The invention belongs to the technical field of bioengineering and
pesticide residue detection, and discloses a method for detecting organophosphorus
pesticide by a two-
enzyme colorimetric method based on immobilized
plant esterase, which comprises the following steps of: (1) respectively mixing every 50 microliters of immobilized
plant esterase with final concentration of 1mg / mL with 50 microliters of organophosphorus standard sample solutions with different concentrations, taking
distilled water as a blank control, and taking
distilled water as a standard control; the preparation method comprises the following steps of: performing incubation at
room temperature, sequentially adding HRP (
horse radish peroxidase), a substrate 1-NA, TMB (tetramethylbenzidine), H2O2 and an
acetic acid buffer solution into a reaction solution, oscillating, incubating, measuring an
absorbance value A659, and calculating an inhibition ratio I; and (2) taking a to-be-detected sample, operating according to the step (1), calculating the inhibition ratio I, and substituting the inhibition ratio I to obtain the concentration of the organophosphorus
pesticide in the to-be-detected sample. The method is simple and rapid to operate, low in cost, good in stability, high in OPs detection sensitivity and wide in detection range, and has a relatively good application prospect in rapid screening and detection of organophosphorus pesticide detection in
fruits and vegetables.