Glyphosate and glufosinate are a class of carcinogenic,non selective and internally suction herbicides,which have received more and more attention from the society.Much work about inspection or identification of glyphosate and glufosinate in the drug related cases are required as poisoning cases of this two herbicides poisoning,suicide and other causes also occur from time to time.To develop a detection method for such herbicides in biological fluids,eg.blood and urine could provide a technical support for the qualitative and detection of cases and provide a reference for clinical toxic analysis and treatment.Therefore,a non derivative high performance liquid chromatography-tandem mass spectrometry was established for the rapid detection of 8 polar pesticides such as glyphosate and glufosinate and their metabolites in biological fluids.8 polar pesticides were separated on a Metrosep A Supp 5 anion column(150 mm×4.0 mm,5 μm),with pure water-200 mmol/L ammonium hydrogen carbonate solution containing 01% ammonia water as mobile phase by gradient elution,and analyzed in negative ion multiple reaction monitoring(MRM) mode.In this study,the chromatographic and mass spectrometry conditions for the instrument were optimized,and the blood and urine of biological fluids were studied.The blood and urine samples were mixed with water and oscillated,then freeze centrifugated with appropriate amount of acetonitrile for 10 min.The supernatant was in turn passed through a Cleanert PEP Plus solid phase extraction column and a 0.22 μm organic micro porous membrane,then collected for analysis.The experimental results showed that there were good linear relationships for 8 polar pesticides in the range of 0.5-50 ng/mL with their correlation coefficients(r2) larger than 0.99.The limits of detection(LOD，S/N≥3) and the limits of quantitation(LOQ，S/N≥10) of this method were in the ranges of 0.08-0.3 ng/mL and 0.3-1 ng/mL,respectively.The matrix effects of the method in blood and urine were in the range of 86.5%-106%，and the recoveries for the target compounds ranged from 81.5% to 114%,with intra day relative standard deviations(intra day RSD,n=6) and inter day RSD of 0.30%-2.8% and 0.50%-5.3%,respectively.With the advantages of simple operation,time saving and labor saving,high sensitivity,good accuracy and good reproducibility,the developed method could be used for the rapid and accurate qualitative and quantitative analysis of 8 polar pesticides,to meet the testing and identification requirements of glyphosate,glufosinate related toxic cases.