Delayed Deproteinization Causes Methodological Errors in Amino Acid Levels in Plasma Stored at Room Temperature or -20°C

Deproteinization has been recognized as a prerequisite for amino acid analysis of plasma samples. For plasma stored at room temperature, delaying deproteinization for 30, 60 or 120 minutes did not result in significant changes in the mean CV (coefficient of variation), which ranged from 4.4 to 5.6%. However the mean CV of aspartic acid, α-aminoadipic acid, alanine and lysine was about 10%. When the plasma was stored frozen at -20°C, the CV was increased at 0 and 120 minutes after thawing, to 12.4% (range, 4.1 to 35.3%) and 8.0% (2.5 to 30.7%), respectively. The concentrations in plasma during storage at room temperature of all the amino acids analyzed showed significant changes. In plasma stored for 30 minutes at room temperature, 17 amino acids increased in concentrations and two decreased. Extending this period to 60 or 120 minutes increased the instability as compare to the reference group. Storing plasma at -20°C for 2 weeks resulted in significantly greater changes in the amino acid concentrations than at room temperature. On extending the storage time at room temperature, after thawing, to 30, 60, and 120 minutes, 21, 20, and all 22 amino acids respectively changed significantly (p<0.01). The present study indicates that methodological errors occur in the concentrations determined for all amino acids when plasma is left at room temperature. The storage of frozen non-deproteinized plasma accompanied more significant changes in most amino acid concentrations and thus should be avoided. Deproteinization should be performed as soon as possible after plasma collection.