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MB Sample ID: SA209275
| Local Sample ID: | Hgd81.4 - No treatment NEG |
| Subject ID: | SU002265 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Combined analysis:
| Analysis ID | AN003568 | AN003569 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Agilent Infinity II | Agilent Infinity II |
| Column | Atlantis dC18 (3.0x100mm,3m,Waters,UK) | Atlantis dC18 (3.0x100mm,3m,Waters,UK) |
| MS Type | ESI | ESI |
| MS instrument type | Triple quadrupole | Triple quadrupole |
| MS instrument name | Agilent 6550 QTOF | Agilent 6550 QTOF |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | area | area |
MS:
| MS ID: | MS003325 |
| Analysis ID: | AN003568 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Quadrupole time-of-flight mass spectrometry (QTOF-MS) conditions An Agilent 6550 QTOF-MS equipped with a dual jet stream electrospray ionisation source was operated in 2GHz mode, over the mass range of 50-1700, in negative and positive polarities. A reference mass correction solution was continually infused at a flow rate of 0.5 mL/min via an external isocratic pump (Agilent, UK) for constant mass correction (see Preparation of reference mass correction solution). Capillary and fragmentor voltages were 4000 V and 380 V, respectively. Desolvation gas temperature was 200 °C with flow rate at 15 L/min. The sheath gas temperature was 300 °C with flow rate at 12 L/min and nebulizer pressure was 40 psi and nozzle voltage 1000 V. Data acquisition rate was 3 spectra/s. Preparation of reference mass correction solution Reference mass correction solution was prepared in 95:5 methanol:water containing 5 mmol/L purine (CAS No. 120-73-0), 100 mmol/L trifluoroacetic acid ammonium salt (TFA, CAS No. 3336-58-1) and 2.5 mmol/L hexakis(1H, 1H, 3H-tetrafluoropropoxy)phosphazine (HP-0921, CAS No. 58943-98-9) (Agilent, Cheadle, UK). Reference ions monitored were: purine (m/z 121.0509) and HP-0921 (m/z 922.0098) (positive polarity) and TFA (m/z 112.9856), purine (m/z 119.0363) and HP-0921 (HP-0921 + formate adduct: m/z 966.0007) (negative polarity). Data acquisition and handling parameters Data were acquired using Acquisition (Build 06.00, Agilent, Cheadle, UK). Quality checks and processing of raw data files (Agilent ‘.d’ files) were performed with Qualitative Analysis software (Build 07.00, Agilent, Cheadle, UK). Extracted ion chromatograms of reference masses were performed to check mass accuracy remained <5 ppm throughout the run and that the reference ion signal did not drop out during the chromatographic run. In addition, to check chromatographic reproducibility binary pump pressure curves for injections across each analytical sequence were overlaid. Mass accuracy and chromatographic reproducibility were acceptable for all experiments performed. Acquired profiling sample data were mined for signals against an established in-house AMRT database of compounds (contains theoretical accurate mass, measured retention time, and empirical formula for 469 intermediary metabolites, MW 72-785) using ‘targeted feature extraction’ with Profinder software (Build 08.00, Agilent, Cheadle, UK). Targeted feature extraction uses the molecular formulae from the AMRT database to extract and group spectral signals (i.e. adducts, isotopes and multimers) that correspond to individual database compounds. Feature extraction employed a window of theoretical accurate mass ±10 ppm and database retention time ±0.3 min. Allowed species were: H+, Na+ and NH4+ (positive polarity) and H- and CHO2- (negative polarity). Dimers were allowed for both polarities. Charge state range was 1-2. Data files were then exported from Profinder as ‘.CEF’ files as a whole batch for each profiling experiment and imported to Mass Profiler Professional (MPP) software for statistical analysis (Build 14.5, Agilent, Cheadle, UK). |
| Ion Mode: | POSITIVE |
| MS ID: | MS003326 |
| Analysis ID: | AN003569 |
| Instrument Name: | Agilent 6550 QTOF |
| Instrument Type: | Triple quadrupole |
| MS Type: | ESI |
| MS Comments: | Quadrupole time-of-flight mass spectrometry (QTOF-MS) conditions An Agilent 6550 QTOF-MS equipped with a dual jet stream electrospray ionisation source was operated in 2GHz mode, over the mass range of 50-1700, in negative and positive polarities. A reference mass correction solution was continually infused at a flow rate of 0.5 mL/min via an external isocratic pump (Agilent, UK) for constant mass correction (see Preparation of reference mass correction solution). Capillary and fragmentor voltages were 4000 V and 380 V, respectively. Desolvation gas temperature was 200 °C with flow rate at 15 L/min. The sheath gas temperature was 300 °C with flow rate at 12 L/min and nebulizer pressure was 40 psi and nozzle voltage 1000 V. Data acquisition rate was 3 spectra/s. Preparation of reference mass correction solution Reference mass correction solution was prepared in 95:5 methanol:water containing 5 mmol/L purine (CAS No. 120-73-0), 100 mmol/L trifluoroacetic acid ammonium salt (TFA, CAS No. 3336-58-1) and 2.5 mmol/L hexakis(1H, 1H, 3H-tetrafluoropropoxy)phosphazine (HP-0921, CAS No. 58943-98-9) (Agilent, Cheadle, UK). Reference ions monitored were: purine (m/z 121.0509) and HP-0921 (m/z 922.0098) (positive polarity) and TFA (m/z 112.9856), purine (m/z 119.0363) and HP-0921 (HP-0921 + formate adduct: m/z 966.0007) (negative polarity). Data acquisition and handling parameters Data were acquired using Acquisition (Build 06.00, Agilent, Cheadle, UK). Quality checks and processing of raw data files (Agilent ‘.d’ files) were performed with Qualitative Analysis software (Build 07.00, Agilent, Cheadle, UK). Extracted ion chromatograms of reference masses were performed to check mass accuracy remained <5 ppm throughout the run and that the reference ion signal did not drop out during the chromatographic run. In addition, to check chromatographic reproducibility binary pump pressure curves for injections across each analytical sequence were overlaid. Mass accuracy and chromatographic reproducibility were acceptable for all experiments performed. Acquired profiling sample data were mined for signals against an established in-house AMRT database of compounds (contains theoretical accurate mass, measured retention time, and empirical formula for 469 intermediary metabolites, MW 72-785) using ‘targeted feature extraction’ with Profinder software (Build 08.00, Agilent, Cheadle, UK). Targeted feature extraction uses the molecular formulae from the AMRT database to extract and group spectral signals (i.e. adducts, isotopes and multimers) that correspond to individual database compounds. Feature extraction employed a window of theoretical accurate mass ±10 ppm and database retention time ±0.3 min. Allowed species were: H+, Na+ and NH4+ (positive polarity) and H- and CHO2- (negative polarity). Dimers were allowed for both polarities. Charge state range was 1-2. Data files were then exported from Profinder as ‘.CEF’ files as a whole batch for each profiling experiment and imported to Mass Profiler Professional (MPP) software for statistical analysis (Build 14.5, Agilent, Cheadle, UK). |
| Ion Mode: | NEGATIVE |
