基于固相萃取-高效液相色谱-串联质谱测定孕妇尿液中8种邻苯二甲酸酯代谢物及新生儿出生结局评价

Studies had suggested possible association between polymer products and phthalate esters exposures in humans, but only a few had studied the level of phthalates that leaches out of these polymers into blood, which would eventually be detectable in urine or breast milk samples of blood transfusion recipients. This study examined the level of diethyl phthalate (DEP), dipropyl phthalate (DPP), di-n-butyl phthalate (DnBP), diethylhexyl phthalate (DEHP), monoethyl phthalate (MEP), monopropyl phthalate (MPP), monobutyl phthalate (MBP), mono-2-ethylhexyl phthalate (MEHP), mono-2-ethyl-5-hydroxyhexyl phthalate (MEHHP) and mono-2-ethyl-5-oxohexyl phthalate (MEOHP) using liquid chromatography tandem mass spectrometry in blood, urine and breast milk samples of mothers who had blood transfusion during intra-partum care. We also examined the level of phthalates in some controls who were mothers who had no transfusion at all during intra-partum care. The transfused mothers had significantly high levels of MEHHP, MEHP, MPP and DEHP in all of their samples and high DBP and MEP in almost all of the samples. Other phthalates were also detected at high concentrations, although relatively low levels of these were detected in all of the control samples. Analysis of variance (ANOVA) showed significant difference between the samples and controls. In this study, a positive association between blood transfusion and phthalate exposure in transfused mothers was established.

To investigate the levels and influencing factors of phthalate internal exposure in pregnant women (gestation age ≤ 16 weeks). During April to June in 2013, 1 020 pregnant women (gestation age ≤ 16 weeks) who had established the maternal care manual were recruited in maternal and child health hospital of Siming District, Xiamen city. Participators were asked to complete a questionnaire to obtain information on socio-demographic characteristics, lifestyle behaviors, and antenatal examination and to provide a urine sample. Finally, 998 pregnant women who provided a urine sample and completed the questionnaire were enrolled. Adopting systematic sampling method, 100 ones were selected randomly among 998 pregnant women. High performance liquid chromatography-electrospray ionization-tandern mass was used to determine the concentration of five phthalate monoesters in each urine, including mono-n-methyl phthalate (MMP), mono-ethyl phthalate (MEP), mono-butyl phthalate (MBP), mono-benzyl phthalate (MBzP), mono-ethylhexyl phthalate (MEHP). Based on the measurements and questionnaire data, multivariate logistic regression was used to analyze the association between the phthalate monoester levels and potential influential factors. The detection rates of MMP, MEP, MBP, MBzP and MEHP in 100 pregnant urine samples were 94%, 93%, 87%, 83%, 99%, respectively. And the urinary median uncorrected concentrations of MMP, MEP, MBP, MBzP and MEHP in 100 urine samples were 20.56, 17.62, 10.15, 2.03, and 5.12 ng/ml, respectively. Specific gravity-corrected concentration were 20.81, 20.36, 12.88, 2.58, 5.00 ng/ml, respectively. The results of multivariate logistic regression analysis indicated that: education degree was negatively associated with urinary concentration of MMP, MEP, MBP, MBzP and MEHP, OR (95% CI) were 0.495 (0.253-0.966), 0.380 (0.191-0.755), 0.379 (0.186-0.774), 0.401 (0.196-0.819), 0.373(0.183-0.762), respectively. Participants who had hair permed and dyed during pregnancy had higher urinary level of MBP and MBzP, OR (95% CI) were 12.867 (1.240-133.525), 15.982 (1.367-186.911), respectively; Participants who use cosmetics during pregnancy had higher urinary level of MEP and MBP, OR (95% CI) were 2.977 (1.012-8.757), 4.440 (1.485-13.272), respectively; plastic bottled water consumption was positively associated with urinary concentrations of MEP and MEHP, OR (95% CI) were 3.780 (1.417-10.083), 2.699 (1.039-7.010), respectively; annual household income was negatively associated with urinary concentration of MMP, OR (95% CI) was 0.597 (0.372-0.959); individuals who took medications during pregnancy had higher urinary level of MEHP than non-takers, OR (95% CI) was 4.853 (1.084-21.732). Pregnant women whose gestation age was less than 16 weeks are generally exposed to phthalate. Phthalate internal exposure levels are significantly associated with most measured factors and the influencing factors with different phthalates internal exposure levels are different.

Phthalate levels in urine of pregnant women and their associated missed abortion risk

Evaluation of exposure to phthalate esters and DINCH in urine and nails from a Norwegian study population

邻苯二甲酸酯(PAEs)因具有改善塑料柔软性和柔韧性的特性而被作为增塑剂广泛应用于各种食品包装材料,然而,其对人类健康的重大危害使得建立准确的分析方法显得至关重要。为了解决油性基质样品前处理费时费力的问题,我们开发了一种基于顶空固相微萃取(HS-SPME)结合气相色谱-质谱(GC-MS)检测的分析方法,用于快速检测植物油基质中的12种PAEs。将苄基化环糊精超交联聚合物(BnCD-HCP)涂敷在不锈钢纤维上制备了一种具有较大比表面积、良好稳定性好和较高重复性的固相微萃取探针,并对其结构、形貌进行了表征。将油脂样品以甲醇改性后以该探针顶空萃取PAEs,优化得到如下最佳萃取参数:萃取时间20 min,萃取温度50 ℃,解吸时间4 min,解吸温度275 ℃。在优化的分析条件下,12种目标PAEs在线性范围内具有良好的线性关系(相关系数(R2)均大于0.99),检出限和定量限分别为0.21~3.74 μg/kg和0.69~12.34 μg/kg,单针的相对标准偏差(RSD)≤11.4%,针间RSD≤13.9%。将此方法应用于大豆油、花生油和葵花油样品中PAEs的检测,加标试验结果表明该方法具有良好的精密度(RSD为1.17%~11.73)和回收率(72.49%~124.43%)。该研究建立的基于BnCD-HCP的HS-SPME方法不需要溶剂萃取和净化等繁琐操作,具有快速、灵敏、准确、环保等优点,为植物油中PAEs的灵敏筛检提供了新的方法和技术。

An enrofloxacin (ENR) molecularly imprinted membrane (MIM) was prepared with a polyvinylidenedifluoride (PVDF) membrane as the carrier, ENR as the dummy template molecule, α-methacrylic acid (MAA) as the functional monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker, and a chloroform-methanol mixture solvent as the porogen. The MIM showed excellent selectivity, high adsorption capacity, and high adsorption rate for ciprofloxacin. Additionally, a method combining molecularly imprinted membrane extraction (MIME) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed and validated for the selective analysis of trace ciprofloxacin residue in milk samples. The sample pretreatment involved only a single step of protein precipitation. Ciprofloxacin showed good linearity in mass concentration range of 0.1-200 μg/L with a high correlation coefficient (r2>0.9996). The limit of detection (LOD, S/N=3) and limit of quantification (LOQ, S/N=10) were 0.02 μg/L and 0.1 μg/L, respectively. The relative standard deviations (RSDs) of interday and intraday precisions ranged from 3.3% to 7.9%. The ciprofloxacin recovery was in the range of 92.6%-119.1%. The results showed that the proposed method is simple and fast, with high accuracy and sensitivity, thus being suitable for the rapid detection of trace ciprofloxacin residue in milk samples.

Objective: To examine the association between the phthalate exposure in the first trimester and fasting blood glucose level or gestational diabetes mellitus (GDM) in the third trimester in pregnant women. Methods: A total of 3 474 pregnant women, receiving their prenatal examination in Ma' anshan Maternal and Child Health-Care Hospital of Anhui province, were selected from May 2013 to September 2014. Questionnaires were used to collect the information about their socio-demographic characteristics, clinical characteristics and GDM diagnostic results in the first, second and third trimesters. Urine samples and fasting venous blood samples were collected. Concentrations of 7 kinds of phthalate metabolites in urine samples were detected by solid phase extraction-high performance liquid chromatography-tandem mass spectrometry (SPE-HPLC-MS/MS), and multiple linear regression model was used for statistical analyses. Logistic regression analysis on the risk of the first trimester phthalate exposure for GDM in the third trimester was conducted. Results: The prevalence of GDM in this study was 12.8%, monomethyl phthalate (MMP), monoethyl phthalate (MEP), mono-n-butyl phthalate (MBP), monobenzyl phthalate (MBzP) and mono-(2-ethyl-5-oxohexyl) phthalate (MEHHP) exposure levels were positively correlated with the fasting blood glucose level in the third trimester (P<0.05), but mono-(2-ethylhexyl) phthalate (MEHP) and mono-(2-ethyl-5-hydroxylhexyl) phthalate (MEOHP) exposure levels were negatively correlated with the fasting blood glucose level in the third trimester (P<0.05). Stratified analysis showed a positive correlation between MEHHP exposure and the third trimester fasting blood glucose level in both normal group and GDM group. However, MMP, MEP, MBP, MBzP, MEHP and MEOHP exposure levels had influences on the third trimester fasting blood glucose level in normal group but not in GDM group. MMP and MBP exposure might increase the risk of GDM, but MEOHP exposure might reduce the risk of GDM. Conclusion: The phthalate exposure in the first trimester might be associated with the fasting blood glucose level in the third trimester, MMP, MEP, MBP, MBzP and MEHHP concentrations were positively associated with the third trimester blood glucose level, MEHP and MEOHP concentrations were negatively associated with the third trimester blood glucose level. Moreover, the effects of different kinds of phthalates might be different.

Young, healthy men with a gene variant that may impair skin barrier function had higher urine levels of certain phthalates than men without the variant, according to a group of Danish researchers.1 The findings, reported in this issue of EHP, suggest that a genetic factor may determine the extent of a person’s exposure to an environmental chemical. “This idea that a certain level of exposure to a chemical can mean something very different depending on your genetic makeup is quite new and important,” says Shanna Swan, a reproductive health scientist at the Mount Sinai School of Medicine in New York, who was not involved in the study. Men with a gene variant that predisposed them to dry skin were more likely to have higher urinary concentrations of phthalates found in skin care products. Small percentages of Europeans and Asians have a genetic mutation that diminishes the production of filaggrin,2 an epidermal protein that helps to lock moisture into the skin and keep out pathogens, allergens, and chemicals. People with filaggrin-depleted skin may be more likely to have allergic skin disorders.3 The researchers hypothesized that these people may also absorb phthalates through the skin at higher rates than people with normal filaggrin production. Certain phthalates—a large group of chemicals found in cosmetics, fragrances, solvents, and plastics—have been associated with markers of decreased testicular function in some human studies4 and altered male genital development in animal research.5 They’ve also been associated with diabetes,6 asthma,7 attention disorders,8 and obesity9 in some epidemiological studies. In the current study, researchers measured levels of phthalate metabolites in the urine of 861 Danish men between the ages of 18 and 22. Sixty-five men, roughly 7.5% of the study participants, carried at least one nonfunctional copy of the filaggrin gene (FLG), indicating diminished filaggrin production. On average, those men had 33% higher urinary concentrations of a metabolite of di-n-butyl phthalate (DnBP) than men with two functional FLG copies. They also higher average levels of metabolites of diisobutyl phthalate (DiBP) and butylbenzyl phthalate (BBzP). These low-molecular-weight phthalates are commonly found in cosmetics and personal care products. However, the average urinary concentration of a metabolite for diethyl phthalate (DEP), the most frequently detected phthalate in personal care products, was not significantly higher in men with FLG mutant alleles. Metabolites of several high-molecular-weight phthalates, which are primarily found in PVC plastics, also were not significantly higher in this group. When the researchers compared concentrations of reproductive hormones and markers of semen quality between the two groups of men, they found no significant differences. It’s unclear from the study why men with FLG mutations tended to have higher levels of some phthalates. It’s possible their skin was more permeable to chemicals they encountered. People with reduced filaggrin production also tend to have drier skin, says lead study author Ulla Nordstrom Joensen, a urologist at Copenhagen University Hospital, so it’s plausible that higher phthalate levels could be the result of this group using more skin care products. “Whatever the explanation, they appear to be more exposed, which is potentially concerning,” Joensen says. In 2008 the U.S. federal government heavily restricted the use of DBP, BBzP, and di(2-ethylhexyl) phthalate in toys and other children’s products, while the European Union has placed similar restrictions on these chemicals.10 For the general population, exposure to high-molecular-weight phthalates appears to result largely from diet, whereas nondietary routes such as use of personal care products and indoor dust and air appear to explain most exposure to low-molecular-weight phthalates.11 Future studies should investigate whether similar associations appear between FLG mutations and phthalate exposures in other population groups, such as women and children, and whether FLG mutation carriers have greater exposure to other environmental chemicals. If it turns out these individuals’ skin is more permeable to phthalates, Joensen says, it’s possible they should pay special attention to dermal chemical exposures in general.

Phthalates are a group of diesters of ortho-phthalic acid (dialkyl or alkyl aryl esters of 1,2-benzenedicarboxylic acid). Higher-molecular-weight phthalates, such as di-2-ethylhexyl phthalate (DEHP), are primarily used as plasticizers to soften polyvinyl chloride (PVC) products, while the lower-molecular-weight phthalates, such as diethyl phthalate (DEP), di-n-butyl phthalate (DBP), and butyl benzyl phthalate (BBzP), are widely used as solvents to hold color and scent in various consumer and personal care products. Phthalates have become ubiquitous environmental contaminants due to volatilization and leaching from their widespread applications, and thus contamination of the environment has become another important source for phthalates in foods in addition to migration from packaging materials. Human exposure to phthalates has been an increased concern due to the findings from toxicology studies in animals. DEHP, one of the important and widely used phthalates, is a rodent liver carcinogen. DEHP, DBP, BBzP, and several phthalate metabolites, such as monobutyl phthalate, monobenzyl phthalate, and mono-(2-ethylhexyl) phthalate, are teratogenic in animals. Since foods are the major source of exposure to phthalates, information on levels of phthalates in foods is important for human exposure assessment. The objective of this review is to identify the knowledge gaps for future investigations by reviewing levels of a wide range of phthalates in a variety of foods, such as bottled water, soft drinks, infant formula, human milk, total diet foods, and others, migration of phthalates from various food-packaging materials, and traditional and new methodologies for the determination of phthalates in foods.

Carbamates are used in broad-spectrum insecticides and herbicides, and have highly efficient, low-residue, and long-lasting characteristics. However, this type of pesticide exerts mutagenic, teratogenic, carcinogenic, and other adverse effects, and its frequent use can exceed the recommended scope and limits. Research on the determination of carbamate pesticides mainly focuses on foods of plant origin and pays less attention to foods of animal origin. The methods for carbamate determination described in the current national standards have complicated operating procedures and low efficiency. Therefore, highly efficient and accurate methods for carbamate detection in milk must be established. In this work, a rapid method based on pass-through solid-phase extraction (SPE) purification coupled with ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the simultaneous determination of 10 carbamate pesticides in liquid milk. The pretreatment and instrument methods were systematically optimized. The milk sample was extracted with acetonitrile, and then purified using a Captiva EMR-Lipid filtration kit. The purified extract was separated on an ACQUITY UPLC BEH C18 column with mobile phase of methanol and 0.1% formic acid aqueous solution in gradient elution. The flow rate was 0.3 mL/min. Column temperature was 35 ℃. Quantitative analysis was performed using the external standard method with matrix matching curves. The 10 carbamate pesticides showed good linear relationships in the mass concentration range of 2-200 μg/L, with correlation coefficients greater than 0.999. The limits of detection (LODs) and quantification (LOQs) for the 10 carbamate pesticides were 0.045-0.23 and 0.15-0.77 μg/kg, respectively. Recovery tests were conducted using the blank-matrix method at three spiked levels of 15, 50, and 100 μg/kg, and good recoveries for the 10 carbamate pesticides were obtained. In particular, the recoveries for the three spiked levels of 15, 50, and 100 μg/kg were 68.7%-93.3% with relative standard deviations (RSDs) of 1.8%-8.0%. The proposed method is efficient, convenient, accurate, and suitable for the rapid detection of the 10 carbamate pesticides in liquid milk. Compared with the conventional NH2 and ENVITM-18 SPE columns used in the national standard determination method, the proposed method demonstrated better purification effects. The recoveries for aldicarb sulfoxide, aldicarb sulfone, methomyl, and carbaryl after purification using the Captiva EMR-Lipid kit increased from 60% to 80%. Thus, the proposed method is suitable for targets with strong polarity and gives measurement results with good repeatability and accuracy.

A method of solid phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS/MS) for simultaneous determination of 54 drug residues (sulfonamides, nitroimidazoles, quinolones, macrolides, lincosamides and praziquantel) in honey was developed. The honey samples were diluted by phosphate buffer solution (pH 8) followed by a further cleanup procedure with an Oasis (HLB) SPE column. The detection was carried out by LC-MS/ MS, using positive-ion electrospray ionization in multiple reaction monitoring (MRM) mode, with one precursor/two product ion transitions for each compound. Isotope dilution internal standard method or external standard method was used to determine the residue contents with a good linear relationship (r > 0.992). The limits of quantification (LOQs, S/N > 10) were 1.0 microg/kg for sulfonamides and nitroimidazoles, 2.0 microg/kg for quinolones and lincosamides, 3.0 microg/kg for macrolides, and 0.3 kg/kg for praziquantel. The recovery ranges were from 32.6% to 114% with the relative standard deviations (RSDs) between 1.3% and 28.9%. As a screening method, the developed procedure is suitable for the determination and confirmation of the drug residues in honey samples.

Analytical characterization of nucleotides and their concentration variation in drinking water treatment process

Girls’age of onset puberty has decreased globally in recentyears. Emerging evidence suggests that exposure to phthalate esters(PAEs) could play a role in this trend. However, the relationshipbetween PAE exposure and pubertal development in Chinese girls remainspoorly understood. The present study aimed to address this issue byinvestigating the link between levels of urinary phthalate ester metabolites(mPAEs) and the risk of early puberty among 546 Chinese girls (6–8years old) over a two-year monitoring program. Poisson regression,restricted cubic spline models, weighted quantile sum regression,and Bayesian kernel machine regression were used to assess both individualand combined effects of PAEs on pubertal development. Significantlinks were found between urinary concentrations of monoisobutyl phthalate(MiBP), monobutyl phthalate (MnBP), mono-(2-ethyl-5-oxohexyl) phthalate,the total of low molecular weight mPAEs, the total of nine commonlydetected mPAEs, and the total of all mPAEs with the risk of earlybreast development (all P-trend < 0.05). Furthermore,levels of MnBP, MiBP, low molecular weight mPAEs, and mono-(2-ethyl-5-oxohexyl)phthalate were positively associated with early menarche. Mixtureeffect analysis revealed incidence rate ratios of 1.64 (95% confidenceinterval (CI): 1.10–2.45) for early breast development and2.33 (95% CI: 1.22–4.45) for early menarche. Short-chain mPAEs,particularly MiBP and MnBP, were identified as the major contributorsto this combined effect. The present study demonstrated that exposureto PAEs, particularly short-chain PAEs, could lead to the early onsetof puberty in Chinese girls.

Phthalates (PAEs) are common endocrine disrupting chemicals (EDCs) that disrupt fetal development. The present study aimed to evaluate the effects of single and coexposure to phthalates in early pregnancy on fetal growth restriction (FGR) by a nested case-control study based on the Guangxi Zhuang Birth Cohort (GZBC). Maternal serum concentrations of seven phthalates in 97 neonates with FGR and 291 matched controls were detected through gas chromatography-mass spectrometry (GC-MS). The associations between phthalates and FGR were analyzed using multiple logistic regression, weight quantile sum (WQS) regression, and Bayesian kernel machine regression (BKMR) models. We found that exposures to butyl-benzyl phthalate (BBP, ORadj = 1.849, 95% CI: 1.080-3.177, Padj = 0.025, Ptrend = 0.046), di (2-ethyl-hexyl) phthalate (DEHP, ORadj = 3.893, 95% CI: 1.305-11.910, Padj = 0.015, Ptrend = 0.098) and dimethyl phthalate (DMP, ORadj = 1.722, 95% CI: 1.089-2.725, Padj = 0.020, Ptrend = 0.002) were significantly positively associated with the risk of FGR, while mono-butyl phthalate (MBP) showed a significant negative association with FGR (ORhigh = 0.192, 95% CI: 0.036-0.795, Padj = 0.033, Ptrend = 0.035) only among girls. The WQS model identified that BBP, di(2-ethyl)phthalate (DEP), DMP, DEHP, di-n-butyl phthalate (DBP), and MBP were highly weighted in the association with FGR. The BKMR model supported the positive association between joint exposure to phthalates and the risk of FGR and identified no significant interaction between the seven phthalates. Overall, maternal exposure to BBP, DEHP, and DMP may cause adverse effects on FGR, especially with combined effects.

Phthalates (PAEs) are widespread persistent organic pollutants and endocrine disruptors. However, the associations between PAE exposure and the risk of miscarriage in humans are unclear, and an insufficient number of studies have evaluated the possible threshold or dose-dependent effects of first trimester PAE exposure on miscarriage risk. Our research measured the levels of mono-methyl phthalate (MMP), mono-ethyl phthalate, mono-isobutyl phthalate, MiBP mono-butyl phthalate (MBP), mono-octyl phthalate, mono-benzyl phthalate, mono(2-ethylhexyl) phthalate, mono(2-ethyl-5-oxohexyl) phthalate, and mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) in maternal urine collected in early gestation between 150 pregnancies ending in miscarriage and 150 pregnancies with live birth. We also estimated the odds ratios (ORs) and 95% confidence intervals (CIs) for miscarriage and each PAE as a continuous variable or quartile. A restrictive cubic splines was used to assess dose-dependent effects after controlling for maternal characteristics (e.g., age, educational level). we identified monotonically increasing dose-dependent effects of MEHHP and MMP on the risk of miscarriage. The largest effect estimates were approximately threefold higher for the highest MBP (OR = 2.57; 95% CI = 1.32-5.01) or MMP quartile (OR = 3.57; 95% CI = 1.82-7.00) and two-fold higher for the highest MEHHP quartile (OR = 2.12; 95% CI = 1.10-4.11). Our research preliminarily obtained possible thresholds of MBP, MEHHP, and MMP which were 18.07, 2.38, and 0.80µg/g Cr for the risk of miscarriage, respectively. First-trimester exposure to MBP, MEHHP, and MMP exceeding certain thresholds increases the risk of miscarriage.