Organotins Remain a Serious Threat to the Indo-Pacific Humpback Dolphins in the Pearl River Estuary.

Marine mammals often accumulate high levels of environmental contaminants, even those that are globally regulated regarding usage, raising concerns about their health status. Here, we conducted the first investigation of tissue distribution, spatiotemporal trends, and potential risks of six organotin compounds (OTs) in Indo-Pacific humpback dolphins (n = 101) from the northern South China Sea during 2003-2021. We detected the highest level of hepatic triphenyltin in these humpback dolphins compared with the results reported in cetaceans globally, and the liver accumulated the highest OT concentrations than other analyzed tissues. Despite the downward trend of butyltins in humpback dolphins after the global ban on the use of OTs as antifouling paints, levels of phenyltins have continued to increase over the past 20 years, suggesting that the other applications of phenyltins in South China remain prevalent. In vitro and in vivo analyses revealed that tissue-relevant doses of OTs could induce agonistic effects on the dolphin peroxisome proliferator-activated receptor γ as a master regulator of lipid homeostasis and altered the dolphin fatty acid profiles. Our results highlight the lipid-disrupting effects of current OT exposure in humpback dolphins and emphasize the need for further efforts to eliminate OT contamination in South China.

Fit-for-Purpose Assessment of QuEChERS LC-MS/MS Methods for Environmental Monitoring of Organotin Compounds in the Bottom Sediments of the Odra River Estuary.

Organotin compounds (OTCs) are among the most hazardous substances found in the marine environment and can be determined by either the ISO 23161 method based on extraction with non-polar organic solvents and gas chromatography analysis or by the recently developed QuEChERS method coupled to liquid chromatography-mass spectrometry (LC-MS/MS). To date, the QuEChERS LC/MS and ISO 23161 methods have not been compared in terms of their fit-for-purpose and reliability in the determination of OTCs in bottom sediments. In the case of ISO 23161, due to a large number of interferences gas chromatography-mass spectrometry was not suitable for the determination of OTCs contrary to more selective determination by gas chromatography with an atomic emission detector. Moreover, it has been found that the derivatization of OTCs to volatile compounds, which required prior gas chromatography determination, was strongly affected by the sediments' matrices. As a result, a large amount of reagent was needed for the complete derivatization of the compounds. Contrary to ISO 23161, the QuEChERS LC-MS/MS method did not require the derivatization of OTC and is less prone to interferences. Highly volatile and toxic solvents were not used in the QuEChERS LC-MS/MS method. This makes the method more environmentally friendly according to the principles of green analytical chemistry. QuEChERS LC-MS/MS is suitable for fast and reliable environmental monitoring of OTCs in bottom sediments from the Odra River estuary. However, determination of di- and monobutyltin by the QuEChERS LC-MS/MS method was not possible due to the constraints of the chromatographic system. Hence, further development of this method is needed for monitoring di- and monobutyltin in bottom sediments.

Low impact leaching agents as remediation media for organotin and metal contaminated sediments.

All over the world, elevated levels of metals and the toxic compound tributyltin (TBT) and its degradation products are found in sediments, especially close to areas associated with shipping and anthropogenic activities. Ports require regular removal of sediments. As a result, large volumes of often contaminated sediments must be managed. The aim of this study was to investigate enhanced leaching as a treatment method for organotin (TBT) and metal (Cu and Zn) contaminated marine sediments. Thus, enabling the possibility to reuse these cleaner masses e.g. in construction. In addition to using acid and alkaline leaching agents that extract the OTs and metals but reduce the management options post treatment, innovative alternatives such as EDDS, hydroxypropyl cellulose, humic acid, iron colloids, ultra-pure Milli-Q water, saponified tall oil ("soap"), and NaCl were tested. Organotin removal ranged from 36 to 75%, where the most efficient leaching agent was Milli-Q water, which was also the leaching agent achieving the highest removal rate for TBT (46%), followed by soap (34%). The TBT reduction accomplished by Milli-Q water and soap leaching enabled a change in Swedish sediment classification from the highest class to the second highest class. The highest reduction of Zn was in HPC leached samples (39% removal) and Cu in EDDS leached samples (33% removal). Although high metal and OT leaching were achieved, none of the investigated leaching agents are sufficiently effective for the removal of both metals and OTs. The results of this study indicate that leaching with ultra-clean water, such as Milli-Q water, may be sufficient to treat TBT contaminated sediments and potentially allow mass reuse.

Ultrasensitive determination of organotin compounds in plastic food packaging and edible oils by sheathless capillary electrophoresis-electrospray ionization-mass spectrometry.

The determination of trace-amount organotins in plastic food packaging materials is of great significance in food safety. However, due to the complexity of organotins and sample treatment processes, it is still a challenging task. Here, we report a method for the sensitive and simultaneous determination of organotins in plastic food packaging materials and edible oils, by utilizing sheathless capillary electrophoresis-electrospray ionization-mass spectrometry. The method of sample pretreatment with ultrasonic extraction and solid phase extraction is used to eliminate interference. The results showed low limits of detection (LODs) of 2 pg mL-1-50 pg mL-1 and excellent inter/intra-day repeatability. Good average recoveries in the range of 80.27% to 108.52% were obtained at three spiked concentrations, with a relative standard deviation less than 8.71%. The successful simultaneous determination of the target analytes will pave the way for further assessment of contamination and migration behaviour of organotins from packaging materials to food, which is of great significance for evaluating and controlling food safety.

Development and Application of a Novel QuEChERS Method for Monitoring of Tributyltin and Triphenyltin in Bottom Sediments of the Odra River Estuary, North Westernmost Part of Poland.

A Quick, Easy, Cheap, Effective, Rugged, and Safe (QuEChERS) extraction method combined with Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) for determination of organotin compounds (OTC) has been newly developed. The novel analytical method was validated and the quality of the results was tested by the use of certificate reference material of freshwater sediment BCR 646. The method was applied in determination of OTC concentration in real samples of bottom sediments collected from the Polish part of Odra River Estuary. The samples came from locations with different anthropogenic impact. Additionally, the extraction recovery of OTC and matrix effect on MS signal response was investigated based on those real environmental samples. It was found that organic compounds and anthropogenic contaminations present in bottom sediments may affect extraction efficiency of the organotin compounds (OTC) and change the matrix effect on MS signal response. The highest concentrations of tributyltin were found in bottom sediments collected from locations in vicinity of the Szczecin harbor and shipyards. The presence of triphenyltin above limit of detection (5 ng TPhT/g of sediment) was observed only in two samples and its concentration was several times lower compared to concentration of tributyltin (from 58 ng/g to 5263 ng/g). In spite of the fact that, the application of TBT-based paints on hull of vessel entering EU ports has been banned by European Commission regulation No. 782/2003 since 2008, the OTC compounds are still present in bottom sediment and pose significant threat to the environment. This threat should be taken into account during dredging of waterways and other hydrotechnical works.

[Determination of Tributyltin and Triphenyltin in Fish and Shellfish Using Accelerated Solvent Extraction and Liquid Chromatography with Tandem Mass Spectrometry].

A determination method for tributyltin (TBT) and triphenyltin (TPT) in fish and shellfish using an accelerated solvent extractor (ASE) and LC-MS/MS was developed. The chromatographic separation was conducted on a Poroshell 120 EC-C18 column using an isocratic mobile phase of 0.1% formic acid in 70% methanol. Sample preparation was performed using ASE at 125℃ with n-hexane and a cleanup using a Florisil cartridge. Internal calibration curves using deuterium-labeled TBT and TPT were employed for quantification. For both TBT and TPT, the calibration curves were linear in the range of 0.2-250 ng/mL, and the method quantification limits were 0.8 ng/g for both TBT and TPT. A National Institute for Environmental Studies certified reference material, No. 15 (adductor muscle of scallop), was analyzed to assess the performance of the developed method. The trueness, relative standard deviations of repeatability, and within laboratory reproducibility of this method, evaluated using a recovery test with four spiked fish species and one shellfish, ranged from 89.3 to 105.3%, 1.0 to 4.5%, and 1.3 to 7.6%, respectively.

Simultaneous determination of three organotin pesticides in fruits and vegetables by high-performance liquid chromatography/tandem mass spectrometry.

RATIONALE: The presence of organotins in the environment affects food safety, making it important to monitor the levels of organotin pesticides (OTPs) in fruit and vegetable samples. METHODS: In the present study, a simple and low cost method for simultaneous determination of three OTPs (azocyclotin, fenbutatin oxide and triphenyltin hydroxide) in vegetable and fruit samples was developed and validated, based on solid-phase extraction and liquid chromatography/tandem mass spectrometry. RESULTS: Extraction with acetonitrile containing 0.2% formic acid positively affected the recoveries of the three OTPs. Moreover, the simultaneous purification of the three OTPs was the most efficient using mixed-mode cation-exchange cartridges and 5.0% ammonium hydroxide in methanol as eluent, and, in this case, mild matrix effects (-9.3% to 21.6%) were obtained for the three OTPs monitored. The developed method reached limits of quantification of 1 µg kg-1 , and linearity was satisfactory, with correlation coefficients >0.995. A fortification study showed that when spiked at 1.0-50.0 µg kg-1 the mean trueness values were from 72.3 to 110.0% in all matrices (three vegetables and three fruits). The intra-day precision was <14.1%, and the inter-day precision (n = 11) was <18.2%. CONCLUSIONS: The proposed method was successfully applied to the simultaneous analysis of three OTPs in vegetables and fruits.

Preliminary studies about the role of physicochemical parameters on the organotin compound dynamic in a South American estuary (Bahía Blanca, Argentina).

This work provides a preliminary study of the destination, mobility, and availability of tributyltin (TBT), dibutyltin (DBT), and monobutyltin (MBT) in contaminated sediments and water column within Puerto Rosales Port, located in the middle zone of the Bahía Blanca Estuary (Argentina). Therefore, this study presents the first comprehensive results of the role of several physicochemical parameters (temperature, pH, Eh, salinity, turbidity, organic matter, chlorophyll, and macronutrients) in behavior of organotin compounds (OTCs) in a marine-coastal ecosystem. The samples were collected seasonally in May, August, and November during 2014. Levels of OTCs were determined in sediments and water column samples by means of gas chromatography-mass spectrometry analysis. Degradation index analyses suggested not recent inputs of TBT at the area of study. However, results submitted a continuous input of TBT into the column water; further, its distribution and degradation pattern were shown to be influenced by salinity, turbidity, particulate organic matter, chlorophyll, and nitrates. These last two parameters, chlorophyll and nitrates, also were very important for sediment samples. Chlorophyll together with high temperatures recorded in the surface sediments triggers biodegradation process of TBT and DBT resulting in high MBT levels while nitrates seemed to promote debutylation process. Furthermore, pH appeared to influence drastically the adsorption/desorption activity of TBT and DBT in sediment. Finally, the Eh obtained suggested a degradation of TBT thanks to the presence of Fe (III) in this compartment. In addition, in fact, the results outlined a possible MBT additional input that contributes to the pollution observed in the study area. Graphical abstract Organotin compounds behavior according to several physicochemical parameters.

Fast and Feasible Ultrasound-Assisted Pretreatment for the Determination of Organotin Compounds in Environmental Samples.

The goal of this study was to improve an already established reference method, such as the one devoted to organotin compounds determination (Reference Method for Marine Pollution Studies, No. 59, UNEP). The proposed upgrade consists of replacing the mechanical shaking by ultrasound energy and applying low temperature throughout the whole procedure. The optimization of the new operational conditions was performed by using a factorial design. Quality control was performed using a certified sediment reference material (PACS-2) for sediments (82.5-97% of recovery) and recoveries on spiked samples for suspended particulate matter (SPM) and mussels (94-100%). The proposed procedure was applied to surface sediment samples, SPM, and native bivalve mollusks (Brachidontes rodriguezii) collected in Bahia Blanca estuary, a very industrialized zone. The relative standard deviation (RSD %) of the environmental samples were less than 7.9%. It is important to note that the proposed procedure reduced the sample pretreatment time about seven times.

Organotin compounds in Brachidontes rodriguezii mussels from the Bahía Blanca Estuary, Argentina.

Levels of tributyltin and its breakdown compounds, including the first record of monobutyltin (MBT) in history for Latin America, were determined in native mussels (Brachidontes rodriguezii) by means of CG-MS, after extraction/derivatization assisted by ultrasound. The samples were collected in 2013 in Bahía Blanca Estuary (Argentina) at 6 sites, which reflect different levels of maritime activities. Total butyltins (TBts = TBT+ DBT+ MBT) levels ranged from 19.64 to 180.57ng Sn g-1 dry weight. According to the Oslo-Paris commission, the results indicated that 73.9% of mussels could be under biological effects risks associated with TBT pollution. In accordance with the calculated bioaccumulation factors, approximately 56% of samples appeared to have accumulated TBT through the sediments. All sampling sites were shown to be impacted by organotin compounds (OTCs) showing variable levels through seasons, which could be related with the variation of the water temperature. Degradation index analyses suggested aged inputs of TBT possibly under a general degradation process at the area of study. In addition, the occurrence of DBT and MBT could not be uniquely attributed to the degradation pathway of the TBT; in fact, results outlined the possible contribution of some punctual and diffuse sources at the area such as proximity to plastic industries, industrial effluents, sewage outlets and domestic wastewaters.

Triphenyltin degradation and proteomic response by an engineered Escherichia coli expressing cytochrome P450 enzyme.

Although triphenyltin (TPT) degradation pathway has been determined, information about the enzyme and protein networks involved was severely limited. To this end, a cytochrome P450 hydroxylase (CYP450) gene from Bacillus thuringiensis was cloned and expressed in Escherichia coli BL21 (DE3), namely E. coli pET32a-CYP450, whose dosage at 1gL-1 could degrade 54.6% TPT at 1mgL-1 within 6 d through attacking the carbon-tin bonds of TPT by CYP450. Sequence analysis verified that the CYP450 gene had a 1214bp open reading frame, encoding a protein with 404 amino acids. Proteomic analysis determined that 60 proteins were significantly differentially regulated expression in E. coli pET32a-CYP450 after TPT degradation. The up-regulated proteins enriched in a network related to transport, cell division, biosynthesis of amino acids and secondary metabolites, and microbial metabolism in diverse environments. The current findings demonstrated for the first time that P450 received electrons transferring from NADH could effectively cleave carbon-metal bonds.

A temporal and spatial monitoring of organotin pollution in a harborside region of Brazil by imposex and ecological quality ratio using Leucozonia nassa.

This study represents the first use of Leucozonia nassa (Mollusca-Gastropoda) in a program of long-term monitoring. Specimens of L. nassa were collected between 2006 and 2014 and analyzed for the occurrence of imposex in two areas in Southeastern Brazil, one control area and another characterized by the presence of many ports and marinas. Imposex was evaluated in nine sites based on the percentage of females exhibiting imposex (I%), relative penis length index and vas deferens sequence index, classical indices to evaluate tributyltin (TBT) impacts. Moreover, this study uses for the first time in a tropical environment the ecological quality ratio (EQR), an approach to verify the imposex levels as an indicator of the impact by organotin compounds according to the EU Water Framework Directive. The results confirm the occurrence of high levels of imposex in L. nassa with all the indices evaluated. Furthermore, the results also indicate that, despite the international ban, antifouling paints based on TBT are still possibly used in Brazil. In addition, this study shows that L. nassa has the characteristics that make it a suitable bioindicator to monitor TBT pollution, and that the use of the EQR may create important data regarding this problem.

Contaminants in Liquid Organic Fertilizers Used for Agriculture in Japan.

To provide an overview of anthropogenic contaminants in liquid organic fertilizers (LOFs), products from four biogas plants in Kyushu, Japan, were analyzed for a wide range of contaminants, including copper, cadmium, tributyltin (TBT), dibutyltin (DBT), perfluorooctane sulfonate, 952 semi-volatile organic compounds, and 89 antibiotics. The highest concentrations of copper (31.1 mg/L) and cadmium (0.08 mg/L) were found in LOFs from the Hita biogas plant. Only ofloxacin and sulfapyridine were detected in total 89 antibiotics screened. TBT, DBT, and perfluorooctane sulfonate were present at low concentrations in the LOFs from all four locations. Among the 952 semi-volatile organic compounds, 78 compounds were detected in at least one sample and were present at concentrations between 1.2 and 139.6 mg/L. On the basis of comparisons with previous studies and quality standards for the use of organic fertilizers, the concentrations of contaminants in the studied LOFs indicate that they might be safe for agricultural purposes.

Fast and effective low-temperature freezing extraction technique to determine organotin compounds in edible vegetable oil.

Most organotin compounds that have been widely used in food packaging materials and production process show serious toxicity effects to human health. In this study, a simple and low-cost method based on high-performance liquid chromatography with inductively coupled plasma mass spectrometry for the simultaneous determination of four organotins in edible vegetable oil samples was developed. Four organotins including dibutyltin dichloride, tributyltin chloride, diphenyltin dichloride, and triphenyltin chloride were simultaneously extracted with methanol using the low-temperature precipitation process. After being concentrated, the extracts were purified by matrix solid-phase dispersion using graphitized carbon black. The experimental parameters such as extraction solvent and clean-up material were optimized. To evaluate the accuracy of the new method, the recoveries were investigated. In addition, a liquid chromatography with tandem mass spectrometry method was also proposed for comparison. The procedures of extracting and purifying samples for the analysis were simple and easy to perform batch operations, also showed good efficiency with lower relative standard deviation. The limits of detection of the four organotins were 0.28-0.59 µg/L, and the limits of quantification of the four organotins were 0.93-1.8 µg/L, respectively. The proposed method was successfully applied to the simultaneous analysis of the four organotins in edible vegetable oil. Some analytes were detected at the level of 2.5-28.8 µg/kg.

Pollutant Concentrations and Toxic Effects on the Red Alga Ceramium tenuicorne of Sediments from Natural Harbors and Small Boat Harbors on the West Coast of Sweden.

This investigation set out to analyze the toxicity of surface sediments in a number of natural harbors and small boat harbors on the west coast of Sweden. This was done with the growth inhibition method with Ceramium tenuicorne. Also, concentrations of copper (Cu), lead (Pb), zinc (Zn), irgarol, organotin compounds, and polycyclic aromatic hydrocarbons (PAHs) in the sediments were analyzed. The small boat harbors were heavily polluted by Cu, Zn, butyltins, and PAHs, and to a lesser extent by Pb. The Cu, Pb, Zn, and butyltins probably originated from their past and/or present use in antifouling paints, whereas the PAHs probably had multiple sources, including boat motor exhausts. The measured toxicity of the sediment was generally related to their Cu, Zn, and butyltin content, although other toxic substances than those analyzed here probably contributed to the toxicity in some of the harbors. The natural harbor sediments contained less pollutants and were less toxic than the small boat harbor sediments. Nevertheless, our data indicate that the boating pressure today may be high enough to produce toxic effects even in natural harbors in pristine areas. The strongest relationship between toxicity and the major pollutants was obtained when the sediment toxicity was expressed as gram wet weight per liter compared with gram dry weight per liter and gram total organic carbon per liter. Hence, for pollutants that can be elutriated with natural sea water, sediment toxicity expressed as gram wet weight per liter appears preferable.

Butyltin and PAH Contamination of Mar del Plata Port (Argentina) Sediments and Their Influence on Adjacent Coastal Regions.

The distribution of butyltins (BTs) and polycyclic aromatic hydrocarbons (PAHs) were studied in surface sediments to assess how relevant is Mar del Plata port (Argentina) as a source of contamination to the surrounding environments. Within the port, TBT concentrations ranged from 24.2 to 150 ng Sn g(-1) and PAHs (Σ16) from 180 to 17,094 ng g(-1). At the surrounding beaches, PAHs were detected at low concentrations and TBT concentrations reached 10.9 ng Sn g(-1). Although those low levels indicate that the Port might not be an important source of contamination to the surrounding beaches, the very low TOC content and the coarse grain size of the beaches sediments could explain the sedimentary levels. The results show a reduction in TBT levels in Mar del Plata port after national and international use restrictions of TBT-based antifouling paints.

Biodegradation of Tributyltin (TBT) by Extremophile Bacteria from Atacama Desert and Speciation of Tin By-products.

Biodegradation of tributyltin (TBT) by four tin resistant Gram negative bacteria isolated from extremely contaminated river sediments in the Atacama Desert in Chile was studied. Moraxella osloensis showed the greatest resistance and degradation capability of TBT, producing less toxic by-products, such as dibutyltin (DBT) and inorganic tin. In 7 days, approximately 80 % of TBT degradation was achieved, generating close to 20 % of DBT as degradation product. The degradation rate constant (k) was 0.022 [day(-1)] and TBT half-life (t1/2) in culture was 4.3 days. Debutylation is stated a probable mechanism of TBT degradation.

Comparison between GC-MS and GC-ICPMS using isotope dilution for the simultaneous monitoring of inorganic and methyl mercury, butyl and phenyl tin compounds in biological tissues.

The aim of this work is to compare simultaneous isotope dilution analysis of organotin and organomercury compounds by gas chromatography-mass spectrometry (GC-MS) and gas chromatography-inductively coupled plasma mass spectrometry (GC-ICP/MS) on certified bivalve samples. These samples were extracted by microwave with tetramethylammonium hydroxide (TMAH). Derivatization with both NaBEt4 and NaBPr4 was evaluated, and analytical performances were compared. Two CRM materials, BCR-710 and CRM-477, were analyzed by both techniques to verify accuracy. A mixed spike containing (201)Hg-enriched methylmercury (MeHg), (199)Hg-enriched inorganic mercury (iHg), (119)Sn-enriched monobutyltin (MBT), dibutyltin (DBT), and tributyltin (TBT) as well as homemade (116)Sn-enriched monophenyltin (MPT), diphenyltin (DPT), and triphenyltin (TPT) was used for the isotope dilution analysis of samples. The two techniques studied were compared in terms of classic analytical parameters: linearity, precision or repeatability (i.e., percent relative standard deviation, RSD%), limit of detection (LOD), and limit of quantification (LOQ), showing excellent linearity, precision below 12% for all analytes, and LOQs of 0.06-1.45 pg for GC-MS and 0.02-0.27 pg for GC-ICP/MS.

Association of placenta organotin concentrations with growth and ponderal index in 110 newborn boys from Finland during the first 18 months of life: a cohort study.

BACKGROUND: Humans are exposed to tributyltin (TBT), previously used as an antifouling paint in ships, mainly through fish consumption. As TBT is a known obesogen, we studied the association of placenta TBT and other organotin compounds (OTCs) with ponderal index (PI) and growth during the first 18 months of life in boys. METHODS: In a prospective Finnish study, 110 placenta samples were collected from mothers of boys born in 1997-1999 with (n = 55) and without (n = 55) cryptorchidism. To account for the original study design, linear regression, weighted for sampling fractions of boys with (121/55) and without (5677/55) cryptorchidism from the total cohort, was used to study the association between placenta OTCs and children's weight, length, growth rates and PI up to 18 months of age. RESULTS: Placenta TBT concentrations were above the limit of quantification (LOQ) in 99% of the samples. However, monobutyltin (MBT), dibutyltin (DBT) and triphenyltin (TPhT) concentrations were below LOQ in 90%, 35% and 57% of samples, respectively. Placenta TBT was positively associated (p = 0.024) with weight gain during the first three months of life, but no other significant associations were observed for weight or length gain. Also, no significant associations between placenta OTC concentrations and child length, weight or PI at any time point were found. CONCLUSIONS: We observed a trend towards higher weight gain from birth to 3 months of age with increasing placenta TBT concentration. These results should be interpreted with caution because obesogenic effects in animal experiments were seen after in-utero TBT exposures to doses that were orders of magnitude higher. Also the number of study subjects included in this study was limited.

Speciation of methyltins by dispersive liquid-liquid microextraction and gas chromatography with mass spectrometry.

Dispersive liquid-liquid microextraction in combination with an in situ derivatization is suggested for methyltin compound sampling and preconcentration from water solutions. The derivatization was carried out with sodium tetraethylborate at pH 3. The effects of extraction and disperser solvents type, volume, and extraction time on the extraction efficiency were investigated. 1,2-Dichlorobenzene was used as an extraction solvent and ethanol was used as a disperser solvent. The calibration graphs for all the analytes were linear up to 2 µg (Sn) L(-1), correlation coefficients were 0.998-0.999, LODs were 0.13, 0.05, and 0.06 ng (Sn) L(-1) for trimethyltin, DMT, and monomethyltin, respectively. Repeatabilities of the results were acceptable with RSDs up to 12.1%. A possibility to apply the proposed method for methyltin compound determination in water samples was demonstrated.