Columns

YMC-Triart Bio C4

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
Features

Useful for separation of intact antibodies under high temperature

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

Good peak shapes with mobile phase suitable for LC/MS

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)

Excellent chemical durability

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