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MB Sample ID: SA273831
Local Sample ID: | nist2_1 |
Subject ID: | SU002827 |
Subject Type: | Mammal |
Subject Species: | Rattus norvegicus |
Taxonomy ID: | 10116 |
Age Or Age Range: | 12 weeks |
Gender: | Male |
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Combined analysis:
Analysis ID | AN004411 | AN004412 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | HILIC | Reversed phase |
Chromatography system | Thermo Dionex Ultimate 3000 | Thermo Dionex Ultimate 3000 |
Column | Waters XBridge BEH Amide (50 x 2.1mm,2.5um) | Thermo Higgins C18 (50 x 2.1mm,3um) |
MS Type | ESI | ESI |
MS instrument type | Orbitrap | Orbitrap |
MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
Ion Mode | POSITIVE | NEGATIVE |
Units | peak area | peak area |
Chromatography:
Chromatography ID: | CH003310 |
Chromatography Summary: | The HILIC column is operated parallel to reverse phase column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6-port switching valves. During operation of HILIC separation method, the MS is operated in positive ion mode and 10 microliters of sample is injected onto the HILIC column while the reverse phase column is flushing with wash solution. Flow rate is maintained at 0.35 mL/min until 1.5 min, increased to 0.4 mL/min at 4 min and held for 1 min. Solvent A is 100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C is 2% formic acid (v/v) in LC-MS grade water. Initial mobile phase conditions are 22.5% A, 75% B, 2.5% C hold for 1.5 min, with linear gradient to 77.5% A, 20% B, 2.5% C at 4 min, hold for 1 min, resulting in a total analytical run time of 5 min. During the flushing phase (reverse phase analytical separation), the HILIC column is equilibrated with a wash solution of 77.5% A, 20% B, 2.5% C. |
Methods Filename: | EmoryUniversity_HRM-QEHF_chromatography_5min_092017_v1.pdf |
Chromatography Comments: | Triplicate injections for each chromatography mode |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Waters XBridge BEH Amide (50 x 2.1mm,2.5um) |
Column Temperature: | 60 |
Flow Gradient: | A= water, B= acetonitrile, C= 2% formic acid in water; 22.5% A, 75% B, 2.5% C hold for 1.5 min, linear gradient to 77.5% A, 20% B, 2.5% C at 4 min, hold for 1 min |
Flow Rate: | 0.35 mL/min for 1.5 min; linear increase to 0.4 mL/min at 4 min, hold for 1 min |
Solvent A: | 100% water |
Solvent B: | 100% acetonitrile |
Chromatography Type: | HILIC |
Solvent C: | 100% water; 2% formic acid |
Chromatography ID: | CH003311 |
Chromatography Summary: | The C18 column is operated parallel to the HILIC column for simultaneous analytical separation and column flushing through the use of a dual head HPLC pump equipped with 10-port and 6- port switching valves. During operation of the C18 method, the MS is operated in negative ion mode and 10 μL of sample is injected onto the C18 column while the HILIC column is flushing with wash solution. Flow rate is maintained at 0.4 mL/min until 1.5 min, increased to 0.5mL/min at 2 min and held for 3 min. Solvent A is 100% LC-MS grade water, solvent B is 100% LC-MS grade acetonitrile and solvent C is 10mM ammonium acetate in LC-MS grade water. Initial mobile phase conditions are 60% A, 35% B, 5% C hold for 0.5 min, with linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3.5 min, resulting in a total analytical run time of 5 min. During the flushing phase (HILIC analytical separation), the C18 column is equilibrated with a wash solution of 0% A, 95% B, 5% C until 2.5 min, followed by an equilibration solution of 60% A, 35% B, 5% C for 2.5 min. |
Methods Filename: | EmoryUniversity_HRM-QEHF_chromatography_5min_092017_v1.pdf |
Chromatography Comments: | Triplicate injections for each chromatography mode |
Instrument Name: | Thermo Dionex Ultimate 3000 |
Column Name: | Thermo Higgins C18 (50 x 2.1mm,3um) |
Column Temperature: | 60 |
Flow Gradient: | A= water, B= acetontrile, C= 10mM ammonium acetate in water; 60% A, 35% B, 5% C hold for 0.5 min, linear gradient to 0% A, 95% B, 5% C at 1.5 min, hold for 3 min |
Flow Rate: | 0.4 mL/min for 1.5 min; linear increase to 0.5 mL/min at 2 min held for 3 min |
Solvent A: | 100% water |
Solvent B: | 100% acetonitrile |
Chromatography Type: | Reversed phase |
Solvent C: | 100% water; 10mM ammonium acetate |