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MB Sample ID: SA057752
Local Sample ID: | 2 |
Subject ID: | SU001005 |
Subject Type: | Fish |
Subject Species: | Megalobrama amblycephala |
Taxonomy ID: | 75352 |
Gender: | Not applicable |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001005 |
Subject Type: | Fish |
Subject Species: | Megalobrama amblycephala |
Taxonomy ID: | 75352 |
Gender: | Not applicable |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
2 | SA057752 | FL010388 | Control group | Experimental variables |
Collection:
Collection ID: | CO000999 |
Collection Summary: | Blood was obtained from the caudal vein using sterile syringes with pre-added anticoagulant solution and then centrifuged (3000×g) at 4°C for 10 min to obtain the serum, which was quickly frozen in liquid nitrogen and stored at -80°C for biochemical assays. |
Sample Type: | Blood (plasma) |
Treatment:
Treatment ID: | TR001019 |
Treatment Summary: | Control diet (CD group) was formulated to contain 27.11±0.38% carbohydrates. The high-carbohydrate diet (HCD group) was formulated to contain over 36.75±0.92% of easily digestible carbohydrates (mostly from flour). The long-term betaine treatment diet (LBD) to contain a high level of carbohydrates (35.64±0.43%) with 1% betaine supplemented. SBD group (short-term betaine treatment diet) was fed a combination of HCD (first twelve weeks) and LBD diets (last four weeks) |
Sample Preparation:
Sampleprep ID: | SP001012 |
Sampleprep Summary: | Relative metabolites and amino acids were determined using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) on the Ultimate3000-API 3200Q TRAP (USA). The analyses were performed by the Beijing Mass Spectrometry Medical Research Co.,Ltd. (Beijing, China). HPLC-MS/MS detail: Appropriate amount of water was added to serum, mixed thoroughly, and centrifuged at 13200rpm for 5min. Supernatant was collected, 100μl of it was mixed with 400μl of methanol thoroughly, centrifuged again, and the resulting supernatant was subjected to the HPLC-MS/MS. The HPLC-MS/MS system consisted of a SRD-3600 Solvent Rack with analytical 6-channel vacuum degasser, a DGP-3600A pump, WPS-3000TSL analytical autosampler, and a tcc-3200 column compartment. Chromatographic separations were performed on an MSLab C18 column (150×4.6mm, 5μm). The mobile phase A was 0.1% formic acid in water, and the organic mobile phase B was 0.1% ammonium formate in acetonitrile with pH=5.8. The solvent was delivered to the column at a flow rate of 1 ml min−1 as follows: 0-1 minutes from A-B (90:10) to A-B (90:10); 1-12 minutes from A-B (90:10) to A-B (30:70); 12-12.1 minutes from A-B (30:70) to A-B (0:100); 12.1-15 minutes from A-B (0:100) to A-B (0:100); 15-15.1 minutes from A-B (0:100) to A-B (90:10); 15.1-20 minutes from A-B(90:10) to A-B (90:10). The conditions for mass spectrometry detection, optimized to obtain the highest signal intensity, were as follows: mode=positive-ion mode; ion spray voltage=5500V; nebulizer gas pressure=55psi; curtain gas pressure=20psi; collision gas pressure=medium; turbo gas temperature=500°C; entrance potential=10V; collision cell exit potential=2V. Nitrogen gas was used as the collision gas in the multiple reaction monitoring mode. The data were processed using Analyst software version 1.5.1 (Applied Biosystems). Metabolites standard solution (Sigma, USA) was subjected to HPLC-MS/MS using for calibration of the system and quantification of metabolites. |
Combined analysis:
Analysis ID | AN001580 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Unspecified |
Column | Unspecified |
MS Type | ESI |
MS instrument type | Triple quadrupole |
MS instrument name | |
Ion Mode | POSITIVE |
Units | ng/ml or umol/L |
Chromatography:
Chromatography ID: | CH001109 |
Chromatography Summary: | The HPLC system consisted of a SRD-3600 Solvent Rack with analytical 6-channel vacuum degasser, a DGP-3600A pump, WPS-3000TSL analytical autosampler, and a tcc-3200 column compartment. Chromatographic separations were performed on an MSLab C18 column (150×4.6mm, 5μm). The mobile phase A was 0.1% formic acid in water, and the organic mobile phase B was 0.1% ammonium formate in acetonitrile with pH=5.8. The solvent was delivered to the column at a flow rate of 1 ml min−1 as follows: 0-1 minutes from A-B (90:10) to A-B (90:10); 1-12 minutes from A-B (90:10) to A-B (30:70); 12-12.1 minutes from A-B (30:70) to A-B (0:100); 12.1-15 minutes from A-B (0:100) to A-B (0:100); 15-15.1 minutes from A-B (0:100) to A-B (90:10); 15.1-20 minutes from A-B(90:10) to A-B (90:10). |
Instrument Name: | Unspecified |
Column Name: | Unspecified |
Flow Gradient: | 0-1 minutes from A-B (90:10) to A-B (90:10); 1-12 minutes from A-B (90:10) to A-B (30:70); 12-12.1 minutes from A-B (30:70) to A-B (0:100); 12.1-15 minutes from A-B (0:100) to A-B (0:100); 15-15.1 minutes from A-B (0:100) to A-B (90:10); 15.1-20 minutes from A-B(90:10) to A-B (90:10). |
Flow Rate: | 1ml/min |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% acetonitrile; 0.1% ammonium formate, pH 5.8 |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001458 |
Analysis ID: | AN001580 |
Instrument Type: | Triple quadrupole |
MS Type: | ESI |
Ion Mode: | POSITIVE |