First inter-laboratory comparison exercise for the determination of anticancer drugs in aqueous samples

The results of an inter-laboratory comparison exercise to determine cytostatic anticancer drug residues in surface water, hospital wastewater and wastewater treatment plant effluent are reported. To obtain a critical number of participants, an invitation was sent out to potential laboratories identified to have the necessary knowledge and instrumentation. Nine laboratories worldwide confirmed their participation in the exercise. The compounds selected (based on the extent of use and laboratories capabilities) included cyclophosphamide, ifosfamide, 5-fluorouracil, gemcitabine, etoposide, methotrexate and cisplatinum. Samples of spiked waste (hospital and wastewater treatment plant effluent) and surface water, and additional non-spiked hospital wastewater, were prepared by the organising laboratory (Jo?ef Stefan Institute) and sent out to each participant partner for analysis. All analytical methods included solid phase extraction (SPE) and the use of surrogate/internal standards for quantification. Chemical analysis was performed using either liquid or gas chromatography mass (MS) or tandem mass (MS/MS) spectrometry. Cisplatinum was determined using inductively coupled plasma mass spectrometry (ICP-MS). A required minimum contribution of five laboratories meant that only cyclophosphamide, ifosfamide, methotrexate and etoposide could be included in the statistical evaluation. z-score and Q test revealed 3 and 4 outliers using classical and robust approach, respectively. The smallest absolute differences between the spiked values and the measured values were observed in the surface water matrix. The highest within-laboratory repeatability was observed for methotrexate in all three matrices (CV?≤?12 %). Overall, inter-laboratory reproducibility was poor for all compounds and matrices (CV 27–143 %) with the only exception being methotrexate measured in the spiked hospital wastewater (CV?=?8 %). Random and total errors were identified by means of Youden plots.     

Biodegradability of the anticancer drug etoposide and identification of the transformation products

Etoposide susceptibility to microbiological breakdown was studied in a batch biotransformation system, in the presence or absence of artificial wastewater containing nutrients, salts and activated sludge at two concentration levels. The primary focus of the present study was to study etoposide transformation products by ultra-high performance liquid chromatography coupled to high-resolution hybrid quadrupole-Orbitrap tandem mass spectrometry (MS/MS). Data-dependent experiments combining full-scan MS data with product ion spectra were acquired to identify the molecular ions of etoposide transformation products, to propose the molecular formulae and to elucidate their chemical structures. Due to the complexity of the matrix, visual inspection of the chromatograms showed no clear differences between the controls and the treated samples. Therefore, the software package MZmine was used to facilitate the identification of the transformation products and speed up the data analysis. In total, we propose five transformation products; among them, four are described as etoposide transformation products for the first time. Even though the chemical structures of these new compounds cannot be confirmed due to the lack of standards, their molecular formulae can be used to target them in monitoring studies.     

Water quality index for agricultural systems in Northwest Uruguay

Agricultural systems have experienced rapid expansion and intensification in the last several decades. In Uruguay, since the beginning of 2000, the most common cropping systems have included soybeans. Currently, this crop is expanding towards lowlands traditionally occupied by rice in rotation with pastures. However, the environmental effects of agricultural intensification and diversification are not well known. Thus, some indices have been proposed to quantify the changes in agricultural production systems and assess water quality. The main goal of this study was to develop a water quality index (WQI) to assess the impacts of the diversification of rice production systems in northwest Uruguay. The study was carried out in an agricultural basin where other summer crops have been incorporated in the rice-pasture sequence. Agriculture intensification and crop diversification indices were calculated using information provided by farmers. Water samples were collected downstream of the production area before crop sowing and after crop harvest (2008–2009 to 2010–2011 and 2016–2017 to 2017–2018). Biochemical oxygen demand, nitrates, total phosphorus, fecal coliforms, and total suspended solids were the variables that mainly explained the effects of the agricultural activities on water quality. The proposed water quality index included these unweighted variables, which allowed for the pre-sowing and post-harvest to be differentiated, as well as the degree of diversification. Therefore, the proposed WQI constitutes a tool that can be used to evaluate the water quality in an agricultural basin. Likewise, it can be used to select agricultural sequences that generate the least possible impacts on the associated water resources.     

Pesticide dissipation curves in peach, pear and tomato crops in Uruguay

Dissipation curves of azoxystrobin and of the neonicotinoids acetamiprid and thiacloprid in peach; azinphos-methyl and carbaryl in pear and azoxystrobin, chlorfenapyr and chlorpyrifos in high-tunnel tomato crops were studied in the Southern region of Uruguay. An analytical methodology based on solid phase extraction (SPE) and detection by High Performance Liquid Chromatography with Diode Array Detector (HPLC/DAD) was used for acetamiprid and thiacloprid. Coupled SPE and detection by Gas Chromatography with Mass Selective Detector (GC/MSD) was used for the detection of azinphos-methyl, azoxystrobin, carbaryl, chlorfenapyr and chlorpyrifos residues. Curves were modeled mathematically with Solver program of Microsoft Excel. The best fit for acetamiprid and thiacloprid in peach was achieved with the exponential model (r(2)=0.961 and 0.944, respectively). In the case of peach fruits there is not a Maximum Residue Limit (MRL) for acetamiprid in the Codex Alimentarius, while 0.5 mg/kg is the value rated for thiacloprid. The MRLs accepted by the European Union (EU) are 0.1 mg/kg for acetamiprid and 0.3 mg/kg for thiacloprid. According to the curves determined in these experiments, thiacloprid residues 10 to 12 days after application (daa) were below the MRLs established by both sources. In the case of acetamiprid, 25 daa would be required, according to the exponential mathematical model, to get residues levels below the MRL values established by the EU. For azinphos methyl in pear, the residues detected were mathematically fitted to an exponential model (r(2)=0.999). According to it, residue levels under the MRL established by the EU (0.05 mg/kg) are gotten in our conditions in 20 daa. In plastic tunnel tomato chlorfenapyr residues were not detected from 16 daa, having the dissipation curve an exponential trend. In the same condition, there was not a decay of the azoxystrobin concentration during a 24-day trial, being it around 0.40 ± 0.05 mg/kg.     

Pesticide dissipation curves in peach,pear and tomato crops in Uruguay*

Dissipation curves of azoxystrobin and of the neonicotinoids acetamiprid and thiacloprid in peach; azinphos-methyl and carbaryl in pear and azoxystrobin, chlorfenapyr and chlorpyrifos in high-tunnel tomato crops were studied in the Southern region of Uruguay. An analytical methodology based on solid phase extraction (SPE) and detection by High Performance Liquid Chromatography with Diode Array Detector (HPLC/DAD) was used for acetamiprid and thiacloprid. Coupled SPE and detection by Gas Chromatography with Mass Selective Detector (GC/MSD) was used for the detection of azinphos-methyl, azoxystrobin, carbaryl, chlorfenapyr and chlorpyrifos residues. Curves were modeled mathematically with Solver program of Microsoft Excel®. The best fit for acetamiprid and thiacloprid in peach was achieved with the exponential model (r²=0.961 and 0.944, respectively). In the case of peach fruits there is not a Maximum Residue Limit (MRL) for acetamiprid in the Codex Alimentarius, while 0.5 mg/kg is the value rated for thiacloprid. The MRLs accepted by the European Union (EU) are 0.1 mg/kg for acetamiprid and 0.3 mg/kg for thiacloprid. According to the curves determined in these experiments, thiacloprid residues 10 to 12 days after application (daa) were below the MRLs established by both sources. In the case of acetamiprid, 25 daa would be required, according to the exponential mathematical model, to get residues levels below the MRL values established by the EU. For azinphos methyl in pear, the residues detected were mathematically fitted to an exponential model (r²=0.999). According to it, residue levels under the MRL established by the EU (0.05 mg/kg) are gotten in our conditions in 20 daa. In plastic tunnel tomato chlorfenapyr residues were not detected from 16 daa, having the dissipation curve an exponential trend. In the same condition, there was not a decay of the azoxystrobin concentration during a 24-day trial, being it around 0.40 ± 0.05 mg/kg.     

Disinfection efficacy on cyst viability of<Emphasis Type="Italic"> Artemia franciscana</Emphasis> (Crustacea),<Emphasis Type="Italic"> Hexarthra fennica</Emphasis> (Rotifera) and<Emphasis Type="Italic"> Fabrea salina</Emphasis> (Ciliophora)

Resting stages (cysts) of Artemia franciscana, Hexarthra fennica and Fabrea salina were exposed for two periods (acute and chronic exposures) to different concentrations of five aquaculture disinfectants (formalin, sodium hypochlorite, potassium permanganate, organic iodine, copper sulphate). The effects of chemical treatments were ranked according to their action on cyst hatching: total inhibition (no cyst hatching), heavy inhibition (significant decrease of cyst hatching), alteration of hatching synchrony, and occurrence of anomalous hatching (death of emerging individuals or birth of malformed organisms). Resting-stage viability was not completely impaired by disinfectant exposures at the ordinary doses used in aquaculture, so that usual treatments are possibly ineffective against the protist and metazoan cysts potentially present in breeding systems. On the other hand, the high resistance shown by cysts suggests the possibility of using these chemicals in live-feed disinfection.     

The inactive temporary component: an unexplored fraction of meiobenthos

Numerous morphotypes and great abundances of resting stages falling within the meiobenthic size-range were found in meiobenthos samples collected in a coastal lagoon of the North Adriatic Sea. Biodiversity, abundances and energetic values of both active and inactive meiobenthic organisms were analysed by the traditional techniques of meiobenthology. Inactive stages were represented by more than 100 morphotypes and formed 52% of the meiobenthos in terms of numbers of individuals. Inactive meiobenthos came mostly from plankton: the second and third most abundant groups of all meiobenthic organisms were the eggs of a rotifer and the cysts of a dinophyte, respectively. Resting stages of plankters fit the definition of temporary meiobenthos, making up the “inactive temporary meiobenthos”, but they are usually not considered in meiobenthology. The diversity and abundance of the meiobenthos is, thus, currently underestimated; this leads to neglect of the role of meiobenthos in several processes linking different ecosystems. It is hypothesised, for instance, that the meiobenthic resting stages of planktonic organisms (besides being the “seeds” for the perpetuation of planktonic communities) might also be the prey of meiofaunal organisms with piercing mouth parts. If this proves correct, a regulatory role might be proposed for the meiofauna in respect to plankters with benthic stages. Received: 10 September 1998 / Accepted: 12 April 1999     

Spatial–temporal pattern of bluefin tuna (Thunnus thynnus L. 1758) gonad maturation across the Mediterranean Sea

Atlantic bluefin tuna (BFT; Thunnus thynnus) is a migrating species straddling the North Atlantic Ocean and Mediterranean Sea. It is assumed that this species is divided into a western and an eastern stock, which spawn in the Gulf of Mexico and the Mediterranean Sea, respectively. To learn more about the reproductive behavior of the eastern BFT stock, we tracked gonadal development in adult fish that were sampled between April and July during three consecutive years (2003–2005). Sampling campaigns were carried out using common fishing methods at selected locations within the Mediterranean Sea, namely Levantine Sea, Malta, and Balearic Islands. An additional sampling point, Barbate, was situated northwest of the Straits of Gibraltar along the Atlantic coast. Morphometric parameters such as the total body mass (M _B) and the weights of the gonads (M _G) were recorded, and the respective gonadosomatic index (GSI) values were calculated. The data collected revealed two important trends: (1) GSI values are higher in fish caught in the eastern rather than the western locations across the Mediterranean Sea, and (2) the GSI reaches maximum values between late May and early June in Levantine Sea (eastern Mediterranean Sea), and only 2 and 4 weeks later in the central (Malta) and western (Balearic Islands) locations, respectively. The advanced gonadal development in BFT correlates well with higher sea surface temperatures. Our findings also distinguish the northern Levantine Sea BFT population (mean M _B 78.41 ± 4.13 kg), and the Barbate BFT population with the greatest M _B (all fish sampled > 100 kg). These data reflect a situation in which the eastern Mediterranean basin may function as a habitat for young BFT, until they gain a larger M _B and are able to move to the Atlantic Ocean. However, the existence of genetically discrete BFT populations in the Mediterranean Sea cannot be ruled out.     

Validation of a multiclass multiresidue method and monitoring results for 210 pesticides in fruits and vegetables by gas chromatography-triple quadrupole mass spectrometry

A rapid, sensitive, accurate and reliable multiresidue method for the identification and quantification of 210 relevant pesticides in four representative fruit and vegetable commodities (tomato, potato, spring onion and orange) has been developed and validated by gas chromatography in tandem with triple quadrupole mass spectrometry. The method has been fully validated and applied to 292 samples from different countries. Prior to instrumental analysis, an extraction procedure based on a sample extraction of multiclass analytes, using the ethyl acetate method was employed. Mass spectrometric conditions were individually optimized for each compound in the selected reaction monitoring (SRM) mode to achieve maximum sensitivity. The pesticides were separated in less than 25 min. This was followed by an exhaustive control of the retention times. The Retention Time Locking Method was applied, working at a constant pressure throughout the analysis. System maintenance was reduced by using a purged capillary flow device that provided backflush capabilities by reversing column flow immediately after elution of the last compound of interest. Istotopically labelled internal standards were employed to improve the quality of the analytical results.     

Screening of heavy metal containing waste types for use as raw material in Arctic clay-based bricks

In the vulnerable Arctic environment, the impact of especially hazardous wastes can have severe consequences and the reduction and safe handling of these waste types are therefore an important issue. In this study, two groups of heavy metal containing particulate waste materials, municipal solid waste incineration (MSWI) fly and bottom ashes and mine tailings (i.e., residues from the mineral resource industry) from Greenland were screened in order to determine their suitability as secondary resources in clay-based brick production. Small clay discs, containing 20 or 40% of the different particulate waste materials, were fired and material properties and heavy metal leaching tests were conducted before and after firing. Remediation techniques (washing in distilled water and electrodialytical treatment) applied to the fly ash reduced leaching before firing. The mine tailings and bottom ash brick discs obtained satisfactory densities (1669–2007 kg/m³) and open porosities (27.9–39.9%). In contrast, the fly ash brick discs had low densities (1313–1578 kg/m³) and high open porosities (42.1–51. %). However, leaching tests on crushed brick discs revealed that heavy metals generally became more available after firing for all the investigated materials and that further optimisation is therefore necessary prior to incorporation in bricks.