Advanced SearchSearch Tips
Delayed Deproteinization Causes Methodological Errors in Amino Acid Levels in Plasma Stored at Room Temperature or -20℃
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Delayed Deproteinization Causes Methodological Errors in Amino Acid Levels in Plasma Stored at Room Temperature or -20℃
Li, Junyou; Piao, Chunxiang; Jin, Huazi; Wongpanit, Kannika; Manabe, Noboru;
  PDF(new window)
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, -aminoadipic acid, alanine and lysine was about 10%. When the plasma was stored frozen at -20, 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 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.
Amino Acid Assay;Plasma Concentration;Deproteinization;Storage;Blood Sample;
 Cited by
Analytic validation of a gas chromatography–mass spectrometry method for quantification of six amino acids in canine serum samples, American Journal of Veterinary Research, 2015, 76, 12, 1014  crossref(new windwow)
Boucher, J. L., C. Charret, C. Coudray-Lucas, J. Giboudeau and L. Cynober. 1997. Amino acid determination in biological fluids by automated ion-exchange chromatography: performance of Hitachi L-8500A. Clin. Chem. 43:1421-1428

Endo, J. and Y. Notsu. 1992. Advance in automated amino acid analysis. J. Clin. 50:69-75

Faurschou, M. and N. Borregaard. 2003. Neutrophol granules and secretory vesicles in inflammation. Microbes Infect. 5:1317-1323 crossref(new window)

Jeffrey, D. H., D. Bethea, K. Ho, S. P. Huang, D. L. Ricci, J. Gregory, S. A. Opiteck and A. Hefta. 2004. An investigation of plasma collection, stabilization, and storage procedures for proteomic analysis of clinical samples. Clin. Proteomics J. 1: 17-31 crossref(new window)

Leon, H. de J. and M. Breuer. 1996. Evaluation of systematic errors due to deproteinization, calibration and storage of plasma for amino acid assay by ion-exchange chromatography. J.Chromatogr. 677:61-68 crossref(new window)

Olek, K., S. Uhlhaas, P. Wardenbach and M. Yamaguchi. 1979. Influence of storing conditions on the amino acid concentration in human serum. J. Clin. Chem. Biochem. 17:599-604

Parvy, P., J. Bardet, D. Rabier and P. Kamoun. 1991. Methodological errors in the amino acid assay in biological fluids. Ann. Biol. Clin. 27:180-182

Peng, Y., J. K. Tews and A. E. Harper. 1972. Amino acid imbalance, protein intake, and changes in rat brain and plasma amino acids. Am. J. Physiol. 222:314-319

Perry, T. L. and S. Hansen. 1969. Technical pitfalls leading to errors in the quantitation of plasma amino acids. Clin.Chim. Acta 25:53-58 crossref(new window)

Robbie, L. and P. Libby. 2001. Inflammation and atherothrombosis. Ann. NY. Acad. Sci. 947:167-179

Sahai, S. and S. Uhlhaas. 1985. Stability of amino acids in human plasma. Clin. Chim. Acta 148:255-259 crossref(new window)

Ayache, S., M. Panelli, F. M. Marincola and D. F. Stroncek. 2006. Effects of storage time and exogenous protease inhibitors on plasma protein levels, Clin. Chem. 126:174-184 crossref(new window)

Sarwar, G. and H. G. Botting. 1993. Evaluation of liquid chromatographic analysis of nutritionally important amino acids in food and physiological samples. J. Chro. 615:1-2 crossref(new window)

Schaefer, A., F. Piquard and P. Haberey. 1987. Plasma amino-acids analysis: effects of delayed samples preparation and of storage. Clin. Chim. Act 164:163-169 crossref(new window)

Spackman, D. H., W. H. Stein and S. Moore. 1958. Automatic recording apparatus for use in the chromatography of amino acids. Anal. Chem. 30:1190-1206 crossref(new window)

Perry, T. L. and S. Hansen. 1969. Technical pitfalls leading to errors in the quantitation of plasma amino acids. Clin. Chim. Acta. 25:53-58 crossref(new window)

Weiss, S. J. 1989. Tissue destruction by neutrophile. N. Engl. J. Med. 320:365-376

Yoneda, T., M. Yoshikawa, A. Fu, K. Tsukaguchi, Y. Okamoto and H. Takenaka. 2001. Plasma levels of amino acids and hypermetabolism in patients with chronic obstructive pulmonary disease. Nutrition 17:95-99 crossref(new window)

Veresegyhazy, T., H. Febel and A. Rimanoczy. 2001. Absorption of leucine, alanine and lysine from the rumen. Acta Vet. Hung. 49:81-86 crossref(new window)