Columns

YMC-Triart Bio C18

YMC-Triart Bio C18, which is a wide-pore column with organic/inorganic hybrid silica, is suitable for separation of peptides, proteins and oligonucleotides. YMC-Triart Bio C18 is also possible to analyze sample materials in a short time, which are strongly retained on conventional C18 columns, due to its low hydrophobicity.

SPECIFICATION
Base organic/inorganic hybrid silica
Particle size 1.9 µm, 3 µm, 5 µm
Pore size 300 Å
Usable pH range 1-12
USP L1
Features

Suitable for separation of peptides and proteins with molecular weight over 10,000

The effects of pore size on separation of peptides and proteins

The widths at half the peak height (WHPH) were compared when peptides and proteins with molecular weight (MW) from 1,859 to 76,000 were separated using Triart Bio C18 and Triart Bio C4 with pore size 300 Å, and Triat C18 with pore size 120 Å. Good peak shape was obtained using Triart Bio C18 and Triart Bio C4, which are wide-pore columns, even in separation of proteins with high MW. On the other hand, peak broadening was observed when using Triart C18 (120 Å) on separation of proteins with MW over 10,000. Wide-pore columns with pore size 300 Å are suitable for separation of proteins with MW over 10,000.

Column

150 X 3.0 mmI.D.

Eluent

A) water/TFA (100/0.1)
B) acetonitrile/TFA (100/0.1)
10-95%B (0-15 min)

Flow rate

0.425 mL/min

Temperature

40ºC

Detection

UV at 220 nm

Injection

4 µL (0.1-0.5 mg/mL)

Sample

γ-Endorphin (MW 1,859), Insulin (MW 5,733),
Lysozyme (MW 14,000), β-Lactoglobulin (MW 18,363),
α-Chymotrypsinogen A (MW 25,656),
BSA (MW 66,000), Conalbumin (MW 76,000)

Separation example of Human growth hormone (MW 22,000)

This example shows that chromatograms of Human growth hormone, a peptide with MW 22,000, which was separated using Triart Bio C18 and Triart Bio C4 with pore size 300 Å, and Triart C18 with pore size 120 Å, respectively. Good peak shape can be obtained on analysis of high MW molecules such as proteins with Triart Bio C18 and Triart Bio C4, wide-pore columns. Excellent separation was achieved using Triart Bio C18 with longer alkyl chain in its bonded phase.

Column

150 X 3.0 mmI.D.

Eluent

A) water/TFA (100/0.1)
B) acetonitrile/TFA (100/0.08)
50-70%B (0-15 min)

Flow rate

0.425 mL/min

Temperature

40ºC

Detection

UV at 220 nm

Injection

8 µL

Sample

Human growth hormone (0.1 mg/mL)

Good peak shape with mobile phase suitable for LC/MS

Triat Bio C18 is suitable for highly sensitive analysis and structural analysis of proteins using LC-MS since it gives good peak shape in mobile phase containing formic acid.

Column

150 X 3.0 mmI.D., 150 X 4.6 mmI.D.

Eluent

A) water/formic acid (100/0.1)
B) acetonitrile/formic acid (100/0.08)
45-65%B (0-15 min)

Flow rate

0.425 mL/min for 3.0 mmI.D.
1.0 mL/min for 4.6 mmI.D.

Temperature

40ºC

Detection

UV at 220 nm

Sample

Human growth hormone (0.1 mg/mL)