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MB Sample ID: SA114174
| Local Sample ID: | POTR_05_vAT |
| Subject ID: | SU001480 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 11-20 years |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001480 |
| Subject Type: | Human |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 11-20 years |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| POTR_05_vAT | SA114174 | FL014389 | SUBCUTANEOUS ADIPOSE TISSUE | Tissue Type |
Collection:
| Collection ID: | CO001475 |
| Collection Summary: | Eleven adolescents 12–20 years of age undergoing bariatric surgery at Cincinnati Children’s Hospital between 2006 and 2012 were offered enrollment in a prospective biospecimen repository protocol (Pediatric Obesity Tissue Repository [POTR]). Sample recruitment and other POTR features have been reported previously (Davidson et al. 2017). Intraoperatively, visceral adipose tissue (vAT) samples from the omentum, abdominal subcutaneous AT (sAT), and liver samples were obtained by the surgeon and processed immediately in an area adjacent to the operating room. All samples were snap-frozen in liquid nitrogen, then stored at −80°C. Plasma was collected pre-operatively after overnight fasting and stored at -80°C. Written informed consent was obtained from participants equal to or above 18 years old or from the parent or guardian if participants were less than 18 years old. The study was approved by the Institutional Review Board at Cincinnati Children’s Hospital. |
| Sample Type: | Adipose tissue |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001495 |
| Treatment Summary: | The objective of the observational study was to evaluate the relationship between adipose and liver tissue POPs and the plasma metabolome. All participants underwent bariatric surgery at the time of tissue collection. No other treatment or intervention was evaluated. |
Sample Preparation:
| Sampleprep ID: | SP001488 |
| Sampleprep Summary: | Tissue POPs concentrations were measured in vAT, sAT and liver tissues collected during surgery. All tissue samples were prepared in batches of 11 study samples and 3 method blanks using a modified version of the QuECHERS method described by (Zamariola et al. 2017). Briefly, 0.2-0.5g of tissue was weighed, placed in an amber glass vial and treated with 3.5mL of LC-MS grade water. Each sample was then spiked with 50μL internal standard solution prepared in 2-proponal that was designed to represent environmental chemicals with a range of physiochemical properties to monitor analysis QA/QC, and included 500 ng/mL [13C6]-Anthracene, [13C12]-PCB28, [DIETHYL-D10]-Chlorpyrifos, [13C12]-PCB101, [13C12]-4,4'-DDE, [13C12]-PCB153, [13C12]-PCB180, [13C12]-PBDE47, [13C10]-Mirex, [13C6]-cis-Permethrin, [13C12]-PBDE99 and [13C12]-PBB153. Following addition of the internal standard solution, the sample was then homogenized for 1 min and placed in a sonicating bath for 10 min. The resulting homogenate was transferred to a 50 mL conical tube containing 10mL acetonitrile, 4000mg MgSO4 and1000mg NaCl, and vortexed for 5 min. After centrifuging, a 1.5mL aliquout was transferred to a cleanup tube containing 50 mg primary and secondary amine exchange material (PSA), 50 mg C18 and 150 mg MgSO4, vortex-mixed for 1 min and centrifuged at max speed for 5 min. From the supernatant, a 1 mL aliquot was transferred to a clean, glass tube and dried completely in a vacuum centrifuge operated at 35°C. The residue was then resuspended in 50μL isooctane and transferred to a GC vial containing a low volume insert and capped with a Teflon lined cap until analysis. |
| Sampleprep Protocol ID: | douglas_walker_Protocol_for_adipose_tissue_exposomics_v3_08Mar2018.pdf |
| Sampleprep Protocol Filename: | douglas_walker_Protocol_for_adipose_tissue_exposomics_v3_08Mar2018.pdf |
| Processing Storage Conditions: | Room temperature |
Combined analysis:
| Analysis ID | AN002349 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Thermo Trace 1310 |
| Column | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
| MS Type | EI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive GC Orbitrap GC-MS/MS |
| Ion Mode | POSITIVE |
| Units | pg/g |
Chromatography:
| Chromatography ID: | CH001721 |
| Chromatography Summary: | Tissue extracts were analyzed using a Thermo Scientific 1310 gas chromatograph connected to a Q Exactive GC Orbitrap GC-MS/MS ultra-high-resolution mass spectrometer and Triplus RSH autosampler. A 2 µL aliquot of extract was injected into an inlet maintained at 250ºC in pulsed split-less mode. The analytes were separated on an Agilent DB-5MSUI capillary column (30m length × 0.25mm inner diameter × 0.25µm film thickness) using high purity helium (99.999% purity) as the carrier gas at a constant flow rate of 1 mL/min. The oven temperature program consisted of an initial temperature of 100ºC for 1 min, increased to 180ºC at 25ºC/min; followed by a temperature ramp to 215ºC at 5ºC/min, and finally increased to 300ºC at 25ºC/min and held for 10 min, resulting in a total run time of 26.6 min. |
| Instrument Name: | Thermo Trace 1310 |
| Column Name: | Agilent DB5-MS (30m x 0.25mm, 0.25um) |
| Flow Rate: | 1 mL/min |
| Injection Temperature: | 250C |
| Internal Standard: | [13C6]-Anthracene, [13C12]-PCB28, [DIETHYL-D10]-Chlorpyrifos, [13C12]-PCB101, [13C12]-4,4'-DDE, [13C12]-PCB153, [13C12]-PCB180, [13C12]-PBDE47, [13C10]-Mirex, [13C6]-cis-Permethrin, [13C12]-PBDE99 and [13C12]-PBB153 |
| Sample Injection: | 2 uL |
| Analytical Time: | 26.6 |
| Oven Temperature: | The oven temperature program consisted of an initial temperature of 100ºC for 1 min, increased to 180ºC at 25ºC/min; followed by a temperature ramp to 215ºC at 5ºC/min, and finally increased to 300ºC at 25ºC/min and held for 10 min |
| Transferline Temperature: | 280 |
| Sample Syringe Size: | 10uL |
| Chromatography Type: | GC |
MS:
| MS ID: | MS002191 |
| Analysis ID: | AN002349 |
| Instrument Name: | Thermo Q Exactive GC Orbitrap GC-MS/MS |
| Instrument Type: | Orbitrap |
| MS Type: | EI |
| MS Comments: | Targeted peak assignment and integration was completed using TraceFinder |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | 250C |
| Ionization: | Postive |
| Ionization Energy: | -70eV |
