Assessment of source water contamination by estrogenic disrupting compounds in China

Detection of estrogenic disrupting compounds (EDCs) in drinking waters around China has led to rising concerns about health risks associated with these compounds. There is, however, a paucity of studies on the occurrence and identification of the main compounds responsible for this pollution in the source waters. To fill this void, we screened estrogenic activities of 23 source water samples from six main river systems in China, using a recombinant two-hybrid yeast assay. All sample extracts induced significant estrogenic activity, with E2 equivalents (EEQ) of raw water ranging from 0.08 to 2.40 ng/L. Additionally, 16 samples were selected for chemical analysis by gas chromatography-mass spectrometry. The EDCs of most concern, including estrone (E1), 17βup-estradiol (E2), 17αup-ethinylestradiol (EE2), estriol (E3), diethylstilbestrol (DES), estradiol valerate (EV), 4-t-octylphenol (4-t-OP), 4-nonylphenols (4-NP) and bisphenol A (BPA), were determined at concentrations of up to 2.98, 1.07, 2.67, 4.37, 2.52, 1.96, 89.52, 280.19 and 710.65 ng/L, respectively. Causality analysis, involving comparison of EEQ values from yeast assay and chemical analysis identified E2, EE2 and 4-NP as the main responsible compounds, accounting for the whole estrogenic activities (39.74% to 96.68%). The proposed approach using both chemical analysis and yeast assay could be used for the identification and evaluation of EDCs in source waters of China.     

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.     

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.     

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.     

辽东湾海水中甾体雌激素的检测及生态风险评价

建立了SPE-丹酰化衍生-UPLC-MS/MS海水中雌激素的高灵敏度分析方法,对辽东湾海水中甾体雌激素的浓度水平及分布进行了调查并对其潜在的生态风险进行了评估.结果表明:辽东湾海水样品中检出雌酮,17β-雌二醇,17α-雌二醇,雌三醇和17α-乙炔基雌二醇的浓度分别为(0.714±0.407),(0.089±0.077),(0.009±0.011),(0.008±0.008),(0.001±0.003)ng/L.雌激素浓度在大辽河入海口区域最高,双台子河入海口次之,大凌河及小凌河入海口区域最低.辽东湾海水中的17β-雌二醇等当量浓度(EEQ-E2β)为0.562±0.327ng/L,导致野生梭鱼雌雄同体发生的概率约为0.83%,不足以解释该海域野生梭鱼雌雄同体的高发生率.为了有效管理该海域的环境安全,有必要对原因物质进行进一步解析.     

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...     

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.     

基于定位栏试验的奶牛粪便中雌激素含量特征

为探究奶牛粪便中雌激素含量特征,采用定位栏试验收集奶牛粪便样,采用超高效液相色谱串联(UPLC-MS/MS)法对粪便中4种天然雌激素(雌酮;17α-雌二醇,17β-雌二醇,雌三醇)和2种人工合成雌激素(17a-炔雌醇,己烯雌酚)的含量进行测定,结果表明:6种雌激素的检出率居于8.33％～98.89％之间,4种天然雌激素...     

AnSBR法处理生活污水中典型环境雌激素的研究

为评价和考察厌氧序批式反应器(AnSBR)对生活污水中9种典型环境雌激素的去除效果和机制,采用固相萃取(SPE)——LC/MS/MS分析方法,分析所选环境雌激素的浓度和污泥吸附量. 结果表明:大豆苷元,4-t-辛基酚(OP),4-n-壬基酚(NP)和染料木素的检出浓度高于其他5种环境雌激素;厌氧污泥对NP,OP,雌酮(E1),17α-乙炔基雌二醇(EE2)和17β-雌二醇(E2)的吸附作用明显;大豆苷元和染料木素的去除率在60%以上;大豆苷元、染料木素和雌三醇(E3)的主要去除机制为生物降解作用,EE2,OP和NP的去除是生物降解、污泥吸附的共同作用.      

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.     

Enzymatic treatment of estrogens and estrogen glucuronide

Natural and synthetic estrogens from sewage treatment systems are suspected to influence the reproductive health of the animals in the rivers.In this article we investigated the enzymatic treatment of three estrogens (estrone,17β-estradiol,and 17α-ethynyletstradiol) by a fungal laccase which oxidize phenolic compounds with dissolved oxygen.The elimination of the estrogenic activities by enzymatic oxidation was demonstrated by medaka vitellogenin assay.In addition,we developed an enzymatic treatment system comprised of β-D-glucuronidase and the laccase for 17β-estradiol 3-(β-D-glucuronide) degradation.The two enzymes eliminated 17β-estradiol 3-(β-D-glucuronide) and the intermediate,17β-estradiol,efficiently.     

水环境中雌激素的酶联免疫快速生物筛选技术研究

采用酶联免疫(ELISA)检测技术,对来自长江口及污水处理厂的水样分析典型雌激素雌二醇(E2)的浓度,将检测结果与化学方法检测数据进行比较验证,以阐明ELISA应用于水环境雌激素快速筛选的可行性。结果表明:ELISA的检测限可低至0.5 ng/L,试验的变异系数<30%,具有较高的灵敏性和良好的重复性;ELISA检测长江口水样中E2浓度范围≤0.7 ng/L,污水处理厂水样为2.5～11.5 ng/L,与液相色谱串联质谱(LC-MS/MS)的检测数据进行比较分析,其相关系数R~2为0.9147,表明2种方法具有良好的相关性;ELISA检测所需的水样量比LC-MS/MS更少,使用96微孔板可同时对43个样品进行检测,缩短了多个样品的分析时间,具有快速简便、经济适用的特点。因此,ELISA作为1种快速、简便、灵敏的雌激素高通量筛选技术,可为环境保护部门开展水环境雌激素物质调查和水体雌激素快速诊断等工作提供技术支持。     

Assessing the estrogenic potency in a Portuguese wastewater treatment plant using an integrated approach

The estrogenic potency of a wastewater treatment plant(WWTP) was evaluated using chemical and biological analyses,which showed that after the station treatment processes some of the selected endocrine disruptor compounds(EDCs) were still present in the treated effuent(e.g.,bisphenol A,alkylphenols,estrone) .Thus,the most common endocrine EDCs were identified and quantified and the overall estrogenicity of the treated effuent assessed by integrating the results.Male goldfish(Carassius auratus) were used as biological indicators in a 28-day experiment.Vitellogenin(Vtg) ,gonadosomatic and hepatosomatic indices,steroids(17β-estradiol and 11-ketotestosterone) and histopathology were biomarkers used in fish to evaluate WWTP treated effuent estrogenicity,in combination with instrumental analyses.The results showed a significant increase(P0.01) in plasma and liver Vtg,which were significantly correlated(r=0.66;P0.01) .The gonadosmatic index was significantly(P0.01) reduced in exposed fish.The steroid analyses revealed significant elevations in 17β-estradiol and depressed 11-ketotestosterone concentrations.The histological examinations show changes in exposed fish gonads,such as regressed testes and in some cases(43%to 75%) the development of ovo-testis in fish exposed to 50% and 100% treated effuent.     

Determination of estrogenic activity in the river Chienti (Marche Region, Italy) by using in vivo and in vitro bioassays

Environmental estrogen-like compounds (i.e. xenoestrogens) are a variety of pollutants, ranging from synthetic to natural occurring molecules, that are found in surface and waste waters over a wide range of concentrations. In aquatic environment, the overall estrogenic activity is often due to the presence of a mixture of chemicals and their degraded products which can induce synergistic effects. Current strategies for monitoring estrogen-like chemicals are based on the use of a battery of in vivo and in vitro ecotoxicological tests. In this regard, the aim of the present work was to carry out a bio-monitoring study for testing estrogenicity of the Chienti river (Marche Region, Italy) by using both an E-screen and a vitellogenin (Vtg) induction assay in juvenile goldfish. Three sites were used for analysis, localized at the mouth (sampling point 1), in the middle (sampling point 2) and at the origin (sampling point 3) of Chienti river. For most of the water samples (i.e. samples collected at sampling points 2 and 3), clear estrogenic activity was detected in the E-screen assay suggesting different proliferation activities in function of the collecting site. In contrast, the Vtg ELISA demonstrated that water samples collected from each sampling point were estrogenic. Overall, we showed for the first time that the estrogenic activities in water samples from the Chienti river were significant in both in vivo and in vitro; we also observed a different sensitivity between bioassays.     

猪粪中典型类固醇雌激素的固相萃取/GC-MS检测

以猪粪中常见且具有较大潜在危害的E1(雌酮),17α-E2(17α-雌二醇),17β-E2(17β-雌二醇)和E3(雌三醇)为研究对象,建立了猪粪中这4种典型类固醇雌激素的GC-MS(气相色谱-质谱)分析检测方法.着重优化了样品的前处理方法,包括对超声提取剂、超声提取时间、固相萃取淋洗剂、衍生化温度和时间的优化选择.结果表明:在优化的试验条件下,猪粪中的典型雌激素E1,17α-E2,17β-E2和E3检出限分别为8.0,2.5,5.1和3.3 μg/kg,平均加标回收率在92.3%～137.4%之间.      

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.     

17α-乙炔基雌二醇的降解及其共基质代谢特性

以一株降解甾体雌激素17α-乙炔基雌二醇(EE2)的鞘氨醇杆菌(Sphingobacterium sp.)JCR5为材料,研究了JCR5降解EE2的适宜接种量、底物利用情况和添加营养物对降解能力的影响.结果表明,在接种量为7%(体积分数)(OD600≈1)时,EE2的降解率最高;JCR5能够利用甾体雌激素(甾酮、17β-雌二醇、雌三醇以及炔雌醇甲醚),避孕药生产中间体以及一些芳烃化合物为唯一碳源生长;添加少量葡萄糖、蛋白胨和酵母粉,均能促进JCR5对EE2的降解.葡萄糖、17β-雌二醇(E2)分别与EE2的共基质代谢试验表明,在共基质条件下,葡萄糖的存在对EE2的降解起到一定的促进作用,而E2的存在对EE2的降解具有一定的抑制作用.利用HPLC检测EE2降解过程发现,在EE2降解的同时伴有未知的EE2代谢产物峰出现.      

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.     

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.      

基于H4IIE细胞株测试间接雌激素效应物质的代谢活化方法

利用鼠肝癌细胞(H4IIE)代谢活化环境样品中间接雌激素效应物质的方法,以标准化合物双酚A和北方某污水处理厂各工艺过程的水样以及南方某水厂自来水样的有机提取物为对象,比较了代谢活化作用对雌激素效应检测结果的影响.研究结果表明,双酚A代谢活化产物的雌激素活性明显高于双酚A;环境样品中存在大量的间接雌激素效应物质,通过细胞代谢活化的方法能够检出其活性.因此,推荐将鼠肝癌细胞代谢活化与重组基因酵母检测的方法相结合,用于环境样品中直接和间接雌激素效应物质的检测.