Return to study ST001181 main page
MB Sample ID: SA082011
Local Sample ID: | 22.07_subj |
Subject ID: | SU001247 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Age Or Age Range: | 16-44 years old |
Gender: | Female |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001247 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Age Or Age Range: | 16-44 years old |
Gender: | Female |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
22.07_subj | SA082011 | FL012455 | 4 | Trimester |
Collection:
Collection ID: | CO001241 |
Collection Summary: | Blood serum samples were collected from nonfasting participants in the CHDS Study during preganancy (trimester 1-3) or early postpartum (trimester 4). The CHDS recruited women residing in the Oakland, California area who were members of the Kaiser Foundation Health Plan and received obstetric care for pregnancies between 1959 and 1967. All serum samples were stored at -20°C at the repository and shipped frozen to the laboratory for further storage at -80°C. |
Sample Type: | Blood (serum) |
Treatment:
Treatment ID: | TR001262 |
Treatment Summary: | Samples were received frozen in aliquouts of <250uL. Prior to analysis, samples were thawed and prepared for HRM analysis using the standard protocols described in the Sample Preparation section. |
Sample Preparation:
Sampleprep ID: | SP001255 |
Sampleprep Summary: | Samples were prepared for metabolomics analysis using established methods(Johnson et al. (2010). Analyst; Go et al. (2015). Tox Sci). Prior to analysis, plasma aliquots were removed from storage at -80 degrees C and thawed on ice. Each cryotube was then vortexed briefly to ensure homogeneity, and 50 microliters was transferred to a clean microfuge tube. Immediately after, the plasma was treated with 100 microliters of ice-cold LC-MS grade acetonitrile (Sigma Aldrich) containing 2.5 microliters of internal standard solution with eight stable isotopic chemicals selected to cover a range of chemical properties. Following addition of acetonitrile, urine was equilibrated for 30 min on ice, upon which precipitated proteins were removed by centrifuge (14,000 rpm at 4 degrees C for 10 min). The resulting supernatant (100 microliters) was removed, added to a low volume autosampler vial and maintained at 4 degrees C until analysis (<22 h). |
Sampleprep Protocol ID: | HRM_SP_082016_01 |
Sampleprep Protocol Filename: | HRM_sample_preparation_082016_01.pdf |
Combined analysis:
Analysis ID | AN001959 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 |
Column | Thermo Accucore C18 (100 x 2.1mm,2.6um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo Q Exactive Orbitrap |
Ion Mode | POSITIVE |
Units | Peak area |
Chromatography:
Chromatography ID: | CH001420 |
Chromatography Summary: | Mass spectral data were collected with a 10-minute gradient on a Thermo QExactive coupled with Dionex Ultimate 3000 (Thermo Fisher, San Diego, CA) set to collect data from m/z 85 to 1275 in positive ionization mode at 70,000 resolution. Three technical replicates were run for each sample using dual chromatography measuring both polarities (positive and negative) with a C18 column (Higgins Analytical, 100x2.1mm, 5uL). The right pump controls the gradient with Solvent A as 100% water, Solvent B as 100% Acetontrile, and Solvent C as 98% water with 2% Formic Acid. The left pump is set up to wash (Acetonitrile with 2% Formic Acid) one of the columns while the other column is running the gradient. |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Thermo Accucore C18 (100 x 2.1mm,2.6um) |
Solvent A: | 100% water(A), 100% acetontrile(B), 100% water; 2% formic acid© |
Solvent B: | 100% water(A), 100% acetontrile(B), 100% water; 2% formic acid(C) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001814 |
Analysis ID: | AN001959 |
Instrument Name: | Thermo Q Exactive Orbitrap |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Mass spectral data were collected with a 10-minute gradient on a Thermo QExactive coupled with Dionex Ultimate 3000 (Thermo Fisher, San Diego, CA) set to collect data from m/z 85 to 1275 in positive ionization mode at 70,000 resolution. Three technical replicates were run for each sample using dual chromatography measuring both polarities (positive and negative) with a C18 column (Higgins Analytical, 100x2.1mm, 5uL). The right pump controls the gradient with Solvent A as 100% water, Solvent B as 100% Acetontrile, and Solvent C as 98% water with 2% Formic Acid. The left pump is set up to wash (Acetonitrile with 2% Formic Acid) one of the columns while the other column is running the gradient. The raw data was processed with Xcalibur 3.0.63 |
Ion Mode: | POSITIVE |