Impact of storage temperature, storage duration, and deproteinization on plasma amino acid concentrations in dogs

Reliability of canine plasma amino acid analysis depends on sample stability which can be influenced by pre-analytical handling techniques, storage temperature, storage time, and deproteinization status. Extrapolating data to dogs from research in other species is limited given discordant methodology and interspecies differences. The present study investigated the effects of deproteinization status (non-deproteinized or deproteinized) and storage temperature (at -20 °C or - 80 °C) on the concentration of 22 canine plasma amino acids during a 300-day storage period. Storage time had a significant effect (p < 0.05) of overall declining concentration of most amino acids. Compared to non-deproteinized samples, deproteinization contributed to overall higher concentrations of cyst(e)ine and glutamic acid, and consistently modified the effect of storage time and temperature on cyst(e)ine, glutamic acid, and glutamine. Compared to -20 °C, storage at -80 °C contributed to a higher concentration of cyst(e)ine and glutamic acid, and modified the effect of storage time on arginine, glutamic acid, glutamine, and tryptophan. Storage time had a consistent, significant effect on amino acid concentrations in canine plasma samples. Although sample deproteinization and low storage temperature modified the effect of storage time, these interactions were variable among analyzed amino acids. Therefore, timely sample analysis is recommended. If delayed sample analysis is inevitable, deproteinization should be performed prior to sample banking to preserve amino acid stability.