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MB Sample ID: SA242785
| Local Sample ID: | Heavy_Pool-P03 |
| Subject ID: | SU002516 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002516 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Heavy_Pool-P03 | SA242785 | FL030397 | Heavy Dansylation | Treatment |
Collection:
| Collection ID: | CO002509 |
| Collection Summary: | Human plasma was collected and pooled by BioIVT, purchased as lot# HMN654203 and stored at -80 C. |
| Sample Type: | Blood (plasma) |
| Storage Conditions: | -80℃ |
| Collection Vials: | K2EDTA |
Treatment:
| Treatment ID: | TR002528 |
| Treatment Summary: | Human subject treatment is not part of this experiment. |
Sample Preparation:
| Sampleprep ID: | SP002522 |
| Sampleprep Summary: | Protein was precipitated from pooled human plasma with ice cold methanol and centrifugation. Pooled human plasma was split for metabolite tagging with either light or heavy (2 carbon-13) dansyl chloride. Tagged aliquots were recombined at known mixing ratios to generate samples for LC-MS data acquisition and subsequent testing of isotopic labeling analysis software. |
| Sampleprep Protocol Filename: | DansylatedHumanPlasma.pdf |
Combined analysis:
| Analysis ID | AN003950 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Vanquish |
| Column | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE |
| Units | unitless L/H ratio |
Chromatography:
| Chromatography ID: | CH002924 |
| Chromatography Summary: | A Thermo Scientific Vanquish Horizon UHPLC System coupled with a Thermo Scientific Q Exactive HF Orbitrap was used to conduct LC-(+)FTMS analysis. A Waters ACQUITY UPLC BEH C18 (1.7 μm, 2.1 x 100 mm) column was used along with the following mobile phases: (A) 0.1% formic acid in water and (B) 0.1% formic acid in acetonitrile. The gradient was 25% B (0 min), 25% B (1.2 min), 99% B (11.2 min), 99% B (16.2 min), 25% B (16.3 min), and 25% B (19 min) to achieve column equilibration. The column was maintained at 40 ºC with flowrate of 400 μL/min. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC BEH C18 (100 x 2.1mm,1.7um) |
| Column Temperature: | 40 |
| Flow Gradient: | 25% B (0 min), 25% B (1.2 min), 99% B (11.2 min), 99% B (16.2 min), 25% B (16.3 min), 25% B (19 min) |
| Flow Rate: | 400 uL/min |
| Solvent A: | 100% water, 0.1% formic acid |
| Solvent B: | 100% acetonitrile, 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003685 |
| Analysis ID: | AN003950 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | A Thermo Scientific Vanquish Horizon UHPLC System coupled with a Thermo Scientific Q Exactive HF Orbitrap was used to conduct LC-(+)FTMS analysis. Ionization for FTMS analysis utilized a HESI source in the positive mode with spray voltage 3 kV, capillary temperature 300 ºC, sheath gas flowrate 35 au, aux gas flowrate 10 au, spare gas flowrate 2 au, max spray current 100 V, probe heater temperature 300 ºC, and S-lens RF level 50 V. FTMS scanning was full MS1 in the range 250 to 1000 m/z with maximum IT 200 ms, AGC target 5x105, and mass resolution 60,000. MS-DIAL v4.9.2 was used to perform peak alignment and annotation. The novel MS-DIAL addon Peak Pair Pruner was used to perform peak pair validation and quantitation with light-to-heavy ratio reporting and corrections for background peaks and isotopic overlap between the light and heavy tags. |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 300 C |
| Ion Spray Voltage: | 3000 V |
| Analysis Protocol File: | DansylatedHumanPlasma.pdf |
