A surface plasmon resonance (SPR) immunoassay using a PH-BSA immobilized sensor chip was developed to measure phosmet in food samples. The in-competitive inhibition assay showed highly sensitive and good specificity of the cross-reactivity with analogue structure pesticides. The biosensor exhibited a linear detection range from 8.0 to 60.0 ng/L of phosmet with a lower detection limit of 1.6 ng/L (S/N = 3). The sensitivity obtained with the present SPR affinity biosensor was significantly higher than most of the sensors reported with different measurement methodologies for phosmet. A recovery test of pesticide quantification in peaches, apples, cabbages, and rapes was also studied. Good recoveries (86.4-102.8%) and coefficients of variation (CVs) (5.1-12.6%) were obtained in all cases. The SPR biosensor assay method was compared with cd-ELISA in terms of analysis time, antibody dosage, recoveries, precision, detection limit, pretreatment, and testing costs, and clear advantages could be seen over the traditional ELISA-based detection systems. The developed SPR method was suitable for the rapid quantitative or qualitative determination of phosmet in agricultural products.