- Communication
In Situ Anion-Generating Molecularly Imprinted Solid-Phase Extraction Coupled with HILIC-MS/MS for Determination of Metanephrines in Low Volume of Plasma
- Antons Podjava and
- Artūrs Šilaks
Metanephrine (MN) and normetanephrine (NMN) are critical biomarkers for neuroendocrine tumors (pheochromocytoma and paraganglioma). Following our previous development of a molecularly imprinted solid-phase extraction (MISPE) sorbent for urine analysis, this study evaluated MISPE coupled with HILIC-MS/MS for determining metanephrines in human plasma. Unlike conventional phases, the novel polymer selectively binds analytes as in situ-generated anions via quaternary alkylammonium groups in hydroxide form, ensuring accurate extraction from just 25 µL of plasma. Validated per U.S. FDA guidelines, the assay showed good intra- and interday precision (CV < 10.8%), accuracy (bias < −10.6%) and excellent linearity (R2 > 0.99) across pathological ranges (184.3–877.8 ng/L for MN; 174.8–923.0 ng/L for NMN), with low relative standard errors (<6.9%). Excellent selectivity was demonstrated in the presence of structurally close analogs (catecholamines, DOPA and its derivatives). Compared with commercial WCX, the sorbent yielded cleaner extracts, significantly reducing the phospholipid interference. Although lower limits of quantification (92.2 ng/L MN; 87.4 ng/L NMN) slightly exceeded healthy upper thresholds, the method has potential for use in specific clinical scenarios with pronounced biomarker elevations: diagnosis of pheochromocytoma/paraganglioma, monitoring post-treatment metanephrine decline, and tracking tumor-induced hypertensive crises in emergencies. This accessible protocol forms a solid foundation for advanced diagnostics.
19 June 2026








