To address the fast detection need towards poisoned patients in clinic,a sensitive,accurate liquid chromatographic-tandem mass spectrometric method was developed here for the qualitative and quantitative determinations of twelve anticoagulant rodenticides in dried blood spots.For the thoroughly punched dried blood spots,conditions influencing the extraction efficiency,including the kind of filter paper card,the water-soaking step and the kind of extraction solvent were investigated.A C18 column was employed as separation phase while ammonium acetate in water(5mmol/L)-ammonium acetate in methanol(5mmol/L) was used as mobile phase by gradient elution.Then the sample was detected using negative electrospray ionization under multiple reaction monitoring mode.The results showed that,with the 903 type filter card as supporting matrix and a water-soaking step,followed by an extraction with methanol solvent containing internal standard for 5min,the extraction efficiencies of the twelve rodenticides ranged from 66% to 115% with their intra-day relative standard deviations less than 15%.Good method validation results were obtained for all the anticoagulant rodenticides.The linearity for pindone was from 20 to 500 ng/mL with a correlation coefficient(r2) of 0.9987,and that for the other eleven analytes were from 5 to 500ng/mL with their r2 of 0.9988-0.9996. The recoveries for all the analytes ranged from 61% to 105%,with the matrix effects of 71%-193% and the intra-day relative standard deviations less than 15%.The established method was accurate,sensitive,rapid and convenient,and was successfully applied in the rapid detection of rodenticides in three clinic blood samples. It provides a new approach for clinical diagnosis and forensic toxicant analysis on anticoagulant rodenticide intoxication accidents.