Online aptamer-based solid-phase extraction coupled with high-performance liquid chromatography-mass spectrometry for the determination of Alzheimer's disease biomarkers
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Updated: Aug 5, 2026
A High Throughput, Multiplexed and Targeted Proteomic CSF Assay to Quantify Neurodegenerative Biomarkers and Apolipoprotein E Isoforms Status
Published on: October 20, 2016
Israel Donizeti de Souza1, Maria Eugenia Costa Queiroz2, Valerie Pichon1
1Department of Analytical, Bioanalytical Sciences and Miniaturization (LSABM) Chemistry, Biology and Innovation (CBI) UMR 8231, ESPCI Paris, PSL University, CNRS, 10 Rue Vauquelin, Paris, 75005, France.
Background:
Reliable quantification of amyloid-β (Aβ) peptides in biological fluids is of major clinical and research interest in Alzheimer disease context. Conventional offline extraction approaches often involve labor-intensive manual steps and high solvent consumption, limiting throughput routine applications, reproducibility, and sustainability. To address these limitations, this study reports the development of an online coupling between a monolithic oligosorbent (mOS) in capillary and high-performance liquid chromatography-mass spectrometry (HPLC-MS) method for selective Aβ peptide analysis.
Results:
A mOS was incorporated online in a set up including a C18 trap column coupled with an HPLC-MS analytical system. Systematic optimization of mOS loading/elution conditions, trapping column desalting/preconcentration, and chromatographic mobile phase composition enabled efficient retention, transfer, and separation of Aβ40 and Aβ42 in a fully automated method. This method achieved lower limits of quantification down to 0.03 ng mL-1 with good precision and accuracy (CV ranging from 1.1 to 6.2% for Aβ40 and from 4.3 to 10.9% for Aβ42). Comparative evaluation of offline and online extraction using the same mOS capillary demonstrated improved reproducibility and enhanced sensitivity for the online configuration. The method's applicability to a controlled CSF-like matrix was demonstrated using artificial cerebrospinal fluid (aCSF) diluted 1:2 (v/v) in binding buffer (BB), yielding recoveries of 60% for Aβ40 and 34% for Aβ42.
Significance:
The online coupling of the mOS capillary with HPLC-MS also represents a strategic advance toward environmentally responsible bioanalysis. Indeed, the superior AGREEprep score (0.7) for the online configuration compared to offline mode (0.59), underscores its alignment with green analytical chemistry principles, reflecting reductions in solvent use, manual intervention, and overall environmental burden.
