ZORBAX Poroshell superficially porous particle technology allows the fastest, most rugged HPLC separation of proteins to date. A 0.25-µm porous silica crust formed around a nonporous 4.5-µm diameter silica core results in extremely hard, spherical, 5-µm ZORBAX Poroshell particles. Molecules diffusing the short distance into and out of the thin porous crust are rapidly separated. While the inherent properties of ZORBAX Poroshell columns make them usable with most modern HPLC instruments, the use of superficially porous particle columns of small diameter (2.1 mm, 1.0 mm, and 0.5 mm i.d.) for the separation of proteins and peptides places certain demands on the instrument configuration. This technical overview briefly addresses temperature flow rate and gradient adjustment and then details several of the more important parameters that can influence separation performance. These include: detector peak- width setting, extra-column volume, and flow-cell volume. Optimization of the Agilent 1100 HPLC System for Superior Results with ZORBAX Poroshell Columns Technical Overview ZORBAX Poroshell Column Features ZORBAX Poroshell 300SB columns come in a vari- ety of internal diameters and phases, all being extremely useful for the fast separation of proteins and peptides. Poroshell particles are extremely hard, 5-µm silica spheres, each consisting of a 0.25-µm porous crust formed around a nonporous 4.5-µm core. See Figure 1. With ZORBAX Poroshell packed columns, protein and
This document provides a technical overview of optimizing the Agilent 1100 HPLC system for the separation of proteins and peptides using ZORBAX Poroshell superficially porous particle technology. It highlights how this technology allows for faster separations with reduced peak broadening compared to totally porous particles, even at higher flow rates. The text explains the mechanism of the Poroshell particle structure, which consists of a porous silica crust around a nonporous core, enabling molecules to diffuse in and out quickly.
| Recommended Flow Rate | 1 or 2 mL/min |
|---|---|
| Maximum Temperature | 80 °C |
The flow rate should be reduced 5-fold to maintain the same relative flow rate (linear velocity).
The columns are stable at temperatures up to 80 °C.
The flow rate should be reduced by a factor of five.