The Research Of Phthalate And Its Metabolites In Human Chorion Tissues

Phthalates are the kinds of chemical which are widely used as industrial compou- nds, and mostly used as plasticizers.Cosmetics like soaps,cosmetics,shampoos and perfumes also contain phthalates.People contact with phthalates mainly through oral, dermal or inhalation routes.The toxicity of phthalates has been widely noticed, especially in human reproductive system. Phthalates can directly or indirectly affect the development of germ cells.Chorion tissues as inchoate human embryo are the base of organogenesis. It has been found that the constant of the phthalate esters in mother's body is related to the anogenital distance of new born baby boy.The investigation about epidemiological displayed that abortion, fetal growth restriction and other adverse pregnancy outcomes are closely related to the exposure of phthalates, but research about the levels of phthalates in embryo chorion tissues has not been reported in china or overseas.This thesis established detection method of phthalate esters and metabolites.It investigated two different methods of the sample pretreatment which were solid-phase extraction and direct extraction of samples,and used high performance liquid chromat- ography to analyze the pollution level of phthalates in human chorion tissues. The objectives included ten materials such as DMP, DEP, DBP, BBP, DEHP and its metabolites MMP, MEP, MBP, MBZP and MEHP.The researches contained using different solvents like methanol, acetonitrile, water, phosphate buffer to extract the materials in the organization, the preparation of exploring phthalate esters in the extraction of the solid phase conditions, and finally choose the method which is solid phase extraction and high performance liquid chromatography.To determinate of phthalate and its metabolites in human chorion tissues, samples were extracted with 3ml phosphate buffers, extracted by HLB cartridge, with 3ml water purification, 2ml acetonitrile, 2ml ether, 1ml ethyl acetate elution, high performance liquid chroma- tography conditions for separation was Hypersil C6H5 column (250 mm×4.6 mm, 5μm), the best UV detector wavelength was 228nm, column temperature was 25℃, the initial flow rate of 0.8ml/min, high pure water (0.1% Acetic acid) - acetonitrile (0.1% acetic acid) gradient elution, the entire process was 25min. The recoveries of standard samples and chorion tissues samples were 71.76%～102.1% and 50.78%～82.78%, relative standard deviation were 0.78%～12.60%, detection limit were 1.89～7.63μg/L and linearity range were 10～4000μg/L.It determined totally 58 individual embryos, containing normal human chorion samples and fetal diapause tissues .MMP, MEP and DEHP which were detected in the human chorion tissues were higher, and reaching 80%. Other detection rates of phthalates were different. MBP detection rate was only 8.62%. MEHP and BBP detction rates were 20%, MBZP detection rate was 46.55%, and DBP detection rate was 68.97%. Concentration range of phthalate and its metabolites in the human chorion tissues were 0.13 ~ 2.04μg/g. The results showed the presence of phthalates in chorion tissues, and that suggested it had been contaminated to phthalates in the early fetal environment. It is likely to cause adverse effects on the future growth and development of fetus, which people should pay serious attetion to.