海口市部分市售蔬菜4-壬基酚、双酚A污染情况初步调查研究

[目的]了解海口市部分蔬菜中4-壬基酚(4-NP)和双酚A(BPA)的污染情况。[方法]到海口市4个城区的农贸市场和超级市场随机获取5种蔬菜,采用GC-MS检测方法进行定性定量检测。[结果]4-NP和BPA在5种蔬菜中均有检出,其中白菜和油菜检出率为100%,含量分别为1.26～8.58μg/kg(4-NP)和0.43～5.31μg/kg(BPA)。[结论]由于4-NP和BPA及其下游产物在我国的广泛应用,有可能已对部分蔬菜造成污染,应引起有关部门的注意。     

气相色谱-质谱法检测血液中15种农药残留

目的建立血液中15种农药的气相色谱-质谱检测方法(GC-MS)。方法取2ml全血,用二氯甲烷萃取浓缩,以古洛糖酸内酯甲醇溶液定容,供装有DB-35MS(30m×0.25mm×0.25μm)毛细管柱的GC-MS定性定量测定。结果二嗪农、阿特拉津、扑草净、丁草胺、联苯菊酯在4～80μg/L浓度范围内,甲拌磷、2,4-D丁酯、甲基对硫磷、马拉硫磷、氯丹、甲氰菊酯在10～200μg/L浓度范围内,α-硫丹、β-硫丹、三氟氯氰菊酯在20～400μg/L浓度范围内,乐果在40～800μg/L浓度范围内,具有良好的线性响应,相关系数(r2)在0.998～1.000之间,各待测物的方法加标回收率为51.3%～113.1%,方法的检出限为0.05～1.00μg/L,方法的定量限为0.20～3.00μg/L,批内精密度在3.17%～22.79%,批间精密度在5.58%～26.13%。结论该方法适用于血液样品中15种农药的含量检测。     

工作场所空气中碘的离子色谱测定法

目的 建立工作场所空气中蒸气态碘的离子色谱测定方法.方法 工作场所空气中的碘蒸气用碱性炭管采集,碘与碱反应生成碘离子.用碳酸氢钠溶液解吸活性炭上的碘离子后进样,经离子色谱柱分离后,用电化学检测器检测,以保留时间定性、峰高或峰面积定量.结果 该法具有很好的回收率和批内、批间精密度,回收率在碘的5、10和15 μg的加标水平的平均回收率分别为115.0%、103.8%和106.4%;5μg的批内和批间精密度分别为3.1%和1.9%,10μg的批内和批间精密度分别为2.0%和1.2%,15μg的批内和批间精密度分别为1.8%和2.5%.该法的最低检出浓度(采集7.5 L空气样品)为0.08 mg/m3,满足职业接触限值检测的要求.结论 该方法适用于工作场所空气中蒸气态碘的浓度测定.     

4-壬基酚、双酚A对幼雌SD大鼠生殖器官的拟雌激素样作用探讨

目的检测4-壬基酚(4-NP)、双酚A(BPA)对幼雌SD大鼠的拟雌激素样作用和内分泌干扰作用。方法以21日龄雌性SD大鼠为实验对象,选取4-NP和BPA为染毒物质,均设高中低剂量,连续喂饲3天,于末次给药后24h处死。以子宫湿重、脏器系数、子宫内膜及平滑肌增生程度和细胞增殖核抗原(PCNA)的蛋白表达作为检测指标,采用单因素方差分析方法进行统计。结果染毒动物实验结果显示,高中剂量的4-NP(200mg/kg和100mg/kg)和BPA(600mg/kg和400mg/kg)能够使染毒动物的子宫湿重及脏器系数明显增加(P<0.01),并且具有一定的剂量-效应关系;4-NP和BPA高、中、低剂量都能够使子宫内膜上皮增高,平滑肌增厚,PCNA蛋白表达增强(P<0.05)。结论动物染毒实验进一步证实了4-NP、BPA作为环境内分泌干扰物的拟雌激素作用及其剂量-效应关系。     

高效液相色谱法测定尿中肌酐的方法改进

目的改进高效液相色谱法测定尿液中肌酐的方法。方法尿样经过离心(转速=10 000 r/min)后,取上清液,用流动相(20 mmol/L乙酸铵,pH=3.20)稀释10倍～500倍后,取10.0μl进样,在流速为1.0 ml/min条件下,经EC-C_(18)反相色谱柱(250 mm×4.6 mm, 4μm,柱温:25℃)分离,采用紫外检测器(VWD)检测(检测波长:236 nm),以保留时间定性,峰面积定量。结果本法尿肌酐在2.0μg/ml～10.0μg/ml浓度内呈线性,相关系数为0.999 6,以3倍噪音值估算方法检出限为0.10μg/ml。在2.0μg/ml～10.0μg/ml内添加低、中、高3个浓度水平,样品加标回收率为99.02%～100.26%;批内精密度为0.55%～2.73%(n=6),批间精密度为1.05%～3.20%(n=6)。结论本法操作简便,环境友好,分析时间短,分离效果好,干扰物质少,灵敏度高,定性定量准确,可以满足尿样中肌酐检测的要求。     

高效液相色谱法测定尿中苯巯基尿酸的方法改进

目的改进高效液相色谱法测定尿液中苯巯基尿酸的方法。方法尿样经过离心后,取上清液,加入水涡旋15 s,硫酸溶液(50%)涡旋30 s并需在10 min内加入氢氧化钾(7.8 mol/L)溶液涡旋30 s。以氯仿和异丙醇(V/V=5∶1)混合液进行液-液提取,提取液经氮气吹干后,甲醇定容注入液相色谱仪,经EC-C18反相色谱柱(150 mm×4.6 mm,4μm)分离,采用紫外检测器(VWD)检测,以保留时间定性,峰面积定量。结果本法苯巯基尿酸在200.0μg/L~1 000.0μg/L浓度内呈线性,相关系数为0.999 4,以3倍噪音值计算方法检出限为2.0μg/L。在200.0μg/L~1 000.0μg/L内添加低、中、高3个浓度水平,样品加标回收率为95.67%~97.74%;批内精密度为0.41%~1.56%(n=6),批间精密度为0.44%~2.52%(n=6)。结论本法简单实用,分析时间短,灵敏度高,定性定量准确,可以满足尿样中苯巯基尿酸的检测要求。     

凝胶渗透色谱与气相色谱-串联质谱法检测蔬菜中15种有机氯农药残留

目的通过优化凝胶渗透色谱净化系统(GPC)及气相色谱-串联质谱(GC-MS/MS)技术参数,建立GPC与GC-MS/MS法检测蔬菜中15种有机氯农药残留的方法。方法蔬菜样品经乙腈溶剂提取,氮吹至近干,上GPC处理净化,样品净化后用GC-MS/MS法进行定性、定量分析。实验中比较2条不同凝胶柱及GPC流动相在不同流速下对净化效果的影响,同时比较2条不同毛细管柱的分离效果,根据实验结果,采用20 mm×400 mm凝胶柱、流量为3 ml/min为净化条件,选用中等极性毛细管柱TR-1701为分析色谱柱。结果本方法中,15种有机氯农药的浓度为0.005μg/ml~0.200μg/ml时,线性相关系数(r)≥0.990,定量限为0.000 04 mg/kg~0.001 60 mg/kg。对韭菜样品按0.005 mg/kg、0.025 mg/kg、0.100 mg/kg 3组水平进行加标试验,方法的精密度(RSD)为2.8%~12.0%,回收率为72.1%~104.1%。结论本方法检测样品的范围广、灵敏度高、定性可靠、定量准确,适合蔬菜中农药多残留的分析。     

双酚A和对-壬基酚雌激素受体的竞争性结合

目的　用雌激素受体竞争性结合试验检测双酚A(BPA)、对 -壬基酚 (p -NP)的雌激素样活性。 方法　将SD大鼠进行人工卵巢去除术 ,饥饿受体 14d后提取子宫胞浆雌激素受体 (ER) ,受体蛋白定量 ,作雌二醇 (E2 )非标记配体饱和分析 ,以确定E2 的IC50 。随后进行BPA和 p -NP的竞争性取代实验 ,观察BPA ,p -NP是否能与3 H -E2竞争性结合大鼠子宫胞浆ER。结果　E2 的IC50 是 1× 10 -9mol/L ,BPA、p -NP的IC50 分别为 2 0 8× 10 -5mol/L和2 6 6R10 -5mol/L ,BPA、p -NP的相对结合亲和力系数 (RBA)分别为 0 0 0 4 8和 0 0 0 38,BPA与ER的结合能力稍高于 p -NP。 结论　BPA和 p -NP结合ER是发挥其雌激素样效应机制中的重要因素     

固相萃取-气质联用法测定蔬菜中氟虫腈和溴虫腈残留

目的建立蔬菜中氟虫腈和溴虫腈残留的固相萃取-气相色谱/质谱分析方法。方法试样经乙腈提取,硫酸镁和氯化钠盐析,通过比较PSA柱、石墨化碳/NH_2柱、中性氧化铝柱、Qu Ch ERS净化管4种小柱,对蔬菜中氟虫腈和溴虫腈的吸附和净化情况,选取PSA固相萃取柱进行净化,GC-MS测定,外标法定量。结果该方法在5μg/L~100μg/L具有良好的线性关系(相关系数均0.999),氟虫腈和溴虫腈的回收率分别为93.1%~99.2%和97.4%~100.0%,相对标准偏差(RSD)分别为3.5%~8.6%、2.8%~6.1%,检出限为1.0μg/kg~2.0μg/kg,定量限为3.0μg/kg~6.0μg/kg。应用该方法对90份蔬菜样品进行分析,检出2份阳性样品。结论该方法的灵敏度、精密度和LOD均符合残留分析要求,具有简便、准确、成本低的特点,适用于蔬菜中氟虫腈和溴虫腈农药残留的检测。     

电感耦合等离子体质谱法测定兔尿样、血浆及组织器官中的总铂

采用电感耦合等离子体质谱法测定兔尿样、血浆和组织器官中的总铂(Pt),兔血浆和尿样适当稀释后直接测定,脏器类样品湿法消解后测定。在0.02～50.00μg/L范围内方法线性良好,相关系数为1.00;尿样精密度良好,批内相对标准偏差2.17%～3.77%,批间相对标准偏差3.54%～4.35%,加标回收率100.90%～103.65%;血浆样品批内相对标准偏差1.57%～2.27%,批间相对标准偏差2.47%～2.64%,加标回收率100.14%～102.30%;组织样品批内相对标准偏差3.47%～4.45%,批间相对标准偏差3.55%～5.61%,加标回收率96.55%～97.60%;兔尿、血浆和脏器样品的方法定量限分别为0.18μg/L、0.20μg/L、0.03μg/g。本方法检出限低,灵敏度高,精密度好,适用于测定血浆、尿样及动物脏器中总铂含量,可为铂类抗癌药物药理、毒理学研究提供技术支持。     

用高压液相色谱法检测罐装食品中类雌激素双酚A和烷基酚

目的 建立不同基质食品中双酚A（BPA）、壬基酚（NP）和辛基酚（OP）等类雌激素的高压液相色谱法（HPLC）,检测市售罐装食品中类雌激素。方法 固体样品经甲醇超声提取、二氯甲烷萃取、浓缩。液体样品的分析采用NH。柱浓缩、富集和净化。HPLC采用WatersXTerra^TM MSC18液相色谱柱分离,甲醇．水作为流动相梯度洗脱。采用荧光检测,激发波长：225nm;发射波长：310nm。结果 建立的方法具有良好的线性关系,蔬菜类罐装食品和方便面中BPA、NP和OP的检出限分别为0．5、0．1和0．1μg/kg,鱼、肉类罐装食品BPA、NP和OP的检出限分别为1、0．5和0．5μg/kg。BPA、NP和OP加标回收率分别为74．9％～95．1％、76．3％～103．6％和72．1％～109．2％;相对标准偏差（RSD）分别为4．98％～11．2％、2．35％～8．88％和5．61％～12．3％。结论 该方法测定罐装食品及方便面中类雌激素（BPA、OP和NP）,具有较高的灵敏度和选择性,结果准确可靠、重现性好。     

4-壬基酚、双酚A对雌性SD幼鼠生殖器官拟雌激素样作用的联合效应

[目的]检测4-壬基酚（4-NP）、双酚A（BPA）对幼雌sD大鼠拟雌激素样作用的联合效应。[方法]选取4-NP、BPA为染毒物质;以21d雌性SD大鼠为实验对象,共分9组,每组8只。分组为：4-NP高、中、低剂量组（200、100、50mg／kg）,BPA高、中、低剂量组（600、400、200mg／kg）,混合组（50mg／kg4-NP＋200mg／kgBPA）,阳性对照组,溶剂对照组。连续喂饲3d,末次给药后24h处死。以子宫湿重、脏器系数、子宫内膜及平滑肌增生程度和细胞增殖核抗原（PCNA）的蛋白表达作为检测指标,采用单因素方差分析方法进行统计。[结果]高、中剂量的4-NP和BPA能够使染毒动物的子宫湿重及脏器系数明显增加（P〈0.01）;4-NP和BPA高、中、低剂量都能够使子宫内膜上皮、平滑肌增厚,PCNA蛋白表达增强（P〈0.05）,并且具有一定的剂量-效应关系。最大无效应剂量的4-NP（50mg／kg）和BPA（200mg／kg）混合喂饲后能够使染毒动物的子宫湿重及脏器系数明显增加;能够使子宫内膜上皮、平滑肌增厚,PCNA蛋白表达增强（P〈0.05）。[结论]4-NP、BPA具有拟雌激素样作用,干扰内分泌系统;两种物质混合后可以发挥联合效应。     

Fate of estrogens and xenoestrogens in four sewage treatment plants with different technologies

The fate and removal of the estrogens 17beta-estradiol (E(2)), estrone (E(1)), and 17alpha-ethynylestradiol (EE(2)) and of the xenoestrogens bisphenol A (BPA), 4-tert-octylphenol (4-t-OP), 4-nonylphenol (4-NP), and nonylphenol mono- and diethoxylate (NPEO1 and NPEO2, respectively) were investigated in four South Australian sewage treatment plants (STPs; plants A-D) with different treatment technologies. The concentrations in the effluent from the two-year survey were similar to those reported in other studies. In the effluent, 4-NP, NPEO1, and NPEO2 had total concentrations up to 8 microg/L, which were much higher than those of BPA and 4-t-OP. Estrone had the highest concentrations among the three estrogens, ranging between 13.3 and 39.3 ng/L, whereas the concentrations for E(2) and EE(2) varied between 1.0 and 4.2 ng/L and between 0.1 and 1.3 ng/L, respectively. The removal rates for the estrogens and xenoestrogens were variable but consistent with the plant performance parameters (biochemical oxygen demand, suspended solids, and ammonia). Considering all the estrogenic compounds analyzed in the present study, plant D, with a series of anaerobic and aerobic lagoons, was the least efficient of the four STPs in the removal of these compounds. The removal rates for 4-NP, NPEO1, and NPEO2 within the plants were 92% for plant A, with conventional activated sludge treatment; 80% for plant B, with two oxidation ditches; 70% for plant C, with three bioreactors; and 64% for plant D, with 10 lagoons in series. Comparatively, the removal of estrogens was lower, with rates ranging between 47 and 68% for E(2) at the four plants. Both E(1) and EE(2) were more persistent during treatment, especially in plants C and D.     

Temporal variability and predictors of urinary bisphenol A concentrations in men and women

BACKGROUND: Bisphenol A (BPA) is used to manufacture polymeric materials, such as polycarbonate plastics, and is found in a variety of consumer products. Recent data show widespread BPA exposure among the U.S. population. OBJECTIVE: Our goal in the present study was to determine the temporal variability and predictors of BPA exposure. METHODS: We measured urinary concentrations of BPA among male and female patients from the Massachusetts General Hospital Fertility Center. RESULTS: Between 2004 and 2006, 217 urine samples were collected from 82 subjects: 45 women (145 samples) and 37 men (72 samples). Of these, 24 women and men were partners and contributed 42 pairs of samples collected on the same day. Ten women became pregnant during the follow-up period. Among the 217 urine samples, the median BPA concentration was 1.20 microg/L, ranging from below the limit of detection (0.4 microg/L) to 42.6 microg/L. Age, body mass index, and sex were not significant predictors of urinary BPA concentrations. BPA urinary concentrations among pregnant women were 26% higher (-26%, +115%) than those among the same women when not pregnant (p > 0.05). The urinary BPA concentrations of the female and male partner on the same day were correlated (r = 0.36; p = 0.02). The sensitivity of classifying a subject in the highest tertile using a single urine sample was 0.64. CONCLUSION: We found a nonsignificant increase in urinary BPA concentrations in women while pregnant compared with nonpregnant samples from the same women. Samples collected from partners on the same day were correlated, suggesting shared sources of exposure. Finally, a single urine sample showed moderate sensitivity for predicting a subject's tertile categorization.     

Estimation of intake level of bisphenol A in Japanese pregnant women based on measurement of urinary excretion level of the metabolite

OBJECTIVE: The daily bisphenol A (BPA) intake level of Japanese pregnant women was surveyed based on the measurement of the urinary excretion level of a BPA metabolite. METHODS: Spot urine samples were collected from 56 pregnant women who visited the gynecology division of a hospital for a routine health check between June and October 2003. The urinary concentrations of the BPA metabolite and creatinine were measured by GC/MS/MS and spectrophotometry, respectively. Daily BPA intake was assumed to be equal to daily excretion. RESULTS: The daily intake of BPA among Japanese pregnant women was estimated to be in the range of < 0.3 to 7.9 microg/day (median < 2.0 microg/day), being consistent with the levels in previous studies for non-pregnant Japanese women. This level was far below the current Acceptable Daily Intake (0.01 mg/kg/day) which was set by the European Commission. The maximum estimated intake per body weight (0.16 microg/kg/day) reached 1/10 of the Lowest Adverse Effect Level of BPA for pregnant mice for a reproductive effect on the offspring (2 microg/kg/day). CONCLUSION: It is desirable to lessen BPA intake from a precautionary viewpoint, particularly in pregnant women.     

A Survey of Alkylphenols,Bisphenols, and Triclosan in Personal Care Products from China and the United States

Exposure of humans to environmental phenolic compounds such as bisphenol A (BPA) and alkylphenols is a matter of concern, due to these compounds’ ubiquitous occurrence and estrogenic potencies. Little is known about the levels of environmental phenolics in personal care products (PCPs). In this study, nonylphenol, two octylphenols, eight bisphenols (BPA and its analogs), and triclosan (TCS) were determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in PCP samples (n = 231) collected from China and the United States (U.S.). The concentrations of 4-n-nonylphenol (4-NP), 4-n-octylphenol (4-OP), 4-tert-octylphenol (4-t-OP), and TCS were in the ranges of <0.5–39,100 [geometric mean (GM): 21.5], <0.5–315 (0.680), <1.0–10,100 (2.69), and <0.5–53,900 (3.03) ng/g, respectively. The GM concentrations of individual bisphenols, including BPA, bisphenol S (BPS), and bisphenol F (BPF), were generally at sub-nanogram per gram levels. No significant differences in concentrations of the target compounds were found among various PCP categories or between China and the U.S. The estimated GM daily intakes of 4-NP, ∑OPs (sum of 4-OP and 4-t-OP), ∑BPs (sum of eight bisphenols), and TCS through dermal absorption from the use of PCPs were 0.932, 0.093, 0.072, and 0.016 μg/day, respectively, for adult Chinese women and 0.340, 0.054, 0.120, and 0.068 μg/day, respectively, for adult U.S. women. Body lotions, face creams, and liquid foundations accounted for the majority (>85 %) of the dermal exposure doses of the target phenolics.     

Effects of bisphenol A on the development, growth, and sex ratio of the housefly Musca domestica

The effects of bisphenol A (BPA) on the life cycle of the housefly Musca domestica were examined. The sex ratio of the imago shifted in favor of males when eggs and larvae were exposed to BPA in culture media at concentrations of 1,000 microg kg(-1) for five generations and 100 microg kg(-1) for seven generations. Notably, at an initial concentration of 100 microg kg(-1), BPA levels in the growth medium decreased 61% at 4 h after egg inoculation, and no BPA was detected after 24 h. Pupal weight increased upon exposure to 100 microg kg(-1) BPA but decreased after exposure to 1,000 microg kg(-1), suggesting highly variable concentration-dependent toxicity. Both the survival ratio of eggs to the third instar larval stage and the ratio of pupae to larvae decreased, indicating that BPA affected both eggs and larvae. A delay in the timing of emergence typically was observed in insects exposed to >100 microg kg(-1) BPA. Compared to the control group, juvenile hormones II and III levels were elevated significantly in larvae between days 4 and 7 in medium spiked with 100 microg kg(-1) BPA. These results suggest that exposure to environmentally relevant concentrations of BPA during the early stages of the housefly life cycle can result in various disorders (pupal weight and sex ratio) that may be a consequence of endocrine disruption.     

The Effects of Three Selected Endocrine Disrupting Chemicals on the Fecundity of Fruit Fly, Drosophila melanogaster

Bisphenol A (BPA), 4-nonylphenol (4-NP) and 4-tert-octylphenol (4-tert-OP) are the endocrine disrupting chemicals (EDCs) that has been shown to exert both toxic and biological effects on living organisms. The present study investigated effects of environmentally relevant concentrations of BPA, 4-NP and 4-tert-OP (0.1, 1 and 10 mg/L) on the fecundity of fruit fly Drosophila melanogaster. In the all exposure groups of BPA, 4-NP and 4-tert-OP, it was found a statistically significant decrease in mean fecundity as compared to the control groups (p < 0.05).