Columns
YMC-Triart Bio C4 is an organic/inorganic hybrid silica based wide-pore column suitable for separation of proteins. YMC-Triart Bio C4 can separate proteins with molecular weight of up to 150,000 such as antibodies under high temperature. YMC-Triart Bio C4 achieves excellent peak shape even when using mobile phase containing formic acid. Therefore it is effective for the high sensitive LC/MS analysis. YMC-Triart Bio C4 is also useful for separation of peptides, oligonucleotides, and small molecules with high hydrophobicity which are difficult to be separated with C18 columns.
| SPECIFICATION | |
| Base | organic/inorganic hybrid silica |
|---|---|
| Particle size | 1.9 µm, 3 µm, 5 µm |
| Pore size | 300 Å |
| Usable pH range | 1-10 |
| USP | L26 |
Intact monoclonal antibody was analyzed at the temperatures between 40°C and 90°C using Triart Bio C4. Good peak shapes were acquired at 70°C and higher while there was no elution at 50°C and lower. Highly durable Triart Bio C4 can perform stable analysis even at 90°C and is suitable for the reversed-phase analysis of antibodies at high temperatures.
| Column | YMC-Triart Bio C4 (3 µm, 300 Å) 150 X 3.0 mmI.D. |
|---|---|
| Eluent | A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 30-60%B (0-15 min), 90%B (15-30 min) |
| Flow rate | 0.4 mL/min |
| Detection | UV at 220 nm |
| Sample | Humanized monoclonal IgG1 |
| Injection | 4 µL |
Below are the chromatograms of monoclonal antibody fragments analysis with mobile phase containing either TFA or formic acid. Triart Bio C4 achieves excellent peak shape even when using mobile phase containing formic acid. Therefore it is ideal for the high sensitive LC/MS analysis.
| Column size | 150 X 2.1 mmI.D |
|---|---|
| Eluent <TFA> | A) water/TFA (100/0.1) B) acetonitrile/TFA (100/0.1) 25-50%B (0-10 min), 90%B (10-12.5 min) |
| Eluent <Formic acid> | A) water/formic acid (100/0.1) B) acetonitrile/formic acid (100/0.1) 20-45%B (0-10 min), 90%B (10-12.5 min) |
| Detection | UV at 280 nm |
| Flow rate | 0.4 mL/min |
| Temperature | 80°C |
| Injection | 4 µL (0.25 mg/mL) |
| Sample | mAb subunit Standard (Waters Corporation) |
Generally, improvement of peptide and protein separation is expected under a high temperature condition using a mobile phase containing TFA, although shorter lifetime of conventional columns becomes a problem under such a condition. Since Triart is based on organic/inorganic hybrid particle, which has its innovative surface modification, it keeps excellent durability even under harsh conditions such as 1% TFA and/or 90°C.
| Test conditions | |
| Column | 5 µm, 150 X 3.0 mmI.D. for C4 5 µm, 50 X 2.0 mmI.D. for C18 |
|---|---|
| Eluent | acetonitrile/water (60/40) |
| Flow rate | 0.4 mL/min for 3.0 mmI.D. 0.2 mL/min for 2.0 mmI.D. |
| Temperature | 37ºC |
| Sample | butyl benzoate |
