天然雌激素雌酮（E1）、17β-雌二醇（17β-E2）和雌三醇（E3）及其结合体，人工合成雌激素17α-乙炔基雌二醇（EE2），以及类雌激素活性物质如双酚A（BPA）等内分泌干扰物在水环境中仅以ng/L的浓度就可以导致鱼类的雌性化等生殖异常化问题。畜禽养殖场是环境雌激素的重要来源之一，因此有必要研究养殖场畜禽的雌激素排放特征，以及雌激素在养殖场的去除。本研究在研究室之前研究的基础上，利用加速溶液萃取（ASE）、液液萃取、HLB固相萃取、NH2柱固相萃取和Fluid固相萃取进行萃取和净化，进一步开发了高基质干扰的畜禽粪便、尿液、养殖场污水和堆肥样品中雌激素自由体（E1、17β-E2、E3和EE2）、雌激素结合体（E1-3G、E1-3S、17β-E2-3G和17β-E2-3S）以及BPA的前处理和LC-MS/MS分析检测方法。可以更全面和较为准确地解释畜禽养殖业的雌激素排放特征，以及在养殖场粪污处理设施中的浓度和去除特征。选取长江三角洲区域的上海市、浙江省，以及山西省作为研究区域，对三个研究区域的15个规模化养殖场（包括奶牛、肉牛、肉猪、羊、蛋鸡和肉鸡）中畜禽粪便产生及畜禽污的处理处置方式进行现场资料调查并采样；在此基础上，对样品中9种化学物质进行浓度测试分析。得出研究区域内典型规模化养殖场中各畜种粪尿、堆肥和污水中雌激素的浓度分布，并比较粪尿中雌激素浓度的畜种、年龄和性别差异，计算畜禽的日雌激素排放量，不同的粪尿处理方式对雌激素的去除效果，以及受纳水体和土壤中的浓度特征。结果显示，不同畜种、性别、生长和生殖阶段的畜禽的雌激素排放特征不同。例如，在长三角畜禽养殖场的调查显示，奶牛的雌激素日排放量为145.23 – 179.27 μg/d，其中的92%是通过粪便；而猪的雌激素日排放量为42.56 – 219.25 μg/d，其中的98%以上是通过尿液。雌激素结合体在奶牛所排放的总结合体中的贡献率为14.6 – 48.8%，对猪所排放的总结合体的贡献率超过了98%。常用的厌氧消化池对雌激素的去除率仅为14.7 – 21.8%，而好氧条件下的高效组合气浮对雌激素的去除率达到了97.6%。好氧条件仅仅提高的是雌激素的自由体的去除率，对结合体的去除没有增强的效果。BPA的去除率为60 – 70%，其去除率也没有收到供氧条件的影响。对上海市一座奶牛场堆肥的不同时间段雌激素含量的研究表明，雌激素的去除率约为70%，BPA在堆肥前后没有明显变化。山西省采用的地面自然堆放的堆肥方式对鲜粪和堆肥成品中的雌激素浓度调查表明，雌激素的去除率为18.7 – 59.6%，BPA在堆肥后浓度升高。在长三角地区污水出水和堆肥成品中，总雌激素的浓度最高达到2695 ± 181 ng/L（厌氧消化出水）和80.8 ± 6.0 ng/g，山西省养殖场堆肥成品中总雌激素的浓度范围为13.0 – 74.0 ng/g，对环境造成了潜在的雌激素风险。另外，养殖场受纳水体和土壤中均有雌激素结合体E1-3S和E2-3S仍有检出，仍有可能转化为自由体产生雌激素活性。

Natural estrogens estrone (E1), 17β-estradiol (17β-E2), estriol (E3) and their conjugated forms, synthetic estrogens like 17α-ethinyl estradiol (EE2), as well as xenoestrogens bisphenol A (BPA) can lead to the feminization of male fish at ng/L levels in the aquatic environment. Livestock farm is one of the main sources of environmental estrogens. Thus, it is meaningful to investigate the estrogen excretions of livestock as well as the removal of estrogens in manure treatment facilities.Based on the previous study of our research group, relatively efficient pretreatment procedures and LC-MS/MS detection methods were developed to analyze free (E1, 17β-E2, E3 and EE2) and conjugated estrogens (E1-3G, E1-3S, 17β-E2-3G and 17β-E2-3S) and BPA in livestock feces, urine, wastewater and compost. Pretreatment procedures include ASE extraction, HLB extraction, NH2 cartridge cleanup, and Florisil cartridge cleanup. The detection of conjugated estrogens made it more comprehensive to explain the source and removal of estrogens in livestock farms.Shanghai City and Zhejiang Province located in Yangtze River Delta, and Shanxi Province located in North China were selected as study area. Fresh excreta and treated manure samples were collected in 15 livestock farms in study area, and 9 targeted compounds were analyzed in these samples. Based on the concentration data, daily estrogen excretions were calculated for different livestock (cow, beef cattle, sheep, swine, boiler and laying chicken), and removal of estrogens in different manure treatment plants were assessed.Results showed that estrogen excretions varied as the function of species, sex, growth and reproductive stages. The daily estrogen excretions of a cow were 145.23 – 179.27 μg/d, among which 92% were through feces. While swine excreted 42.56 – 219.25 μg/d of estrogens daily, and more than 98% of them were through urine. Estrogen conjugates contributed 14.6 – 48.8% to the total estrogen excreted by cow, and contributed more than 98% to the total estrogen excreted by swine.Normally used anaerobic digesters removed 14.7 – 21.8% of estrogens, and combined air floatation removed 97.6% of estrogens at aerobic conditions. Removal efficiencies of BPA were 60 – 70%. Aerobic conditions could only increase the removal efficiencies of free forms and had little effect on removal of conjugated forms or BPA. By investigating estrogen levels in compost at different stages in a livestock farm in Shanghai, we concluded that estrogens were removed by about 70%, and BPA levels barely changed during composting. Natural composting by just piling on ground used in Shanxi Province removed estrogens by 18.7 – 59.6%, and BPA levels rose after composting.In compost products and wastewater effluent, total estrogen concentrations were up to 80.8 ± 6.0 ng/g and 2695 ± 181 ng/L (anaerobic digester effluent) in Shanghai and Zhejiang Province, and in Shanxi Province, estrogen concentrations in compost products were up to 13.0 – 74.0 ng/g, posing potential estrogenic risk to the environment. In the river water and soil receiving treated manure, E1-3S and E2-3S were still discovered, which still might turn into free forms with estrogenic activity.