Return to study ST000972 main page
MB Sample ID: SA058604
| Local Sample ID: | S7_3 |
| Subject ID: | SU001011 |
| Subject Type: | Other |
| Subject Species: | Papio hamadryas |
| Taxonomy ID: | 9557 |
| Age Or Age Range: | 18 |
| Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001011 |
| Subject Type: | Other |
| Subject Species: | Papio hamadryas |
| Taxonomy ID: | 9557 |
| Age Or Age Range: | 18 |
| Gender: | Male |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| S7_3 | SA058604 | FL010438 | GC-Orbitrap-MS | Platform Type |
Collection:
| Collection ID: | CO001005 |
| Collection Summary: | All procedures involving animals were reviewed and approved by the Texas Biomedical Research Institute’s Institutional Animal Care and Use Committee and conducted in AAALAC approved facilities. For this study, we utilized a healthy adult male olive baboon (Papio hamadryas) maintained as part of the baboon colony at the Southwest National Primate Research Center, located on the campus of the Texas Biomedical Research Institute, San Antonio, Texas. The male baboon used in this study was 18 yrs. old. The baboon had been raised and maintained on a standard monkey chow diet (high complex carbohydrates; low fat) prior to the fasting blood collection. All procedures involving animals were reviewed and approved by the Texas Biomedical Research Institute’s Institutional Animal Care and Use Committee (IACUC). Freshly collected serum samples were stored in aliquots at -80 C until analysis. |
| Sample Type: | Blood (serum) |
| Collection Location: | Southwest National Primate Research Center, San Antonio, Texas, USA |
| Collection Frequency: | 1 |
| Collection Duration: | NA |
| Volumeoramount Collected: | 40 mL |
| Storage Conditions: | -80℃ |
| Collection Tube Temp: | 4 C |
| Additives: | None |
Treatment:
| Treatment ID: | TR001025 |
| Treatment Summary: | None |
Sample Preparation:
| Sampleprep ID: | SP001018 |
| Sampleprep Summary: | Aliquots of serum (30 µL) samples were subjected to sequential solvent extraction once each with 1 mL of acetonitrile: isopropanol: water (3:3:2) and 500 µL of acetonitrile: water (1:1) mixtures at 4 C.22 Adonitol and d4-succinic acid (both 5 µL from 10 mg/ml stock) were added to each aliquots as two internal standards prior to the extraction. The pooled extracts (~ 1500 µL) from the two steps were dried under vacuum at 4 C prior to chemical derivatization. Dummy extractions performed on blank tubes served as extraction blanks to account for background (extraction) noise and other sources of contamination. Six (S1, S2, S3, S6, S7, S8) samples were then sequentially derivatized with methoxyamine hydrochloride (MeOX) and 1% TMCS in N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA) as described elsewhere.23,24 Steps involved addition of 10 μL of MeOX (20 mg mL-1) in pyridine incubated under shaking at 55 °C for 60 min followed by trimethylsilylation at 60 °C for 60 min after adding 90 μL MSTFA. |
| Processing Method: | Fiehn et al., 2008 |
| Processing Storage Conditions: | On ice |
| Extraction Method: | Fiehn et al., 2008 |
| Extract Enrichment: | None |
| Extract Cleanup: | None |
| Extract Storage: | -80℃ |
| Sample Derivatization: | Methoxyamination + silylation (MSTFA) |
| Sample Spiking: | Adonitol, d4-succinic acid |
Combined analysis:
| Analysis ID | AN001592 | AN001593 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | GC | GC |
| Chromatography system | Leco Pegasus 4D GC | Thermo Trace 1310 |
| Column | Restek Rtx-5Sil (30m x 0.25mm,0.25um) | Thermo Scientific TraceGOLD TG-5SILMS (30m x 0.25 mm,0.25um) |
| MS Type | EI | EI |
| MS instrument type | GC x GC-TOF | Orbitrap |
| MS instrument name | Leco Pegasus 4D GCxGC TOF | Thermo Q Exactive Orbitrap |
| Ion Mode | POSITIVE | POSITIVE |
| Units | Arbitrary units | Arbitrary units |
Chromatography:
| Chromatography ID: | CH001117 |
| Chromatography Summary: | 1µL of derivatized sample into a split/splitless (SSL) injector at 250 °C using a splitless injection on a Thermo Scientific™ TRACE™ 1310 GC. Helium carrier gas at a flow rate of 1 mL/min was used for separation on a Thermo Scientific™ TraceGOLD™ TG-5SILMS 30 m length × 0.25 mm i.d. × 0.25 µm film thickness column. The initial oven temperature was held at 70 °C for 4 min, followed by an initial gradient of 20 °C/min ramp rate. The final temperature was 320 °C and held for 8 min. |
| Chromatography Comments: | 2D GC |
| Instrument Name: | Leco Pegasus 4D GC |
| Column Name: | Restek Rtx-5Sil (30m x 0.25mm,0.25um) |
| Flow Rate: | 1 ml/min |
| Injection Temperature: | 250 |
| Internal Standard: | Adonitol, d4-succinic acid |
| Sample Injection: | 1 uL |
| Running Buffer: | Helium |
| Transferline Temperature: | 250 |
| Randomization Order: | Yes |
| Chromatography Type: | GC |
| Chromatography ID: | CH001118 |
| Chromatography Summary: | Samples were injected in splitless mode using an autosampler (VCTS, Gerstel™, Linthicum, MD, USA) consisting of an Agilent© 7890 B gas chromatograph (Agilent Technologies, Palo Alto, CA, USA) in line with a Pegasus ® 4D ToF-MS instrument (Leco Corp., San Jose, CA, USA) equipped with an electron impact (EI) ionization source. Injection temperature was set at 250 °C (front inlet) and the helium (carrier gas) flow rate was set to 1 mL min-1. Separation on the GC was achieved using two columns, a primary Rxi®-5Sil MS capillary column (Cat. No. 13623-6850, Restek, Bellefonte, PA, USA) (30 m × 0.25 mm × 0.25 μm) in line with a secondary Rxi®-17Sil capillary column (Cat. No. 40201-6850, Restek, Bellefonte, PA, USA) (2 m × 0.15 mm × 0.15 μm). The temperature program for the primary column started isothermal at 70 °C for 1 min followed by a 6 °C min-1 ramp to 310 °C and a final 11 min hold at 310 °C. The secondary oven temperature was programmed with an offset of 5°C whereas the modulator temperature offset was 15° C relative to the first oven temperature. The modulation temperature (second-dimension separation time) was 4 s divided into a hot and cold pulse times of 0.60 s and 1.4 s, respectively between the two stages. |
| Instrument Name: | Thermo Trace 1310 |
| Column Name: | Thermo Scientific TraceGOLD TG-5SILMS (30m x 0.25 mm,0.25um) |
| Injection Temperature: | 250 |
| Internal Standard: | Adonitol, d4-succinic acid |
| Sample Injection: | 1 uL |
| Running Buffer: | Helium |
| Transferline Temperature: | 250 |
| Randomization Order: | Yes |
| Chromatography Type: | GC |
MS:
| MS ID: | MS001470 |
| Analysis ID: | AN001592 |
| Instrument Name: | Leco Pegasus 4D GCxGC TOF |
| Instrument Type: | GC x GC-TOF |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
| Fragment Voltage: | -70 eV |
| Fragmentation Method: | EI |
| Helium Flow: | 1 ml/min |
| Ion Source Temperature: | 250 |
| Mass Accuracy: | Unit resolution |
| Scan Range Moverz: | 40-600 |
| Scanning: | 200 scans/s |
| MS ID: | MS001471 |
| Analysis ID: | AN001593 |
| Instrument Name: | Thermo Q Exactive Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
| Fragment Voltage: | -70 eV |
| Fragmentation Method: | EI |
| Helium Flow: | 1 ml/min |
| Ion Source Temperature: | 250 |
| Mass Accuracy: | 60,000 resolution (FWHM at m/z 200 |
| Scan Range Moverz: | 50-650 |
