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Clinical Chemistry 37: 728-733, 1991;
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Clinical Chemistry, Vol 37, 728-733, Copyright © 1991 by American Association for Clinical Chemistry

Quantifying intermediary metabolites in whole blood after a simple deproteinization step with sulfosalicylic acid

K Khan, E Blaak and M Elia
MRC Dunn Nutrition Unit, University of Cambridge, U.K.

We assessed the reproducibility, recovery, and stability of several circulating metabolites--glucose, pyruvate, lactate, alanine, glutamate, glutamine, 3-hydroxybutyrate, acetoacetate, and glycerol--in the presence of sulfosalicylic acid (SSA), which was used to deproteinize blood. The assays, which involved reactions linked to NADH/NAD+, were carried out at 37 degrees C and measured at 355 nm with a Cobas-Bio centrifugal analyzer. The intra- and interbatch CVs were less than 2.1%, except for the interbatch CV for 3-hydroxybutyrate at low concentration (15-30 mumol/L), which was 5.4%. Analytical recovery of metabolites added to blood ranged from 96.4% to 103.0%. Of the metabolites studied, all were stable at -20 degrees C for 90 days in the SSA-blood extract, except for glutamine and acetoacetate, which progressively decreased with time. We conclude that these nine circulating metabolites can be satisfactorily measured after a single deproteinizing step with SSA. This single-step procedure has several advantages over many of the currently used methods.


The following articles in journals at HighWire Press have cited this article:


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N. Hiscock and B. K. Pedersen
Exercise-induced immunodepression- plasma glutamine is not the link
J Appl Physiol, September 1, 2002; 93(3): 813 - 822.
[Abstract] [Full Text] [PDF]


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Copyright © 1991 by the American Association for Clinical Chemistry.