Hybrid QuEChERS–Micro-Solid Phase Extraction for the Determination of Chloramphenicol in Honey by HPLC-MS/MS: Method Validation and Matrix-Effect Assessment

An HPLC-MS/MS method combining modified QuEChERS-based extraction with micro-solid-phase extraction (MSPE) using C18P-PVDF microspin filters was developed and validated for chloramphenicol (CAP) determination in honey. At validation levels of 0.10, 0.15, and 0.30 µg kg−1, apparent recoveries ranged from 102.1% to 105.1%, repeatability (RSDr) from 4.5% to 9.5%, and within-laboratory reproducibility (RSDwR) from 6.0% to 12.5%. The decision limit (CCα) was 0.123 µg kg−1, below the reference point for action of 0.15 µg kg−1, while the estimated limit of detection (LOD; not experimentally verified) was 0.01 µg kg−1.. Matrix-effect assessment across 20 honey samples showed responses within the 80–120% range for the proposed procedure and the SPE and QuEChERS comparators. No CAP was detected in the authentic honey samples. The developed workflow provides a practical, validated alternative for routine CAP determination in honey.