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MB Sample ID: SA059727

Local Sample ID:179
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
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