A simple, miniaturized microplate chemiluminescence assay for determination of methyl parathion (MP) was developed in 384-microwell plates. Zirconia (ZrO2) was added in microwell for adsorption of acetylcholinesterase (AChE). The developed assay is based on inhibition of AChE by MP. A good dynamic range 0.008-1,000 ng/mL was obtained for MP with limit of detection 0.008 ng/mL. Intrabatch and interbatch reproducibility for miniaturized assay was obtained with % RSD up to 3.07 and 5.66, respectively. In 384 well plate formats, 70 samples were simultaneously analyzed within 20 min with assay volume of 41.5 mu L. The application of developed assay was extended for MP remediation. Column containing ZrO2 was utilized for remediation where MP was selectively adsorbed. Under optimized condition, adsorption of MP on ZrO2 was found to be 98-99% with 2-h contact time in real water samples. Adsorption of MP on ZrO2 column followed by quantification using developed bioassay provides a novel approach to monitor remediation. The applicability of assay was successfully extended for determination of MP in water samples after removal through ZrO2.