Introduction <p>Meningiomas are mainly benign brain tumors, but they can evolve to higher grades. The phenomena of these changes are not well-known. Therefore, more basic research is needed. This study attempted to assess the lipidome profile in meningiomas harboring different NF2 mutation statuses (wildtype and mutated). Solid-phase microextraction (SPME) probes were used to sample and extract the metabolites and reduce the invasiveness of lipidomic analysis.</p> Objectives <p>This study aimed to select the set of lipids distinguishing meningiomas with different genotypes using two chromatography methods (hydrophilic interaction chromatography (HILIC) and reversed-phase chromatography (RPLC) in two ionization modes.</p> Methods <p>Brain tumors were obtained during neurosurgical procedures. Then, sampling using SPME fibers was performed directly after the lesion excision. After collecting the whole batch of samples, desorption using an isopropanol-methanol solution was performed. Subsequently, instrumental analysis was carried out using liquid chromatography coupled with high-resolution mass spectrometry. The remaining part of the lesion was stored as paraffin tissue blocks, and then genetic testing was performed to determine the presence of mutations in the NF2 gene.</p> Results <p>Genetic profiling of meningiomas revealed that most lesions had a mutation in the NF2 gene. A wide range of analytes was extracted from the studied tumors using SPME probes. A set of 34 lipids was selected as crucial metabolites in tumor differentiation. A combination of analytes detected in more than one analysis mode demonstrated higher sensitivity and specificity compared to the individual models and increased the differentiation of mutant and wildtype samples.</p> Conclusions <p>SPME coupled liquid chromatography and mass spectrometry, can be successfully applied to the screening of lipids in meningiomas with different NF mutation statuses.</p> Graphical abstract <p></p>