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MB Sample ID: SA317511
Local Sample ID: | Con_14 |
Subject ID: | SU003036 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Age Or Age Range: | infant |
Gender: | Not applicable |
Human Race: | Pacific asian |
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Subject:
Subject ID: | SU003036 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Age Or Age Range: | infant |
Gender: | Not applicable |
Human Race: | Pacific asian |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
Con_14 | SA317511 | FL037816 | Con | Factor |
Collection:
Collection ID: | CO003029 |
Collection Summary: | The study population consisted of 158 six-month-old infants (46 healthy infants, 30 only AD group, and 82 with combined AD and FA) involved in the Cohort for Childhood Origin of Asthma and Allergic Diseases (COCOA) (reference: BMC Pulm Med 2014, 14:109.) |
Sample Type: | Feces |
Treatment:
Treatment ID: | TR003045 |
Treatment Summary: | no treatment |
Sample Preparation:
Sampleprep ID: | SP003042 |
Sampleprep Summary: | ~10 mg of the freeze-dried feces was homogenized well with an internal standard solution (200 μl of 1 μM C17 S1P solution). Then, 300 μl of chloroform, 200 μl of methanol, 50 μl of H2O, and 10 μl of 10N NaOH were added to each solution, and vortex well. The aqueous layer was collected after centrifugation, where S1P was separated from other lipids under alkaline condition. 50 uL of H2O was added to the remaining solution and the aqueous layer was collected again after centrifugation. Then, S1P was re-extracted into the chloroform phase under acidic conditions using 400 μl of chloroform and 20 μl of 10M HCl. This step was repeated three times. The chloroform was evaporated, and the dried samples were reconstituted with methanol prior to LC-MS/MS analysis. |
Combined analysis:
Analysis ID | AN004793 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 |
Column | Phenomenex Jupiter C4 (50 × 1.0 mm, 5um) |
MS Type | ESI |
MS instrument type | Orbitrap |
MS instrument name | Thermo LTQ XL |
Ion Mode | NEGATIVE |
Units | pmol/mg |
Chromatography:
Chromatography ID: | CH003624 |
Chromatography Summary: | Ultimate 3000-gradient |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Phenomenex Jupiter C4 (50 × 1.0 mm, 5um) |
Column Temperature: | 35 |
Flow Gradient: | 10% B for 0 min, 10–90% B for 5 min, 90% B for 10 min, 90–10% B for 0.1 min, and 10% B for 4.9 min |
Flow Rate: | 300 μL/min |
Solvent A: | 0.1% formic acid in H2O |
Solvent B: | 0.1% formic acid in MeOH |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS004539 |
Analysis ID: | AN004793 |
Instrument Name: | Thermo LTQ XL |
Instrument Type: | Orbitrap |
MS Type: | ESI |
MS Comments: | Selected ion monitoring (SRM) was uesd in the negative ion mode and the extracted ion chromatogram corresponding to the specific transition (phosphate ion, PO3-, m/z 78.9585) for S1P was used for quantification. Calibration range was 0.01 – 10 μM with R2 > 0.99. Data analysis was performed by using Xcalibur 2.2 software. Ultimate3000 |
Ion Mode: | NEGATIVE |