BACKGROUND: Lycii Fructus and its raw juice are widely consumed but may be contaminated with pesticide residues, posing health risks. Traditional methods for pesticide residue detection are often labor-intensive and time-consuming. OBJECTIVE: This study aims to develop a rapid, automated method for screening pesticide residues in Lycii Fructus and its raw juice using a combination of micro solid-phase extraction (μ-SPE) and gas chromatography-quadrupole-time-of-flight mass spectrometry (GC-Q-TOF/MS). METHODS: An automated sample clean-up platform (PAL-RTC) was integrated with μ-SPE technology for sample preparation. Matrix-matched external standards were used for quantification, and method validation was conducted to compare μ-SPE with dispersive solid-phase extraction (d-SPE). Performance parameters including linearity, limit of quantitation (LOQ), recovery rates, and relative standard deviations (RSD) were evaluated. RESULTS: 84.5% of the pesticides showed strong linearity (R2 >
0.99) over the concentration range of 2-1000 μg/L. LOQ for 91.4% of pesticides was below 20 μg/kg, with recovery rates between 70% and 120% and RSD ≤ 20%. Screening detection limits (SDLs) were between 1-20 μg/kg, with 96.8% of pesticides having SDLs below 5 μg/kg. The μ-SPE method demonstrated superior reproducibility at the low spiking level (10 μg/kg), detecting 415 pesticides, compared to 369 for d-SPE. Analysis of 100 Lycii Fructus and 50 raw juice samples revealed the presence of 24 pesticides, including 3 restricted types. CONCLUSION: The μ-SPE method, integrated with PAL-RTC and GC-Q-TOF/MS, offers a more efficient and accurate approach for detecting pesticide residues in Lycii Fructus and its raw juice compared to traditional methods, reducing labor and improving reproducibility. HIGHLIGHTS: Compared to the d-SPE method, the μ-SPE method integrated with PAL-RTC demonstrated better reproducibility and stability at low spiking levels, significantly enhancing the efficiency of sample clean-up.