Hydrophobic covalent organic framework as a solid-phase microextraction coating for high-throughput quantification of...

This study successfully synthesized a hydrophobic covalent organic framework (COF) material and applied it as solid-phase microextraction (SPME) coating for the detection of endocannabinoids N-arachidonoylethanolamine (AEA) and 2-arachidonoylglycerol (2-AG) in rat plasma. Under optimized extraction conditions, a high-throughput sample pretreatment method was established by integrating the prepared hydrophobic COF-SPME fibers with a 96-well plate platform, coupled with liquid chromatography-tandem mass spectrometry for analysis. The method enables the extraction of 96 samples within 25 min, significantly improving sample processing efficiency. In methodological validation, AEA and 2-AG showed good linearity in the ranges of 0.025-4.0 ng mL^-1 and 0.1-10.0 ng mL^-1, respectively, with correlation coefficients R² ≥ 0.9988. The limits of detection for both analytes were in the range of 0.008-0.020 ng mL^-1, recoveries ranged from 103.93% to 117.97%, and relative standard deviations were below 10.97%. When applied to plasma samples from Parkinson's Disease (PD) model rats, the results indicated significant alterations in AEA and 2-AG levels compared to the control group. This study demonstrates that the established method is suitable for rapid and highly sensitive detection of endocannabinoids in complex biological samples, providing a valuable analytical tool and reference for investigating the pathological mechanisms of PD.