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MB Sample ID: SA166515
| Local Sample ID: | LP22_plasma_B_M |
| Subject ID: | SU001866 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 25(IQR 22-31) |
| Gender: | Male and female |
| Human Nutrition: | Participants were given a banana (~100g) |
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Subject:
| Subject ID: | SU001866 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 25(IQR 22-31) |
| Gender: | Male and female |
| Human Nutrition: | Participants were given a banana (~100g) |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| LP22_plasma_B_M | SA166515 | FL019832 | Pre | Group |
Collection:
| Collection ID: | CO001859 |
| Collection Summary: | Measurements and samples were collected on two days separated by 4 weeks. Samples were drawn at the same time on the two experimental days to control for circadian rhythms. A baseline blood sample (Pre) was collected, the cannula was closed with a screw cap and protected with an elastic bandage. Participants were given a banana (~100g) then completed a 90-minute outdoor run at local time 09:00 to 10:30 am. . Immediately after completing the 90-minute run, a time 0 blood sample was collected (Time 0). After 60 minutes of recovery (1130 am local time), a time 60 blood sample was collected (Time 60). |
| Sample Type: | Blood (plasma) |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001879 |
| Treatment Summary: | Participants were given a banana (~100g) then completed a 90-minute outdoor run at local time 09:00 to 10:30 am, maintaining an effort of 75%-80% of their heart rate. A baseline blood sample (Pre) was collected before the run. Immediately after completing the 90-minute run, a time 0 blood sample was collected (Time 0). After 60 minutes of recovery (1130 am local time), a time 60 blood sample was collected (Time 60). |
Sample Preparation:
| Sampleprep ID: | SP001872 |
| Sampleprep Summary: | Plasma samples (90 µl) were mixed with 5 µl of commercial internal standards and 5 µl of custom-synthesized 13C labeled standards and then incubated for 10 min at room temperature to permit small molecules and vitamins in the internal standards to associate with plasma binding proteins. The metabolites were extracted with 400 µl of cold (-20 °C) methanol: acetonitrile (50:50, v/v), yielding a final concentration of methanol: acetonitrile: water (40:40:20, v/v/v). The samples were then vortexed vigorously, incubated on crushed ice for 10 min, and then centrifuged at 16,000 g for 10 min at 4 °C. The supernatants containing the extracted metabolites and internal standards were transferred to labeled cryotubes and stored at -80 °C prior to LC-MS/MS. |
| Processing Storage Conditions: | On ice |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN002901 | AN002902 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | Reversed phase |
| Chromatography system | Shimadzu 20AD | Shimadzu 20AD |
| Column | Shodex NH2P-40 2E (250 x 2mm,4um) | Raptor Biphenyl (150 × 2.1 mm,2.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole |
| MS instrument name | ABI Sciex 5500 QTrap | ABI Sciex 5500 QTrap |
| Ion Mode | UNSPECIFIED | UNSPECIFIED |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH002151 |
| Chromatography Summary: | Ten µL of the extract was injected into the column through a CTC PAL autosampler. The samples were separated in HILIC mode using a polymer-based NH2 column (250 × 2 mm, 4 µm) (Asahipak NH2P-40 2E, Shodex, Japan). The optimal LC conditions were as follows: Mobile phase A: 95% H2O with 20 mM (NH4)2CO3 and 5% ACN, pH 9.8. Mobile phase B: 100% ACN. The gradient was: 0 - 3.5 min 95% B, 3.6 - 8 min 85% B, 8.1 - 13 min 75% B, 14 - 30 min 0% B, 31 - 41 min 95% B, 41.1 min stop. The flow rate was 200 µL/min, and the column temperature was held at 25 ºC. |
| Instrument Name: | Shimadzu 20AD |
| Column Name: | Shodex NH2P-40 2E (250 x 2mm,4um) |
| Column Pressure: | 900 psi - 2400 psi |
| Column Temperature: | 25 |
| Flow Gradient: | Constant |
| Flow Rate: | 200 µL/min |
| Injection Temperature: | 4 |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium acetate, pH 9.8 |
| Solvent B: | 100% acetonitrile |
| Oven Temperature: | 25 |
| Sample Loop Size: | 10 |
| Sample Syringe Size: | 100 |
| Chromatography Type: | HILIC |
| Chromatography ID: | CH002152 |
| Chromatography Summary: | Ten µL of the extract was injected into the column through a CTC PAL autosampler. The samples were separated in the RP model using a Raptor Biphenyl column (150 × 2.1 mm, 2.7 µm) (Restek, PA). The LC conditions were as follows: Mobile phase A: 90% H2O with 0.1% formic acid and 10% MEOH, pH 4.0. Mobile phase B: MEOH-IPA (50:50, v/v) with 0.1% formic acid. The gradient was: 0 - 2 min 10% B, 2.1 - 4 min 40% B, 4 -12 min, linear ramping up to 100% B, 12 – 18 min 100% B, 19 - 24 min 10% B, 24.1 min stop. The flow rate was 250 µL/min, and the column temperature was controlled at 40 ºC. |
| Instrument Name: | Shimadzu 20AD |
| Column Name: | Raptor Biphenyl (150 × 2.1 mm,2.7um) |
| Column Pressure: | 2800 psi - 4200 psi |
| Column Temperature: | 40 |
| Flow Gradient: | Constant |
| Flow Rate: | 250 µL/min |
| Injection Temperature: | 4 |
| Solvent A: | 90% water/10% methanol; 0.1% formic acid, pH 4.0 |
| Solvent B: | 50% methanol/50% isopropanol; 0.1% formic acid |
| Oven Temperature: | 40 |
| Sample Loop Size: | 10 |
| Sample Syringe Size: | 100 |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002693 |
| Analysis ID: | AN002901 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | The MS/MS detection was performed using electrospray ionization (ESI) and advanced scheduled multiple reaction monitoring (MRM) with dynamic windowing and fast polarity switch. The ESI source conditions were set as follows: electrospray voltage of -4500 V for negative mode and 5500 V for positive mode, source temperature of 500 °C, curtain gas of 30, ion source gas 1, and gas 2 of 35 psi, respectively.Peaks were manually inspected and integrated using Multiquant 3.0.2. The peak area was exported to excel for further analysis. |
| Ion Mode: | UNSPECIFIED |
| Collision Energy: | Compound-dependent |
| Ion Source Temperature: | 500 |
| Ion Spray Voltage: | -4500 V for negative and 5500 V for positive |
| Ionization: | ESI |
| Mass Accuracy: | 0.1 Da |
| MS ID: | MS002694 |
| Analysis ID: | AN002902 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | The MS/MS detection was performed using electrospray ionization (ESI) and advanced scheduled multiple reaction monitoring (MRM) with dynamic windowing and fast polarity switch. The ESI source conditions were set as follows: electrospray voltage of -4500 V for negative mode and 5500 V for positive mode, source temperature of 500 °C, curtain gas of 30, ion source gas 1, and gas 2 of 35 psi, respectively.Peaks were manually inspected and integrated using Multiquant 3.0.2. The peak area was exported to excel for further analysis. |
| Ion Mode: | UNSPECIFIED |
| Collision Energy: | Compound-dependent |
| Ion Source Temperature: | 500 |
| Ion Spray Voltage: | -4500 V for negative and 5500 V for positive |
| Ionization: | ESI |
| Mass Accuracy: | 0.1 Da |
