<p>In this study, a robust LC-HRMS (QTOF) method was developed for the simultaneous determination of ultrashort- and short-chain perfluoroalkyl substances (PFAS) in complex plant matrices using a porous graphitic carbon column. The method enabled sensitive and selective detection of seven PFAS (C2–C10) in 18 min, including trifluoroacetic acid (TFA), which is known to exhibit poor retention on conventional C18 phases. The retention behavior of the analytes was systematically investigated under isocratic and gradient conditions, revealing a non-linear dependence on methanol content. Based on these findings, an optimized gradient elution program was proposed, ensuring excellent separation and peak shape even when injecting methanol-rich plant extracts—one of the advantages of porous graphitic carbon columns. Calibration curves for all analytes demonstrated good linearity (<i>R</i>² &gt; 0.99) over relevant concentration ranges with accuracy within ± 15%. The method showed good recovery (71–103%) and precision (RSD &lt; 7%) in rapeseed and burr medic methanolic extracts. Application to real plant samples exposed to PFAS-contaminated media (10&#xa0;µg/mL) revealed substantial accumulation of ultrashort-chain PFAS, especially TFA and perfluoropropionic acid (PFPrA). These results support the potential of plant-based remediation strategies and highlight the importance of monitoring short- and ultrashort-chain PFAS in environmental and agricultural systems.</p> Graphical Abstract <p></p>