Next Article in Journal
The Effect of Nordic Walking Training Combined with Vitamin D Supplementation on Postural Control and Muscle Strength in Elderly People—A Randomized Controlled Trial
Previous Article in Journal
Ecohydraulogical Characteristic Index System of Schizopygopsis younghusbandi during Spawning Periods in the Yarlung Tsangpo River
 
 
Search for Articles:
Title / Keyword
Author / Affiliation / Email
Journal
Article Type
 
 
Advanced Search
 
Section
Special Issue
Volume
Issue
Number
Page
 
Journals
IJERPH
Volume 15
Issue 9
10.3390/ijerph15091950
ijerph-logo
Submit to this Journal Review for this Journal Propose a Special Issue
► Article Menu

Article Menu

share Share announcement Help format_quote Cite question_answer Discuss in SciProfiles thumb_up
...
Endorse textsms
...
Comment
first_page
settings
Order Article Reprints
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

Temporal Trends of Urinary Phthalate Concentrations in Two Populations: Effects of REACH Authorization after Five Years

by
Giovanna Tranfo
,
Lidia Caporossi
*,
Daniela Pigini
,
Silvia Capanna
,
Bruno Papaleo
and
Enrico Paci
Department of Occupational and Environmental Medicine, Epidemiology and Hygiene, INAIL-National Institute for Insurance against Accidents at Work, 00078 Monteporzio Catone, Italy
*
Author to whom correspondence should be addressed.
Int. J. Environ. Res. Public Health 2018, 15(9), 1950; https://doi.org/10.3390/ijerph15091950
Submission received: 11 July 2018 / Revised: 30 August 2018 / Accepted: 31 August 2018 / Published: 6 September 2018
(This article belongs to the Section Environmental Health)
Browse Figure
Versions Notes

Abstract

:
Phthalates are widely used in the industrial manufacture of many products. Some phthalates have shown reproductive toxicity in humans, acting as endocrine disruptors, so they were included in the authorization process defined in Reg. CE 1907/2006 (REACH). Two groups of population were recruited, before and after the inclusion of some phthalates in the authorization list in REACH: the first group of 157 volunteers was studied in 2011 and the second, 171 volunteers, in 2016. Each subject completed a questionnaire about personal lifestyle, working activities and use of chemical products. The main urinary metabolites of five phthalates were analyzed by HPLC/MS/MS: mono(2-ethylhexyl)phthalate (MEHP) and mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) for di(2-ethylhexyl)phthalate (DEHP) exposure; monoethylphthalate (MEP) for diethylphtahate (DEP); monobenzylphthalate (MBzP) for butylbenzylphtahalate (BBP) and dibenzylphthalate (DBzP), mono-n-butylphthalate (MnBP) for butylbenzylphtahalate (BBP) and di-n-butylphthalate (DnBP). The results show a significant difference for all metabolites between the two periods, with the exception of MEP in women. The comparison of the two sets of results shows a decrease in urinary metabolites excretion from 2011 to 2016, statistically significant for the three phthalates included in Annex XIV of REACH. DEP, not currently included in the list for authorization, maintains a constant presence in the daily life of the population, particularly for women.

1. Introduction

The diesters of phthalic acid are a group of similar molecules, widely used since the 1930s in many commercial industrial product manufacturing processes, the most important of which is the production of plastics. The inclusion of phthalates in rigid polymers (such as PVC) allows the production of more flexible plastics. They are also used in construction materials, vinyl flooring, food packaging and medical devices. This widespread use results in a possible human exposure to phthalates [1].
Phthalates having a smaller molecular structure and lower molecular weight are more soluble in water and are therefore used as industrial solvents, in the composition of cosmetics and pharmaceuticals, or as insecticides; this is the case, for example, of diethyl phthalate (DEP) and di-n-butyl phthalate (DnBP) [2,3]. In particular, DEP is the phthalate of choice used in the cosmetic and personal hygiene products industry, both for adults and for children/infants [4], while DnBP is usually used as excipient in pharmaceuticals products [5,6], and it was present in cosmetics, particularly nail care products for adults [7]; in Europe this last use is forbidden by the EU cosmetics directive, Dir. 76/768/EEC, where other forbidden phthalates were also listed: di(2-ethylhexyl)-phthalate (DEHP), benzylbutylphthalate (BBP), bis2-methoxyethylphthalate, n-pentylisopentyl-phthalate, di-n-pentylphthalate, diisopentylphthalate and di (n-butyl) phthalate (DnBP). Historically, DEHP is the phthalate produced in the largest quantities around the world, about 2 million tons per year [8]. Nevertheless, DEHP has been recently replaced in numerous production cycles by diiso-nonylphthalate (DiNP), including the production of flexible PVC, because of its dangerous characteristics and in particular its toxicity for human reproduction [9]. In the scientific literature, some authors have shown that also infants are exposed to some phthalates, possibly through breast milk, infant formulae and baby food [10,11,12]. Furthermore the possibility of phthalate migration from packaging to food or beverages represent a possible risk of contamination [13,14] and therefore of human exposure.
Some phthalates act as endocrine disruptors, particularly interacting with estrogen receptors and interfering on hormone homeostasis. In particular, DnBP and DEHP have shown to cause antiandrogenic effects in in vivo experiment [15], with decreased testosterone concentrations and sperm production [16,17].
In addition to the toxic effects for the endocrine system, effects on kidney and liver have been highlighted [17,18], as well as the possibility that exposure to high concentrations of phthalates may affect the etiology of certain tumors [19], associated with higher fetal death or childhood malformations [17,20].
The exposure of the general population to these substances was assessed using biomonitoring studies both in the United States and in Europe [21,22,23]. Since 1979, in Europe, the legislation started to protect the more vulnerable consumers from this type of exposure by means of the European directive 79/769/CEE, in which the maximum permitted concentrations of six phthalates in toys and children products were stated; these indications currently remain as restrictions.
Presently, the European Regulation CE 1907/2006, named Registration, Evaluation and Authorization of Chemicals (REACH), that contains these restrictions, also includes some phthalates in a list of “substances of high concern”. We report in Table 1 the list of phthalates that are on the candidate list for authorization and/or in the Annex XIV of REACH, with details about the date of inclusion and the reason for it. This action will gradually bring to the substitution of these chemicals in the involved industrial processes, with alternative substances, even if it’s not simple to find chemicals with the same industrial characteristics and lower or no health effects. At the moment in fact there’s a heated scientific debate in Europe about the safety of replacement chemicals.
Phthalates are not persistent molecules from an environmental point of view, as they tend to degrade easily; at a biochemical level, similarly, they rapidly produce the respective monoester metabolites, which follow a mainly urinary excretory pathway. High molecular weight molecules trend to further react giving oxidative products [24], and/or partially undergo glucuronidation reactions and are excreted mainly in the urine, and, to a lesser extent, in the feces [25]. The environmental contamination levels that are reported in the scientific literature are justified by a continuous production of the phthalates and wide dispersion, resulting in their presence in dust, soil, indoor and outdoor air.
In order to evaluate the internal dose of phthalates, biomonitoring is the preferred strategy as it offers the advantages of integrating exposure through all routes (e.g., inhalation, dermal, ingestion). Furthermore the urinary metabolites analysis permit to carry out a more precise measurement with respect to the parent compounds and, even more important, the risk of accidental contamination of samples during collection, storage and analysis (due to the use of plastic devices and to the environmental contamination) is greatly reduced [26,27]. Therefore, the urinary metabolites became the most used biological indicators of exposure, both for general population and for particular group of subjects [28,29,30,31]. The specific metabolic route of each phthalate, and the corresponding monoester metabolite, is related to the size of the aryl and/or alkyl chains present in the molecular structure.
The aim of the present study was to evaluate the exposure due to lifestyle to five phthalates, namely DEHP, DEP, DnBP, BBP and dibenzylphthalate (DBzP), by means of the analysis of the respective metabolites in urine samples collected with a five year gap (2011–2016) in two population groups. We compared the two series of biomonitoring data, gathered before and after the phthalates were inserted into annex XIV of REACH, in order to understand if the application of the authorization process could bring to a real reduction of the general population exposure.
For the present study DEHP, DEP, DnBP, BBP, DBzP were selected, due to their human toxicity but also to their wide presence in living environments. DEHP, DnBP and BBP are included in Annex XIV, but not DEP and DBzP. We considered DEP because it is largely used in cosmetics and cleaning and care products, and DBzP, that is an ingredient of drug capsules, as its metabolite, MBzP, is also the metabolite of BBP. We report in Table 2 the list of all the phthalates considered in the present study, with details about their main urinary metabolites and their presence in processes and products, as well as possible exposure sources [32].

2. Materials and Methods

We ensured the respect for human dignity, protected the value, rights and interests of the participants to the research project according to Declaration of Helsinki and following the European and Italian Law about ethical principles and data protection/management and privacy.
For these purposes, all the enrolled subjects were volunteers, of both sexes, adult workers of different tasks in several workplaces, not belonging to vulnerable populations, not minors, without diagnosed pathologies, able to perform their working task according to their occupational physician. All subjects gave their informed consent to participate to the study. They were tested in an occupational medicine clinic, during the periodic health surveillance for the prevention of risks for safety and health at work. They represent a general population sample, from professionals graduated to blue-collar workers. The sampling strategy included a preliminary phase of illustration of the project, with adhesion through the signing of the informed consent by the subjects meeting the inclusion criteria, then the definition of the medical examination time and contextual collection of the spot urine samples.
We enrolled the first sample of population in 2011, consisting in 157 volunteers (83 women and 74 men) [39] and the second sample in 2016, 171 volunteers (111 women and 60 men). All of them were living in Central Italy. Each subject was asked to complete a questionnaire for collecting information about personal lifestyle, food habits, smoking, drug use, working activities, hobbies and use of chemical products.
In order to identify possible sources of exposure we examined also the following variables: the habit to store fat foods in plastic containers, because is documented that a migration between plastic containers and food is greater and faster in case of fat food [40], and the habit of eating fish, especially if rich in fat like mackerel, salmon and eel, and a regular intake of canned foods [41].
The inclusion criteria were:
  • Healthy volunteers not occupationally exposed to phthalates;
  • Subjects having creatinine concentrations between 0.3 and 3 g/L [42] because when the urine is too diluted or concentrated this could affect the accuracy of the analytical results [43,44].
Urine samples were collected in sterile polypropylene containers, and, after being transported in our laboratory at room temperature, quickly frozen, and stored at −20 °C until analysis. The analysis was carried out using the procedure previously published [31], using HPLC/MS/MS in order to determine the urinary concentrations of the following metabolites: mono(2-ethylhexyl)phthalate (MEHP), mono(2-ethyl-5-hydroxyhexyl)phthalate (MEHHP), monoethylphthalate (MEP), mono-benzylphthalate (MBzP), and mono-n-butylphthalate (MnBP). We summed the molar concentrations of the two DEHP metabolites to calculate the concentration related to DEHP exposure.
Statistical analysis was performed with the software SPSS® 19.0 (IBM® Corporation, New York, NY, USA). To study the distribution of the data the Shapiro–Wilk test was used, with a significance of 0.05. The application of the Shapiro- Wilk test indicated that the urinary phthalates’ value were not normally distributed (p < 0.05), and therefore non-parametric tests were applied for the evaluation of the difference between the two periods. The urinary concentrations of phthalates were analyzed to determine possible differences between the two groups of data using the Mann–Whitney, nonparametric test, with a significance of 0.05.

3. Results

In Table 3 the population characteristics and the information collected with the questionnaire are reported separately for the two biomonitoring campaigns.
In Table 4 the Limit of Detection for each metabolite and the respective detection frequency over the LOD for all samples (both campaigns) are reported.
The comparison between the two studies is graphically reported in Figure 1, stratified by gender, in terms of median values.
The analytical results of the urinary phthalate metabolites for all the volunteers and for the two campaigns are reported in Table 5 and Table 6. The results presented in Table 5 and Table 6 show a statistically significant difference (p < 0.05) for all metabolites, both for men (median value, in µg/g creatinine, of: MEP 49.9 vs. 21.6; MnBP 37.6 vs. 0.0; MBzP 4.8 vs. 0.0; ∑DEHP metabolites 14.1 vs. 3.1) and for women (median value, in µg/g creatinine, of: MnBP 38.8 vs. 0.0; MBzP 7.0 vs. 0.5; ∑DEHP metabolites 15.6 vs. 4.5) between the two periods, with the exception of MEP in women (median value 73.1 vs. 49.9 µg/g creatinine); the group whose samples were taken in the more recent period presents an overall lower urinary metabolites concentration of phthalates than the other group.
With regard to the dosage of the MEP, it should be remembered that the DEP is one of the phthalates, among those taken into consideration, which is not subject to the REACH authorization process. This means that DEP is still widely used in everyday products, many of which are cosmetics and personal hygiene products; this probably justifies a higher concentration of the MEP in the urine of women and a not statistically significant reduction in the five years’ time lapse.
A comparison study between the two genders was also conducted, within the same period of recruitment and no significant differences emerged in 2011, while in 2016 MEP and MBzP were significantly (Mann-Whitney, p < 0.05) higher in women than in men (median value, in µg/g creatinine of: MEP 49.9 vs. 21.6; MBzP 0.5 vs. 0.0).
Besides, some considerations about the possible sources of exposure were done using the data from the questionnaires. The application of a statistical test to smoking and non-smoking subjects (all data) did not show any significant difference (Kruskall–Wallis > 0.05). The same test, performed between subjects consuming canned food at least weekly and all the others, showed higher concentrations of MEHHP (median value, in µg/g creatinine: 16.0 vs. 3.2) in people consuming canned food, in the first group of subjects (enrolled in 2011); the molar sum of MEHHP and MEHP urinary concentrations (as they are both metabolites of the same phthalate) are, on average, higher in people using plastic containers to store fatty foods, in both groups (median value, in µg/g creatinine, in 2011: 18.7 vs. 4.6; in 2016 3.9 vs. 0.2).

4. Discussion

Our results suggest that non-occupational exposures to phthalates of the studied subjects, without doubt, occurred in the considered geographical area; the exposure concentrations are quite similar to those measured in different countries for some metabolites. We report in Table 7 and Table 8 a comparison of urinary concentrations of phthalate metabolites found in different populations, stratified for gender.
More numerous studies on women than on men can be found in the literature, and papers that focus on both sexes sometimes showed results without a difference between genders [45]; this can be a problem for the interpretation of data, particularly because gender is an important variable for exposure.
Looking for scientific articles reporting temporal trends in phthalate exposure we can found a decreased concentration, in the last decade, both in USA [46,47] and in Europe [48,49], with values decreasing from 17% to 42%; the reason for it was indicated in the legislative activity in western countries.
In particular it was observed that exposure to the “alternative chemicals” was increasing while the exposure to the original compounds was lowered [45,47,49,50]. The questionnaire allowed us to highlight some of the possible common sources of exposure to phthalates in everyday life, particularly the use of plastic containers for the storage of fat foods and eating canned foods [41].

4.1. Strengths and Limitations of the Study

The strength of the present study is certainly to have proposed a biomonitoring survey of exposure to phthalates in Italy, for a general population group. Currently, phthalate dosing studies, in Italy, are aimed to investigate specific correlations with pathologies [55,56], selecting samples of specific subjects, mainly children [55,56,57,58] and pregnant [59] or menopausal [60] women, and not to a more general exposure characterization. A second strength is to produce data, useful for the scientific debate, to confirm the temporal trends of phthalate in Europe, after the REACH application.
Considering the rate of metabolism and excretion of phthalates, the individual concentration can show daily and/or weekly variability, which a sampling of spot urine cannot evaluate. Testing a single spot urine sample may therefore represent a limitation of the study. Other limitations are showing results only from a population of working adults, with a range of age 28–50, and enrolling subjects living in a specific geographic area, that make the generalization of results to all the population, also in considerations of possible regional variability, largely approximate. Concluding, it is certainly desirable to increase the number of subjects recruited in order to be able to extend the data and increase their significance.

5. Conclusions

The comparison of the two sets of results clearly shows the dramatic decrease in urinary metabolites excretion from 2011 to 2016, for the three phthalates included in Annex XIV of REACH.
The analytical method performances render it suitable for the quantitative determination of the analytes in urine samples of the general population.
DEP, not currently included in the candidate list for authorization, maintains a constant presence in the daily life of the population; this is more important for the female gender, with high probability in consideration of the larger use of cosmetics and products for personal hygiene.
It is important to underline how DEP is included in the European classification of endocrine disruptors in category 1, therefore as a “substance of high concern” with regards of possible endocrine interference for humans. This suggests that this phthalate will certainly require to be further studied and reconsidered by the legislators to assess the possible risk for human health, particularly for women, and consequently it will be needed to reduce the exposure sources in the near future.
Future research should be aimed to increase the number of enrolled subjects for a stronger statistical significance of the data and to allow the identification of the possible exposure sources for the general population.
We think that the downtrend in the urinary excretion of phthalate metabolites is clearly a consequence of their less widespread use and of the lower environmental contamination of the last few years, reflecting the application of the authorization process to some phthalates starting from 2013, and confirming the efficacy of REACH legislation.

Author Contributions

Conceptualization and project administration, G.T.; Methodology and data curation, E.P. and D.P.; Formal Analysis, data curation, S.C.; Writing Original draft preparation and writing, review and editing, L.C., E.P.; Supervision, B.P.; Validation, E.P.

Funding

This research received no external funding.

Conflicts of Interest

The authors declare no conflict of interest.

References

  1. Schettler, T. Human exposure to phthalates via consumer products. Int. J. Androl. 2006, 29, 134–139. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  2. Taylor, J.; Ingerman, L.; Gray, D.A.; Little, S.; Amata, R. Toxicological Profile for n-butylphthalate. Available online: https://www.atsdr.cdc.gov/toxprofiles/tp.asp?id=859&tid=167 (accessed on 31 August 2018).
  3. Richardson, M.; Bosh, S.; Swarts, S.; Llados, S.; Gray, F. Toxicological Profile for di(2-ethylexyl)phthalate (DEHP). Available online: https://www.atsdr.cdc.gov/toxprofiles/tp.asp?id=684&tid=65 (accessed on 31 August 2018).
  4. Houliham, J.; Brody, C.; Schwan, B. Not too Pretty; Phthalates, Beauty Products & the FDA. Available online: https://noharm-asia.org/sites/default/files/documents-files/110/Not_Too_Pretty.pdf (accessed on 20 June 2018).
  5. Nassan, F.L.; Coull, B.A.; Skakkebaek, N.E.; Andersson, A.M.; Williams, M.A.; Minques-Alarcon, L.; Krawets, S.A.; Hall, J.E.; Hait, E.J.; Korzenik, J.R.; et al. A crossover-crossback prospective study of dibutyl-phthalate exposure from mesalamine medications and serum reproductive hormones in men. Environ. Res. 2018, 160, 121–131. [Google Scholar] [CrossRef] [PubMed]
  6. Nassan, F.L.; Coull, B.A.; Skakkebaek, N.E.; Williams, M.A.; Dadd, R.; Minques-Alarcon, L.; Krawets, S.A.; Hait, E.J.; Korzenik, J.R.; Moss, A.C.; et al. A crossover–crossback prospective study of dibutyl-phthalate exposure from mesalamine medications and semen quality in men with inflammatory bowel disease. Environ. Int. 2016, 95, 120–130. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  7. Wormuth, M.; Scheringer, M.; Vollenweider, M.; Hungerbuhler, K. What are the sources of exposure to eight frequently used phthalic acid esters in Europeans? Risk Anal. 2006, 26, 803–824. [Google Scholar] [CrossRef] [PubMed]
  8. Lorz, P.M.; Towae, F.K.; Enke, W.; Jäckh, R.; Bhargava, N.; Hillesheim, W. Phthalic Acid and Derivatives. In Ullmann’s Encyclopedia of Industrial Chemistry, 7th ed.; Wiley-VCH: Weinheim, Germany, 2007. [Google Scholar]
  9. Lyche, J.L.; Gutleb, A.C.; Bergman, A.; Eroksen, G.S.; Murk, A.J.; Ropstad, E.; Saunders, M.; Skaare, J.U. Reproductive and developmental toxicity of phthalates. J. Toxicol. Environ. Health B Crit. Rev. 2009, 12, 225–249. [Google Scholar] [CrossRef] [PubMed]
  10. Main, K.M.; Mortensen, G.K.; Kaleva, M.M.; Boisen, K.A.; Damgaard, I.N.; Chellakooty, M.; Schmidt, I.M.; Suomi, A.M.; Virtanen, H.E.; Petersen, J.H.; et al. Human breast milk contamination with phthalates and alterations of endogenous reproductive hormones in infants three months of age. Environ. Health Perspect. 2006, 114, 270–276. [Google Scholar] [CrossRef] [PubMed]
  11. Petersen, J.H.; Breindahl, T. Plasticizers in total diet samples, baby food and infant formulae. Food Addit. Contam. 2000, 17, 133–141. [Google Scholar] [CrossRef] [PubMed]
  12. Mortensen, G.K.; Main, K.M.; Andersson, A.M.; Leffers, H.; Skakkebæk, N.E. Determination of phthalates monoesters in human milk, consumer milk, and infant formula by tandem mass spectrometry (LC-MS-MS). Anal. Bioanal. Chem. 2005, 382, 1084–1092. [Google Scholar] [CrossRef] [PubMed]
  13. Bosnir, J.; Puntaric, D.; Skes, I.; Klaric, M.; Simkic, S.; Zoric, I. Migration of phthalates from plastic products to model solutions. Coll. Antropol. 2003, 27, 23–30. [Google Scholar] [PubMed]
  14. Becker, K.; Seiwert, M.; Angerer, J.; Heger, W.; Koch, H.M.; Nagorka, R.; Rosskamp, E.; Schlüter, C.; Seifert, B.; Ullrich, D. DEHP metabolites in urine of children and DEHP in house dust. Int. J. Hyg. Environ. Health 2004, 207, 409–417. [Google Scholar] [CrossRef] [PubMed]
  15. Jarfelt, K.; Dalgaard, M.; Hass, U.; Borch, J.; Jacobsen, H.; Ladefoged, O. Antiandrogenic effects in male rats perinatally exposed to a mixture of di(2-ethylhexyl)phthalate and di(2-ethylhexyl)adipate. Reprod. Toxicol. 2005, 19, 505–515. [Google Scholar] [CrossRef] [PubMed]
  16. McKee, R.H.; Butala, J.H.; David, R.M.; Gans, G. NTP center for the evaluation of risks to human reproduction reports on phthalates: Addressing the data gaps. Reprod. Toxicol. 2004, 18, 1–22. [Google Scholar] [CrossRef] [PubMed]
  17. Hauser, R.; Calafat, A. Phthalates and human health. Occup. Environ. Med. 2005, 62, 806–818. [Google Scholar] [CrossRef] [PubMed]
  18. Soto, B.D.; Lacroix, G.M. The E-screen assay as a tool to identify estrogens: An update on estrogenic environmental pollutants. Environ. Health Perspect. 1995, 103, 113–122. [Google Scholar] [CrossRef] [PubMed]
  19. Lopez-Carrillo, L.; Hernandez-Ramirez, R.U.; Calafat, A.M.; Torres-Sanchez, L.; Galvan-Portillo, M.; Needham, L.L.; Ruiz-Ramos, R.; Cebrian, M.E. Exposure to phthalates and breast cancer. Risk in Northern Mexico. Environ. Health Perspect. 2010, 118, 539–544. [Google Scholar] [CrossRef] [PubMed]
  20. National Toxicology Program, Center for the Evaluation of Risks to Human Reproduction. NTP-CERHR Expert Panel Report on di-Isononylphthalate. Available online: http://cerhr.niehs.nih.gov/news/index.html (accessed on 31 August 2018).
  21. Koch, H.M.; Rossbach, B.; Drexler, H.; Angerer, J. Internal exposure of the general population to DEHP and other phthalates determination of secondary and primary phthalate monoester metabolites in urine. Environ. Res. 2003, 93, 177–185. [Google Scholar] [CrossRef]
  22. Barr, D.B.; Silva, M.J.; Kato, K.; Reidy, J.A.; Malek, N.A.; Hurtz, D.; Sadowski, M.; Needham, L.L.; Calafat, A.M. Assessing human exposure to phthalates using monoesters and their oxidized metabolites as biomarkers. Environ. Health Perspect. 2003, 111, 1148–1151. [Google Scholar] [CrossRef] [PubMed]
  23. Silva, M.J.; Barr, D.B.; Reidy, J.A.; Malek, N.A.; Hodge, C.C.; Caudill, S.P.; Brock, J.W.; Needham, L.L.; Calafat, A.M. Urinary levels of seven phthalate metabolites in the U.S. population from the National Health and Nutrition Examination Survey (NHANES) 1999–2000. Environ. Health Perspect. 2004, 112, 331–338. [Google Scholar] [CrossRef] [PubMed]
  24. Silva, M.J.; Samandar, E.; Preau, J.L.J.; Needham, L.L.; Calafat, A.M. Urinary oxidative metabolites of di(2-ethylhexyl)phthalate in humans. Toxicology 2006, 219, 22–32. [Google Scholar] [CrossRef] [PubMed]
  25. Silva, M.J.; Barr, D.B.; Reidy, J.A.; Kato, K.; Malek, N.A.; Hodge, C.C.; Hurtz, D.; Calafat, A.M.; Needham, L.L.; Brock, J.W. Glucuronidation patterns of common urinary and serum monoester phthalate metabolites. Arch. Toxicol. 2003, 77, 561–567. [Google Scholar] [CrossRef] [PubMed]
  26. Latini, G. Monitoring phthalate exposure in humans. Clin. Chim. Acta 2005, 361, 20–29. [Google Scholar] [CrossRef] [PubMed]
  27. Swan, S.H. Environmental phthalate exposure in relation to reproductive outcomes and other health endpoints in humans. Environ. Res. 2008, 108, 177–184. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  28. Blount, B.C.; Silva, M.J.; Caudill, S.P.; Needham, L.L.; Pirkle, J.L.; Sampson, E.J.; Lucier, G.W.; Jackson, R.J.; Brock, J.W. Levels of seven urinary phthalate metabolites in a human reference population. Environ. Health Perspect. 2000, 108, 979–982. [Google Scholar] [CrossRef] [PubMed]
  29. Koch, H.M.; Drexler, H.; Angerer, J. Internal exposure of nursery-school children and their parents and teachers to di(2-ethylhexyl)phthalate (DEHP). Int. J. Hyg. Environ. Health 2004, 207, 15–22. [Google Scholar] [CrossRef] [PubMed]
  30. Adibi, J.J.; Perera, F.P.; Jedrychowski, W.; Camann, D.E.; Barr, D.; Jacek, R.; Whyatt, R.M. Prenatal exposures to phthalates in New York City and Krakow, Poland. Environ. Health Perspect. 2003, 111, 1719–1722. [Google Scholar] [CrossRef] [PubMed]
  31. Tranfo, G.; Caporossi, L.; Paci, E.; Aragona, C.; Romanzi, C.; De Carolis, C.; De Rosa, M.; Capanna, S.; Papaleo, B.; Pera, A. Urinary phthalates monoesters concentration in couples with infertility problems. Toxicol. Lett. 2012, 213, 15–20. [Google Scholar] [CrossRef] [PubMed]
  32. European Chemical Agency. Chemicals Database. Available online: https://echa.europa.eu (accessed on 20 June 2018).
  33. Kurata, Y.; Shimamura, N.; Katoh, M. Metabolite profiling and identification in human urine after single oral administration of DEHP. J. Toxicol. Sci. 2012, 37, 401–414. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  34. Kim, J.H.; Lee, S.; Shin, M.Y.; Kim, K.N.; Hong, Y.C. Risk assessment for phthalate exposures in the elderly: A repeated biomonitoring study. Sci. Total Environ. 2018, 618, 690–696. [Google Scholar] [CrossRef] [PubMed]
  35. Koch, H.M.; Christensen, K.L.; Harth, V.; Lorber, M.; Brüning, T. Di-n-butyl phthalate (DnBP) and diisobutyl phthalate (DiBP) metabolism in a human volunteer after single oral doses. Arch. Toxicol. 2012, 86, 1829–1839. [Google Scholar] [CrossRef] [PubMed]
  36. Eigenberg, D.A.; Bozigian, H.P.; Carter, D.E.; Sipes, I.G. Distribution, excretion, and metabolism of butylbenzyl phthalate in the rat. J. Toxicol. Environ. Health 1986, 17, 445–456. [Google Scholar] [CrossRef] [PubMed]
  37. Frederiksen, H.; Skakkebaek, N.E.; Andresson, A.M. Metabolism of phthalates in humans. Mol. Nutr. Food Res. 2007, 7, 899–911. [Google Scholar] [CrossRef] [PubMed]
  38. World Health Organization (WHO), International Programme on Chemical Safety (IPCS). Concise International Chemical Assessment Document (CICADS) n. 52: Diethyl Phthalate 2003. Available online: http://www.inchem.org/documents /cicads/cicads/cicad52.htm (accessed on 17 April 2008).
  39. Tranfo, G.; Papaleo, B.; Caporossi, L.; Capanna, S.; De Rosa, M.; Pigini, D.; Corsetti, F.; Paci, E. Urinary metabolite concentrations of phthalate metabolites in Central Italy healthy volunteers determined by a validated HPLC/MS/MS analytical method. Int. J. Hyg. Environ. Health 2013, 216, 481–485. [Google Scholar] [CrossRef] [PubMed]
  40. Jarošova, A.; Bogdanovičova, S. Phthalates in meat products in dependence on the fat content. Potravinarstvo Slovak J. Food Sci. 2016. [Google Scholar] [CrossRef]
  41. David, F.; Sandra, P.; Tienpont, B.; Vanwallengem, F.; Ikonomou, M. Analysis of Phthalates in Biota (Vegetation, Milk, Fish). In The Handbook of Environmental Chemistry; Staples, C.A., Ed.; Springer: Fairfax, VA, USA, 2003. [Google Scholar]
  42. David, R.M. Commentary regarding the article by Koch et al.: An estimation of the daily intake of di(2-ethylhexyl)phthalate (DEHP) and other phthalates in the general population. Int. J. Hyg. Environ. Health 2004, 207, 75–76. [Google Scholar] [CrossRef] [PubMed]
  43. World Health Organization. Biological Monitoring of Chemical Exposure in the Workplace: Guidelines; WHO: Geneva, Switzerland, 1996. [Google Scholar]
  44. The European Workplace Drug Testing Society (EWDTS). European Guidelines for Workplace Drug Testing in Urine. Available online: http://www.ewdts.org/ewdts-guidelines.html (accessed on 26 September 2016).
  45. Smerieri, A.; Testa, C.; Lazzeroni, P.; Nuti, F.; Grossi, E.; Cesari, S.; Montanini, L.; Latini, G.; Bernasconi, S.; Papini, A.M.; et al. Di-(2-Ethylhexyl) Phthalate Metabolites in Urine Show Age-Related Changes and Associations with Adiposity and Parameters of Insulin Sensitivity in Childhood. PLoS ONE 2015, 10, e0117831. [Google Scholar] [CrossRef] [PubMed]
  46. Göen, T.; Dobler, L.; Koschorreck, J.; Müller, J.; Wiesmüller, G.A.; Drexler, H.; Lolossa-Gehring, M. Trends of the internal phthalate exposure of young adults in Germany—Follow up of a retrospective human biomonitoring study. Int. J. Hyg. Environ. Health 2011, 215, 36–45. [Google Scholar] [CrossRef] [PubMed]
  47. Latini, G.; Gallo, F.; Dipaola, L.; De Angelis, S.; Olivieri, A. Pre-plus postnatal exposures to di-(2-ethylhexyl)-phthalate and thyroid dysfunction in prematurely born children. J. Endocrinol. Investig. 2014, 37, 97–98. [Google Scholar] [CrossRef] [PubMed]
  48. Centers for Disease Control and Prevention (CDC). Fourth National Report on human exposure to environmental chemicals. Department of Health and Human Services. Available online: https://www.cdc.gov/biomonitoring/pdf/fourthReport_UpdatedTables_Volume1_Jan2017.pdf (accessed on 10 July 2018).
  49. Testa, C.; Nuti, F.; Hayek, J.; De Felice, C.; Chelli, M.; Rovero, P.; Latini, G.; Papini, A.M. Di-(2-ethylhexyl) phthalate and autism spectrum disorders. ASN Neuro 2012, 4, 223–229. [Google Scholar] [CrossRef] [PubMed] [Green Version]
  50. Latini, G.; Massaro, M.; Mandich, A. Prenatal plus postnatal exposures to phthalates and child health risks. J Biol. Res. 2011, 84, 60–62. [Google Scholar] [CrossRef]
  51. Fromme, H.; Bolte, G.; Koch, H.M.; Angerer, J.; Boehmer, S.; Drexler, H.; Mayer, R.; Liebl, B. Occurrence and daily variation of phthalate metabolites in the urine of adult population. Int. J. Hyg. Environ. Health 2007, 210, 21–33. [Google Scholar] [CrossRef] [PubMed]
  52. Berman, T.; Celnikier, D.H.; Calafat, A.M.; Needham, L.L.; Amitai, Y.; Wormser, U.; Richter, E. Phthalate exposure among pregnant women in Jerusalem, Israel: Results of a pilot study. Environ. Int. 2009, 35, 353–357. [Google Scholar] [CrossRef] [PubMed]
  53. Zota, A.R.; Calafat, A.M.; Woodruff, T. Temporal trends in phthalate exposure: Findings from the National Health and Nutrition examination survey, 2001–2010. Environ. Health Perspect. 2014, 122, 235–241. [Google Scholar] [CrossRef] [PubMed]
  54. Calafat, A.M.; Valentin-Blasini, L.; Ye, X. Trends in Exposure to chemicals in personal care and consumer products. Curr. Environ. Health Rep. 2015, 2, 348–355. [Google Scholar] [CrossRef] [PubMed]
  55. Latini, G.; De Felice, C.; Presta, G.; Del Vecchio, A.; Paris, I.; Ruggieri, F.; Mazzeo, P. Exposure to Di(2-ethylhexyl)phthalate in Humans during Pregnancy. Biol. Neonate 2003, 83, 22–24. [Google Scholar] [CrossRef] [PubMed]
  56. Luisi, S.; Latini, G.; De Felice, C.; Sanseverino, F.; Di Pasquale, D.; Mazzeo, P.; Petraglia, F. Low serum concentrations of di-(2-ethylhexyl)phthalate in women with uterine fibromatosis. Gynecol. Endocrinol. 2006, 22, 92–95. [Google Scholar] [CrossRef] [PubMed]
  57. Luginbühl, M.; Weinmann, W. Creatinine in urine—A method comparison. Drug Test. Anal. 2017, 9, 1537–1541. [Google Scholar] [CrossRef] [PubMed]
  58. Huang, P.C.; Kuo, P.L.; Guo, Y.L.; Liao, P.C.; Lee, C.C. Association between urinary phthalate monoesters and thyroid hormones in pregnant women. Human Reprod. 2007, 22, 2715–2722. [Google Scholar] [CrossRef] [PubMed]
  59. Gyllenhammar, I.; Glynn, A.; Jönsson, B.A.G.; Lindh, C.H.; Darnerud, P.O.; Svensson, K.; Lignell, S. Diverging temporal trends of human exposure to bisphenols and plastizisers, such as phthalates, cuased by substitution of legacy EDCs? Environ. Res. 2017, 153, 48–54. [Google Scholar] [CrossRef] [PubMed]
  60. Ye, X.; Pierik, F.H.; Hauser, R.; Duty, S.; Angerer, J.; Park, M.M.; Burdorf, A.; Hofman, A.; Jaddoe, V.W.V.; Mackenbach, P.J.; et al. Urinary metabolite concentrations of organophosphorous pesticides, bisphenol A, and phthalates among pregnant women in Rotterdam, the Netherlands: The generation R study. Environ. Res. 2008, 108, 260–267. [Google Scholar] [CrossRef] [PubMed]
Ijerph 15 01950 g001 550
Figure 1. Comparison between periods, median data of urinary concentrations of phthalates’ metabolites in women and in men (MEP: monoethylphthalate; MnBP: mono-n-butylphthalate; MBzP: monobenzylphthalate; MEHP: mono(2-ethylhexyl)phthalate; MEHHP: mono(2-ethyl-5-hydroxyhexyl)phthalate).
Figure 1. Comparison between periods, median data of urinary concentrations of phthalates’ metabolites in women and in men (MEP: monoethylphthalate; MnBP: mono-n-butylphthalate; MBzP: monobenzylphthalate; MEHP: mono(2-ethylhexyl)phthalate; MEHHP: mono(2-ethyl-5-hydroxyhexyl)phthalate).
Ijerph 15 01950 g001
Table
Table 1. Phthalates of “high concern” and information about the authorization due for Reg. CE 1907/2006.
Table 1. Phthalates of “high concern” and information about the authorization due for Reg. CE 1907/2006.
SubstanceAcronymousIn List for Authorization fromIn Annex XIV of REACH fromCause for Being in the List
Di-(2-ethylhexyl)-phthalateDEHP28 October 200821 August 2013Endocrine disrupters
Toxic for reproduction
Di-(n-butyl) phthalateDnBP28 October 200821 August 2013Endocrine disrupters
Toxic for reproduction
Benzylbutyl phthalateBBP28 October 200821 August 2013Endocrine disrupters
Toxic for reproduction
Diisobutyl phthataleDiBP13 October 201021 August 2013Endocrine disrupters
Toxic for reproduction
Diisopentyl phthalateDIPP19 December 201204 January 2019Toxic for reproduction
Dipentyl phthalateDPP19 December 201204 January 2019Toxic for reproduction
n-Pentylisopentyl phthalatenPiPP19 December 201204 January 2019Toxic for reproduction
Bis(2-Methoxyethyl) phthalateDMEP19 December 2011-Toxic for reproduction
Table
Table 2. List of the phthalates considered in the present study with details on their main metabolites and use.
Table 2. List of the phthalates considered in the present study with details on their main metabolites and use.
SubstanceAcronymousMain Metabolites 1 Where You Can Find It
Di-(2-ethylhexyl)-phthalateDEHPMEHP (6%)
MEHHP (33–49%)
5-oxo-MEHP (14%)
[22,33,34]		Production of PVC 2 and vinyl chloride resins, where it is added to plastics to make them flexible. Adhesives and sealants, arts, crafts and hobby materials. Building/construction materials, electrical and electronic products, fabric, textile and leather products, paints and coatings, plastic and rubber products, playground and sporting equipment.
It’s a possible food contaminant for indirect contact.
Di-(n-butyl)phthalateDnBPMnBP (84%) [35]Production of plastics to help make it soft and flexible. Shower curtains, raincoats, food wraps, bowls, car interiors, vinyl fabrics, floor tiles, and other products. Adhesives and sealants, explosive materials, floor coverings, ink, toner and colorant products. Plastic and rubber products, excipient in drugs.
Benzylbutyl phthalateBBPMnBP (44%)
MBzP (16%) [36]		Adhesive and sealants, floor coverings
Dibenzyl phthalateDBzPMBzP (%not defined) [37]Ingredient in drugs for: disorders of the urinary system, prostate, bladder; dermatological disorders; skeletal disorders; antipsoriatics; arthritis, arthrosis, antiasthmatics, muscular and neuromuscular system disorders, nervous system disorders and many other type of drugs.
Diethyl-phthalateDEPMEP (70%) [38]Odor agents, plasticizers, adhesives and sealants, air care products, automotive care products, cleaning and furnishing care products, ink, toner and colorant products, laundry and dishwashing products, paints and coatings, personal care products, plastics and rubber products
1 MEHP: mono(2-ethylhexyl)phthalate, MEHHP: mono(2-ethyl-5-hydroxyhexyl)phthalate, MEP: monoethylphthalate, MBzP: monobenzylphthalate, MnBP: mono-n-butylphthalate, 5-oxo-MEHP: 5-oxo mono(2-ethylhexyl)phthalate; 2 PVC: polyvinyl chloride.
Table
Table 3. Questionnaire results.
Table 3. Questionnaire results.
Characteristics20112016
Women
(n = 83)		Men
(n = 74)		Women
(n = 111)		Men
(n = 60)
Age (SD 1)36.5 (7.2)40.4 (7.3)42.4 (8.3)41 (9.2)
Smoking (% current)15.732.439.347.5
Regular and occasional alcohol intake (%)37.358.15890.2
Area of residence (%)
Urban73.571.663.457.4
Rural21.720.336.826.2
Coast4.88.16.316.4
Other--3.5-
Use of plastic containers for fat food storage (%)
Never59.054.159.355.2
Daily10.89.512.37.1
weekly15.717.615.414.2
monthly14.518.813.023.5
Use of canned foods at least weekly (%)31.331.125.923.0
Eating fat fish at least weekly (%)26.525.75868.9
Job (%)
Office/school35.728.630.823.3
Trade26.932.312.615.5
Craftsman/manual worker1.29.54.317.4
Cleaning man/woman12.02.726.48.2
Hairdresser/beautician3.61.47.5
Armed forces1.212.2 18.9
Healthcare/Laboratory18.113.512.510.1
Other0.1-5.96.6
1 SD: standard deviation.
Table
Table 4. Limits of Detection (LOD) and detection frequencies.
Table 4. Limits of Detection (LOD) and detection frequencies.
LODs (µg/L) MEP 1MnBP 2MBzP 3MEHHP 4MEHP 5
1.03.00.21.01.0
2011201620112016201120162011201620112016
MenN 6 < LOD221915403346049
% 7 > LOD7168991010045959010018
WomenN 6 < LOD0110992380133585
% 7 > LOD10090100119866100885723
1 MEHP: mono(2-ethylhexyl)phthalate; 2 MnBP: mono-n-butylphthalate; 3 MBzP: monobenzylphthalate; 4 MEHHP: mono(2-ethyl-5-hydroxyhexyl)phthalate; 5 MEP: monoethylphthalate; 6 N: number of samples <LOD; 7 %: detection frequencies.
Table
Table 5. Results of urine analysis of women (µg/g creatinine).
Table 5. Results of urine analysis of women (µg/g creatinine).
MEP 4MnBP 5MBzP 6MEHP 7 + MEHHP 8
2011 12016 22011 12016 22011 12016 22011 12016 2
Average (SD 3)297.7 (881.1)187.7 (321.0)54.5 (55.7)1.3 (5.3)15.0 (21.9)1.3 (3.7)20.9 (17.0)5.9 (5.6)
Median73.149.938.80.07.00.515.64.5
5th percentile6.40.04.80.00.70.03.60.8
95th percentile1177.9781.3163.06.472.85.161.416.9
1 83 women in 2011; 2 111 women in 2016; 3 SD:standard deviation; 4 MEP: monoethylphthalate; 5 MnBP: mono-n-butylphthalate; 6 MBzP: monobenzylphthalate; 7 MEHP: mono(2-ethylhexyl)phthalate; 8 MEHHP: mono(2-ethyl-5-hydroxyhexyl)phthalate.
Table
Table 6. Results of urine analysis of men (µg/g creatinine).
Table 6. Results of urine analysis of men (µg/g creatinine).
MEP 4MnBP 5MBzP 6MEHP 7 + MEHHP 8
2011 12016 22011 12016 22011 12016 22011 12016 2
Average (SD 3)142.7 (243.6)97.7 (218.1)57.7 (62.6)1.4 (7.2)14.5 (27.8)0.9 (2.2)21.1 (26.0)5.7 (7.1)
Median49.921.637.60.04.80.014.13.1
5th percentile4.20.07.30.00.80.03.80.0
95th percentile637.9406.7145.64.851.73.062.120.1
1 74 men in 2011; 2 60 men in 2016; 3 SD: standard deviation; 4 MEP: monoethylphthalate; 5 MnBP: mono-n-butylphthalate; 6 MBzP: monobenzylphthalate; 7 MEHP: mono(2-ethylhexyl)phthalate; 8 MEHHP: mono(2-ethyl-5-hydroxyhexyl)phthalate.
Table
Table 7. Comparison among biomonitoring median data in different geographical areas—women (µg/g creatinine).
Table 7. Comparison among biomonitoring median data in different geographical areas—women (µg/g creatinine).
Country Italy [This Study]Italy [This Study]Taiwan [51]Israel [52]USA [46]USA [46]Sweden [48]the Netherlands [53]Germany [54]
Sampling year201120162005–200620061999–20002011–20122009–201420042005
N 83 11176191326122917810027
phthalates
MEP 273.149.968.0140.5123.0 151.8 124.3112.0 1-
MnBP 338.80.0195.045.928.6 19.8 142.743.2 146.8
MBzP 47.00.53.79.611.0 15.87 18.768.9 17.6
∑DEHP 5 met.15.64.560.831.33.36 11.70 111.421.2 14.3
1 geometric mean; 2 MEP: monoethylphthalate; 3 MnBP: mono-n-butylphthalate; 4 MBzP: monobenzylphthalate; 5 DEHP: di(ethylhexyl)phthalate.
Table
Table 8. Comparison among biomonitoring median data in different geographical areas—men (µg/g creatinine).
Table 8. Comparison among biomonitoring median data in different geographical areas—men (µg/g creatinine).
Country Italy [This Study]Italy [This Study]USA [46]USA [46]Germany [54]
Sampling year201120161999–20002011–20122005
N 74601215125823
phthalates
MEP 249.921.692.8 135.8 1-
MnBP 337.60.017.3 17.61 141.4
MBzP 44.80.09.14 14.5 15.1
∑DEHP 5 met.14.13.12.89 11.41 14.3
1 geometric mean; 2 MEP: monoethylphthalate; 3 MnBP: mono-n-butylphthalate; 4 MBzP: monobenzylphthalate; 5 DEHP: di(ethylhexyl)phthalate.

Share and Cite

MDPI and ACS Style

Tranfo, G.; Caporossi, L.; Pigini, D.; Capanna, S.; Papaleo, B.; Paci, E. Temporal Trends of Urinary Phthalate Concentrations in Two Populations: Effects of REACH Authorization after Five Years. Int. J. Environ. Res. Public Health 2018, 15, 1950. https://doi.org/10.3390/ijerph15091950

AMA Style

Tranfo G, Caporossi L, Pigini D, Capanna S, Papaleo B, Paci E. Temporal Trends of Urinary Phthalate Concentrations in Two Populations: Effects of REACH Authorization after Five Years. International Journal of Environmental Research and Public Health. 2018; 15(9):1950. https://doi.org/10.3390/ijerph15091950

Chicago/Turabian Style

Tranfo, Giovanna, Lidia Caporossi, Daniela Pigini, Silvia Capanna, Bruno Papaleo, and Enrico Paci. 2018. "Temporal Trends of Urinary Phthalate Concentrations in Two Populations: Effects of REACH Authorization after Five Years" International Journal of Environmental Research and Public Health 15, no. 9: 1950. https://doi.org/10.3390/ijerph15091950

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop