This application note presents a method for separating peptides using acetonitrile-free gradient elution on SepTech ST150 10-C18. In response to the global shortage of acetonitrile, alternative solvents such as ethanol have been investigated, and the performance of SepTech ST150 10-C18 is compared to three leading competitor materials. The results demonstrate the superior resolution of peptides achieved by SepTech ST150 10-C18 under acetonitrile-free mobile phase conditions.
Agilent Technologies investigated alternative solvents for reversed phase HPLC analysis of peptides due to global shortages in acetonitrile. This document evaluates ethanol as a viable substitute, comparing its performance against acetonitrile and three leading competitor columns.
The study utilizes the SepTech ST150 10-C18 column with a mixture of four peptides (Oxytocin, Angiotensin II, Angiotensin I, and Insulin). Results indicate that while ethanol provides similar solvent strength to acetonitrile, it alters selectivity, causing partial co-elution between insulin and angiotensin I. Additionally, ethanol exhibits higher UV absorbance at 220 nm compared to acetonitrile.
The column has a particle size of 10 micrometers.
The procedure specifies a flow rate of 1.0 mL/min.