苯并芘对稀有鮈鲫的内分泌干扰效应研究

为评价典型有机污染物对长江三峡库区上游特有珍稀鱼类的环境风险,本实验以稀有鮈鲫为对象,研究了低剂量多环芳烃(PAHs)暴露而引起的内分泌干扰效应.将成年稀有鮈鲫暴露于不同浓度(0、0.1、0.3、1.0和3.0μg·L-1)的苯并芘(Ba P)中28 d,测定了体质系数、性激素含量、下丘脑-垂体-性腺(HPG)轴和肝脏中与繁殖相关基因的表达.结果显示:在雌鱼中,Ba P暴露后,睾酮(T)的含量显著升高,雌二醇(E2)的水平显著下降并改变了T/E2的比值;在雄鱼中,T和E2的含量都显著下降.雌雄鱼性激素的改变可能与类固醇合成相关基因(Cyp17、Cyp19a)表达量的改变有关.Ba P暴露显著抑制了雌雄鱼肝脏中卵黄蛋白原(VTG)基因的表达.以上检测到的激素或者基因表达的改变主要发生在高剂量(3.0μg·L-1)Ba P暴露下,而长江上游库区水体中多环芳烃含量较低(100 ng·L-1),因此推测,库区存在的多环芳烃不会对三峡库区上游水体中的鱼类产生内分泌干扰效应.     

A screening assay for thyroid hormone signaling disruption based on thyroid hormone-response gene expression analysis in the frog Pelophylax nigromaculatus

Amphibian metamorphosis provides a wonderful model to study the thyroid hormone (TH) signaling disrupting activity of environmental chemicals, with Xenopus laevis as the most commonly used species. This study aimed to establish a rapid and sensitive screening assay based on TH-response gene expression analysis using Pelophylax nigromaculatus, a native frog species distributed widely in East Asia, especially in China. To achieve this, five candidate TH-response genes that were sensitive to T3 induction were chosen as molecular markers, and T3 induction was determined as 0.2 nmol/L T3 exposure for 48 hr. The developed assay can detect the agonistic activity of T3 with a lowest observed effective concentration of 0.001 nmol/L and EC50 at around 0.118–1.229 nmol/L, exhibiting comparable or higher sensitivity than previously reported assays. We further validated the efficiency of the developed assay by detecting the TH signaling disrupting activity of tetrabromobisphenol A (TBBPA), a known TH signaling disruptor. In accordance with previous reports, we found a weak TH agonistic activity for TBBPA in the absence of T3, whereas a TH antagonistic activity was found for TBBPA at higher concentrations in the presence of T3, showing that the P. nigromaculatus assay is effective for detecting TH signaling disrupting activity. Importantly, we observed non-monotonic dose-dependent disrupting activity of TBBPA in the presence of T3, which is difficult to detect with in vitro reporter gene assays. Overall, the developed P. nigromaculatus assay can be used to screen TH signaling disrupting activity of environmental chemicals with high sensitivity.     

超高效液相色谱-串联质谱法同时分析地下水中24种内分泌干扰物

建立了基于超高效液相色谱-串联质谱法同时测定地下水中9种雌激素、5种雄激素、3种肾上腺皮质激素、4种酚类和3种非甾类激素共24种内分泌干扰物的方法。在活化后的MCX固相萃取柱中加入水样,再加入pH=5的水溶液淋洗,随后用碱化乙腈(4.5%,体积分数)洗脱。收集洗脱液并用氮气吹至近干后,用甲醇定容。采用Poroshell 120 Bonus-RP色谱柱(2.7 μm,100 mm×2.1 mm),以0.1%氨水溶液-乙腈为流动相进行梯度洗脱,实现液相分离。质谱检测采用MRM模式进行采集,通过正负离子模式切换同时进行测定,使用内标法定量。检测结果显示,目标物在线性范围内,决定系数R²均大于0.995,方法检出限为0.05~2.00 ng/L,加标回收率为68.8%~108%。该方法灵敏度高、准确度好,具有较强的实用价值。     

壬基酚的环境生物地球化学研究进展及对新污染物管理的建议

近年来,我国大气环境、水环境和土壤环境质量得到一定的改善,“碧水蓝天”已经成为常态.但随着持久性有机污染物、内分泌干扰物、抗生素和微塑料等新污染物在环境中不断被检出,新污染物正逐步受到广泛的关注,壬基酚作为一种典型的内分泌干扰物也备受研究人员的关注.系统地概括了我国水体中壬基酚的环境行为和暴露水平,并基于风险商法和联合概率曲线法对壬基酚可能造成的生态风险进行了评估.结果表明,壬基酚对水生生物的毒性效应主要包括急性毒性、生长发育毒性、雌激素效应和繁殖毒性;壬基酚在我国主要流域水体中普遍存在,其浓度平均值在60～1 000 ng·L^-1,浓度最高值可达4 628 ng·L^-1;基于风险商法和联合概率曲线法的风险评估结果表明,壬基酚对我国主要流域里的水生生物均存在一定的风险.最后,总结了目前比较常用的壬基酚处理处置和风险管控技术,比较了国际上内分泌干扰物的监管方法,针对我国在新污染物环境管理中存在的问题,提出了针对性政策建议,研究结果可以为我国新污染物管理和管控提供参考.     

Fate of bisphenol A during treatment with the litter-decomposing fungi Stropharia rugosoannulata and Stropharia coronilla

Bisphenol A is an endocrine disrupting compound, which is ubiquitous in the environment due to its wide use in plastic and resin production. Seven day old cultures of the litter-decomposing fungus Stropharia coronilla removed the estrogenic activity of bisphenol A (BPA) rapidly and enduringly. Treatment of BPA with purified neutral manganese peroxidase (MnP) from this fungus also resulted in 100% reduction of estrogenic activity, as analyzed using a bioluminescent yeast assay, and in the formation of polymeric compounds. In cultures of Stropharia rugosoannulata, estrogenic activity also quickly disappeared but temporarily re-emerged in the further course of cultivation. LC-MS analysis of the extracted estrogenic culture liquid revealed [M−H]⁻ ions with m/z values of 219 and 235. We hypothesize that these compounds are ring fission products of BPA, which still exhibit one intact hydroxyphenyl group to interact with estrogen receptors displayed by the yeast.     

城市污水处理厂内分泌干扰物浓度分布和去除规律

采集南方某城市3个污水处理厂进出水及各工艺单元的水样,通过固相萃取-衍生化过程对水样进行预处理,并用气相色谱-质谱联用仪定量检测水样中的17α-雌二醇(17α-E2)、雌酮(E1)、雌二醇(E2)、雌炔醇(EE2)、雌三醇(E3)、辛基酚(OP)、正壬基酚(NP)和双酚A(BPA)等8种内分泌干扰物(EDCs).连续4个季度的分析结果显示,3个污水厂进出水及各工艺单元中均有EDCs检出,EE2和BPA是含量最高的两种EDCs,出水中最高浓度分别达到498.9ng.L-1和2652.5ng.L-1,其他6种EDCs浓度均不超过100ng.L-1;生物处理工艺对EE2、E3、NP、OP和BPA有较高的去除率,平均去除率约为57%～85%,但对于E1、E2和17α-E2的去除率较低且不稳定;一级处理工艺对EDCs仅有低于18%的去除率;紫外辐照消毒工艺对EDCs的去除没有明显效果;EDCs浓度季节差异显著,由于雨水的稀释作用,旱季浓度约为雨季的2.5～7.8倍.本文对于了解我国南方地区污水处理工艺中EDCs类物质的分布和转化特点有一定的参考意义.     

17β-雌二醇在水体中的分布水平及去除方法研究现状

17β-雌二醇(E2)是一种常见的天然雌激素,普遍存在于各类水体环境,在水环境分布水平已经达到干扰鱼类内分泌系统的水平,被认为是作用最强烈、最具潜在影响的环境内分泌干扰物之一。文章重点介绍了17β-雌二醇在水体环境中的分布水平及去除方法,指出由于传统的污水处理工艺和净水工艺无法将其完全去除,增加深度处理工序已经迫在眉睫。针对E2的特点,现有水处理工艺宜增加能够生成强氧化剂.OH的工序。     

Broad range analysis of endocrine disruptors and pharmaceuticals using gas chromatography and liquid chromatography tandem mass spectrometry

Endocrine disrupting compounds (EDCs) and pharmaceuticals and personal care products (PPCPs) have been globally detected in impacted natural waters. The detection of trace quantities of EDCs and PPCPs in the environment is of great concern since some of these compounds have known physiological responses at low concentrations. EDCs can have a wide range of polarities, acidic and basic moieties, and exist in trace quantities, which often requires numerous complex extractions, large sample collection volumes, and multiple instrumental analyses. A comprehensive method has been developed allowing for the analysis of 58 potential EDCs in various water matrices using a single solid-phase extraction (SPE) of a 1 L sample with subsequent analyses using both gas chromatography and liquid chromatography, each coupled with tandem mass spectrometry (GC–MS/MS and LC–MS/MS). Instrument detection limits ranged between 0.12–7.5 pg with corresponding method reporting limits of 1–10 ng l⁻¹ in water. Recoveries for most compounds were between 50% and 112% with good reproducibility (RSD 6–22%).     

应用重组孕激素基因酵母测定饮用水中内分泌干扰物的方法

研究了用重组孕激素受体基因酵母测定饮用水中内分泌干扰物的方法,并利用该种方法检测了南方某水厂不同处理工艺过程水样对孕激素受体活性的抑制水平.结果表明,重组孕激素受体基因酵母能够与孕激素专一性的结合,诱导产生明显的剂量-效应关系,EC₅₀值为0.5 nmol/L,具有较高灵敏度;环境内分泌干扰物五氯酚和壬基酚具有孕激素受体抑制活性,其IC₅₀值分别为2.4μmol/L和3.7μmol/L;重组孕激素受体基因/报道基因的酵母技术是一种筛选和定量分析具有孕激素受体抑制活性的内分泌干扰物的快速、有效方法.结合固相萃取的前处理技术,重组孕激素受体基因酵母对水厂不同处理工艺水样检测出明显的孕激素受体抑制活性,抑制率均大于58%,表明重组孕激素受体基因酵母检测技术能够快速监测和鉴别水样中具有抑制孕激素受体活性的物质.