Page 26 - Shimadu Consumables Catalog

Bio LC Columns

Shim-pack Scepter C4

Design for biomolecules separations

Shim-pack Scepter C4 is a wide-bore C4 column that uses an Bonded Phase Butyl Groups
organic silica hybrid base which is suitable for separation Particle Size 1.9μm, 3μm, 5μm
of proteins such as monoclonal antibodies. The column can Pore Size 30 nm
be used for analysis of proteins with a molecular weight End-capping Yes
of up to approximately 150,000 and is suitable for analysis pH Range 1 to 10
of antibodies as it is very stable even under acidic or high- USP Code L26
temperature conditions because an organic silica hybrid base
is used. It is also effective for high-sensitivity analysis on LC/MS as it provides good peak shapes even when using a formic acid
mobile phase with low ion paring effect. In addition, it is effective for separation of highly-hydrophobic proteins, nucleotides and
small molecules that are hard to separate using a regular C18 column.

Analysis Examples

Analysis of Proteins Analysis of Monoclonal Antibodies

Peaks
1. Ribonuclease A
(bovine pancreas)
2. Cytochrome C
(horse heart)
3. Insulin (bovine pancreas)
4. BSA
5. α-Chymotrypsinogen A
(bovine pancreas)
NIST mAb
Bevacizumab
Adalimumab
Rituximab
Trastuzumab

Conditions Conditions
Column : Shim-pack Scepter C4 (5 μm, 150 x 3.0 mm I.D.) Column : Shim-pack Scepter C4 (1.9 μm, 50 x 2.1 mm I.D.)
(P/N: 227-31181-04) (P/N: 227-31175-03)
Mobile Phase : A) Water/TFA (100/0.1) Mobile Phase : A) Water/TFA (100/0.1)
B) Acetonitrile/TFA (100/0.1) B) Acetonitrile/TFA (100/0.1)
20-60%B (0-27 min), 90%B (27-35 min), 20%B 25-45%B (0-10 min)
(35-50 min) Flow Rate : 0.4 mL/min
Flow Rate : 0.4 mL/min Col. Temp. : 80 ºC
Col. Temp. : 70 ºC Detection : UV at 280 nm
Detection : UV at 220 nm Injection Vol. : 2 µL
Injection Vol. : 10 µL (0.25 – 0.50 mg/mL) Column Sample Conc. : 0.2 mg/mL

Pg A-22