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MB Sample ID: SA042726
Local Sample ID: | 36916 |
Subject ID: | SU000807 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Species Group: | Human |
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Sample Preparation:
Sampleprep ID: | SP000814 |
Sampleprep Summary: | For extraction of metabolome, 10 mg of tissue was homogenized in 1:4 ice cold water:methanol mixture containing equimolar mixture of 11 standard compounds [Epibrassinolide, [D3] Testosterone (mass difference from endogenous Testosterone = 3 Da), [15N] Anthranilic acid (mass difference from endogenous Anthranilic acid =1 Da), Zeatine, Jasmonic acid, Gibberelic acid, [D4] Estrone (mass difference from endogenous Estrone =4 Da), [15N]-Tryptophan (mass difference from endogenous Tryptophan =1 Da), [D4] Thymine (mass difference from endogenous Thymine =4 Da), [13C] Creatinine (mass difference from endogenous Creatinine =1 Da) and [15N] Arginine (mass difference from endogenous Arginine =1 Da)]. This was followed by sequential addition of ice cold chloroform and water in 3:1 ratio and separation of the organic (methanol and chloroform) and aqueous solvents (water:methanol:chloroform:water; ratio 1:4:3:1). The aqueous extract was de-proteinized using a 3 KDa molecular filter (Amicon Ultracel -3K Membrane, Millipore Corporation, Billerica, MA) and the filtrate containing metabolites was dried under vacuum (Genevac EZ-2plus, Gardiner, NY). Prior to mass spectrometry, the dried extract was resuspended in identical volume of injection solvent composed of water:methanol (50:50) and subjected to liquid chromatography (LC) mass spectrometry. |