Metabolomics Workbench : NIH Data Repository

MB Sample ID: SA057747

Local Sample ID:	21
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:

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.

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.