Return to study ST000405 main page
MB Sample ID: SA019463
Local Sample ID: | Pool_AllNMR_4 |
Subject ID: | SU000426 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | female |
Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000426 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | female |
Species Group: | Human |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Pool_AllNMR_4 | SA019463 | FL004743 | PoolAll | Group |
Pool_AllNMR_4 | SA019463 | FL004743 | - | Vaginal Bleeding |
Collection:
Collection ID: | CO000420 |
Collection Summary: | Serum samples collected in the second trimester from pregnant women |
Sample Type: | Blood |
Blood Serum Or Plasma: | serum |
Treatment:
Treatment ID: | TR000440 |
Treatment Summary: | no treatment |
Sample Preparation:
Sampleprep ID: | SP000433 |
Sampleprep Summary: | Sample Preparation Prior to Biocrates p180 Kit Plate Analysis: Serum samples were thawed on ice and pooled QC samples were prepared. Samples (study and pooled QC samples) were then diluted, vortexed, and transferred into properly labeled 2 mL Lo-Bind eppendorf tubes and dried on a lyophilizer overnight. The residue was reconstituted with 30 µL of PBS buffer. Samples were then vortexed and then centrifuged at room temperature and at 16,000 rcf for 4.0 minutes. All study samples and pooled QC samples were splitted into two AbsoluteIDQ p180 Kit-96 wells plates (Plate 1502 and 1503). Biocrates Plate Preparation: The Biocrates p180 kit-96 wells plates were prepared following the AbsoluteIDQ p180 Kit metabolomics procedure. Briefly, an internal standard mix was added to 95 of the 96 wells. Next, zero samples, QC standards and calibration standards were added to their corresponding wells. The study samples and pooled QC Samples were then added to the appropriate wells and dried for 30 minutes under nitrogen flow. The plate was derivatized using a 5% Phenylisothiocyanate (PITC) solution in (1:1:1) Ethanol:Pyridine:Water and, then, incubated for 20 minutes followed by a drying step under nitrogen flow. An extraction solvent (5 mM ammonium acetate in methanol) was added to all wells. The plate was then vortexed and filtrated by centrifugation. After centrifugation, 150 µL was removed and transferred to a second 96-well plate (LC-MS/MS plate). This second plate was diluted with 150 µL of HPLC grade water for a subsequent LC-MS/MS assay (analysis of amino acids and biogenic amines). All the wells in the original plate was diluted with 400 µL of flow injection analysis (FIA) Running Solvent for a FIA-MS analysis (lipids, acylcarnitines, and hexose analysis). |
Combined analysis:
Analysis ID | AN000645 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 1100 |
Column | Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um) |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | ABI Sciex API 4000 QTrap |
Ion Mode | UNSPECIFIED |
Units | area |
Chromatography:
Chromatography ID: | CH000469 |
Chromatography Summary: | Biocrates P180 analysis combines a RP for positive mode MS , a flow injection for both pos and neg mode MS |
Instrument Name: | Agilent 1100 |
Column Name: | Agilent Eclipse XDB-C18 (100 x 3.0mm,3.5um) |
Column Pressure: | 30-120 bar for RP and 4-50 bar for FIA |
Column Temperature: | 50 c fro RP and RT for FIA |
Flow Rate: | 0.5 ml/min for RP and 0.2 ml/min for FIA |
Sample Injection: | 10 ul |
Solvent A: | 100% water; 0.2% formic acid |
Solvent B: | 100% acetonitrile; 0.2% formic acid for RP; 100% methanol with Biocrates FIa Solvent |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS000577 |
Analysis ID: | AN000645 |
Instrument Name: | ABI Sciex API 4000 QTrap |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
MS Comments: | Biocrates combines a pos and negative mode for analysis |
Ion Mode: | UNSPECIFIED |
Analysis Protocol File: | KIT2-LC-5404 |