Ultra-high performance liquid chromatography tandem mass spectrometry analysis of UV filters in marine mussels (Mytilus galloprovinciallis) from the southern coast of Spain
Graphical abstract
Introduction
UV filters are organic compounds with aromatic structures capable of absorbing solar radiation. Therefore, many personal care products (PCPs) include different families of these substances such as camphor or benzophenone derivatives, among others. UV filters are able to absorb solar radiation UV-A (320–400 nm) and UV-B (280–320 nm) [1], [2], [3]. 4-Methylbenzylidene camphor (4-MBC) is one of the most widely used UV filter. This compound cannot be completely removed by the traditional wastewater treatment process. Some studies show that in a Chinese treatment plant, the removal efficiency of this compound is 40% [4], [5]. Benzophenones (BPs), on the other side, are a family of UV filter compounds which have a high lipophilic character and are able to enter the body and bioaccumulate easily [6], [7], [8], [9], [10].
In recent years, there has been increasing concern about sun exposure and the associated risks. Thus, the consumption and use of products which contain UV filters for sun protection, including sunscreens, shampoos or hair dyes as well as plastics, paints or packaging of outdoor materials, has increased considerably [3]. Many UV filters are recognized as emerging pollutants and constitute a risk to human health and the environment. Toxicological studies have demonstrated their estrogenic effects and toxicity for reproduction and development, even when they are found in low concentrations, being considered endocrine disrupting chemicals (EDCs) [5]. EDCs are exogenous compounds capable of causing adverse effects on the health of an organism and its offspring by interfering with the normal functioning of the endocrine system. Within this category, UV filters have received special attention due to their high estrogenic activity, their widespread use and their ubiquitous presence in marine environments. The high occurrence and persistence, in which they are found, represent a potential danger for the organisms that inhabit these environments. An example is the alarming impact they have on coral reefs, inducing genotoxic alterations and rapid bleaching. For this reason, countries such as Hawaii or Florida have taken measures, and have banned the sale of sun creams containing benzophenone-3 (BP-3), one of the most widely used UV filters [2], [3], [11], [12], [13].
The entry of UV filters into the marine environment can be direct, through recreational activities of bathers on beaches and swimming pools, and the consequent release of these compounds through the skin; but also can be indirect, through discharges from wastewater treatment plants, laundry or urine. However, the primary cause is their extensive use in the manufacture of household and cosmetic products and in the textile industry [14], [15], [16]. The efficiency of removal of these compounds depends on several factors such as the operational conditions, the technology used as well as the type of treatment being hardly completely eliminated [17], [18]. UV filters pose a great health risk since they are able to enter into trophic chain and biomagnificate [16]. Many of these compounds tend to accumulate in tissues of aquatic organisms because they have several benzene rings and therefore, high lipophilicity. In addition, they pose high Kow values and they are very stable against biotic degradation [2]. The European Union has regulated the maximum concentration of these compounds in sunscreens and other PCPs, which must be between 0.1 and 10% [19]. Thus, according to Regulation (EC) No 1223/2009 of the European Parliament and of the Council of 30 November 2009 on cosmetic products, the maximum content allowed in commercial products is up to 4% for the case of 4- MBC and 6% for BP-3 [19].
Some studies have demonstrated the accumulation of these compounds in aquatic organisms. Mussels have deserved our attention since they are very abundant in nature and have an important role in food chain due the human consumption. Mussels are filter feeders capable to retaining complex mixtures of contaminants, being bioindicators of high interest. They present some characteristics such as stress resistance, sessile behaviour or tolerance to changes in salinity, and they retain and accumulate pollutants found in the sea with higher concentration levels [2], [12], [20], [21]. In fact, on the coasts of Norway, Portugal and France, concentration levels of contaminants > 800–1000 ng g−1 have been determined in cod, shrimp or mussels [3]. However, according to the literature consulted, there are few studies on the quantification of UV filters compounds in marine organisms such as mussels, and on the evaluation of the concentrations found according to the characteristics of the sampling area. Furthermore, none of them cover a large number of UV filters, and there is no data available in relation to the benzophenone family. The objectives of this study have been: (i) develop an UHPLC-MS/MS method for the determination of UV filters in mussels trough an efficient extraction process capable of achieving low LODs, high recoveries and without interferences from other compounds; (ii) evaluate the presence of these compounds UV filters on the coast of Granada (Spain); and (iii) interpret and compare the results with previously reported available data, and evaluate the evolution.
Section snippets
Chemicals and reagents
Analytical grade of standards: BP-1 (≥99%), BP-2 (≥97%), BP-3 (≥98%), BP-3d5 (≥99%), BP-8 (≥98%), 4-OH-BP (≥98%) and 4-MBC (≥98%) were obtained from Sigma-Aldrich (Madrid, Spain). BP-6 (≥98%) was purchased from Alfa Aesar (Massachusetts, MA, USA). Chemical structures of compounds are shown in supplementary material (Table S1). Ultrapure water was purified by a Milli-Q Plus® purification system (Millipore, Madrid, Spain). LC-MS grade methanol (MeOH) was obtained by VWR Prolabo Chemicals
Extraction procedure optimization
A deep study was conducted on the most influential variables that affect the digestion and extraction process of mussel samples. The influence of solvent type, acid concentration or number of extraction cycles were investigated. Aliquots of 0.5 g of fortified blank samples (100 ng g−1) were used.
Conclusions
Although mussels have been previously studied to evaluate the bioaccumulation of pollutants, in relation to UV-filters the information available on these organisms is scarce. An UHPLC-ESI-MS/MS method has been developed and validated for the analysis of UV filters in wild mussels from five areas on the coast of Granada, Spain. UAE followed by a dispersive phase clean up with C18 is proposed, being the sample treatment simple and fast. The method provided acceptable validation data: high
CRediT authorship contribution statement
M.C. Gómez-Regalado: Formal analysis, Investigation, Methodology, Writing – original draft, Writing – review & editing. L. Martín-Pozo: Conceptualization, Formal analysis, Data curation, Writing – original draft, Writing – review & editing. S. Cantarero-Malagón: Formal analysis, Investigation, Methodology, Writing – original draft. F. Hidalgo: Formal analysis, Investigation, Methodology, Writing – original draft. A. Zafra-Gómez: Conceptualization, Formal analysis, Funding acquisition,
Declaration of Competing Interest
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
Acknowledgements
The authors are grateful to all those who contributed to the collection of the samples. They also thank the Framework of the Program Operating FEDER Andalusia (2014-2020) from the Andalusia Government, Spain (Project No. B1‐FQM‐073‐UGR18). L. Martín-Pozo expresses her gratitude for the contract from the Youth Employment Operational Program, co-funded by the Regional Government of Andalusia, Spain and the European Social Fund (ESF).
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2022, Science of the Total EnvironmentCitation Excerpt :However, for its congener P. canaliculus (Gmelin, 1791) concentrations exceeded 2000 ng g−1 l.w. in Lyttelton Harbour, Christchurch, New Zealand (Emnet et al., 2020). BP-1 showed minimal concentrations in mussels M. galloprovinciallis, organisms with a slow rate of biotransformation because intake exceeds metabolism, or involve other metabolic pathways (Gómez-Regalado et al., 2021). 4-MBC bioaccumulated in the species M. galloprovincialis and the uptake of waterborne 4-MBC was very rapid, and after only 24 h of exposure to 1 μg L−1, the tissular concentrations were 418 μg kg−1 d.w. (Vidal-Liñán et al., 2018).
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2022, Science of the Total EnvironmentCitation Excerpt :The trace amount of UV-filters was observed even in the soils collected from the protected forest (Vecchiato et al., 2021), which demonstrates that the pollution by these chemical compounds has extrapolated in the more urbanized regions. In addition to that, the increasing number of studies that detected OU-VFs at present in marine organisms such as clams, oysters, and intertidal gastropods (Nakata et al., 2009), corals (Tsui et al., 2017; Mitchelmore et al., 2021; Moeller et al., 2021), loggerhead turtles (Cocci et al., 2020), mussels (Bachelot et al., 2012; Picot-Groz et al., 2018; Vidal-Liñán et al., 2018; Gómez-Regalado et al., 2021), dolphins (Gago-Ferrero et al., 2013) macroalgae (Cunha et al., 2015) and, fish (Kim et al., 2011; Molins-Delgado et al., 2018; Gimeno-Monforte et al., 2020; Peng et al., 2020; Huang et al., 2021a,b). Despite this, several gaps about the (eco) toxicological potential of these compounds still need to be filled.