Return to study ST001257 main page
MB Sample ID: SA091387
Local Sample ID: | S31-105 Pre-Po |
Subject ID: | SU001325 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001325 |
Subject Type: | Human |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Male and female |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
S31-105 Pre-Po | SA091387 | FL013167 | Urine | Sample type |
S31-105 Pre-Po | SA091387 | FL013167 | urine | Food type |
S31-105 Pre-Po | SA091387 | FL013167 | Pre Pork | Pre or Post and food |
Collection:
Collection ID: | CO001319 |
Collection Summary: | Twenty-four-hour urine collections were obtained before and near the end of each of the two DASH-style diet periods. A total of 76 urine samples were used for untargeted metabolomics analysis, including 38 collected while the 19 participants consumed their typical diets (19 before each intervention period) and 38 while the DASH-style diet was consumed (19 during each intervention period). For simplicity, these are referred to as pre- and post-diet urine samples. Food was purchased from a local grocery store for metabolomic analyses. All four types of meat included in the diet (pork, chicken, fish, and beef loin) were analyzed. Fruits and vegetables, if not pre-washed, were washed with tap water and prepared with inedible parts (i.e. leaves or peels) removed. Meat and eggs were cooked as instructed to match prescribed cooked weight. |
Sample Type: | Urine |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001340 |
Treatment Summary: | During the 18-week study period, participants initially consumed their self-chosen, unrestricted (typical) diets for two weeks. Participants were then randomly assigned to consume a DASH-style diet with either lean pork or chicken and fish as the predominant sources of dietary protein for six weeks. This time period was followed by a four-week washout period during which participants again consumed their typical diets prior to consuming the DASH-style diet containing the other predominant protein sources. Samples utilized for the current research consisted of 12 individual foods consumed during the DASH-style diet and urine samples collected prior to each intervention and during the final two weeks of each controlled feeding period. |
Sample Preparation:
Sampleprep ID: | SP001333 |
Sampleprep Summary: | Individual food and urine samples were stored at -80 °C prior to sample preparation. Individual foods (including apple juice) were individually dried using a FreeZone 2.5 plus lyophilizer (Labconco, Kansas City, MO, USA). Dried samples were then divided and macerated using a chilled bio-pulverizer (BioSpec Products, Bartlesville, OK). Approximately 50 mg of each sample was measured into chilled microcentrifuge tubes and stored at -80o C prior to further processes and analysis. 10 µL of coffee was diluted with 90 µL of water, vortexed, aliquotted and stored at -80° C until analysis. An aliquot of coffee was also reserved for neat (i.e. unprocessed) analysis, and stored at -80° C until analysis. Freeze-dried foods were removed from -80° C and allowed to thaw on ice. To each sample, 480 μl of chilled methanol, 10 µl each of a hydrophilic spike mix, hydrophilic positive control mix, hydrophobic spike mix and hydrophobic positive control mix were added (see Supplemental Table S1). Samples were gently vortexed for 10 seconds, placed at -80o C for 60 minutes, and then centrifuged 0o C at 18,000 x g for 15 minutes to facilitate protein precipitation. Supernatants were transferred to new microcentrifuge tubes and dried using vacuum centrifugation at 45o C for approximately 60 minutes. Each sample was suspended in 50 µl of 95:5 LC/MS grade water-acetonitrile and gently vortexed for 30 seconds. Following a quick centrifugation, each sample was then divided into replicate 20 µl aliquots, while 10 µl was removed from each sample to generate a pooled QC sample. Samples were stored at -80° C until analysis. Urine samples were analyzed neat (i.e. without any preparation) except that authentic standards were spiked into urine samples to monitor variability in instrument response or batch effects. |
Processing Storage Conditions: | Described in summary |
Combined analysis:
Analysis ID | AN002086 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Agilent 6520 |
Column | Agilent Zorbax Rapid Resolution HT (RRHT) SB-AQ (100 x 2.1mm,1.8um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Agilent 6520 QTOF |
Ion Mode | POSITIVE |
Units | Abundance (Counts Log2 Transformed) |
Chromatography:
Chromatography ID: | CH001523 |
Instrument Name: | Agilent 6520 |
Column Name: | Agilent Zorbax Rapid Resolution HT (RRHT) SB-AQ (100 x 2.1mm,1.8um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001937 |
Analysis ID: | AN002086 |
Instrument Name: | Agilent 6520 QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
MS Comments: | Agilent 6520 Time-of-Flight (TOF-MS) with dual electrospray ionization (ESI) source, scan rate 2.21 spectra/second, mass range 50-1700 m/z, gas temperature 325 °C, gas flow 12.0 L/min, nebulizer 30 psi, skimmer 60 V, capillary voltage 4000 V, fragmentor 120 V, reference masses 121.050873 and 922.009798 (Reference mix, Agilent Technologies,). |
Ion Mode: | POSITIVE |