Reversed-phase HPLC is used for the quantitative analysis of proteins. Proteins are separated based on the difference of hydrophobicity. Sample preparation is necessary when analyzing biological samples to reduce the affect from matrix.
Purification of biological samples using revers-phase solid phase extraction requires acetonitrile or methanol contained solvent for analyte elution. However, there is a case high recovery can’t be obtained due to a few factors: slow partition, precipitation by high concentration of organic solvent and strong adsorption to packing material.
In this note, MonoSpin, uniquely suited for handling small amount samples, is used to purify the proteins. During the process, changes in selectivity and recovery are seen due to the usage of different kind of organic solvent.