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MB Sample ID: SA216357
| Local Sample ID: | Plasma17-01_22_1_2864 |
| Subject ID: | SU002336 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 39.48 +-10.0 |
| Weight Or Weight Range: | 29.09 +-4.76 |
| Gender: | Male and female |
| Human Inclusion Criteria: | Overweight (BMI>=25), otherwise healthy, between 18 and 60 years, no chronic disease, willingness to observe Ramadan Fasting |
| Human Exclusion Criteria: | Medication use within 1 week, bariatric surgery within 6 months, or any weight reduction regimen within 1 month of the commencement of Ramadan |
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Subject:
| Subject ID: | SU002336 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 39.48 +-10.0 |
| Weight Or Weight Range: | 29.09 +-4.76 |
| Gender: | Male and female |
| Human Inclusion Criteria: | Overweight (BMI>=25), otherwise healthy, between 18 and 60 years, no chronic disease, willingness to observe Ramadan Fasting |
| Human Exclusion Criteria: | Medication use within 1 week, bariatric surgery within 6 months, or any weight reduction regimen within 1 month of the commencement of Ramadan |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Plasma17-01_22_1_2864 | SA216357 | FL026093 | Pre-Fasting | Treatment |
Collection:
| Collection ID: | CO002329 |
| Collection Summary: | A total of 4 mL of blood was then collected from each subject into a sterile container. The samples were stored immediately at –80 ºC for long-term storage until further metabolomics analysis. |
| Sample Type: | Blood (plasma) |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR002348 |
| Treatment Summary: | Our prospective study was executed during Ramadan (from June 2016 to July 2016, corresponding to Ramadan month in the 1438 Hijri of the lunar calendar), where the daily fasting period covered nearly 15 hours. Data were collected 1 week before Ramadan (pre-fasting or baseline) and after completing 28–30 days of Ramadan (at the end of Ramadan). During Ramadan, fasting people abstain from food and drink (including water) and do not smoke from dawn to sunset. We compared the studied variables for each participant before and during Ramadan, meaning each participant served as their control. Participants did not receive any nutritional recommendations or physical activity advice at any stage during this study. The Islamic laws of Ramadan excuse females from fasting during Ramadan while their menstrual period; therefore, the fasting days for female participants were about 23–25 days. |
| Human Fasting: | Ramadan Intermittent Fasting |
Sample Preparation:
| Sampleprep ID: | SP002342 |
| Sampleprep Summary: | For analysis, an aliquot of thawed plasma sample was transferred into a microcentrifuge tube and cold methanol added at 3:1 v/v (i.e., 30 μL sample, add 90 μL cold methanol), vortexed and allowed to sit at –20ºC for two hours. Next, samples were centrifuged at 20,817 x g for 15 min at 4ºC and the supernatant transferred to a new microcentrifuge tube. Usually, the transferred volume was three times that of the original sample (i.e., for a 30 μL sample, add 90 μL cold methanol, then transfer 90 μL supernatant). The samples were dried using a Speed vac at 30 – 40°C and the dried sample stored in a –80ºC freezer for further use or dissolved in solvent for LC-MS/MS analysis. Samples were resuspended in the starting solvent (0.1% formic acid) where the volume was three times the original plasma volume, i.e., 90 μL for 30 μL serum/plasma aliquot. |
| Processing Storage Conditions: | Described in summary |
| Extract Storage: | 4℃ |
Combined analysis:
| Analysis ID | AN003676 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Bruker Elute |
| Column | Hamilton Intensity Solo 2 C18 |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Bruker timsTOF |
| Ion Mode | POSITIVE |
| Units | AU |
Chromatography:
| Chromatography ID: | CH002726 |
| Chromatography Summary: | Samples were chromatographically separated using a Hamilton® Intensity Solo 2 C18 column (100 mm x 2.1 mm x 1.8 µm) and eluted using 0.1% formic acid in water (A) and 0.1% formic acid in ACN (B) using the following gradient: at a flow rate of 0.250 ml/min 1% B from 0-2 min, then gradient elution to 99% B from 2-17 min, held at 99% B from 17-20 min, then re-equilibrated to 1% B from 20-30 min using a flow rate of 0.350 ml/min. The autosampler temperature was set at 8℃ and the column oven temperature at 35℃. |
| Instrument Name: | Bruker Elute |
| Column Name: | Hamilton Intensity Solo 2 C18 |
| Column Temperature: | 35 |
| Flow Gradient: | 1%B to 99%B in 15 min |
| Flow Rate: | 250 uL/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Oven Temperature: | 35C |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS003427 |
| Analysis ID: | AN003676 |
| Instrument Name: | Bruker timsTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Auto MS/MS mode with 0.5 second cycle time. The ESI source with dry nitrogen gas was 10 L/min, and the drying temperature was equal to 220℃ with nebulizer gas pressure set to 2.2 bar. The capillary voltage of the ESI was 4500 V and the Plate Offset 500 V. MS acquisition scan was set from 20-1300 m/z and the collision energy to 7 eV. |
| Ion Mode: | POSITIVE |
