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MB Sample ID: SA271277
| Local Sample ID: | pARDS_49_FFA |
| Subject ID: | SU002807 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002807 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| pARDS_49_FFA | SA271277 | FL035201 | pARDS | Factor |
Collection:
| Collection ID: | CO002800 |
| Collection Summary: | This study retrospectively analyzed the samples of the ASAN sepsis registry obtained from March 2011 to February 2018, along with the non-ARDS controls. This study was approved by the Asan Institutional Review Board (IRB No. 2019-1017). |
| Sample Type: | Blood (serum) |
Treatment:
| Treatment ID: | TR002816 |
| Treatment Summary: | Human serum was stored at deep freezer until analysis. For metabolome analysis, human serum was prepared to extract metabolites using conventional liquid-liquid exrtaction method known as BD method. |
Sample Preparation:
| Sampleprep ID: | SP002813 |
| Sampleprep Summary: | Sample solutions were prepared by using commonly used liquid-liquid extaction procedure known as Bligh/Dyer mthod with minor modifications.(Can.J.Biochem.Physiol. 37:911-917) Briefly, 400 μL of chloroform/methanol (1/2, v/v) was added to each sample solution and mixed well. The solution was centrifuged for 15 min at 14000 rpm. After centrifugation, a thick precipitate containing macromolecules was found between the aqueous upper layer and the organic lower layer. Polar metabolites were contained in the upper aqueous phase. The collected volume from each layer were generally the same, however sometimes any specific sample had thicker interface between organic and aqueous layer, which resulted in little variation of the recovered volumes. However, internal standards added prior to sample preparation should correct this variation. The aqueous phases were dried under vacuum and stored at -20℃ until further analysis. The dried matter from the aqueous solutions was reconstituted with 50 μL of H2O/MeOH (50/50 v/v) prior to liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis. For amino acids, 10 μL out of the total 50 μL reconstituted aqueous phase was used for chemical derivatization of amino acids using phenylisothiocyanate.For S1P and free fatty acids, separate serum samples were used. Details can be found in attached method file. |
Combined analysis:
| Analysis ID | AN004379 | AN004380 | AN004381 | AN004382 |
|---|---|---|---|---|
| Analysis type | MS | MS | MS | MS |
| Chromatography type | Reversed phase | Reversed phase | Reversed phase | GC |
| Chromatography system | Agilent 1290 | Agilent 1290 | Thermo Dionex Ultimate 3000 | Agilent 7890A |
| Column | Phenomenex Synergi Fusion-RP (100 x 4.6mm,4um) | Agilent ZORBAX Eclipse Plus C18 (50 x 2.1mm,1.8um) | Phenomenex Jupiter C18 (150 x 2mm,5um) | Agilent HP5-MS (30m x 0.25mm, 0.25 um) |
| MS Type | ESI | ESI | ESI | EI |
| MS instrument type | QTRAP | QTRAP | Orbitrap | Single quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap | Thermo LTQ XL | Agilent 5975C |
| Ion Mode | NEGATIVE | POSITIVE | NEGATIVE | POSITIVE |
| Units | nM | uM | ug/uL | area ratio |
Chromatography:
| Chromatography ID: | CH003284 |
| Chromatography Summary: | Metabolites related to energy metabolism |
| Instrument Name: | Agilent 1290 |
| Column Name: | Phenomenex Synergi Fusion-RP (100 x 4.6mm,4um) |
| Column Temperature: | 23 |
| Flow Gradient: | Hold at 0% B for 5 min, 0% to 90% B for 2 min, hold at 90% for 8 min, 90% to 0% B for 1 min, then hold at 0% B for 9 min |
| Flow Rate: | 70 μL/min except for 140 μL/min during minutes 7–15 |
| Solvent A: | 100% water; 5mM ammonium acetate |
| Solvent B: | 100% methanol; 5 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003285 |
| Chromatography Summary: | Amino acids |
| Instrument Name: | Agilent 1290 |
| Column Name: | Agilent ZORBAX Eclipse Plus C18 (50 x 2.1mm,1.8um) |
| Column Temperature: | 50 |
| Flow Gradient: | Hold at 0% B for 0.5 min, 0–95% B for 5 min, 95% B for 1 min, and 95–0% B for 0.5 min, then hold at 0% B for 2.5 min |
| Flow Rate: | 500 μL/min |
| Solvent A: | 100% water; 0.2% formic acid |
| Solvent B: | 100% acetonitrile; 0.2% formic acid |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003286 |
| Chromatography Summary: | S1P |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Phenomenex Jupiter C18 (150 x 2mm,5um) |
| Column Temperature: | 35°C |
| 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: | 100% water; 0.1% formic acid |
| Solvent B: | 100% methanol; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH003287 |
| Chromatography Summary: | Free fatty acids (FFA) |
| Instrument Name: | Agilent 7890A |
| Column Name: | Agilent HP5-MS (30m x 0.25mm, 0.25 um) |
| Column Temperature: | - |
| Flow Gradient: | - |
| Flow Rate: | - |
| Solvent A: | - |
| Solvent B: | - |
| Chromatography Type: | GC |
MS:
| MS ID: | MS004128 |
| Analysis ID: | AN004379 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | QTRAP |
| MS Type: | ESI |
| MS Comments: | Analyst1.5.2 |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004129 |
| Analysis ID: | AN004380 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | QTRAP |
| MS Type: | ESI |
| MS Comments: | Analyst1.5.2 |
| Ion Mode: | POSITIVE |
| MS ID: | MS004130 |
| Analysis ID: | AN004381 |
| Instrument Name: | Thermo LTQ XL |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | Xcalibur, SRM |
| Ion Mode: | NEGATIVE |
| MS ID: | MS004131 |
| Analysis ID: | AN004382 |
| Instrument Name: | Agilent 5975C |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | Chemstatation, SRM |
| Ion Mode: | POSITIVE |
