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MB Sample ID: SA059660
Local Sample ID: | 136 |
Subject ID: | SU001017 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Age Or Age Range: | >30 |
Gender: | Male and female |
Human Inclusion Criteria: | Patients from the Hospital das Clínicas Outpatient Clinics ≥30 years-old and with at least two measurements of creatinine (with a minimum interval of 3 months) ≥1.6 mg/dL for men and ≥1.4 mg/dL for women were considered potential candidates. |
Human Exclusion Criteria: | Patients attending oncology, psychiatry, urology, HIV/AIDS, viral hepatitis and glomerulonephritis services were excluded. The remaining candidates were then contacted by phone and invited to participate if no exclusion criteria were met (hospitalization or acute myocardial infarction in the last 6 months, autoimmune diseases, pregnancy, psychiatric diseases, ongoing chemo or immunosuppressive therapy, ongoing renal replacement therapy, glomerulonephritis, HIV/AIDS infection, hepatitis C and B, and any previous organ transplantation). |
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Sample Preparation:
Sampleprep ID: | SP001024 |
Sampleprep Summary: | 70uL os serum was thawed on ice and added 300uL of acetronile, isopropanol and water solution (3:3:2 v/v) followed by spike of 5uL of d27 myristic acid (3mg/ml) as internal standard. Samples were vortexed and centrifuged to 15800xg during 15 minutes at 4C. 320uL of supernatante were transferred to a new tube and dried in speedvac during 18 hours. Residues were suspended in 50µL methoxyamine in pyridine solution (40mg/mL), 3µL of FAME (Fatty acid methyl ester ) was added, and the mixture was vortexed for 3 min. This methoximation reaction was performed at room temperature for 16h, followed by trimethylsilylation for 1h adding 100µL MSTFA (N-methyl-N-trimethylsilyltrifluoroacetamine) with 1% TMCS (trimethylchlorosilane) (Sigma-Aldrich). After derivatization, 1µL of this derivative was used for Gas Chromatography Mass Spectrometry (GC/MS) analysis. Storage condition: Desiccator at 4 degree |
Processing Method: | Protein Precipitation followed by metabolite extraction |
Processing Storage Conditions: | On ice |
Extraction Method: | Acetonitrile:Isopropanol: water(3:3:2 v/v) |
Extract Storage: | Described in summary |
Sample Resuspension: | Methoxyamine in pyridine |
Sample Derivatization: | MSTFA + 1% TMCS |
Sample Spiking: | 5uL of d27 Myristic Acid (3mg/mL) and FAME |