Related Experiment Video
Updated: Apr 15, 2026
Absolute Quantification of Aβ1-42 in CSF Using a Mass Spectrometric Reference Measurement Procedure
Published on: March 21, 2017
Importance and impact of preanalytical variables on Alzheimer disease biomarker concentrations in cerebrospinal fluid
Nathalie Le Bastard1, Peter Paul De Deyn2, Sebastiaan Engelborghs3
1Reference Center for Biological Markers of Dementia (BIODEM), Laboratory of Neurochemistry and Behavior, Institute Born-Bunge, University of Antwerp, Antwerp, Belgium; Current affiliation: Fujirebio Europe, Ghent, Belgium;
Background:
Analyses of cerebrospinal fluid (CSF) biomarkers (β-amyloid protein, total tau protein, and hyperphosphorylated tau protein) are part of the diagnostic criteria of Alzheimer disease. Different preanalytical sample procedures contribute to variability of CSF biomarker concentrations, hampering between-laboratory comparisons. The aim of this study was to explore the influence of fractionated sampling, centrifugation, freezing temperature, freezing delay, and freeze-thaw cycles on CSF biomarker analyses.
Methods:
We studied fractionated sampling in sequential aliquots of lumbar CSF. Centrifuged and noncentrifuged samples from the same fraction were compared. CSF samples were subjected to different protocols (liquid nitrogen, -80 °C, and -20 °C; 24 h at 2-8 °C; and 24 and 48 h at room temperature). To study the influence of freeze-thaw cycles, samples were thawed up to 4 times and refrozen at -80 °C. CSF was collected in polypropylene tubes. We measured CSF biomarker concentrations with commercially available single-analyte Innotest assays.
Results:
CSF biomarker concentrations from non-blood-contaminated samples are not influenced by centrifugation or fractionated sampling. Freezing temperature and delayed storage can affect biomarker concentrations; freezing of CSF samples at -80 °C as soon as possible after collection is recommended. Consecutive freezing and thawing of CSF samples up to 3 times had little effect.
Conclusions:
Temperature of freezing, delay until freezing, and freeze-thaw cycles significantly influence CSF biomarker concentrations, stressing the need for standard operating procedures for preanalytical sample handling. The differences observed in this study are, however, relatively small, and the impact on the clinical value of these CSF biomarkers needs to be determined.
