DBP: Contamination in Water and Proposed Regulations By: Helen Davoodi, Camilla Westberg and Antonio F. Machado Department of Environmental and Occupational Health California State University, Northridge Abstract DBP is a chemical used in consumer products. Due to its high production volume and leachability, it is found at low levels in air, water and soil throughout the world. Exposure occurs through ingestion, inhalation and dermal contact. It acts as an endocrine disruptor in animals and humans, and can be especially dangerous to aquatic organisms. Currently in the U.S., DBP is only regulated in some children’s products. Introduction Di-n-butyl phthalate (DBP): •Man-made chemical used as plasticizer to add flexibility, durability, and transparency.1,3 •Used in solvents, varnishes, time-released pharmaceuticals, food packaging and personal care products. •DBP is not covalently bound to the product in which it is used and is therefore easily released into the environment.2 •Endocrine-disrupting chemical due to its anti-estrogenic and anti-androgenic activities. 4 •Can make up to 40% of the plastic volume5 •Reference dose: 0.1 mg/kg bw/day •Carcinogenicity: Class D – not classifiable6 Chemical Properties •Oily liquid that is colorless to faint yellow. •Formula: C16H22O4 •Molecular weight: 278.4 g/mol •Melting point: -35oC •Boiling point: 340oC •Metabolite: Mono-N-butyl phthalate (MBP) Occurrence in Environmental Matrices Ecotoxicology •DBP is one of the most common phthalates detected in environmental samples.13 •Air is the primary element to which DBP is emitted.13 •DBP is prevalent in aquatic environments due to its ubiquitous use.15 •DBP enters water from industrial facilities, wastewater treatment plants, consumer products, and incorrect disposal.15 •The majority of DBP dissolves rapidly in water, the remainder binds to suspended materials.11 •U.S. data for DBP in waters is limited and/or >20 years old. •There is less bioconcentration of DBP/MBP in aquatic organisms with more sophisticated metabolic systems: algae > crustaceans > insects > fish.11 •Exposure to aquatic organisms occurs via gill respiration, dermal contact and by eating contaminated food. •Is as an anti-estrogen in female fish, affecting the ovaries and leading to reduced fecundity. 15 Location Surface water (µg/L) Sediments (mg/kg) Sewage sludge (µg/L) Germany14 0.12-8.8 0.06-2.08 0.2-10.4 Humans •Majority of humans are exposed to DBP. •Exposure occurs primarily through ingestion, but also inhalation and to a limited extent dermal contact. 18 •Example of DBP concentrations in drinking water:19 Country Tap Water (µg/L) Bottled Water (µg/L) China 0.6 54.6 Portugal 0.52 0.35 Tchek Republic 0.05 0.2 Remediation of Water 1. Ultra-Violet (UV) Irradiation at 254 nm •Reduces DBP concentrations in water and wastewaters. •pH levels 5-9 have the longest lag time, but highest efficiency. •Can be used in acidic, neutral and basic environments. •End products include phthalic acid, ketones, and alcohol. 16 2. Bacterial Degradation •Microorganisms are capable of using DBP as their food source. •Sphingobium sp. can break down DBP in water containing <4% NaCl, making it a good candidate for remediating salty wastewater.10 3. Ozonation •Ozone (O3) degrades DBP directly or indirectly by interaction with the O3 molecule or the hydroxyl radicals as O3 breaks down. •O3 can degrade <90% of DBP. Increased efficiency at higher temperatures and low dose of humic substances. 17 •Example of DBP/MBP levels in new mothers in Sweden (values are similar to studies conducted in Germany and the U.S.):20 Body Fluid DBP conc. MBP conc. (average) (average) 2.8 ng/mL 1.2 ng/mL Urine n/a 46 ng/mL Blood 1.2 ng/mL 1.8 ng/mL Breast milk •There is a negative correlation between MBP in urine and testosterone levels in human males. DBP may target the pituitary gland and disrupt the negative feedback mechanism for testosterone production. 18 Health Effects and Proposed Mechanisms The following effects are from experimental levels of DBP exposures in laboratory settings. Exact mechanisms are unknown. Suggested mechanisms are indicated by arrows. 1. Prenatal exposure in male rats: 2. Exposure in pubertal male rats: •Infertility as adults •Fewer offspring later in life •Decreased seminal vesicles •Reduced steroidogenesis •Reduced anogenital distance2 •Chryptorchidism •Hypospadia5 •Damaged /reduced number of Sertoli cells and loss of spermatogenesis 5, 7 •reduced weight of testes and epididymis •histological changes in the testes •increased expression of heterogeneous nuclear ribonucleoprotein A2/B1 •increased expression of vimentin proteins •reduced superoxide dismutase •increased malondialdehyde •altered luteinizing hormone levels (LH) •reduced sperm quality •Reduced sperm count •Impacted motility and viability •Deformed sperms 2 Increased follicle-stimulating hormone (FSH) 7 Regulators •Formation of vacuoles •Leydig cell (LC) hyperplasia •Increased reactive oxygen species7 Seminiferous tubular degeneration2,5 Reduced estrogen and androgen receptor 2 Less inhibin Figure 1: Overview of seminiferous tubule in mammals. LC: Estradiol testosterone •Reduced testosterone •Increased estradiol7 Lipid peroxidation of epididymal tubules Impaired sperm motility8 The combined effect of reduced testosterone, fewer Sertoli cells, damaged LCs, increased FSH and LH, reduced estrogen and androgen receptor may lead to fewer spermatogenic cells and reduced spermatogenesis.7 3. Exposure in female mice: Regulations Impacts ovarian antral follicles: •Increased apoptosis, possibly due to reduced B-cell leukemia/lymphoma 2 levels. •Reduced 17β-estradiol accumulation. •Alters cell cycle regulators.9 •Banned in children’s toys and certain childcare articles in the U.S., E.U., and Canada.21, 22, 23 • Banned in cosmetics and restricted in food packaging in the E.U. 22 •Currently not regulated in drinking water in the U.S. •Included in CERCLA’s Priority List of Hazardous Substances. 24 •Reduced Cyc D9 •Halted progression/ accumulation of follicles in G1 phase. •Reduced Cyc E •Increased p219 •Reduced Cyc A and B9 •Suppressed/delayed growth in follicles9 •Reduction of follicles in S phase9 Fate and Transport •DBP can be removed by hydrolysis, photolysis and biodegradation.10 DBP 11 •Gram positive/ negative bacteria and actinomycetes can use DBP as MBP + Alcohol a carbon and energy source. •Degradation rate is slowed at low temperatures & nutrient poor Phthalic Acid (PA) environments. + Alcohol •Fresh water or marine water does not impact degradation Aerobic Anaerobic rate (<30 days). •Hydrolysis half-life is 22 years. 11 Dihydrophthalate PA-CoA •In humans:12 Protochatecuate Glucoronide Intestine: Body: Benzoyl-CoA DBP hydrolyzed MBP oxidized conjugates Urine: excreted Acetate + CO2 + pyruvate/succinate DBP’s effects on humans at environmentally relevant levels are still not clear. Further studies are necessary to decipher how it affects humans and through what mechanisms. The main exposure route is through ingestion, but drinking water is not the main contributor. Instead of regulating DBP in water, current regulations for DBP in childcare products should be expanded to include consumer products, primarily those that are marketed to teenagers and women of childbearing age. Regulating DBP in consumer products will lead to lowered production of DBP and will potentially reduce human exposure and the likelihood of environmental releases. As a result, such regulations would offer protection for both humans and aquatic organisms. In theory, limiting the widespread use of DBP would reduce clean up costs in the future. Ongoing water monitoring is required to evaluate if proposed regulations are effective. References Figure 2: Overview of cell cycle. Multi-step process Conclusion Multi-step process Acetate + CO2 + H2 1. Swan, Shanna. 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