Development Of Urine Heavy Metal Profiling Methods And Its Application In The Study Of Idiopathic Male Infertility

Infertility has been a major scientific issue that needs to be addressed in reproductive health research.Many heavy metals have been confirmed to pass through the blood testosterone,and their harm to male reproductive health has become a hotspot.However,so far,the study of health effects of heavy metals is still limited to single or some kinds of heavy metals,it is difficult to objectively and comprehensively screen the main heavy metal exposure that endangers human health.Therefore,this study focused on the influence of heavy metal exposure to male reproduction,using the exposure profiling strategy.In the current dissertation,we established a method to study the exposure profiling of heavy metals in urine by optimizing the sample pretreatment method and we optimized and evaluated it using a series of methodological methods.The method was not only accurate,high precision,but also had the characteristics of simple operation and small sample demand.This method was applied to the study of Idiopathic male infertility.It was found that the increase of Co and Mo were found to be toxic to male.These studies provided evidence of the epidemiological evidence and a new understanding of the potential male reproductive toxicity of heavy metals and provided scientific data for the development of a reasonable range of reference values for heavy metals in adult males.Part ? The Development and Evaluation of the ICP-MS Based Urine Heavy Metal Profiling MethodObjective To establish the urine heavy metal profiling detection technology and to realize the overall analysis of heavy metals in urine.Methods The urine heavy metal profiling method was established by the method of sample pretreatment,standard curve preparation and internal standard selection using ICP-MS.The urine heavy metal profiling method was evaluated by linearity,accuracy and precision.Results Among the 62 elements,the detection limit ranged from 0.00003 ?g/L to 0.78768 ?g/L,the linear range ranged from 0.005 ?g/L to 200 ?g/L(0.01 to 200 mg/L for macro elements),the recovery rate ranged from 85 to 110%,the relative standard deviation was less than 12%.Conclusion Compared with the traditional quantitative analysis of elements,this urine profiling method can detect 62 elements at the same time.In addition,this method is simple with a small urine sample volume and has high-throughput and unbiased characteristics.Part ? Study on the Association Between the Urine Heavy Metal Profiling and Idiopathic Male InfertilityObjective Using the established heavy metal element profiling method,the urine samples of idiopathic male infertility and normal control were analyzed to find the differences between the heavy metals.Methods Morning urine samples from idiopathic male infertility cases(596)and controls(366)were selected to analyze the urine heavy metal profiling.The multidimensional pattern recognition model was established,and the reliability of the difference elements was verified by the rank sum test and the regression analysis,finding the different heavy metals in male infertility and identifying the interaction between heavy metal exposure and male infertility.Results In the rank sum test,the concentrations of Ti,Mn,Co,Cu,Se,Sr,Zr,Mo,Ag,Yb,Ta and Hg in the urine of idiopathic male infertility patients were significantly different from those of in the healthy control group(P<0.05).The elements Mo,Se,Ag,Co and Hg were contributors to the differences in the multivariate statistical model OPLS-DA.As a result of the regression analysis,the elements Co,Sr and Mo were related to the occurrence of idiopathic male infertility.By the combination of single dimensional analysis and multidimensional analysis,the raise of Co and Mo in male urine would increase the risk of the occurrence of idiopathic male infertility.In addition,the interaction between Co and Mo was discovered.Conclusion Based on the large sample analysis,we firstly discovered Co and Mo exposure were associated with male infertility and Co and Mo exposure had an interaction to the risk of the occurrence of male infertility.