Table 3.
LC conditions for HILIC and reverse phase MS.
| A: LC conditions for HILIC-WAX LC-MS for HS/CS disacchrides | |||
|---|---|---|---|
| Time (min) | Flow rate μL/min | A | B |
| 0 | 0.6 | 15 | 85 |
| 5 | 0.6 | 15 | 85 |
| 20 | 0.6 | 30 | 70 |
| 22 | 0.6 | 42 | 58 |
| 25 | 0.6 | 50 | 50 |
| 40 | 0.6 | 85 | 15 |
| 45 | 0.6 | 85 | 15 |
| 48 | 0.6 | 15 | 85 |
| 55 | 0.6 | 15 | 85 |
| 1. Set up mobile phase solution using LC-MS grade solvents: A: 50 mM Ammonium formate pH 4.5 B: 95% acetonitrile / 5% Water Weak wash: 90% acetonitrile / 10% Water Strong wash: 80% water / 20% acetonitrile Seal Wash:90% water / 10% acetonitrile 2. Set up gradient conditions as below for HILIC-WAX analysis | |||
| 3B| LC conditions for HILIC MS for N-glycan analysis using Amide-80 | |||
|---|---|---|---|
| Time (min) | Flow rate μL/min | A | B |
| 0 | 0.6 | 20 | 80 |
| 5 | 0.6 | 20 | 80 |
| 35 | 0.6 | 80 | 20 |
| 50 | 0.6 | 80 | 20 |
| 52 | 0.6 | 20 | 80 |
| 70 | 0.6 | 20 | 80 |
| 1. Set up mobile phase solution using LC-MS grade solvents: A: 50 mM Ammonium formate pH 4.5 B: 95% acetonitrile / 5% Water Weak wash: 80% acetonitrile / 20% Water Strong wash: 80% water/ 20% acetonitrile Seal Wash:90% water/ 10% acetonitrile 2. Set up gradient conditions as below for HILIC analysis | |||
| 3C| LC conditions for reverse phase MS for peptide analysis using C18 waters BEH | ||||
|---|---|---|---|---|
| Time (min) | Flow rate μL/min | A | B | curve |
| 0 | 0.5 | 98 | 2 | 6 |
| 1 | 0.5 | 98 | 2 | 6 |
| 91 | 0.5 | 60 | 40 | 6 |
| 93 | 0.5 | 60 | 40 | 6 |
| 96 | 0.5 | 2 | 98 | 6 |
| 101 | 0.5 | 2 | 98 | 6 |
| 103 | 0.5 | 98 | 2 | 6 |
| 120 | 0.5 | 98 | 2 | 6 |
| 1. Set up mobile phase solution using LC-MS grade solvents: A: 50mM Ammonium formate pH 4.5 B: 95% acetonitrile / 5% Water Weak wash: 80% acetonitrile / 20% Water Strong wash: 80% water/ 20% acetonitrile Seal Wash:90% water/ 10% acetonitrile 2. Set up gradient conditions as below for a 120min reverse phase analysis. | ||||