Analyses were performed using the instrumentation described in the Mass spectrometric analysis (link) section. Then, 1 μl of synthetic samples or metabolome extract were separated using a water-acetonitrile gradient on an Agilent Zorbax Hilic Plus column (150 × 2.1 mm, particle size 1.8 μm) maintained at 40 °C with a flow rate of 0.3 ml min−1. Solvent A was 0.1% formic acid in water and solvent B was 0.1% formic acid in acetonitrile. Analytes were separated using a gradient profile as follows: 2 min (95% B) → 20 min (50% B) → 20 min (50% B) lvent A, 0.1% formic acid in water; solvent B, 0.1% samples was analyzed in ESI+ mode, m/z range 100 to 700.
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HILIC-MS-based Metabolomics Analysis
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Corresponding Organization :
Other organizations : Cornell University, Howard Hughes Medical Institute, Columbia University
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Variable analysis
independent variables
- Gradient profile (2 min 95% B -> 20 min 50% B -> 20 min 50% B)
- Flow rate (0.3 ml min^-1)
- Column (Agilent Zorbax Hilic Plus, 150 × 2.1 mm, 1.8 μm particle size)
- Column temperature (40 °C)
- Metabolite separation and detection
- Sample volume (1 μl)
- Solvents (Solvent A: 0.1% formic acid in water, Solvent B: 0.1% formic acid in acetonitrile)
- Mass spectrometry ionization mode (ESI+ mode)
- Mass spectrometry scan range (m/z 100 to 700)
- Synthetic samples
- Metabolome extract
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