Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA027767

Local Sample ID:	POS_Lip_S_38_LSRH-L6
Subject ID:	SU000550
Subject Type:	Cells
Subject Species:	Homo sapiens
Taxonomy ID:	9606
Species Group:	Human

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Sample Preparation:

Sampleprep ID:	SP000557
Sampleprep Summary:	Cell pellets were resuspended in volume of ice cold Lipid Extractions Solvent (50 µg/mL) based on biomass. Contents were transferred to new, pre-labeled 2.0 mL LoBind tube with 10-15 ceramic beads, and homogenized using 2 pulses at 2,000 rpm for 30 sec on a MagNA Lyser. Volume of HPLC-grade water (with 0.02 mg/mL Tryptophan-d5 in water) was added based on biomass. Samples were allowed to sit at room temperature for 10 minutes, then centrifuged at 16,000 rcf for 10 min at 10˚C. For lipidomics, transferred the maximum clean volume of the lower lipid-rich DCM layer to a new, pre-labeled LoBind tube and a 375 µL aliquot was then transferred to a new 1.5 mL, LoBind tube for analysis. An additional 125 µL aliquot of each study sample was transferred to a 7.0 mL glass vial to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (375 µL each), and the remaining was used for 1 Equilibrium sample (column conditioning). Samples were frozen at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in ACN/IPA/H2O (65:30:5 v/v/v), vortexed at 4,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 225 µL of the supernatant was transferred to pre-labeled autosampler vials and 10 µL was injected into OrbiTrap Velos.

Sampleprep ID:

SP000557

Sampleprep Summary:

Cell pellets were resuspended in volume of ice cold Lipid Extractions Solvent (50 µg/mL) based on biomass. Contents were transferred to new, pre-labeled 2.0 mL LoBind tube with 10-15 ceramic beads, and homogenized using 2 pulses at 2,000 rpm for 30 sec on a MagNA Lyser. Volume of HPLC-grade water (with 0.02 mg/mL Tryptophan-d5 in water) was added based on biomass. Samples were allowed to sit at room temperature for 10 minutes, then centrifuged at 16,000 rcf for 10 min at 10˚C. For lipidomics, transferred the maximum clean volume of the lower lipid-rich DCM layer to a new, pre-labeled LoBind tube and a 375 µL aliquot was then transferred to a new 1.5 mL, LoBind tube for analysis. An additional 125 µL aliquot of each study sample was transferred to a 7.0 mL glass vial to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (375 µL each), and the remaining was used for 1 Equilibrium sample (column conditioning). Samples were frozen at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in ACN/IPA/H2O (65:30:5 v/v/v), vortexed at 4,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 225 µL of the supernatant was transferred to pre-labeled autosampler vials and 10 µL was injected into OrbiTrap Velos.