Phthalate esters, commonly used as plasticizers, show anti-androgenic activity and cause male reproductive malformation in experimental animals. However, the effects of prenatal exposure to phthalate esters in humans have not been extensively studied. The purpose of this study was to examine the relationship between prenatal exposure to phthalate esters and the anogenital distance (AGD) as a reproductive endpoint in human male newborns. Spot urine samples were collected from 111 Japanese pregnant women after obtaining their informed consent. Seven urinary phthalate ester metabolites were determined by high performance liquid chromatography-tandem mass spectrometry. Urinary isoflavones concentrations were measured as possible covariates because their oestrogenicities and high exposure levels among Japanese have the potential to affect male genital development. Birth outcomes and AGD, the distance from the centre of the anus to external genitalia, were measured for their male newborns. In a multiple regression model, the log-transformed mono-2-ethylhexyl phthalate concentration (specific gravity-corrected) was negatively significant, and maternal smoking status was positively significant, in explaining anogenital index (AGI) when potential covariates were controlled for. Urinary isoflavones did not significantly contribute to AGI in any models. Our results suggest that prenatal exposure to di(2-ethylhexyl) phthalate affects reproductive development in human males.
Objectives: Concerns over dietary exposure to bisphenol A (BPA), an endocrine disruptor, have been raised because BPA is contained in resins and plastics commonly used for the preservation of food and beverages. The purpose of the present study was to assess daily intake levels of BPA in a group of male subjects by measuring total urinary BPA (free BPA plus BPA released by treatment with β-glucuronidase), as well as determining intra-individual variation in BPA excretion.Methods: Twenty-four-hour urine was collected from 5 subjects for 5 consecutive days for the evaluation of between-day variation in urinary BPA excretion and from 36 male subjects for the estimation of the level of daily BPA intake. BPA in the urine samples was measured by GC/MS/MS following enzymatic hydrolysis of BPA glucuronate, solid phase extraction, and derivatization.Results: A large between-day variation was found over 5 days for the daily excretion of urinary BPA in the 5 subjects. The daily excretion of urinary BPA was distributed log-normally in the 36 male subjects, with the median value being 1.2 µg/day (range: <0.21-14 µg/day), which was far below the Tolerable Daily Intake (0.01 mg/kg bw) recommended by a scientific committee in the European Commission in 2002. However, the maximum estimated intake per body weight (0.2 µg/kg/day) was only one order of magnitude lower than the reported lowest level for reproductive/behavioral effects in pregnant mice (2 µg/kg/day).Conclusions: Measuring urinary BPA in urine is a suitable approach for estimating short-term BPA intake levels in individuals and/or estimating the average exposure level of populations. Urine analyses will be increasingly important in the human health risk assessment of BPA.
Objectives Our objectives were (1) to evaluate whether single spot urine is suitable media for longer-term phthalate esters exposure assessment, and (2) to estimate intake level of phthalate esters of Japanese pregnant women using urinary metabolites as an indicator of prenatal exposure level in their offspring. Methods We analyzed nine metabolites (MMP, MEP, MnBP, MBzP, MEHP, MEOHP, MEHHP, MINP, MnOP) of seven phthalate esters in spot urine samples from 50 pregnant women by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Using four urine samples collected from each of 12 subjects from 50 pregnant women within 5-12 weeks, we compared intra-and interindividual variation in urinary metabolites by calculation of intraclass correlation coefficient (ICC). We estimated daily intakes of 50 pregnant women from their urinary metabolite concentrations.Results ICCs for seven phthalate metabolite concentrations in single spot urine samples were: MMP (0.57), MEP (0.47), MnBP (0.69), MBzP (0.28), MEHP (0.51), MEHHP (0.43), and MEOHP (0.41) in 12 pregnant women. Phthalate ester metabolites had high detection rates in 50 subjects. The mean daily intake ranged from 0.01 to 2 lg/kg per day. The daily intake levels in all subjects were lower than corresponding tolerable daily intake (TDI) set by the European Food Safety Authority (EFSA), though maximum value for DnBP of 6.91 lg/kg per day accounted for 70% of TDI value. Conclusions Higher ICCs indicated that phthalate metabolite levels in single spot urine could reflect longer-term exposure to the corresponding diesters of subjects. Although the current exposure level was less than TDIs, further studies and exposure monitoring are needed to reveal the toxicity of phthalate esters to sensitive subpopulation.
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