Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA151181

Local Sample ID:	16P_016
Subject ID:	SU001717
Subject Type:	Human
Subject Species:	Homo sapiens
Taxonomy ID:	9606

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Sample Preparation:

Sampleprep ID:	SP001723
Sampleprep Summary:	Extraction of plasma lipids is based on the “Maytash'' method [1] which was subsequently modified. Extraction is carried out using a bi-phasic solvent system of cold methanol, methyl tert-butyl ether (MTBE), and water. In more detail, cold methanol (225 µL) containing a mixture of odd chain and deuterated lipid internal standards [lysoPE(17:1), lysoPC(17:0), PC(12:0/13:0), PE(17:0/17:0), PG(17:0/17:0), sphingosine (d17:1), d7-cholesterol, SM(17:0), C17 ceramide, d3-palmitic acid, MG(17:0/0:0/0:0), DG(18:1/2:0/0:0), DG(12:0/12:0/0:0), and d5-TG(17:0/17:1/17:0)] is added to a 20 µL sample aliquot, which is placed into a 1.5 mL Eppendorf tube, and the tube is vortexed for 10 s. Then, 750 µL of cold MTBE containing CE(22:1) (internal standard) are added, followed by vortexing for 10 s. and shaking for 6 min. at 4ºC. Phase separation is induced by adding 188 µL of mass spec-grade water. After vortexing for 20 s. The sample is centrifuged at 14,000 rpm for 2 min. The upper organic phase is collected in two 300 µL aliquots. One is stored at -20ºC as a backup and the other is evaporated to dryness in a SpeedVac. Dried extracts are resuspended using a mixture of methanol/toluene (9:1, v/v) (60 µL) containing an internal standard [12-[[(cyclohexylamino)carbonyl]amino]-dodecanoic acid (CUDA)] used as a quality control.

Sampleprep ID:

SP001723

Sampleprep Summary:

Extraction of plasma lipids is based on the “Maytash'' method [1] which was subsequently modified. Extraction is carried out using a bi-phasic solvent system of cold methanol, methyl tert-butyl ether (MTBE), and water. In more detail, cold methanol (225 µL) containing a mixture of odd chain and deuterated lipid internal standards [lysoPE(17:1), lysoPC(17:0), PC(12:0/13:0), PE(17:0/17:0), PG(17:0/17:0), sphingosine (d17:1), d7-cholesterol, SM(17:0), C17 ceramide, d3-palmitic acid, MG(17:0/0:0/0:0), DG(18:1/2:0/0:0), DG(12:0/12:0/0:0), and d5-TG(17:0/17:1/17:0)] is added to a 20 µL sample aliquot, which is placed into a 1.5 mL Eppendorf tube, and the tube is vortexed for 10 s. Then, 750 µL of cold MTBE containing CE(22:1) (internal standard) are added, followed by vortexing for 10 s. and shaking for 6 min. at 4ºC. Phase separation is induced by adding 188 µL of mass spec-grade water. After vortexing for 20 s. The sample is centrifuged at 14,000 rpm for 2 min. The upper organic phase is collected in two 300 µL aliquots. One is stored at -20ºC as a backup and the other is evaporated to dryness in a SpeedVac. Dried extracts are resuspended using a mixture of methanol/toluene (9:1, v/v) (60 µL) containing an internal standard [12-[[(cyclohexylamino)carbonyl]amino]-dodecanoic acid (CUDA)] used as a quality control.