Quantitative Detection Of 5-HT₃ Receptor Antagonists In Plasma By SPE-HPLC-MS/MS

Selective three 5-HT₃ receptor antagonists,including ondansetron?OND?,granisetron?GRA?and tropisetron?TRO?,are effective and potent antiemetic drugs in the prevention of vomiting and nausea resulting from surgery and cancer chemotherapy.An essential task for the investigation of the pharmacokinetics,bioavailability and metabolism during the therapeutic treatment is to quantify the concentrations of 5-HT₃ receptor antagonist in biological fluids.As a consequence,development of reliable approaches to monitor 5-HT3receptor antagonist in complex matrices is of significance.However,with the complexity of the sample matrix,it is still a very arduous task.Before the analysis of chromatography-mass spectrometry,a high efficient and high selective pretreatment method is developed to effectively remove the matrix effect and not to affect the detection of actual samples.In this paper,the solid phase extraction?SPE?pretreatment technique combined with a triple quadrupole liquid-mass spectrometer was used to optimize and establish a quantitative method for the determination of three strong alkaline 5-HT₃ receptor antagonists,i.e.ondansetron?OND?,granisetron?GRA?and tropisetron?TRO?in goat plasma.A high efficiency and selectivity SPE method for simultaneous extraction of OND,GRA and TRO was developed.A SPE method combined with high performance liquid chromatography?HPLC?tandem mass spectrometry?MS/MS?was established for the simultaneous determination of OND,GRA and TRO in goat plasma.The samples were extracted by precipitation protein and Silica SPE column.The HPLC separations were performed on a Thermo Accucore HILIC column?2.6?m,100 mm×2.1 mm?protected by a Thermo Accucore HILIC guard column?2.6?m,10 mm×2.1 mm?.Gradient elution was carried out using methanol-ammonium formate?10mM?-formic acid?0.1%,vol.%?as mobile phase.Multiple reaction monitoring?SRM?mode using electrospray ionization and external standard method was used for MS/MS quantification.The experiment data indicates that the linear correlation coefficient of the three 5-HT₃ receptor antagonists between 0.2 ng/mL²0ng/mL was greater than 0.99 with a quantitative detection limit of 0.2 ng/mL.Quality control samples were selected from goat plasma for low,medium and high concentrations.The precision of the intra-and inter-day assay for OND,GRA and TRO were 1.84-6.23%and3.89-5.31%,2.63-6.29%and 3.76-5.31%,1.99-5.67%and 2.64-4.70%respectively.The accuracy of the intra-and inter-day assay for OND,GRA and TRO were 89.15-97.39%and89.46-95.17%,91.08-100.82%and 91.24-99.47%,92.30-100.74%and 94.21-97.90%respectively.The mean absolute recoveries were 103-150%,115-121%,and 98-141%for OND,GRA and TRO respectively.Furthermore,the mean process efficiency values of silica SPE were 98-135%,92-124%,and 72-109%for OND,GRA and TRO respectively.SPE-HPLC-MS/MS method is suitable for quantitative determination of OND,GRA,and TRO in plasma.