A Method Comparison For The Simultaneous Determination Of 4 Antidepressive Drugs In Human Urine By HPLC And GC/MS

Objectives1. Explore the best extraction separation method of four antipressive drugs including Fluoxetine、Paroxetine、Sertraline and Venlafaxine in human urine.2. Establish gas chromatography/mass spectrometry(GC/MS) of four antipressive drugs including Fluoxetine、Paroxetine、 Sertraline and Venlafaxine in human urine.3. Establish high performance liquid chromatography(HPLC) of four antipressive drugs including Fluoxetine、 Paroxetie、Sertraline and Venlafaxine in human urine.4. Validate the most suitable analysis method by comparing two methods and provide the method data for antipressant clinical detection and forensic toxicological analysis.MethodsLiquid-Liquid Extraction(LLE) and Solid Phase Extraction(SPE) were used as extraction separation methods. GC/MS method and HPLC method were used on study. Total ion current retention time comparison method and mass spectrogram matching method were used as qualitative analysis method of GC/MS method. Retention time comparison method was used as qualitative analysis method of HPLC method. Internal standard method was used as quantitative analysis method.ResultsThe recovery values for samples extracted by LLE were ranged from 57.23% to 71.27% in the GC/MS method and from 64.22% to 71.19% in the HPLC method, for samples extracted by SPE were ranged from 71.84% to 82.17% in the GC/MS method and from 75.29% to 80.84% in the HPLC method.In the GC/MS method, retention times of Fluoxetine, Venlafaxine, Sertraline and Paroxetine were expressed at 4.679min,6.163min,8.015min and 9.687min. The calibration curves for Fluoxetine, Venlafaxine and Paroxetine were linear within the range of 20gμ/ml～500μg/ml, for Sertraline was linear within the range of 20μg/ml-2000μg/ml, R2> 0.997. LODs and LOQs of the analytes were found to be between 0.5μg/ml～1μg/ml and 1μg/ml～20μg/ml. Accuracies of the analytes were found to be between 84.90% and 94.16%. Intra-day precisions and Inter-day precisions of the analytes were found to be between 4.82%～8.28% and 5.86%～12.30%.In the HPLC method, retention times of Fluoxetine, Venlafaxine, Sertraline and Paroxetine were expressed at 7.999min,3.959min,8.159min and 6.603min. The calibration curves for Fluoxetine and Paroxetine were linear within the range of 10μg/ml-500μg/ml, for Venlafaxine was linear within the range of 5μg/ml～500μg/ml, for Sertraline was linear within the range of 5μg/ml～2000μg/ml with LLE and within the range of 10μg/ml～2000μg/ml with SPE, R2> 0.997. LODs and LOQs of the analytes were found to be between 0.5μg/ml-1μg/ml and 1μg/ml～10μg/ml. Accuracies of the analytes were found to be between 89.64% and 99.32%. Intra-day precisions and Inter-day precisions of the analytes were found to be between 1.96%～5.84% and 4.95%～8.33%.ConclusionA GC/MS method and a HPLC method of the determination of four antipressive drugs including Fluoxetine、Paroxetine、 Sertraline and Venlafaxine in human urine were established, which could also used to analyze one of the four antipressive drugs. SPE should be chosen to extract the analytes by comparing two extraction separation methods.With the comparison of two analysis methods, if the aim of study was qualitative analysis, the GC/MS method should be chosen. If the aim of study was quantitative analysis, the HPLC method should be chosen, which had a higher accuracy and a higher precision than the GC/MS method.