The Legacy L8 column is a single-mode, ion-exchange column with aminopropylsilane (AP) groups chemically bonded to totally porous silica gel micro-particles. The Legacy L8 column meets the UPS standard for the L8 designation, which states that the column consists of an essentially monomolecular layer of aminopropylsilane chemically bonded to totally porous silica gel support, 3-10μm diameter. The AP groups allow for both cation-exclusion and hydrophilic interaction chromatography (HILIC).

If you would like to learn more about SIELC’s line of Legacy columns, you can download the brochure here.

Below, we have shared a select sample of methods showcasing the retention capabilities of the Legacy L8 column.

Short-chain peptides can often be highly polar, and when they include lysine moieties, they can be very basic as well. Short-chain lysine-based peptide oligomers have been found to have their best peak shape in HILIC/cation-exclusion mode. Figure 1 shows how these oligomers of different sizes (one made from 3 lysine moieties and one made from 6) can be retained on a Legacy L8 column using a gradient mobile phase consisting of MeCN, water, and phosphoric acid as a buffer.

 

Polylysine, significantly longer-chain polymers made out of many more lysine units, are easily produced by batcerial fermentation. Since any given sample can have differently sized polymers, it is often difficult to obtain a single, sharp peak. Figure 2 shows, however, that a relatively sharp peak can be obtained on a Legacy L8 column with a gradient mobile phase consisting of MeCN, water, and phosphoric acid as a buffer.