| In the practice of forensic science,cases of poisoning or even death due to overdose of Phosphodiesterase type 5 inhibitors（PDE5is）are often encountered.At present,few studies on PDE5 is have been reported in forensic science toxicology identification,and the existing detection methods cannot meet the need for simultaneous and rapid analysis of multiple trace PDE5 is in vivo.Therefore,it is of great importance to establish and optimize the detection methods for PDE5 i drugs in biological samples.In this paper,we investigated the screening methods for four common PDE5 i drugs and their metabolites in blood and urine.The main elements are as follows.1.the research progress of PDE5 is in forensic science was outlined,and the abuse,hazards,pre-treatment methods,detection methods and analytical methods of common PDE5 i drugs and their metabolites were introduced.2.A UPLC-MS/MS analytical method was developed for the detection of four common PDE5 i drugs and their metabolites.An ACQUITY UPLC HSS T3（100 mm×2.1mm,1.8 μm）column with mobile phases A: 0.1% formic acid acetonitrile and B: 0.1%formic acid-2 mmol/L ammonium formate aqueous solution was used for gradient elution,and the target compounds were detected in positive ion mode with electrospray ionization source in dynamic multiple reaction monitoring mode.3.A UPLC-MS/MS method was developed for the determination of four common PDE5 is and their metabolites in the blood.Using the precipitated protein method,the detection limits of the 16 targets were 0.1-10 ng/m L,the quantification limits were 0.25-20ng/m L,the correlation coefficients R were greater than 0.99,the recoveries were75.5%-110.9%,the matrix factors were 2.1%-20.2%,the intra-day and inter-day precision were less than 15%,the accuracy was within ±15%,the precision at the limit of quantification was less than 20%,and The accuracy was within ±20%,and the stability of the 14 targets was good to meet the requirements of quantitative analysis.4.A UPLC-MS/MS method was established for the determination of four common PDE5 is and their metabolites in urine.Using the precipitated protein method,the detection limits of 16 targets were 0.1-10 ng/m L and the quantification limits were 0.25-20 ng/m L,with the correlation coefficients R greater than 0.99,the recoveries were 90.0%-114.0%,the matrix factors-10.0%--0.6%,the intra-day and inter-day precision were less than 15%,the accuracy was within ±15%,the precision at the limit of quantification was less than20% The accuracy was within ±20%,and the stability of the 14 targets was good to meet the requirements of quantitative analysis.5.An animal experimental model was established,in which rats were gavaged with clinical therapeutic doses of PDE5 is,and the blood and the urine were collected 2 h later and analyzed according to the established method.The results showed that 4 prodrugs and10 metabolites were detected in blood and urine,which verified the feasibility and validity of the method.The phase II metabolites of tadalafil were characterized by ultra performance liquid chromatography-high resolution mass spectrometry,and the detection method established in this study can provide a reference for the determination of PDE5 is poisoning cases. |
