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The analysis of small molecules in biological samples is frequently hindered by the presence of protein and various enzyme activities. Many bioassays require removal of protein from samples prior to analysis. Perchloric acid (PCA) precipitation is one of the most extensively used deproteinization protocols since it not only removes most of the protein present in the sample but it also functions to stabilize many of the small molecule analytes.
PCA deproteinization has been successfully used in the preparation of samples prior to quantitation of an array of small molecules, including glycogen, ATP, cAMP, glutathione, antioxidants, etc.
Samples are now deproteinized, neutralized and PCA has been removed. The samples may now be used directly for the relevant assays.