四溴双酚A（Tetrabromobisphenol A，TBBPA）是一种重要的溴系阻燃剂（Brominated Flame Retardants，BFRs），广泛应用于电子电器、建筑纺织等行业，随着阻燃行业的发展和对阻燃产品消费量的增加，TBBPA大量进入环境和生物体内。目前关于TBBPA对环境的污染及人体健康的影响引起了广泛关注，其中TBBPA在生物体的分布、代谢特征研究对准确评价TBBPA健康风险具有重要作用。为了更全面地了解长期摄入TBBPA后生物体的内暴露和代谢特征，本文通过动物模型实验，研究了经口摄入条件下TBBPA在生物体的内暴露水平特征以及可能的生理损伤，并采用代谢组学技术分析了生物体经口摄入TBBPA后血清中内源性代谢产物的变化，初步筛选出具有潜在指示作用的代谢标志物。具体研究内容和主要结果如下：（1）采用超高效液相色谱-串联三重四级杆质谱法（UPLC-MS/MS）结合同位素稀释技术建立了大鼠血清、尿液、粪便样品中TBBPA含量的测定方法。优化了不同生物样品的前处理方法，在保证方法准确性的同时，也使其更适用于大批量样品的快速处理。血清、尿液、粪便样品的加标回收率均值分别达到为88.39%、86.56%、90.57%，相对标准偏差均在10%以下，方法的灵敏度和准确性较好。（2）建立动物实验模型，对Wistar大鼠进行为期90天、6组不同剂量浓度的TBBPA亚慢性暴露实验，获得大鼠血清样品72个、粪便样品792个、尿液样品792个。利用所建立的检测方法，定量分析样本中TBBPA浓度。通过分析发现，随着暴露剂量的增加，大鼠血清、尿液和粪便3类生物样品中TBBPA浓度均逐渐增加；随着暴露时间的推移，大鼠尿液和粪便2类生物样品中TBBPA浓度呈峰型波动变化；随着TBPPA的持续暴露，不同剂量梯度上，大鼠尿液和粪便2类生物样品中TBBPA浓度都趋于一个稳定值，且该数值与暴露剂量有较好的线性关系。（3）大鼠代谢TBBPA的主要途径为粪便排泄，其24h代谢量占总暴露剂量的11~56%；经尿液的排泄量则较小，24h尿液代谢量小于总暴露剂量的0.03%。大鼠经口暴露TBBPA 90天后，血清中TBBPA占总暴露剂量的0.01~0.03%，表明TBBPA的生物利用度较低。主要原因是TBBPA在生物体内半衰期较短，限制了生物积累；另外长期暴露TBBPA条件下，生物体通过机体调节形成一定的代谢应对机制，能最大限度的排泄身体内TBBPA；同时TBBPA经口进入生物体后吸收能力较差。（4）大鼠代谢TBBPA过程中，具有较强的性别差异，雄鼠相对于雌鼠，有较高的血清TBBPA浓度、较低的尿液和粪便TBBPA浓度，说明TBBPA更易在雄鼠体内蓄积，这可能与不同性别生物体内的激素调节机制不同有关。（5）采用气相色谱-飞行时间质谱（GC-TOF-MS）法对对照组、TBBPA低剂量组和TBBPA高剂量组暴露下的大鼠血清进行代谢组学检测，分析经口摄入TBBPA大鼠血清内源性代谢产物的变化，结果显示大鼠经口摄入TBBPA后血清代谢产物变化明显，TBBPA暴露降低了血清中脂肪酸类物质浓度。通过分析随暴露剂量升高而发生变化的代谢产物数量与程度，初步筛选出9-十八碳烯酸、软木酸、羧基丁酸可作为具有潜在指示作用的生物标志物。

Tetrabromobisphenol A (TBBPA) is one of the most widely used brominated flame retardants (BFRs) in the industries of electronic appliances, construction and textile.With the development of the flame retardant industry and the increase of TBBPA consumption, significant amount of TBBPA poured into the environment and organism.Currently, environmental scientists have paid more attention to the environmental pollution of TBBPA and its negative health effects to human. TBBPA metabolism and the distribution patterns in organism play an important role in accurate assessment of the potential health risk.In order to more fully understand the exposure and metabolic characteristics of TBBPA, animal models were created. Based on the models the exposure characteristics, physiological damages and metabolism characteristics of TBBPA were analyzed in vivo after long-term oral administration, and biomarkers were screened initially. The contents and the main results were shown as follows.(1) For the determination of TBBPA in serum, urine and feces，the methods were developed and optimized by using ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). The optimized methods were more suitable to process large numbers of samples. The spiked recoveries of TBBPA in serum, urine and feces varied from 83~93% with the relative standard deviation of <10%. The data of quality control indicated the accuracy and precision for the determination of target compounds in serum, urine and feces. These showed this method had a good sensitivity, accuracy and reproducibility.(2) A repeated oral dose study in Wistar rats was conducted. The study set up 6 dose levels and performed as a 90-day repeat dose subchronic toxicity. Through the experiment, 72 serum samples, 792 urine samples and 792 feces samples were obtained. The samples were determined with the established methods. The quantitative analysis showed that, the concentration of TBBPA in the biological samples increased gradually with the increase of exposure dose. The concentration of TBBPA in urine and feces samples showed peak fluctuations with the exposure time. As continued exposure to TBPPA, the concentrations of TBBPA in urine and feces samples tended to be stable values, and these values had good linear relationship with exposure doses.(3) Fecal excretion was the main way for Wistar rats to excrete TBBPA, and about 11~56% of the dose was found in the feces for 24 h. Cumulative urine excretion was very low, urine excretion was less than 0.03% for 24 h. The content of TBBPA in serum accounted for 0.01~0.03% of dose after a 90-day repeated oral dose administration, which indicated biological utilization efficiency of TBBPA was low. The main reasons were that the half-life of TBBPA in vivo was short and bioaccumulation was limited. In the condition of long-term exposure to TBBPA, the organism formed a certain metabolic coping mechanisms, and body excreted the TBBPA as much as possible through these mechanisms. Meanwhile, the absorption capacity of TBBPA was poor. (4) A strong gender differences in Wistar rats were found in metabolizing process of TBBPA. Male rats had higher serum concentrations of TBBPA, but lower urine and feces concentrations of TBBPA than female rats. These results showed that male rats were easier to accumulate the TBBPA than female rats, which may be related to the different hormones regulatory mechanisms in different genders.(5) Based on gas chromatography-time of flight-mass spectrometer (GC-TOF–MS), the serum metabolites of rats were detected in control group, low dose group and high dose group after a 90-day repeated TBBPA oral dose administration. The results showed that there was a significant change in serum metabolites of rats exposed to TBBPA, and the concentrations of fatty acids in serum were reduced after TBBPA intake. Through metabonomics research, 9- octadecenoic acid, hydroxybutyric acid and suberic acid were screened initially as biomarkers.