邻苯二甲酸(2-乙基己基)酯对雄性小鼠的生殖毒性

为探讨邻苯二甲酸(2-乙基己基)酯[di(2-ethylhexyl)phthalate,DEHP]对小鼠的生殖毒性.将30只成年健康SPF级雄性昆明小鼠随机分成5组,分别为阴性对照(玉米油)组和低(1 250 mg/kg)、中(2 500 mg/kg)、高剂量(5000mg/kg)DEHP染毒组以及阳性对照(环磷酰胺40 mg/kg)组,每组6只.除阳性对照组采用腹腔染毒外,其余各组采用经口灌胃方式进行染毒,染毒容量为0.02 ml/g,每天1次,连续染毒5d.于首次染毒5周后,采用一般生殖毒性实验方法对精子计数和精子存活率、畸形率、活动率进行测定.结果显示,与阴性对照组比较,各剂量DEHP染毒组小鼠精子计数和精子存活率均较低;而中、高剂量DEHP染毒组精子畸形率较高,活动率较低,差异均有统计学意义(P＜0.05).随着DEHP染毒剂量的升高,小鼠精子计数和活动率、精子存活率均呈下降趋势,精子畸形率呈上升趋势.提示DEHP对雄性小鼠具有一定的生殖毒性.     

邻苯二甲酸二(2-乙基己基)酯对小鼠子宫发育的毒性作用

目的 研究邻苯二甲酸二(2-乙基己基)酯[di- (2-ethylhcxyl)phthalate,DEHP]对未成年小鼠子宫发育的毒性作用.方法 将60只未成年(3周龄)清洁级ICR雌性小鼠随机分为4组,分别为对照(玉米油)组和低、中、高剂量DEHP染毒组(100、400、1600 mg/kg),每组15只.采用灌胃方式进行染毒,每天1次,每周5d,连续6周.测定小鼠子宫内膜及其上皮的厚度.结果 与对照组比较,中、高剂量DEHP染毒组小鼠子宫内膜上皮厚度显著下降,差异均有统计学意义(P＜0.05);而各剂量DEHP染毒小鼠子官内膜厚度无显著改变.且随着DEHP染毒剂量的升高,小鼠子宫内膜上皮的厚度呈下降趋势.结论 在本实验条件下,DEHP染毒未成年雌性小鼠影响了子宫的发育,对子宫发育具有毒性作用.     

邻苯二甲酸二(2-乙基己基)酯对青春前期雄性大鼠的生殖毒性

目的探讨邻苯二甲酸二(2-乙基己)酯(di-2-ethylhexyl phthalate,DEHP)对青春前期雄性大鼠的生殖毒性。方法将32只健康21日龄清洁级雄性SD大鼠按体重随机分为4组,分别为溶剂对照(玉米油)组和250、500、1 000 mg/kg DEHP染毒组,每组8只。采用灌胃方式进行染毒,染毒容量为10 ml/kg,每天1次,连续染毒4周。采用CASA精子分析系统分析精子活力相关指标。结果与对照组比较,1 000 mg/kg DEHP染毒组大鼠的精子密度、精子活动率、活动精子密度、最大侧摆幅度、直线运动的精子密度及A级精子百分率均较低,而D级精子百分率较高;250 mg/kg DEHP染毒组平均直线运动速度、精子运动的向前性均较低;500、1 000 mg/kg DEHP染毒组直线运动的精子个数均较低,差异均有统计学意义(P0.05);而各剂量DEHP染毒组B、C级精子比例和平均曲线运动速度、平均路径速度、平均侧摆幅度、平均鞭打频率、运动的直线性、运动的摆动性、平均移动角度、直线运动精子活率均无明显改变。结论 DEHP对青春前期雄性大鼠精子活力产生明显的毒性作用,从而影响雄性的生殖功能。     

邻苯二甲酸二(2-乙基己基)酯、氯氰菊酯单独及联合染毒对青春前期雄性大鼠生殖发育的不良影响

目的探讨邻苯二甲酸二(2-乙基己基)酯(DEHP)和氯氰菊酯(CYP)单独及联合染毒对青春前期雄性大鼠生殖发育的不良影响。方法选择SPF级3周龄雄性SD大鼠24只,随机分为4组,分别为对照组(喂饲玉米油)、DEHP染毒组(500mg/kg)、CYP染毒组(80mg/kg);DEHP、CYP联合染毒组(DEHP 500mg/kg+CYP 80mg/kg),每组6只。采用经口灌胃方式染毒,每天1次,连续染毒30天。于末次染毒24小时后处死动物,测定大鼠体重、睾丸湿重,并计算睾丸系数;测定血清睾酮水平;制备睾丸病理组织切片,光镜观察其组织学改变,电镜观察生殖细胞超微结构;测定睾丸标志酶乳酸脱氢酶(LDH)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)和琥珀酸脱氢酶(SDH)的活性。结果与对照组相比,DEHP、CYP单独及联合染毒组睾丸下降时间和包皮分离时间明显延迟,肛门生殖器间距明显缩短(P<0.05或0.01);DEHP单独染毒组睾丸重量及睾丸系数明显降低,血清睾酮水平显著下降(P<0.05),睾丸匀浆中ALP、ACP和LDH活性明显增高,SDH活性显著下降(P<0.01);DEHP、CYP单独及联合染毒组睾丸病理组织学、生殖细胞超微结构均可见明显异常。结论 DEHP、CYP单独及联合染毒对青春前期雄性大鼠均具有明显的生殖毒性作用,可引起生精细胞、支持细胞和间质细胞的变性、坏死及功能障碍,从而导致雄性生殖系统发育和功能的显著异常。其中,DEHP单独染毒呈现主效应,DEHP、CYP联合染毒对大鼠的生殖毒性未呈现明显交互影响。     

邻苯二甲酸二(2-乙基己基)酯及其代谢产物MEHP对体外大鼠卵泡发育的影响

目的利用大鼠腔前卵泡体外培养方法研究邻苯二甲酸二(2-乙基己基)酯(DEHP)及其代谢产物邻苯二甲酸单(2-乙基己基)酯(MEHP)对卵泡体外发育的影响。方法机械性分离大鼠腔前卵泡,单个卵泡在96孔板中连续培养。MEHP设2.5、5、10、20、40和80μg/ml6个浓度,DEHP设33.7、67.5、125、250、500、1000nmol/L6个浓度,同时设二甲基亚砜(DMSO)溶剂对照和阴性对照。每组16～20个卵泡,3次重复。隔天换1/2液,在培养第10天诱导排卵,观察卵泡体外发育情况。结果DEHP对第11天卵泡存活率、有腔形成率、异常卵泡形成率和卵丘-卵母细胞复合体(COCs)排出率无明显的影响,与溶剂对照组差异无显著性(P0.05)。第2天暴露10μg/ml以上剂量的MEHP48h后,第11天卵泡存活率明显下降(54.76%±3.37%),与对照组(91.57%±1.32%)比较差异有显著性(P0.05),剂量与卵泡存活率之间存在一定相关性(R2=0.92);COCs排出(24.99%±3.95%)明显减少,与对照组(52.78%±8.45%)比较差异有显著性(P0.05);异常卵泡出现率(45.24%±3.37%)明显升高,与对照组比较差异有显著性(P0.05);暴露20μg/mlMEHP有腔形成率(46.18%±1.67%)减少,与对照组(85.91%±5.03%)比较差异有显著性(P0.05),且有明显剂量-反应关系。结论MEHP可以抑制体外大鼠腔前卵泡的生长发育。     

孕期及哺乳期邻苯二甲酸二(2-乙基己基)酯经口染毒对雄性仔鼠生殖发育的影响

目的观察邻苯二甲酸二(2-乙基已基)酯(DEHP)对雄性仔鼠的生殖发育毒性作用。方法采用围生期毒性试验的研究方法,将清洁级SD大鼠随机分为阴性对照组(玉米油)和4个DEHP染毒组(125、250、500、1000mg/kg),采用宫内及哺乳期[受孕后第2天～仔鼠出生后第21天(PND21)]经口灌胃的方式染毒。记录比较雄性仔鼠肛殖距、不同发育阶段雄性仔鼠体重、睾丸、附睾重量、仔鼠出生后第50天(PND50)附睾尾精子总数、精子活率及精子畸形数。结果 125～500mg/kg组不同发育阶段仔鼠体重随染毒剂量增加而增加,500mg/kg组PND21体重与对照组比较,差异有统计学意义(P0.01),1000mg/kg组体重均低于同发育阶段对照组,PND21体重与对照组比较,差异具有统计学意义(P0.05)。1000mg/kg组不同发育阶段仔鼠睾丸重量明显降低,与对照组比较,差异有统计学意义(P0.01),附睾重量与对照组比较差异均无统计学意义(P0.05)。仔鼠出生后4天肛殖距不同程度缩短,除125mg/kg组外,其余各剂量组与对照组比较,差异均有统计学意义(P0.01)。PND50雄性仔鼠精子总数、精子活率均不同程度降低,精子畸形率不同程度增高,1000mg/kg组与对照组比较,差异有统计学意义(P0.05)。结论母鼠孕期及哺乳期暴露DEHP可影响雄性仔鼠生长、生殖发育。     

邻苯二甲酸二(2-乙基己)酯对初断乳雄性小鼠的毒性作用

选用ICR初断乳雄性小鼠,按体重随机分为对照组和250、500、1 000 mg/kg邻苯二甲酸二(2-乙基己)酯(DEHP)染毒组,连续染毒灌胃8周,观察各组小鼠的一般状况、血液指标、脏器系数及病理组织学检查。结果显示,DEHP能损伤小鼠肝脏和睾丸,且睾丸损伤较明显。     

邻苯二甲酸二(2-乙基己基)酯生物降解研究进展

邻苯二甲酸二 (2 乙基己基 )酯是一种污染严重的有毒难降解有机物 ,自然降解能力很弱 ,降解过程主要为生物降解。运用微生物对其进行生物降解有着十分广阔的前景     

邻苯二甲酸二(2-乙基己基)酯毒性作用

邻苯二甲酸二(2 乙基己基)酯(DEHP)作为聚氯乙烯(PVC)等塑料制品的增塑剂,可增加塑料的弹性和韧性,被广泛应用于塑料工业。DEHP在塑料中是以游离的形式存在的,在重量上可达 30%~50%[1],很容易进入环境。作为环境污染物,可以在地表水、地下水、饮用水、空气、土壤及动、植物体内广泛检测到。目前的研究发现其毒性作用主要是作为内分泌干扰物,表现为生殖生长毒性、肝肾毒性及血液和生化方面的改变。随着DEHP产量的逐年增加,其在环境中的浓度也逐渐升高,对人体的毒性作用也越来越受到人们的关注[2]。目前对其健康效应方面的评价尚处…     

Metabolites of the plasticizer di (2-ethylhexyl) phthalate in urine samples of workers in polyvinylchloride processing industries

Summary Little is known about occupational exposure to the plasticizer di(2-ethylhexyl)phthalate (CAS number 117-81-7), a compound widely used in polyvinylchloride (PVC) plastics. We have studied the uptake of DEHP in workers by determining the concentrations of four metabolites of DEHP in urine samples, i.e., mono(2-ethylhexyl)phthalate (MEHP), mono (5-carboxy-2-ethylpentyl)phthalate, mono(2-ethyl-5-oxohexyl)phthalate, and mono(2-ethyl-5-hydroxyhexyl)phthalate. In addition DEHP concentrations in the air were determined by personal air sampling. Nine workers in a PVC boot factory exposed to a maximum of 1.2 mg/m³ DEHP showed an increase in the urinary concentrations of all four metabolites over the workshift. These results were obtained on both the first and the last day of the workweek. With the exception of MEHP, the increases in the concentrations of the metabolites during a workday were statistically significant. Six workers from a PVC cable factory exposed to a maximum of 1.2 mg/m³ DEHP showed a one-to fourfold increase in the concentrations of the four metabolites over the workshift, but these increases were not statistically significant. These results indicate that measurement of DEHP metabolites in urine samples may be of use for monitoring the occupational exposure to DEHP.     

Determination of four metabolites of the plasticizer di (2-ethylhexyl) phthalate in human urine samples

Summary A method for biological monitoring of exposure to the plasticizer di(2-ethylhexyl)phthalate (DEHP) is described. In this method the four main metabolites of DEHP [i.e., mono (2-ethylhexyl) phthalate (MEHP), mono (5-carboxy-2-ethylpentyl)phthalate, mono(2-ethyl-5-oxohexyl)phthalate, and mono(2-ethyl-5-hydroxyhexyl)-phthalate] are determined in urine samples. The procedure includes enzymatic hydrolysis, ether extraction, and derivatization with triethyloxonium tetrafluoroborate. Analysis is performed by gas chromatography electron impact mass spectrometry. The detection limit for all four metabolites is less than 25 g/l urine. The coefficient of variation based on duplicate determinations of urine samples of workers occupationally exposed to DEHP was 16% for MEHP (mean concentration 0.157 mg/l) and 6% -9% for the other three metabolites (mean concentrations 0.130-0.175 mg/1). The method described here was used to study DEHP metabolism in man. Most persons excrete mono(2-ethyl-5-oxohexyl)-phthalate and mono (2-ethyl-5-hydroxyhexyl)phthalate as a (glucuronide) conjugate. Mono (5-carboxy-2-ethyl-pentyl)phthalate is mainly excreted in free form, while for MEHP a large interindividual variation in conjugation status was observed. Of the four metabolites quantified, 52% are products of a ((-l)-hydroxylation reaction of MEHP [i.e., mono (2-ethyl-5-oxohexyl)phthalate and mono (2-ethyl-5-hydroxyhexyl)phthalate], 22% is the product of a -hydroxylation reaction of MEHP [i.e., mono (5-carboxy-2-ethylpentyl)phthalate], and 26% is not oxidized further (i.e., MEHP). A good correlation is obtained when the amount of MEHP -hydroxylation products is compared with the amount of MEHP (-1)hydroxylation products in urine samples. When the internal dose of DEHP has to be established we recommend that the levels of all four metabolites of DEHP be studied in urine samples.     

塑料拆解地区水环境中DEHP检测

目的 监测某塑料拆解地区水环境中邻苯二甲酸二(2-乙基己基)酯(DEHP)的污染水平及探讨其可能的来源,为保护当地环境、促进居民健康提供基础数据。方法 用固相萃取和气相色谱联用技术检测河水、井水、塘水、无组织排放水和饮用水中DEHP水平。结果 各类水样中均检出DEHP,检出率为100%。河流污染断面DEHP平均浓度2.05μg/L,为清洁断面平均浓度(0.32μg/L)的6.4倍。暴露区井水(14.20μg/L)和塘水(135.68μg/L)DEHP的平均浓度分别为对照组井水(0.79μg/L)和塘水(0.37μg/L)的18和367倍。暴露区无组织排放水样中DE-HP的检出范围在0.36~161.86μg/L。结论 塑料拆解地区井水、塘水和无组织排放水样中均检出DEHP,且DEHP的平均浓度均超过中国《生活饮用水卫生标准》(GB5479-2006)和《地表水环境质量标准》(GB3838-2002)的要求,应引起重视。     

Di(2-ethylhexyl)phthalate (DEHP) exposure of voluntary plasma and platelet donors

Each year thousands of healthy volunteers undergo apheresis procedures to donate blood components and safe lives. However, many disposables used in apheresis contain di(2-ethylhexyl)phthalate (DEHP). This way, donors are exposed to DEHP, which is a reproductive and developmental toxicant in animals and a suspected endocrine modulator in humans. We quantified the DEHP exposure of six plasma donors, six discontinuous-flow platelet donors and six continuous-flow platelet donors by determining three specific metabolites in urine (5OH-MEHP: mono(2-ethyl-5-hydroxyhexyl)phthalate; 5oxo-MEHP: mono(2-ethyl-5-oxo-hexyl)phthalate and MEHP: mono(2-ethylhexyl)phthalate). We found maximum concentrations in urine samples after the discontinuous-flow plateletpheresis procedure with 826 microg/l for 5OH-MEHP, 774 microg/l for 5oxo-MEHP and 266 microg/l for MEHP (mean of the six volunteers). Metabolite excretions were found to be significantly (p<0.0001) higher for both plateletpheresis techniques compared to plasmapheresis and controls. Continuous-flow plateletpheresis led to significantly higher (p<0.0001) excretions than discontinuous-flow plateletpheresis. Mean absolute DEHP exposures were 1.2 mg for discontinuous- and 2.1 mg for continuous-flow plateletpheresis. Exposure for plasmapheresis (0.37 mg) was in the range of the controls (0.41 mg). Mean DEHP doses for both plateletpheresis techniques (18.1 and 32.3 microg/kg/day) were close to or exceeded the reference dose (RfD) of the US EPA and tolerable daily intake (TDI) value of the EU on the day of the apheresis. Therefore, margins of safety might be insufficient to protect especially young men and women in their reproductive age from effects on reproductivity. At present, discontinuous-flow devices should be preferred to avert conceivable health risks from plateletpheresis donors. Strategies to avoid DEHP exposure of donors during apheresis need to be developed.     

Species and inter-individual differences in metabolic capacity of di(2-ethylhexyl)phthalate (DEHP) between human and mouse livers

Objectives

This study was conducted to assess inter-species and inter-individual differences in the metabolism of di(2-ethylhexyl)phthalate (DEHP) in humans and mice.

Methods

The activities of four DEHP-metabolizing enzymes [lipase, UDP-glucuronocyltransferase (UGT), alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH)] were measured in the livers of 38 human subjects of various ages and in eight 129/Sv male mice.

Results

Microsomal lipase activity was significantly lower in humans than in mice. The V _max/K _m value in humans was one-seventh of that in mice, microsomal UGT activity in humans was a sixth of that in mice, and cytosolic ALDH activity for 2-ethylhexanal in humans was one-half of that in mice. In contrast, ADH activity for 2-ethylhexanol was twofold higher in humans than in mice. The total amount of DEHP urinary metabolites and the concentration of mono(2-ethylhexyl)phthalate (MEHP) were much higher in intact mice than in the U.S. general population based on data reported elsewhere, regardless of the similar estimated DEHP intake between these mice and the human reference population. However, mono(2-ethyl-5-oxo-hexyl)phthalate (5oxo-MEHP) and mono(2-ethyl-5-carboxypentyl)phthalate (5cx-MEPP) levels were higher in the latter than in the former. Of note, inter-subject variability in the activities of all enzymes measured was 10–26-fold.

Conclusion

The inter-individual variation in the metabolism of DEHP in humans may be greater than the difference between mice and humans (inter-species variation), and both may affects the risk assessment of DEHP.     

高效液相色谱法测定化妆品中邻苯二甲酸二丁酯和邻苯二甲酸二(2-乙基己基)酯的含量

目的：建立了高效液相色谱法测定化妆品中邻苯二甲酸二丁酯（DBP）和邻苯二甲酸二（2-乙基己基）酯（DE-HP）含量的方法。方法：采用固相萃取法处理样品，淋洗试剂为甲醇：乙醚=1：19。甲醇和水作为流动相，二极管阵列检测器测定DBP和DEHP。结果：DBP测定的线性范围为5．0～200μg／ml，线性相关系数为0．9993，DEHP测定的线性范围为5．5～220μg／ml，线性相关系数为0．9997。DBP的回收率范围在84％～93％，相对标准偏差为3．2％～4．8％。DE-HP的回收率范围在98％～105％，相对标准偏差为2．8％～4．7％。结论：该方法灵敏度高，结果准确而且稳定。     

气相色谱-质谱法测定饮用水中19种农药残留及邻苯二甲酸二（2-乙基己基）酯

目的 建立饮用水中19种农药残留及邻苯二甲酸二（2-乙基己基）酯的气相色谱-质谱测定方法。方法 水样用 1∶1正己烷-二氯甲烷混合溶剂萃取、旋蒸浓缩后进样分析。以保留时间、特征离子定性,外标法定量。结果 方法线性范围为0.05～5 mg/L,相关系数为0.9953～0.9998,检出限为0.0009～0.007 mg/L,回收率为86.4%～101.9%,相对标准差为1.8%～8.5%。结论 本方法操作简单、准确快速、灵敏度高、精密度高,满足饮用水中农药和邻苯二甲酸二（2-乙基己基）酯残留的测定要求。     

青春期前邻苯二甲酸二(2-乙基)己酯暴露对雌性大鼠生殖发育及过氧化物酶体增殖剂激活受体的影响

目的探讨邻苯二甲酸二(2-乙基)己酯(DEHP)暴露对青春期前雌性大鼠性发育及生殖内分泌功能的影响及其可能机制。方法将40只健康3周龄雌性SD大鼠,随机分成对照组(玉米油)和3个实验组,每组10只。实验组按50、150、500mg/kg DEHP经口灌胃,连续染毒28天。观察阴道开口、乳房发育、第一次发情周期的日龄及体重,于末次染毒24小时后进行阴道涂片,确定动情间期,处死动物。Real-time PCR测定卵巢组织相关基因表达水平;ELISA法检测血清中卵泡刺激素(FSH)、黄体生成素(LH)、雌二醇(E2)、孕酮(P4)及睾酮(T)水平;通过病理学观察卵巢组织的变化;免疫组织化学法测定卵巢组织中PPARγ的表达。结果 500mg/kg组阴道开口日龄提前,150、500mg/kg组阴道开口时体重增加(P<0.05);150、500mg/kg组芳香化酶(P450Arom)mRNA表达与对照组比较,显著下降;各剂量组与对照组相比,T水平明显下降,150、500mg/kg组FSH、E2水平明显减少,而LH水平明显升高(P<0.05);150、500mg/kg组闭锁卵泡明显增多、黄体数目明显减少;150、500mg/kg组卵泡颗粒细胞和黄体颗粒细胞中PPARγ阳性光密度相对量明显高于对照组及50mg/kg组(P<0.05)。结论青春期前DEHP暴露对雌性大鼠性发育及生殖内分泌功能产生影响,其作用机制可能与激活PPARs有关。     

Internal exposure of nursery-school children and their parents and teachers to di(2-ethylhexyl)phthalate (DEHP)

Di(2-ethylhexyl)phthalate (DEHP) is the main plasticizer for polyvinyl chloride (PVC) products. It has become widely spread in our environment and among people. DEHP is suspected to be responsible for endocrine-disruptor-like effects in mankind. Children are probably most susceptible to these endocrine effects. In this study we determined the internal exposure of nursery school children (aged 2-6 years) to DEHP and compared it to their parents' and teachers' exposure. The DEHP-metabolites mono(2-ethyl-5-hydroxyhexyl)phthalate (5OH-MEHP), mono(2-ethyl-5-oxo-hexyl)phthalate (5oxo-MEHP) and mono(2-ethylhexyl)phthalate (MEHP) were determined in first morning urine. The sum of the three DEHP metabolites in children's and in adults' urine was 90.0 and 59.1 micrograms/l respectively (median values; p = 0.074). Concentrations of the secondary metabolites 5OH-MEHP (median: 49.6 vs. 32.1 micrograms/l; p = 0.038) and 5oxo-MEHP (median: 33.8 vs. 19.6 micrograms/l; p = 0.015) were significantly higher in children than in adults. MEHP concentrations were low both in adults and children (median: 6.6 micrograms/l vs. 9.0 micrograms/l). Creatinine adjusted values should more accurately reflect the dose taken up with respect to body weight when comparing children with adults. Total creatinine adjusted DEHP metabolites in urine were significantly higher in children than in adults (median values: 98.8 vs. 50.9 micrograms/g creatinine; p < 0.0001). This also applied to the concentrations of both secondary metabolites 5OH-MEHP (55.8 vs. 28.1 micrograms/g creatinine; p < 0.0001) and 5oxo-MEHP (38.3 vs. 17.2 micrograms/g creatinine; p < 0.0001). Creatinine corrected concentrations for the monoester MEHP in children and adults were very similar (8.7 vs. 8.6 micrograms/g creatinine; p = 0.908). Based on the sum of the three determined metabolites we estimated the DEHP dose (in microgram/kg body-weight) taken up by children to be about twice as high as the dose taken up by adults. Routes of the ubiquitous exposure to DEHP remain indistinct. In children's urine the mean relative ratios of MEHP to 5OH-MEHP to 5oxo-MEHP were 1 to 7.1 to 4.9, in adults they were 1 to 3.4 to 2.1. This might indicate an enhanced oxidative metabolism in children. To date no information on the biological activity and toxicity of oxidative metabolites of DEHP is available. Since these are the major metabolites of DEHP toxicological data on these metabolites is urgently needed.