Neonicotinoids (NEOs) are used as pesticides to combat plant-damaging insects, but they can also pose risks to human health, potentially causing cancer, infertility, and congenital disabilities. Therefore, monitoring food residues is essential to protect human health and maintain ecological balance. This study developed and validated a new method for measuring residues of clothianidin, acetamiprid, imidacloprid, dinotefuran, nitenpyram, thiamethoxam, and thiacloprid in wheat. A STRATA XPRO column (Solid-Phase Extraction) was employed to remove interferents and concentrate the NEOs, and a reverse-phase high-performance liquid chromatography equipped with a Kinetex C18 column and Diode Array Detector (DAD) for dosage was also used. The NEO recoveries using the STRATA XPRO cartridge were compared to those obtained with the commonly used QuEChERS method. The STRATA XPRO technology was faster and provided better recoveries for clothianidin (97.7-98.2%) compared to QuEChERS (69.8-80.3%). The accuracy profile strategy, recommended by the Commission of the Société Française des Sciences et Techniques Pharmaceutiques (SFSTP), was used to validate the entire analytical procedure. The method demonstrated recoveries between 70% and 110%, a robust linear relationship between observed and predicted values (R² = 0.999), a limit of quantification higher than the maximum concentration of NEOs to be determined, an accuracy below 10%, (meeting the European Commission’s requirements), and a minimum of 95% of outcomes within the acceptance limits of ±15%. The low cost, along with the speed (~40 min ) and simplicity of operation, recommend its application for routine analyses.