Framingham, MA, USA). Processing settings in Lipidview™ were set at a mass tolerance of 0.5 Da, with a minimum intensity of 0.1% and a minimum signal-to-noise ratio of 4. Smoothing and deisotoping of lipid species were enabled. Lipid species were identified from target lists (see attached protocol files), and peak area for each detected lipid species was then exported. These data underwent further processing in R, including background subtraction and relative quantification from internal standards. Lipid nomenclature follows recommendations for the level of molecular detail known (as per Liebisch et al. 2013 J Lipid Res); and in the present study we report lipids as class (e.g., PC or SM) followed by the total number of carbons and carbon-carbon double bonds present within the fatty acids separated by a colon (e.g., a PC with 34 carbons and 1 double bond as PC 34:1). At the level of identification available by the technique used in this study some ambiguity exists between isobaric PC species containing either odd-chain or ether-linked fatty acid species, and in the absence of further structural detail we chose to report such species as ether-linked only where overlap exists.
Ion Mode:POSITIVE
Analysis Protocol File:MWB_Lipidview_target_list.txt
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