Development Of Simultaneous Analysis Of Contaminant And Multiple Biomarkers And Preliminary Application In Evaluation For Endocrine Disrupting Chemicals

Endocrine disrupting chemicals(EDCs)are exogenous substances those can interfere with hormone synthesis,release,transport,metabolism,binding,effect and elimination in organisms.They affect the function of endocrine system by interfering with hormone behavior.Most of EDCs are easy to accumulate and enrich in the environment,thus causing long-term harm to organisms.Certain EDCs can also affect the normal hormone actions in human body even at very low levels.EDCs exist widely in various products,such as pesticides,cosmetics,plastics,flame retardants,food additives,textiles,building materials and so on.In recent decades,the incidence of endocrine disruptors-related diseases has increased significantly such as breast cancer,prostate cancer,testicular cancer,diabetes,obesity and declining fertility.Measuring the changes of biomarkers such as hormones and proteins is an effective method in the study of endocrine disruption.Radioimmunoassay,GC-MS and LC-MS are commonly used for hormone determination,while ELISA and Western-Blot are commonly used for protein analysis.There are some shortcomings in these analytical methods,such as cumbersome sample preparation,low throughput,easy to be interfered,expensive and so on.LC-MS/MS is very suitable for analysis of endocrine disruption because of its high sensitivity,specificity,high throughput and easy automation.In this study,a method based on LC-MS/MS was developed to simultaneously analyze EDCs and various endocrine disrupting effects.The preliminary application of the method was then carried out.Bisphenol S was selected as targeted compound and mice were used as exposure model.Five macromolecule proteins including gonadotropin-releasing hormone(GnRH),androgen receptor(AR),thyroglobulin(Tg),adiponectin(ADP),resistin(RETN),and two small molecule hormones including tesosterone(T)and thyroxine(T4)were used as biomarkers to indicate the effects of bisphenol S on gonadal system(GnRH,AR,T),thyroid system(Tg,T4)and adipose endocrine system(ADP,RETN)in mice.The macromolecule protein ADP and small molecule hormone T4 were detected in mice serum exposed to bisphonol S.However,the other biomarkers were not found in the mice due to the great difference between the properties of macromolecule and small molecule,the loss in pretreatment and the limitation of existing conditions.Then,three pesticides including coumaphos,profenofos and paclobutrazol were selected as targeted compounds and mice were used as exposure model.Eight small molecular hormones testosterone(T),androstenedione(ADION),progesterone(P4),hydroxyprogesterone,thyroxine(T4),triiodothyronine(T3),cortisol and cortisone were selected as biomarkers to indicate the endocrine disruption effects of the pesticides on androgen(T,ADION),progesterone((P4?hydroxyprogesterone),thyroid hormone(T4,T3)and adrenocortical hormone(cortisol,cortisone).Furthermore,the integrated biomarker response(IBR)was employed to comprehensively analyze the endocrine disrupting effects of the pesticides.