The document introduces the advantages of ZORBAX Poroshell packings in the HPLC separation of large molecules such as proteins. This surface porous particle can make the separation faster. At high pH, peptides and small proteins can also be separated by using Extend-C18-bonded Poroshell particles. The high pH may be desirable for improving selectivity, solubility, ease of downstream processing, or to change signal-to-noise and charge-state in LC/MS. By extending the gradient time, it is possible to achieve even higher resolution in typical run times of 10 to 20 min.
This document describes the use of ZORBAX Poroshell 300Extend-C18 superficially porous particles for achieving extremely rapid HPLC separations of large molecules like proteins and peptides. The text highlights that these particles facilitate fast separations due to the rapid movement of biomolecules into and out of the thin porous layer. High pH conditions are noted as beneficial for improving selectivity, solubility, and downstream processing, or for changing signal-to-noise and charge-state in LC/MS.
| Column dimensions (Totally Porous) | 2.1 x 150 mm, 3.5 um |
|---|---|
| Detection wavelength | 215 nm |
| Injection volume | 0.25 uL |
| Flow rate (Poroshell) | 1.0 mL/min |
| Flow rate (Totally Porous) | 0.2 mL/min |
| Run time (Poroshell) | 4 min |
| Run time (Totally Porous) | 40 min |
It permits up to five times faster separation of peptides and proteins over columns using totally porous silica packing.
High pH may be desirable for improving selectivity, solubility, ease of downstream processing, or to change signal-to-noise and charge-state in LC/MS.
A ten-fold reduction in run time is demonstrated, taking 40 minutes on totally porous material versus 4 minutes on Poroshell.