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MB Sample ID: SA162329
Local Sample ID: | 32 |
Subject ID: | SU001801 |
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: | SU001801 |
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 |
---|---|---|---|---|
32 | SA162329 | FL019177 | restricted | Diet |
32 | SA162329 | FL019177 | high | Wght_birth_cat |
Collection:
Collection ID: | CO001794 |
Collection Summary: | In both balance periods, at d4 between 0900 and 1000 h three 1.5 ml urine samples per pig were taken from the buckets with urine collected from 0800 h that morning. Before taking the urine samples, the urine was mixed. The samples were stored at -80°C pending analyses. |
Sample Type: | Urine |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR001814 |
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. LBW: Low birth weight; HBW: High birth weight; LEBV: Low estimated breeding value for protein deposition; HEBV: High estimated breeding value for protein deposition. |
Sample Preparation:
Sampleprep ID: | SP001807 |
Sampleprep Summary: | In a 96-well plate, the urine samples (112.5 µL) were diluted with H2O (112.5 µL) and 25 µL acetonitrile (100%, ACN) 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. The samples were incubated for 20 min at 4°C and subsequently centrifuged (3700 rpm, 25 min, 4°C). The supernatant, 100 µL per well, was transferred to a 200 µL 96-well plate and a protective film was welded on the plate using a heat sealer, and the plate was centrifuged at 3700 rpm, 4°C for 25 min prior to the LC-MS analysis. Samples were kept in the autosampler at 10°C, and the injection volume was 3 µl. |
Combined analysis:
Analysis ID | AN002808 | AN002809 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Reversed phase | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
Column | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
MS Type | ESI | ESI |
MS instrument type | QTOF | QTOF |
MS instrument name | Bruker Impact HD | Bruker Impact HD |
Ion Mode | NEGATIVE | POSITIVE |
Units | peak area | peak area |
Chromatography:
Chromatography ID: | CH002075 |
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 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 urine samples was as follows: 0-8 min, linear gradient from 5-70% B; 8-8.5 min, linear gradient from 70-100% B; 8.5-9.5 min, 100% B and return to initial conditions in 0.2 min. The column was re-equilibrated at 5% B for 2 min at the beginning of each run and the flow rate was set to 400 µL/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: | MS002603 |
Analysis ID: | AN002808 |
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: | NEGATIVE |
MS ID: | MS002604 |
Analysis ID: | AN002809 |
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 |