Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA128459

Local Sample ID:	S19
Subject ID:	SU001599
Subject Type:	Mammal
Subject Species:	Mus musculus
Taxonomy ID:	10090
Genotype Strain:	C57BL/6
Age Or Age Range:	8 weeks
Gender:	Male and female
Animal Animal Supplier:	Jackson Labs (Bar Harbor, ME USA)

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Sample Preparation:

Sampleprep ID:	SP001607
Sampleprep Summary:	Blood lipids were isolated for lipidomic analysis according to the Bligh and Dyer method (Bligh and Dyer 1959). The lipidomics was performed at the University of Georgia (Athens, GA). Briefly, blood samples designated for lipidomics were suspended in 1.25 ml of methanol and 1.25 ml of chloroform. Tubes were vortexed for 30 s, allowed to sit for 10 min on ice, centrifuged (300 x g; 5 min), and the bottom chloroform layer was transferred to a new test tube. The extraction steps were repeated three times and the chloroform layer combined. A commercial mix of SPLASH Lipidomix internal standards (Avanti Polar Lipids, Inc.) were spiked into each sample. SPLASH Lipidomix Mass Spec standards includes all major lipid classes at ratios similar to that found in human plasma. The collected chloroform layers were dried under nitrogen, reconstituted with 50 µl of methanol: chloroform (3:1 v/v), and stored at 80ºC until analysis. Lipid content was quantified by determining the level of inorganic phosphorus using the Bartlett Assay (Bartlett 1959).

Sampleprep ID:

SP001607

Sampleprep Summary:

Blood lipids were isolated for lipidomic analysis according to the Bligh and Dyer method (Bligh and Dyer 1959). The lipidomics was performed at the University of Georgia (Athens, GA). Briefly, blood samples designated for lipidomics were suspended in 1.25 ml of methanol and 1.25 ml of chloroform. Tubes were vortexed for 30 s, allowed to sit for 10 min on ice, centrifuged (300 x g; 5 min), and the bottom chloroform layer was transferred to a new test tube. The extraction steps were repeated three times and the chloroform layer combined. A commercial mix of SPLASH Lipidomix internal standards (Avanti Polar Lipids, Inc.) were spiked into each sample. SPLASH Lipidomix Mass Spec standards includes all major lipid classes at ratios similar to that found in human plasma. The collected chloroform layers were dried under nitrogen, reconstituted with 50 µl of methanol: chloroform (3:1 v/v), and stored at 80ºC until analysis. Lipid content was quantified by determining the level of inorganic phosphorus using the Bartlett Assay (Bartlett 1959).