Determination of estrogens and estrogenic activities in water from three rivers in Tianjin, China

Studies on estrogenic disrupting compounds(EDCs) occurrence and identification of main responsible compounds in river water discharged into the sea are of significance.In the present research,we screened estrogenic activities of 10 river water samples from 3 main rivers discharged into Bohai Sea in Tianjin using a recombinant two-hybrid yeast assay and chemical analysis by gas chromatography-mass spectrometry.All sample extracts induced significant estrogenic activity,with 17β-estradiol equivalents(EEQ) of raw water ranging from 5.72 to 59.06 ng/L.Six most concerned EDCs in the river water samples including estrone,17β-estradiol,17α-ethinylestradiol,estriol,diethylstilbestrol and estradiol valerate were determined,with their concentrations up to 50.70,31.40,24.40,37.20,2.56,and 8.47 ng/L,respectively.Through causality analysis by comparing the EEQ values of yeast assay and chemical analysis,17α-ethinylestradiol and 17β-estradiol were identified as the main contributors to the estrogenic effects of the river samples,accounting for the whole estrogenic activities(62.99% to 185.66%),and estrogen antagonistic compounds might presented in the heavy polluted water samples.The proposed approach using both chemical analysis and bioassay could be used for identification and evaluation of the estrogenic activity of EDCs in river water.     

Distribution and seasonal variation of estrogenic endocrine disrupting compounds, N-nitrosodimethylamine, and N-nitrosodimethylamine formation potential in the Huangpu River, China

Detection of estrogenic endocrine disrupting compounds(EDCs) and N-nitrosodimethylamine(NDMA) in drinking water has led to rising concerns. There are, however, a paucity of studies on the distribution and seasonal variation of NDMA and NDMA formation potential(NDMA-FP) in natural waters, especially in China. For EDCs, limited studies have investigated the distribution and seasonal variation of estrone(E1), 17β-estradiol(E2), estriol(E3), 17α-ethinylestradiol(EE2), technical-nonylphenols(t-NP), and bisphenol A(BPA) in Shanghai. In this study, water samples were collected from 11 sampling sites along the Huangpu River in 2012. The distribution and seasonal variation of EDCs, NDMA, and NDMA-FP were investigated. The results showed that all of the 11 sampling sites were contaminated by the target compounds. Compared with E2 and EE2, higher E1 and E3 concentrations were usually detected in the Huangpu River. The values of 17β-estradiol equivalents(EEQ) suggest a high possibility of endocrine effects on exposed organisms in the Huangpu River. NDMA-FP and t-NP were the dominant contaminants among the eight target compounds. The detection rates of the target compounds and their concentrations were both higher in dry seasons than in wet seasons. Higher concentrations of target compounds were observed in urban sampling sites near drainage outlets, and also in suburban sampling sites where intensive livestock farming and farmlands were located.     

Profile and removal of endocrine disrupting chemicals by using an ER/AR competitive ligand binding assay and chemical analyses

An estrogen receptor (ER)/androgen receptor (AR) ligand competitive binding assay (ER/AR-binding assay) and chemical analyses were used to evaluate the endocrine disrupting chemicals (EDCs) behavior of two municipal wastewater treatment plants (WWTPs) (K and S). In the influents, estrone (E1), androsterone (A), androstenedione (AD), BPA (bisphenol A), NP (nonylphenol) and daidzein (DZ) were detected in high amounts with subsequent 24 h-average concentrations of 350, 1000, 29, 1300, 3900, and 5700 ng/L in K-WWTP and of 310, 620, 59, 1600, 2600, and 8400 ng/L in S-WWTP. The estrogenic (androgenic) activity as 17 -estradiol (E2) equivalents (EEQ) or testosterone (Te) equivalents (TEQ) was consequently 620 ng E2/L (570 ng Te/L) and 580 ng E2/L (800 ng Te/L) for the two WWTPs. The removal e ciencies of the above mentioned sole target chemicals were 51%–100% for K-WWTP and 55.6%–100% for S-WWTP. The removal e ciencies of EEQ were about 73% for both WWTPs, while the removal e ciencies of TEQ were 62.1% for K-WWTP and 98.4% for S-WWTP. In addition, chemical-derived EEQ were about 1.2%–52.4% of those by ER-binding assay for K-WWTP and the corresponding ratios were 1.3%–83.3% for S-WWTP, while chemical derived TEQ were less than 3% of values measured by the AR-binding assay for both WWTPs.     

AO与AAO工艺去除雌激素效能对比及分析

为探讨不同污水处理工艺对雌激素类有机物的处理效能,本研究设计并同时运行了AO和AAO两种污水处理工艺,在稳定运行条件下,比较了两种工艺对各种常规水质指标和雌激素的去除状况.试验结果表明,AAO系统对污染物去除效能明显优于AO系统.AAO对COD、TN、NH4+-N、TP及TOC的去除效能分别比AO高出7.55%、20....     

北京市城市污水雌激素活性的研究

采用重组酵母雌激素筛检法(YES法)对北京市3个城市污水处理厂工艺流程污水样品的雌激素活性进行了评价,并采用GC/MS分析了样品中8种内分泌干扰物(EDCs),以进一步阐明城市污水雌激素活性的变化.结果表明,污水处理厂能较好地降低污水雌激素活性,降低率为82.2%～97.0%,但出水仍具有一定的雌激素活性,雌二醇(E2)当量浓度(EEQ)为2.6～16.0 ng/L.GC/MS检测显示,污水处理厂并不能完全去除目标化合物,出水中平均浓度最低的是17α-E2,为13.5 ng/L,最高的是BPA,为106.4 ng/L;出水的雌激素活性主要来自类固醇雌激素.污水厂的出水排放具有潜在的环境风险,在污水处理过程中,应特别关注对类固醇雌激素的去除.     

Assessment of estrogen disrupting potency in animal foodstuffs of China by combined biological and chemical analyses

Food has been documented as one of major routes for human exposure to environmental estrogens（EEs）, but information on the occurrence of EEs in animal foodstuffs is still scarce.This study analyzed estrogenic activity in 16 types of animal foodstuffs（n = 142） collected from four cities（Wuhan, Guangzhou, Wenzhou and Yantai） of China by combined yeast estrogen screen（YES） bioassay and chemical analysis. By bioassay, all samples＇ extracts were found to induce estrogenic activities and the bioassay-derived 17β-estradiol equivalent（EEQbio） ranged from 8.29 to 118.32 ng/g. In addition, the samples were analyzed by liquid chromatography coupled to tandem mass spectrometry for further chemical analysis. 17β-Estradiol was found in all samples in this survey at levels of 0.44 to 15.04 ng/g.All samples had 33.1% detection rate of 17α-ethinylestradiol（EE2）, and the maximum concentration was 2.80 ng/g. Bisphenol A and 4-nonylphenols were detected in 83.8% and83.1% of samples, with concentrations up to 12.56 ng/g and 35.76 ng/g, respectively.However, the concentrations of estrone, diethylstilbestrol and 4-t-octylphenol were found to be below the limit of detection. A comparison of EEQbiomeasured from the YES assay and EEQchemcalculated from chemical analysis showed good correlation（R2= 0.84）. Based on the results, the YES assay can be used as a rapid pre-screening method for monitoring the levels of estrogenic activity in large numbers of animal foodstuffs, and chemical analysis used in combination can be used for the identification of specific EEs.     

罗时江中5种环境内分泌干扰物的分布特征

以典型的农业面源污染河流——大理洱海罗时江作为研究对象,采用固相萃取、超声萃取和气相色谱质谱联用技术,对河流水体和表层沉积物样品中5种EDCs〔环境内分泌干扰物,包括BPA(双酚A)、E1(雌酮)、E2(17β-雌二醇)、EE2(乙炔基雌二醇)和E3(雌三醇)〕进行分析测定.结果表明,罗时江水体中普遍存在EDCs,其中ρ(BPA)为1.01～43.64 ng/L,ρ(E1)为2.54～20.53 ng/L,ρ(E2)为nd～23.18 ng/L,ρ(EE2)为1.16～13.74 ng/L,ρ(E3)为nd～26.55 ng/L.各采样点EDCs污染通量结果显示,右所、邓川2个乡镇对EDCs污染贡献量最为显著,之后由于湿地截留以及河流的自净作用EDCs污染通量有所削减.罗时江对洱海的入湖EDCs贡献量为1.35～3.70μg/s.在城镇下游处沉积物中E1和E3的富集倍数均为各采样点中最高,这与河流水体中的污染通量变化趋势一致,进一步说明该处污染负荷的加剧,并且说明EDCs会在污染发生时于下游就近沉积.沉积物中w(E3)最高(1.48～28.44μg/kg),其次为w(E1)(0.36～2.95μg/kg),E2因迅速降解不易富集.      

重组酵母法测评Fenton反应中激素活性变化关系

环境内分泌干扰物的污染已成为环境领域广泛关注的研究领域.文章利用重组基因酵母菌检系统评测雌二醇(E2)在非均相Fenton光催化高级氧化降解过程中的激素活性变化.色谱质谱解析中间产物的结构,很好地解释了该非均相Fenton催化氧化过程不仅可以降低E2浓度,而且可以降低母体和中间产物的雌激素活性,对水体的安全处理具有重要...     

土壤净化槽对农村污水中内分泌干扰物的去除效果

以土壤净化槽污水处理系统为研究对象,采用固相萃取-色谱质谱检测方法,分析了云南洱海流域农村污水中3种天然雌激素类内分泌干扰物〔E1(雌酮)、E2(雌二醇)和E3(雌三醇)〕和2种人工合成内分泌干扰物〔EE2(炔雌醇)和BPA(双酚A)〕的质量浓度与去除效果. 结果表明,洱海流域农村污水中天然雌激素的质量浓度远高于人工合成内分泌干扰物,E1与E2为污水中天然雌激素内分泌干扰物的主要成分,占87.1%~99.2%. 土壤净化槽对天然雌激素类内分泌干扰物、EE2和BPA的净化能力分别为1.42~168.21、0.64和3.48mg/(m2·a). E1、E2和E3在土壤中的平均残留率分别为4.3%、6.3%和3.8%,其中90%以上的雌激素物质在土壤净化槽系统中被降解或转化. 土壤净化槽对天然雌激素、EE2和BPA的平均去除率分别为70.3%、63.6%和77.8%,对常规污染物COD_Cr(69.8%)、TP(88.7%)、TN(57.2%)和NH₃-N(88.7%)的去除效果也较好. 土壤净化槽可以有效去除村落污水中的内分泌干扰物、有机物以及氮磷污染物,对削减村镇面源污染、减少入湖污染负荷以及改善流域内的水环境质量具有重要作用.      

污水处理厂出水中雌激素活性物质浓度与生态风险水平

再生水在环境和景观水体利用中,由微量有毒有害污染物,特别是内分泌干扰物引起的长期生态风险备受关注. 以再生水生态风险控制为目的,对城市污水处理厂出水(再生水)中雌激素活性物质浓度分布情况进行了研究,归纳了8种雌激素活性物质的出水浓度水平,比较分析了各物质的雌激素活性和生态风险. 结果表明:8种物质的质量浓度分布在ng/L～μg/L,其中ρ(类固醇)最低,为nd～50 ng/L;ρ(酚类)及ρ(酞酸酯类)主要在μg/L水平,个别高达40 μg/L. 雌激素活性分析和生态风险评价结果表明,污水处理厂出水(再生水)中3类雌激素活性物质的雌激素活性和生态风险顺序均为类固醇物质＞酚类物质＞酞酸酯类物质. 城市污水再生处理厂应优先控制乙炔基雌二醇(EE2)、雌酮(E1)和雌三醇(E3)等3种雌激素活性物质.      

4-壬基酚臭氧氧化中雌激素活性变化的影响因素

采用有效的重组基因酵母检测方法,研究了不同影响因素下水中典型内分泌干扰物4-壬基酚(4-NP)在臭氧氧化过程中雌激素活性的变化规律.结果表明,臭氧能迅速氧化4-NP,并有效地去除其雌激素活性;臭氧氧化过程中,水中高臭氧浓度、低pH值、低悬浮物含量、低·OH抑制剂浓度以及适量的腐殖酸浓度均有利于降低4-NP的雌激素活性.     

珠江河流胶体中的典型内分泌干扰物

采用切向流超滤、固相萃取、气相色谱-质谱联用等分离和分析方法,研究了珠江2条河流中典型内分泌干扰物(EDCs)在胶体相中的含量分布、胶体/水相间的分配作用.结果发现:胶体中4-壬基酚(NP)、双酚A(BPA)、雌酮(E1) 的含量范围分别为23.2~108ng/L、2.3~97.6ng/L、n.d.(未检出)~0.32ng/L,平均值分别为70,31.4,0.3ng/L;17β-雌二醇(E2)、己烯雌酚(DES)、17α-乙炔雌二醇(EE2)、雌三醇(E3)则未在胶体样品中检出.除石龙外,其余各采样点表、底水层间的胶体EDCs浓度水平无显著差异.NP和BPA含量与胶体有机碳(COC)含量、紫外吸收强度(UV254)均呈正相关,说明COC是控制酚类雌激素在胶体相中分布的一个重要因素,NP、BPA和COC之间的相互作用与胶体有机碳的芳香性有关.计算发现,珠江河水中约21%~67% 的 NP、4%~74% 的BPA、24%~26% 的E1存在于胶体相,NP和BPA在胶体/水相间的有机碳归一化分配系数(Kcoc)分别为10 (5.69±0.50)、10 (5.51±0.77),高出各自的悬浮颗粒物/水分配系数(Koc)1个数量级,表明胶体的强吸附能力对EDCs在水环境介质间分配、迁移、转化发挥了重要作用.     

活性污泥中典型内分泌干扰物的分析方法

对代表烷基酚类的典型内分泌干扰物(EDCs)叔辛基酚、辛基酚、壬基酚、双酚A,和代表类固醇的典型内分泌干扰物17α-乙炔基雌二醇和雌三醇等6种物质,建立了活性污泥水相与固相中的SPE-GC/MS的同时分析方法。对比分析了三种不同的固相萃取浓缩小柱的效果,发现具有梯度极性的Bond Elut-Plexa小柱对所选6种内分泌干扰物萃取回收效果较好。通过所建立的方法对6种EDCs在污水和污泥中的浓度进行了分析,检测回收率达80.5%和84.8%以上。并通过该分析方法对上海某污水净化水厂曝气池中污水和污泥浓度进行了有效分析。     

珠江口典型河段内分泌干扰物的空间分布及风险评价

选择珠江口典型河段为研究对象,调查上覆水和沉积物中5种典型内分泌干扰物(EDCs)4-辛基酚(4-t-OP)、4-壬基酚(4-NP)、双酚A(BPA)、三氯生(TCS)和三氯卡班(TCC)的空间分布特征,并利用风险商值法(RQ)对其上覆水及沉积物进行风险评价.结果表明,5种EDCs在珠江河流上覆水和沉积物样品中均有检出,水体中4-t-OP、4-NP、BPA、TCC、TCS的水体质量浓度分别为:49.19~512.82、112.66~717.31、23.54~2189.88、1.90~120.99、1.51~54.75 ng·L~(-1).沉积物中4-t-OP、4-NP、BPA、TCC、TCS的沉积物质量分数分别为:56.60~1606.56、69.52~6524.35、18.55~522.45、1.54~179.66、0.71~130.92 ng·g-1.5种EDCs在沉积物和上覆水中的空间分布基本遵循上游到下游逐渐减少的趋势.4-t-OP、4-NP、BPA、TCC和TCS的风险商(RQ)在上覆水和沉积物中的范围分别为0.03~4.20和0.026~149.28.对生态健康产生极大压力.沉积物来源、粒度分布和有机质含量均对EDCs在沉积物中的富集有影响,且主要影响因素为沉积物有机质含量.     

南方某水厂处理工艺过程中内分泌干扰物的变化规律

对南方某水厂处理工艺过程中内分泌干扰效应进行了研究.将水源水和各处理工艺的出水用固相萃取方法富集后,按不同极性洗脱,得到从非极性到极性3个组分,对总提取物和各分级富集组分分别进行重组基因酵母检测.探讨了该水厂不同处理工艺过程中内分泌干扰物的变化规律.结果表明:水源水中该类污染物浓度最高,相当于0.25 pmol/L雌二醇当量,主要存在于非极性组分(正己烷洗脱组分)中.各处理工艺段水样都存在一定程度的内分泌干扰物效应.经处理后,非极性组分内分泌干扰物效应降低,极性组分内分泌干扰物效应逐渐增加,但总地内分泌干扰物效应逐渐递减,现有工艺对雌激素类物质有较好的去除效果,去除率达到83%.研究表明,重组基因酵母检测技术结合水样的固相萃取、三步纯化分级前处理方法可以快速、有效地筛选和定量分析水样中未知内分泌干扰物及其总体效应.     

太湖流域宜溧河酚类内分泌干扰物的空间分布及风险评价

以NP(nonylphenol,壬基酚)、4-t-OP(4-t-octylphenol,辛基酚)和BPA(bisphenol A,双酚A)为目标物质,研究酚类EDCs(内分泌干扰物)在太湖流域宜溧河地表水体和悬浮物中的空间分布特征及风险评价. 结果表明,太湖流域宜溧河地表水体中ρ(NP)、ρ(4-t-OP)和ρ(BPA)分别为156.2～434.0、11.8～19.4和89.8～353.8 ng/L,悬浮物中w(NP)、w(4-t-OP)和w(BPA)分别为520.0～9 818.2、52.0～454.5和39.0～2 454.5 ng/g. 流域内生活及工业污水排放可能是宜溧河流域水体中酚类EDCs的主要来源,ρ(NP)与ρ(NO₃-)呈显著正相关(R=0.860,P<0.01). 宜溧河水系中酚类EDCs在水相-悬浮物相的有机碳标准分配系数为4.14～6.41,表明悬浮颗粒物的吸附是水体中EDCs迁移的一个重要途径. 研究区域中NP、4-t-OP和BPA的入湖量分别为506、57.9和297 kg/a. 风险评价结果表明,该地区部分河段ρ(EDCs)具有潜在的生态风险.      

梭鱼和青鳉鱼ERα介导的雌激素活性比较研究

为从物种雌激素受体(ERα)敏感性差异的角度探索辽东湾野生梭鱼雌雄同体高发生率的一个可能原因,从梭鱼肝脏cDNA中扩增出长1113 bp的梭鱼ERα(sERα) DEF区片段,并以此构建pGBT9-sERα DEF 质粒,将其和pGAD424-TIF2质粒依次转入Y190酵母细胞中,建立了sERα物种特异性的酵母双杂交筛选系统.在此系统中测试了4种环境中主要的雌激素雌酮(E1)、17β-雌二醇(E2)、雌三醇(E3)及17α-炔雌醇(EE2)对梭鱼ERα的诱导活性,半数雌激素活性浓度(EC50)为3.7、2.4、1698.7、1.2nmol/L,分别是青鳉鱼ERα活性的4.6、2.3、3.4、3.3倍,说明梭鱼ERα对E1、E2、E3、EE2具有较高的敏感性,这为解释辽东湾梭鱼雌雄同体的高发生现象提供了重要线索.     

酚类化合物雌激素效应的比较研究

采用重组人雌激素受体基因片段(hER)及共激活因子(GRIP1)双杂交酵母检测体系,检测9种酚类物质的类雌激素和抗雌激素效应.结果表明,所检测的化合物中有3种同时表现出雌激素诱导与抑制活性;2种只表现出雌激素诱导活性;1种只表现出雌激素抑制活性;3种均没有雌激素诱导与抑制活性.结构分析表明:酚类中产生雌激素活性的物质具有取代基位于对位且邻位没有大的取代基团的结构;烷基在对位的酚类物质也能显示出雌激素拮抗活性.所采用的双杂交酵母检测体系适用于外源物质的雌激素活性检测,具有较高的灵敏度;结合外源物的雌激素诱导与抑制活性,用于分析污染物的体内作用方式.     

壬基酚对鲫鱼(Carassius auratus)的雌激素效应研究

实验研究了环境雌激素4-壬基酚(NP)对鲫鱼(Carassiusauratus)的剂量-效应关系。雄性鲫鱼腹腔注射不同剂量的17β-雌二醇(E2)(质量分数分别为0 01,0 10,1 00mg kg(注射剂量 鱼体重,下同))和4-壬基酚(NP)(质量分数分别为1,10,50,100,200mg kg),用ELISA方法测定鲫鱼血浆中卵黄蛋白原含量。结果显示,与对照相比,在注射后14d内,0 01mg kgE2处理组和1 00mg kgNP处理组鲫鱼血浆卵黄蛋白原含量没有显著差异;其他各处理组在注射后8d,血浆卵黄蛋白原含量显著升高(P<0 05),且与暴露浓度呈依赖关系;注射后14d,10,50,100和200mg kgNP处理组的含量依次是对照组的10,59,340和435倍。研究表明,NP对鲫鱼具有明显的雌激素效应。      

Competitive sorption between 17α-ethinyl estradiol and bisphenol A/4-n-nonylphenol by soils

The sorption of 17α-ethinyl estradiol(EE2),bisphenol A(BPA),and 4-n-nonylphenol(NP) in single systems and the sorption of EE2 with different initial aqueous concentrations of BPA or NP were examined using three soils.Results showed that all sorption isotherms were nonlinear and fit the Freundlich model.The degree of nonlinearity was in the order BPA(0.537-0.686) > EE2(0.705-0.858) > NP(0.875-0.0.951) in single systems.The isotherm linearity index of EE2 sorption calculated by the Freundlich model for Loam,Silt Loam and Silt increased from 0.758,0.705 and 0.858,to 0.889,0.910 and 0.969,respectively,when BPA concentration increased from 0 to 1000 μg/L,but the effect of NP was comparably minimal.Additionally,EE2 significantly suppressed the sorption of BPA,but insignificantly suppressed that of NP.These findings can be attributed to the difference of sorption affinity of EE2,NP and BPA on the hard carbon(e.g.,black carbon) of soil organic matter that dominated the sorption in the low equilibrium aqueous concentration range of endocrine-disrupting chemicals(EDCs).Competitive sorption among EDCs presents new challenges for predicting the transport and fate of EDCs under the influence of co-solutes.