A new reverse phase media, Agilent ZORBAX RRHD 300SB-C18 1.8µm, was used for the separation of a typical protein biopharmaceutical, insulin.
The document details a method for separating and analyzing oxidized insulin chains (Chain A and Chain B) from bovine pancreas. It utilizes the Agilent ZORBAX Rapid Resolution High Definition (RRHD) column technology, which features 1.8 µm StableBond particles designed for high-pressure UHPLC systems.
The study demonstrates that this media provides robust separations under acidic conditions required for reversed-phase analysis of proteins. Key findings include rapid separation times (under five minutes), excellent reproducibility after 200 consecutive injections, and the ability to resolve various insulin isoforms and heat-degradation products.
| Pore Size | 300 Å |
|---|---|
| Max Pressure | ~1200 bar |
| Flow Rate | 1.0 mL/min |
The standardized conditions used a flow rate of 1.0 mL/min.
Yes, the StableBond technology allows for robust separations under acidic conditions containing trifluoroacetic acid or formic acid.
The column demonstrated excellent reproducibility after 200 consecutive injections of insulin without cleaning.