Pre-analytical stability of novel cerebrospinal fluid biomarkers

Stability of the cerebrospinal fluid (CSF) composition under different pre-analytical conditions is relevant for the diagnostic potential of biomarkers. Our aim was to examine the pre-analytical stability of promising CSF biomarkers that are currently evaluated for their discriminative use in various neurological diseases. Pooled CSF was aliquoted and experimentally exposed to delayed storage: 0, 1, 2, 4, 24, 72, or 168 h at 4 degrees C or room temperature (RT), or 1-4 months at - 20 degrees C; or up to 7 freeze/thaw (f/t) cycles, before final storage at - 80 degrees C. Eleven CSF biomarkers were screened using immunoassays, liquid chromatography, or enzymatic methods. Levels of neurogranin (truncP75), chitinase-3-like protein (YKL-40), beta-site amyloid precursor protein cleaving enzyme 1 (BACE1), acetylcholinesterase (AChE) enzymatic activity, theobromine, secreted protein acidic and rich in cysteine-like 1 (SPARCL-1) and homovanillic acid (HVA) levels were not affected by the applied storage conditions. 3-Methoxy-4-hydroxyphenylglycol (MHPG) levels linearly and strongly decreased after 4 h at RT ( - 10%) or 24 h at 4 degrees C ( - 27%), and with 6% after every f/t cycle. 5-Methyltetrahydrofolate (5MTHF) ( - 29% after 1 week at RT) and 5-hydroxyindoleacetic acid levels (5-HIAA) ( - 16% after 1 week at RT) were reduced and 3,4-dihydroxyphenylacetic acid (DOPAC) levels ( + 22% after 1 week at RT) increased, but only after > 24 h at RT. Ten out of eleven potential CSF novel biomarkers showed very limited change under common storage and f/t conditions, suggesting that these CSF biomarkers can be trustfully tested under the pre-analytical conditions present across different cohorts.