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MB Sample ID: SA069393
Local Sample ID: | S5_2 |
Subject ID: | SU001076 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Genotype Strain: | NA |
Age Or Age Range: | NA |
Gender: | Male |
Human Race: | NA |
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Sample Preparation:
Sampleprep ID: | SP001083 |
Sampleprep Summary: | Serum samples (30 µL) were subjected to sequential solvent extraction once each with 1 mL of acetonitrile: isopropanol: water (3:3:2, v/v) ratio and 500 µL of acetonitrile: water (1:1, v/v) ratio mixtures at 4 C.22 Adonitol and d4-succinic acid (both 5 µL from 10 mg/ml stock) were added to each aliquots as two internal standards prior to the extraction. The pooled extracts (~ 1500 µL) from the two steps were dried under vacuum at 4 C prior to chemical derivatization. Dummy extractions performed on blank tubes served as extraction blanks to account for background (extraction solvent, derivatization reagents) noise and other sources of contamination (septa, liner, column, vials, handling etc.). Blanks were only used to see that no carry overs occurred during randomized run orders and to manually filter out contaminating chemicals from the combined list of features. Samples were then sequentially derivatized with methoxyamine hydrochloride (MeOX) and 1% TMCS in N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA) as described elsewhere.23,24 Steps involved addition of 10 μL of MeOX (20 mg/mL) in pyridine incubated under shaking at 55 °C for 60 min followed by trimethylsilylation at 60 °C for 60 min after adding 90 μL MSTFA. |
Sampleprep Protocol ID: | Fiehn et al., 2008 |
Sampleprep Protocol Comments: | NA |
Processing Method: | Fiehn et al., 2008 |
Processing Storage Conditions: | On ice |
Extraction Method: | Fiehn et al., 2008 |
Extract Enrichment: | None |
Extract Cleanup: | None |
Sample Resuspension: | NA |
Sample Derivatization: | Methoxyaminatin + silylation (MSTFA) |
Sample Spiking: | Adonitol |
Subcellular Location: | NA |