Low MW proteins and peptides on BioSep2000(2)

LC Conditions

Column

BioSep 5 µm SEC-s2000 145 Å, LC Column 300 x 7.8 mm, Ea

Brand

Biosep

Part No

00H-2145-K0

Phase Name

SEC-s2000

Elution Type

Isocratic

Mobile Phase

A: 45% Acetonitrile, 0.1% TFA

Gradient Profile

Step No.

Time(min)

%A

%B

1

0

100

0

Flow Rate

1.00 mL/min

Temperature

22°C

Detection

UV-Vis Abs.-Variable Wave.(UV) @ (22°C)

Detection Info

Sample Notes

Application ID # 18945: Low MW proteins and peptides on BioSep 2000 Application Topic: Low MW Separation of Peptides by GFC Unlike protein GFC separations of proteins using buffers that resemble physiological conditions (10mm PO4, 150mm NaCl, pH 7.4); peptide GFC separations require different conditions to get good low molecular weight resolution. The use of acetonitrile and the weak ion pairing buffer TFA (0.1%) minimize secondary interactions between peptides and the stationary phase leading to sharper peaks and better resolution in low molecular ranges. BioSep 2000 is the recommended media for such separations as it has the smallest pore size of the GFC media. In this example application different peptides from 300 Daltons all the way up to 18 Kilodaltons are injected on a BioSep 2000 300 x 7.8 mm column and overlaid to show the separation range of the column using these mobile phase conditions. Note that peptide generally elute according to their molecular weights under such "partially-denaturing" mobile phase conditions, however in some cases unique peptide sequences size differently than the expected by GFC.