Occurrence of cyclophosphamide and epirubicin in wastewaters by direct injection analysis–liquid chromatography–high-resolution mass spectrometry

Background, aim, and scope

According to the high incidence of cancer worldwide, the amount of cytostatic drugs administered to patients has increased. These compounds are excreted to wastewaters, and therefore become potential water contaminants. At this stage, very little is known on the presence and elimination of cytostatic compounds in wastewater treatment plants (WWTP). The aim of this study was to develop a liquid chromatography?Chigh-resolution mass spectrometry (LC?COrbitrap?CMS) method for the determination of cyclophosphamide and epirubicin in wastewaters. These compounds represent two outmost used cytostatic agents.

Materials and methods

Extraction and analytical conditions were optimized for cyclophosphamide and epirubicin in wastewater. Both solid-phase extraction using Oasis 200?mg hydrophilic?Clipophilic balanced (HLB) cartridges and direct injection analysis were evaluated. Mass spectral characterization and fragmentation conditions were optimized at 50,000 resolving power (full width at half maximum, m/z 200) to obtain maximum sensitivity and identification performance. Quality parameters (recoveries, limits of detection, and repetitivity) of the methods developed were determined, and best performance was obtained with direct water analysis of the centrifuged wastewater. Finally, this method was applied to determine the presence of cyclophosphamide and epirubicin in wastewaters from a hospital effluent, an urban effluent, and influents and effluents from three WWTP.

Results and discussion

Cyclophosphamide and epirubicin were recovered after 50?mL preconcentration on solid-phase extraction 200?mg Oasis HLB cartridges (87% and 37%, respectively), and no breakthrough was observed by extracting 500?mL of water. Limits of detection were of 0.35 and 2.77?ng/L for cyclophosphamide and epirubicin, respectively. On the other hand, direct injection of water spiked at 1???g/L provided recoveries of 107% for cyclophosphamide and 44% for epirubicin and limits of detection from 3.1 to 85?ng?L^?1, respectively. The analysis of wastewaters using direct injection analysis revealed the presence of cyclophosphamide and epirubicin in WWTP influents and hospital and urban effluents at levels ranging from 5.73 to 24.8???g?L^?1.

Conclusions

The results obtained in this study demonstrate the capability of LC?COrbitrap?CMS for accurate trace analysis of these very polar contaminants. This method permitted to identify cyclophosphamide and epirubicin in wastewaters and influents of WWTP, but no traces were detected in WWTP effluents. The methodology herein developed is sensitive and robust and applicable for screening of a large number of samples since no preconcentration is needed.     

In utero and post-natal accumulation of organochlorine compounds in children under different environmental conditions

A study of intake of organochlorine compounds (OCs) in children both in utero and at the age of four years has been performed in two cohorts. One encompasses children born in Menorca Island between 1997 and 1998, and the other those born between 1997-1999 in Ribera d'Ebre, an in-land industrial-agricultural area of Catalonia. Comparison of the OC concentrations in serum samples from both populations, including those obtained from cord blood and blood collected at four years, provides information on the influence of local pollution sources in the accumulation of these compounds. Statistically significant differences (p < 0.01) have been found for hexachlorobenzene, indicating that chronic airborne contamination to this compound in Ribera d'Ebre involved higher in utero exposure, which increased in the first four years of growth. Similarly, in the cohort of Menorca, higher in utero exposure to PCBs was observed and this increased subsequently in the first four years of growth. Other compounds encompassing temporal contamination episodes such as gamma-HCH involved higher in utero exposure but strongly diminished in the first four years of growth. Overall, it can be concluded that local chronic pollution by OCs has a direct effect in children living in the surrounding areas. This influence is reflected in higher exposure, both in utero and in the first years of growth, that significantly stands out over the background contamination due to the ubiquity of these compounds.     

Environmental strategy and performance in small firms: a resource-based perspective

In spite of the widespread recognition of the important roles that small and medium sized enterprises (SMEs) play in most economies, limited research has focused on their impacts on the natural environment and the strategies such enterprises adopt to reduce these impacts. It is usually assumed that SMEs lack the resources to implement proactive environmental strategies that go beyond minimum regulatory compliance. In this study of 108 SMEs in the automotive repair sector in Southern Spain, we found that SMEs undertake a range of environmental strategies from reactive regulatory compliance to proactive pollution prevention and environmental leadership. These strategies are associated with three organizational capabilities: shared vision, stakeholder management, and strategic proactivity, hypothesized based on the unique strategic characteristics of SMEs--shorter lines of communication and closer interaction within the SMEs, the presence of a founder's vision, flexibility in managing external relationships, and an entrepreneurial orientation. We also found that firms with the most proactive practices exhibited a significantly positive financial performance.     

Synthesis and Characterization of Lactic Acid Oligomers: Evaluation of Performance as Poly(Lactic Acid) Plasticizers

The use of fully bio-based and biodegradable materials for massive applications, such as food packaging, is an emerging tendency in polymer research. But the formulations proposed in this way should preserve or even increase the functional properties of conventional polymers, such as transparency, homogeneity, mechanical properties and low migration of their components to foodstuff. This is not always trivial, in particular when brittle biopolymers, such as poly(lactic acid) (PLA), are considered. In this work the formulation of innovative materials based on PLA modified with highly compatible plasticizers, i.e. oligomers of lactic acid (OLAs) is proposed. Three different synthesis conditions for OLAs were tested and the resulting additives were further blended with commercial PLA obtaining transparent and ductile materials, able for films manufacturing. These materials were tested in their structural, thermal and tensile properties and the best formulation among the three materials was selected. OLA with molar mass (M_n) around 1,000 Da is proposed as an innovative and fully compatible and biodegradable plasticizer for PLA, able to replace conventional plasticizers (phthalates, adipates or citrates) currently used for films manufacturing in food packaging applications.     

Roadless and Low-Traffic Areas as Conservation Targets in Europe

With increasing road encroachment, habitat fragmentation by transport infrastructures has been a serious threat for European biodiversity. Areas with no roads or little traffic (“roadless and low-traffic areas”) represent relatively undisturbed natural habitats and functioning ecosystems. They provide many benefits for biodiversity and human societies (e.g., landscape connectivity, barrier against pests and invasions, ecosystem services). Roadless and low-traffic areas, with a lower level of anthropogenic disturbances, are of special relevance in Europe because of their rarity and, in the context of climate change, because of their contribution to higher resilience and buffering capacity within landscape ecosystems. An analysis of European legal instruments illustrates that, although most laws aimed at protecting targets which are inherent to fragmentation, like connectivity, ecosystem processes or integrity, roadless areas are widely neglected as a legal target. A case study in Germany underlines this finding. Although the Natura 2000 network covers a significant proportion of the country (16%), Natura 2000 sites are highly fragmented and most low-traffic areas (75%) lie unprotected outside this network. This proportion is even higher for the old Federal States (western Germany), where only 20% of the low-traffic areas are protected. We propose that the few remaining roadless and low-traffic areas in Europe should be an important focus of conservation efforts; they should be urgently inventoried, included more explicitly in the law and accounted for in transport and urban planning. Considering them as complementary conservation targets would represent a concrete step towards the strengthening and adaptation of the Natura 2000 network to climate change.     

Alkylphenols and polycyclic aromatic hydrocarbons in eastern Mediterranean Spanish coastal marine bivalves

This paper reports the first results on alkylphenol pollution in edible bivalves from the Spanish coast. Two sampling campaigns (July 2006 and July 2007) were carried out to determine the concentration of nonylphenol (NP), octylphenol (OP), and eight polycyclic aromatic hydrocarbons (PAHs) in wild mussels (Mytilus galloprovincialys) and clams (Donax trunculus) at 14 sampling sites along the eastern Mediterranean Spanish coast. The results show that NP is the predominant alkylphenol, being the port of Valencia the most polluted area (up to 147 ??g/kg wet weight in clams). Moving away from the ports the concentration of NP in bivalves decreased. OP concentration was below its detection limit in most of the studied areas and its maximum concentration (6 ??g/kg w/w) was measured in clams from the port of Sagunto. The presence of low levels of PAHs was observed in most of the studied areas. The total PAHs concentration (i.e., sum of the eight measured PAHs) achieved a maximum value of 10.09 ??g/kg w/w in the north coast of Valencia city. The distribution pattern of the individual PAHs showed that both pollution sources petrogenic and pyrolytic were present in the sampled areas. Fluoranthene was the most abundant PAH in mussels while benzo(b)fluoranthene in clams. The maximum concentration of 10 ??g/kg w/w for benzo(a)pyrene established by the European Commission was never reached, indeed sampled bivalves showed concentrations 10 times lower than this reference value. Thus, they can be considered safe for human consumption. Despite the low contamination levels, the results show an overall pollution of bivalves by alkylphenol and PAHs as well as an increment in the number of polluted areas from 2006 to 2007. Thus, periodical sampling campaigns should be carried out to monitor the long-term tendency of these toxic and persistent pollutants.     

Integrative assessment of coastal pollution in a Ría coastal system (Galicia, NW Spain): correspondence between sediment chemistry and toxicity

Elutriate embryo-larval bioassays with sea-urchins (Paracentrotus lividus) and ascidians (Ciona intestinalis) were conducted concurrently with trace metal analyses as part of an integrative assessment of sediment pollution at Ría de Pontevedra (Galicia, NW Spain). High metal contents in sediments were found in localised areas from the inner part of the estuary indicating a clear anthropogenic influence. In particular, very high Cu, Zn and Pb levels were found at sites P2 and P3, which were also the most toxic to the embryo-larval bioassays. Sediment quality guidelines were used to help in the ecological interpretation of sediment chemistry data and to identify pollutants of concern. Cu and Zn in P3 were consistently above the effects range median (ERM) values, which seem to be good predictors of toxicity to sea-urchin and ascidian embryos. A toxic unit approach, based on published EC(50) values and metal levels in elutriates, was used to assess the harmful ecological effects associated to sediment chemistry. Toxicity detected in P3 may be explained on the basis of the toxic unit model; however, the high toxicity detected at P2 may be attributable not only to the metals quantified in the analyses but also to unmeasured organic pollutants. Multidimensional scaling applied independently to the toxicology and chemistry data resulted in a good agreement between both types of configurations. Moreover, the Mantel test revealed a significant correlation (r(M)=0.481; p=0.019) between metal concentrations and toxicity data profiles, supporting the correspondence between configurations.     

Newborns and low to moderate prenatal environmental lead exposure: might fathers be the key?

This study is part of the BioMadrid Project, a bio-monitoring study designed to assess pollutants in the environment surrounding children born in the Madrid region. Our aim in this report is to evaluate the association between prenatal lead exposure and fetal development using three biological samples (maternal and paternal blood lead at around 34 weeks of gestation as well as cord blood lead levels), three biomarkers of effect in cord blood peripheral lymphocytes (micronucleus in binucleated cells, nucleoplasmic bridges, and nuclear buds), and different anthropometrical characteristics at birth. Maternal and cord blood lead were not associated with newborn measurements or genotoxicity biomarkers. In contrast, increases in father blood lead were coupled with lower weight (mean difference (MD), ?110.8 g; 95 % confidence intervals (95%CI), ?235.6 to 6.00; p?<?0.10) and shorter abdominal (MD, ?0.81 cm; 95%CI, ?1.64 to 0.00; p?<?0.05) and cephalic (MD, ?0.32 cm; 95%CI, ?0.65 to 0.00; p?<?0.05) circumferences at birth as well as with the presence of nucleoplasmic bridges (odds ratio, 1.03; 95%CI, 1.00 to 1.06; p?<?0.05) and nuclear buds (odds ratio, 1.02; 95%CI, 0.99 to 1.04; p?<?0.10). These associations were mainly confined to female babies, in whom paternal lead was also inversely associated with length. Our results support the hypothesis that paternal lead exposure may be affecting the development of newborns.     

BIOAMBIENT.ES study protocol: rationale and design of a cross-sectional human biomonitoring survey in Spain

Human biomonitoring (HBM), defined as the measurement of concentrations of chemicals or of their metabolites in human biological matrices, is considered the method of choice for determining internal exposures in individuals. HBM is part of environmental exposure surveillance systems in several countries. In 2007, the Ministry of Agriculture, Food and Environment promoted BIOAMBIENT.ES project, a national-level HBM study on environmental pollutants carried out in Spain to estimate levels of heavy metals, persistent organic pollutants, and other substances on the Spanish active workforce. BIOAMBIENT.ES is a nationwide cross-sectional study, with a stratified cluster sampling designed to cover all geographical areas, sex and occupational sectors, and aimed to obtain a representative sample of the Spanish workforce. Participants were recruited among people residing in Spain for 5 years or more, which underwent their annual occupational medical check-up in the health facilities of the Societies for Prevention of IBERMUTUAMUR, MUTUALIA, MC-PREVENCIÓN, MUGATRA, UNIMAT PREVENCIÓN, and PREVIMAC (March 2009–July 2010). A total of 1,892 subjects fulfilled the criteria for inclusion, donated biological samples (1,880 blood, 1,770 urine, and 577 hair) and completed a short self-administrated epidemiological questionnaire on environmental and lifestyle-related exposures. Additionally, clinical information from participant’s health exams was obtained. This project will provide a first overview of the body burden of selected pollutants in a representative sample of the Spanish-occupied population. This information will be useful to establish reference values of the studied population and, eventually, to evaluate temporal trends and the effectiveness of environmental and health policies.