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MB Sample ID: SA245560
| Local Sample ID: | MT_20211020_023 |
| Subject ID: | SU002543 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002543 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| MT_20211020_023 | SA245560 | FL030803 | Qstd;H2O(1;64)_rep1 | Factor_Dilutionseries |
Collection:
| Collection ID: | CO002536 |
| Collection Summary: | Vegetable juice and Commercially Pooled Plasma Sample used. |
| Sample Type: | Blood (plasma) |
Treatment:
| Treatment ID: | TR002555 |
| Treatment Summary: | None |
Sample Preparation:
| Sampleprep ID: | SP002549 |
| Sampleprep Summary: | A stepwise dilution (Serial dilution) of vegetable juice and standard pooled human plasma (Qstd) in the opposite direction was used for sample preparation process. Samples were prepared using vegetable juice and Qstd in the ratio of 1024:1, 256:1, 64:1, 16:1, 4:1, 1:1, 1:4, 1:16, 1:64, 1:256 and 1:1024. Along with the 11 serial dilution samples, 100% vegetable juice and 100% plasma were also included as samples. All samples were used in triplicates. Metabolites extraction were carried out by protein precipitation technique using extraction solvent, acetonitrile:methanol (8:1, v/v) containing 0.1% formic acid and isotope labelled Trimethyl-13C3]-caffeine, [13C5]-L-glutamic acid, [15N2]-Uracil, [15N,13C5]-L-methionine, [13C6]-D-glucose and [15N]-L-tyrosine as spike-in controls. 30 ul of Qstd was taken and 60 ul of extraction solvent was added. Extraction blanks were also prepared to remove features of non-biological origins. All samples were vortexed and incubated with shaking at 1000 rpm for 10 min at 4 ºC followed by centrifugation at 4 ºC for 15 min at 15,000 rpm. The supernatant was transferred into mass spec vials and 2 ul injected into UHPLC-MS. |
Combined analysis:
| Analysis ID | AN004003 | AN004004 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Thermo Accucore 150 Amide (50 x 2.1mm, 2.6um) | Thermo Hypersil GOLD (50 x 2.1mm, 3um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Orbitrap ID-X tribrid | Thermo Orbitrap ID-X tribrid |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | m/z values | m/z values |
Chromatography:
| Chromatography ID: | CH002956 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Thermo Accucore 150 Amide (50 x 2.1mm, 2.6um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0.55 ml/min: 0-0.1 min: 0% B, 0.10-5.0 min: 98% B, and 5 min for cleaning and equilibration of column. |
| Flow Rate: | 0.55 ml/min |
| Solvent A: | 95% acetonitrile/5% water; 0.1% acetic acid; 10 mM ammonium acetate |
| Solvent B: | 50% acetonitrile/50% water; 0.1% acetic acid; 10 mM ammonium acetate |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002957 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Thermo Hypersil GOLD (50 x 2.1mm, 3um) |
| Column Temperature: | 45 |
| Flow Gradient: | 0-0.1 min: 0% B, 0.10-1.9 min: 60% B, 1.9-5.0 min: 98% B, and 5 min cleaning and column equilibration |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | 10 mM ammonium acetate in acetonitrile:water (95:5, v/v) with 0.1% acetic acid |
| Solvent B: | 10 mM ammonium acetate in acetonitrile:water (95:5, v/v) with 0.1% acetic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003751 |
| Analysis ID: | AN004003 |
| Instrument Name: | Thermo Orbitrap ID-X tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | data processing using asari v1.9.2 |
| Ion Mode: | POSITIVE |
| MS ID: | MS003752 |
| Analysis ID: | AN004004 |
| Instrument Name: | Thermo Orbitrap ID-X tribrid |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | data processing using asari v1.9.2 |
| Ion Mode: | NEGATIVE |
