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MB Sample ID: SA162261
| Local Sample ID: | Plasma_pos_86 |
| Subject ID: | SU001800 |
| Subject Type: | Mammal |
| Subject Species: | Sus scrofa |
| Taxonomy ID: | 9823 |
| Age Or Age Range: | 0-125 days |
| Weight Or Weight Range: | 1-80 kg |
| Gender: | Male |
| Animal Animal Supplier: | Swine Innovation Centre Sterksel, the Netherlands |
| Animal Housing: | Metabolism cages |
| Animal Light Cycle: | 12h light / 12h darknes |
| Animal Feed: | Experimental diets (see publication) |
| Animal Water: | Ad libitum |
| Animal Inclusion Criteria: | Birth weight |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001800 |
| Subject Type: | Mammal |
| Subject Species: | Sus scrofa |
| Taxonomy ID: | 9823 |
| Age Or Age Range: | 0-125 days |
| Weight Or Weight Range: | 1-80 kg |
| Gender: | Male |
| Animal Animal Supplier: | Swine Innovation Centre Sterksel, the Netherlands |
| Animal Housing: | Metabolism cages |
| Animal Light Cycle: | 12h light / 12h darknes |
| Animal Feed: | Experimental diets (see publication) |
| Animal Water: | Ad libitum |
| Animal Inclusion Criteria: | Birth weight |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Plasma_pos_86 | SA162261 | FL019173 | restricted | Diet |
| Plasma_pos_86 | SA162261 | FL019173 | low | Wght_birth_cat |
Collection:
| Collection ID: | CO001793 |
| Collection Summary: | At the end of each feeding regime, blood samples were collected from the jugular vein. Per sampling moment, two 9-mL plasma tubes (Vacuette, Greiner Bio-One, Kremsmünster, Austria) per pig were filled and allowed to clot for 1 h at room temperature. Plasma was collected after centrifugation for 15 min at 2,000 ∙ g and was stored at -80°C pending analyses. |
| Sample Type: | Blood (plasma) |
| Collection Method: | Jugular vein puncture |
| Collection Frequency: | At the end of feeding period |
| Volumeoramount Collected: | 2 x 9 ml blood |
| Storage Conditions: | -80℃ |
| Collection Vials: | Vacuette |
Treatment:
| Treatment ID: | TR001813 |
| Treatment Summary: | At an age of 14 weeks, 10 LBW-LEBV (BiW: 1.07 + 0.09 kg; EBV: -2.52 + 3.97 g/d, compared to an average crossbred pig with a protein deposition of 165 g/d), 10 LBW-HEBV (BiW: 1.02 + 0.13 kg; EBV: 10.47 + 4.26 g/d), 10 HBW-LEBV (BiW: 1.80 + 0.13 kg; EBV: - 2.15 + 2.28 g/d), and 10 HBW-HEBV (BiW: 1.80 + 0.15 kg; EBV: 11.18 + 3.68 g/d), male growing pigs (Synthetic boar x (Dutch Landrace x Large White)) were allotted to the experiment. The pigs were individually housed in metabolism cages (1.80 x 0.80 m) at a room temperature of 22oC. They were subjected to N- balance measurements in two sequential periods of 5 d using a restricted feeding regime. After a 6-d adaptation period to the metabolism cages, pigs were adapted for 5 days to the experimental diets before the start of the first 5-d balance period. Pigs were assigned to a protein adequate (A) or protein restricted (R, 70% of A) regime in a change-over design. |
Sample Preparation:
| Sampleprep ID: | SP001806 |
| Sampleprep Summary: | Plasma samples (150 µL) were pipetted into a 96-well plate and 450 µL acetonitrile containing an internal standard mix of glycocholic acid (glycine-1-13C) and p-chlorophenylalanine to a final concentration of 0.01 mg/mL, was added per well. The samples were mixed immediately and placed at 4°C for 10 min for protein precipitation. After centrifugation (3700 rpm for 25 min at 4°C), the supernatant was transferred to a filter plate fixed on top of a 96-well collection plate in a manifold with a pressure gauge. Vacuum was then applied to the filter plate, and the solvent containing plasma components was collected into the 96-well collection plate. The collection plate was placed in a vacuum centrifuge and the samples were evaporated to dryness (ca. 2.5 h, 805 × g and 30°C). A volume of 150 µL of water/acetonitrile/formic acid (95/5/0.1%) was added to the dried extracts in the collection plate. The dried collection plate was then sealed with a piece of aluminum laminate (#186002789, Waters), centrifuged (3700 rpm for 25 min at 4°C), and kept in the auto-sampler at 10°C for the UPLC-QTOF/MS analysis. The injection volume was 3 µl. |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN002807 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 RS |
| Column | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Bruker Impact HD |
| Ion Mode | POSITIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH002074 |
| Chromatography Summary: | Chromatographic separation was performed on a Dionex UltiMate 3000RSL Binary UHPLC System (Thermo Scientific Dionex, Sunnyvale, CA) equipped with a HSS T3 C18 UHPLC column, 1.8 µm, 100x2.1 mm (Waters Corporation, Milford, MA). The column was maintained at 30°C. Samples were kept in the autosampler at 10°C, and the injection volume for both sample types was 3 µl. The mobile phases were 0.1% formic acid in Milli-Q water (A) and 0.1% formic acid in acetonitrile (B). The gradient program for the plasma samples was as follows: 0-12 min, linear gradient from 5-100% B; 12-13 min, 100% B and return to initial conditions in 0.2 min. In all cases, the column was re-equilibrated at 5% B for 2 min at the beginning of each run. The flow rate was 0.4 ml/min |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 30 |
| Flow Rate: | 0.4 ml/min |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% acetonitrile; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002602 |
| Analysis ID: | AN002807 |
| Instrument Name: | Bruker Impact HD |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | The eluent was introduced into the mass spectrometer by electrospray ionization, with capillary set in the positive and negative mode to 4500 and 3600 V, respectively. End-plate offset voltage was set to 500 V. Nitrogen was used as both nebulizer and drying gas with a gas pressure of 1.8 bar. Drying gas temperature and flow were 200 °C and 8.0 L/min, respectively. Spectra were acquired over the scan range of 50−1000 m/z. A solution of lithium formate clusters (5 mM) (water/isopropanol/formic acid in a 50:50:0.2 v/v/v ratio) was injected prior to each chromatographic run as an external calibrant. |
| Ion Mode: | POSITIVE |
