Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA251340

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Subject:

Factors:

Local Sample ID	MB Sample ID	Factor Level ID	Level Value	Factor Name
KO3n_14	SA251340	FL032015	Knock-out	Genotype
KO3n_14	SA251340	FL032015	Neg	Treatment

Collection:

Collection ID:	CO002593
Collection Summary:	Samples were collected and aliquots representing the same EV counts were dried under nitrogen, and resuspended in 2:2:1 acetonitrile:methanol:H2O (v/v) (for polar metabolites, eg. fatty acids, phospholipids) or in 100% isopropanol (for less-polar metabolites, eg. ceramides). The samples were mixed on an orbital shaker (360 rpm) for 1 min at room temperature before the LC/MS analysis.
Sample Type:	Cultured cells

Treatment:

Treatment ID:	TR002612
Treatment Summary:	Myotube Treatment with hMEC Generated FA-sEV: hMECs grown to confluence were serum-starved and treated with OA:BSA (100µM:50µM) or BSA (50µM controls). The sEVs were isolated from media collected over 48h and particle number and protein content determined.

Sample Preparation:

Sampleprep ID:	SP002606
Sampleprep Summary:	Samples were collected and aliquots representing the same EV counts were dried under nitrogen, and resuspended in 2:2:1 acetonitrile:methanol:H2O (v/v) (for polar metabolites, eg. fatty acids, phospholipids) or in 100% isopropanol (for less-polar metabolites, eg. ceramides). The samples were mixed on an orbital shaker (360 rpm) for 1 min at room temperature before the LC/MS analysis.

Combined analysis:

Chromatography:

Chromatography ID:	CH003047
Methods Filename:	LC_Method.pdf
Instrument Name:	Agilent 1290 Infinity II
Column Name:	Waters CORTECS UPLC C18 (150 x 2.1mm,1.6um)
Column Temperature:	60
Flow Gradient:	0-2 min, 30% B; 17 min, 75% B; 20 min, 85% B; 23-26 min, 100% B; 26 min, 30% B followed by a re-equilibration phase of 5 min.
Flow Rate:	.25ml/min
Solvent A:	A: 60% acetonitrile, 40% water, 0.1% formic acid, 10 mM ammonium formate, 2.5 mM medronic acid
Solvent B:	90% 2-propanol, 10% acetonitrile, 0.1% formic acid, 10 mM ammonium formate
Chromatography Type:	Reversed phase

Chromatography ID:	CH003048
Methods Filename:	LC_Method.pdf
Instrument Name:	Thermo Vanquish
Column Name:	Waters CORTECS HILIC (100 x 2.1mm,1.6um)
Column Temperature:	45
Flow Gradient:	0 to 1 min, 90% B; 12 min, 35% B; 12.5 to 14.5 min, 25% B; 15 min, 90% B followed by a re-equilibration phase of 4
Flow Rate:	.25ml/min
Solvent A:	A: 95% water, 5% acetonitrile, 20 mM ammonium bicarbonate, 0.1% ammonium hydroxide solution (25% ammonia in water), 2.5 μM medronic acid
Solvent B:	B: 95% acetonitrile, 5% water, 2.5 μM medronic acid
Chromatography Type:	HILIC

MS: