Systematic suspect screening and identification of sulfonamide antibiotic transformation products in the aquatic environment.

The formation of environmental sulfonamide transformation products (TPs) has only been investigated for very selected compounds, and their transformation pathways are incompletely elucidated. Given their homologous series, it is likely that similar transformation reactions occur for all sulfonamides. It has recently been demonstrated for sulfamethoxazole (SMX) that its derivative TPs retain antibiotic activity. Unfortunately, TP reference standards are often not available and a number of TPs are still unknown. Therefore, in the present study, a generic scheme was developed to predict 29 potential TPs of sulfonamide antibiotics via identification of the major transformation and breakdown reactions. Mass shifts were calculated for each transformation allowing for a suspect screening using LC/QTOF. Based on the structural elucidation of the parent sulfonamide product ion spectra, five characteristic product ions could be predicted for each of 15 derivative TPs. The predictions were confirmed with reference standards of SMX TPs as well as with data of TPs of further sulfonamides reported in literature. In a final step, samples of activated sludge biotransformation experiments were screened for suspect TPs of two sulfonamides. In total, 13 ecotoxicologically relevant TPs could be tentatively identified by use of the product ion predictions and LC/QTOF.

Intralaboratory validation of a fast and sensitive UHPLC/MS/MS method with fast polarity switching for the analysis of lipophilic shellfish toxins.

This paper shows the results of an intralaboratory validation of a fast method for the determination of lipophilic shellfish toxins working under acidic conditions using ultra-high performance LC (UHPLC) with MS/MS. Fourteen lipophilic marine toxins and domoic acid were acquired with fast polarity switching. Whereas azaspiracids (AZAs), pecenotoxins, 13-desmethyl spirolide C (SPX1), and gymnodimine were analyzed in the positive mode, yessotoxins (YTXs) were measured in negative mode. The okadaic acid (OA) group compounds were analyzed in both positive and negative ionization modes, and the accuracy of the results for both were compared. When using dynamic multiple reaction monitoring (MRM) in fast polarity switching, LODs were lower and reproducibility and linearity were better compared to static MRM. The UHPLC separation allowed for higher sample throughput in routine use. Compared to the previously used HPLC/MSIMS method, LODs were improved up to a factor of 10 in mussel extract. Matrix effects were evaluated by comparing standards prepared in solvent with matrix-matched calibrations in blank mussel extract. For accurate quantification matrix-matched calibrations were used when analyzing reference mussel materials, providing recoveries for OA, Dynophysis toxins (DTX)1, DTX2, YTX, AZA1, and SPX1 between 80 and 120% with RSDs below 8% over a 3-day validation procedure.

Development and validation of a (semi-)quantitative UHPLC-MS/MS method for the determination of 191 mycotoxins and other fungal metabolites in almonds, hazelnuts, peanuts and pistachios.

A multi-target method for the determination of 191 fungal metabolites in almonds, hazelnuts, peanuts and pistachios was developed. The method includes all mycotoxins regulated in the European Union and mycotoxins regularly found in food. After extraction with an acidified acetonitrile water mixture, the raw extract was diluted and injected directly into the UHPLC-MS/MS system. In two chromatographic runs, analysis was performed in positive and in negative ionisation mode. The method was in-house validated for the most important 65 analytes in these four commodities. Apparent recoveries between 80 and 120% were obtained for about half of the analyte-matrix combinations. Good repeatabilities (standard deviations < 10%) were achieved for the vast majority (83%) of all cases. Only in 6% of all combinations did the standard deviations exceed 15%. Matrix effects, arising during electrospray ionisation, significantly influenced the determination. For instance, signal suppression was observed for several early-eluting analytes and also signal enhancement up to 295% for physcion in peanuts was determined. Concerning extraction recovery, 94% of the analyte-matrix combinations showed values higher than 50%. Lower limits of quantification ranged between 0.04 µg kg(-1) for enniatin B3 in peanuts and 500 µg kg(-1) for HC toxin in hazelnuts. Additionally, the applicability of the developed method was demonstrated through the analysis of 53 naturally contaminated nut samples from Austria and Turkey. Overall, 40 toxins were quantified; the most frequently found mycotoxins were beauvericin (79%), enniatin B (62%) and macrosporin (57%). In the most contaminated hazelnut sample, 26 different fungal metabolites were detected.

Stable isotope dilution assay for the accurate determination of mycotoxins in maize by UHPLC-MS/MS.

A fast, easy-to-handle and cost-effective analytical method for 11 mycotoxins currently regulated in maize and other cereal-based food products in Europe was developed and validated for maize. The method is based on two extraction steps using different acidified acetonitrile­water mixtures. Separation is achieved using ultrahigh-performance liquid chromatography (UHPLC) by a linear water­methanol gradient. After electrospray ionisation, tandem mass spectrometric detection is performed in dynamic multiple reaction monitoring mode. Since accurate mass spectrometric quantification is hampered by matrix effects, uniformly [(13)C]-labelled mycotoxins for each of the 11 compounds were added to the sample extracts prior to UHPLC-MS/MS analysis. Method performance parameters were obtained by spiking blank maize samples with mycotoxins before as well as after extraction on six levels in triplicates. The twofold extraction led to total recoveries of the extraction steps between 97% and 111% for all target analytes, including fumonisins. The [(13)C]-labelled internal standards efficiently compensated all matrix effects in electrospray ionisation, leading to apparent recoveries between 88% and 105% with reasonable additional costs. The relative standard deviations of the whole method were between 4% and 11% for all analytes. The trueness of the method was verified by the measurement of several maize test materials with well-characterized concentrations. In conclusion, the developed method is capable of determining all regulated mycotoxins in maize and presuming similar matrix effects and extraction recovery also in other cereal-based foods.

Electrochemical reduction of the iodinated contrast medium iomeprol: iodine mass balance and identification of transformation products.

Potentiostatic-controlled electrochemical reduction of iomeprol was used to deiodinate iomeprol (IMP), a representative of the iodinated X-ray contrast media. The reduction process was followed by product analysis with liquid chromatography-electrospray ionization-tandem mass spectrometry and ion chromatography-inductively coupled plasma-mass spectrometry. The identification is mainly based on the interpretation of the mass fragmentation. The product analysis showed a rather selective deiodination process with the successive occurrence of IMP-I, IMP-2I, IMP-3I, and a transformation product (TP), respectively. The TP was formed from IMP-3I by a further cleavage of an amide bond and release of a (C=O)CHOH group from the side chain of IMP. The iodine mass balance on the basis of IMP and iodide showed a gap of about 26% at the beginning of the electrolysis process which could be completely closed by taking the intermediates IMP-I and IMP-2I into consideration. This means that the major intermediates and the TPs were considered and that the reduction process is a rather selective one to remove organically bound iodine from X-ray contrast media. An attractive application area would be the electrochemical deiodination of X-ray contrast media in urine of patients or hospital effluents.

Swimming pool water--fractionation and genotoxicological characterization of organic constituents.

Swimming pool water treatment in general includes flocculation, sand filtration, and subsequent disinfection with chlorine. The continuous chlorination and input of organic material by bathers in combination with recirculation of the pool water leads to an accumulation of disinfection by-products (DBPs) in the water. Several DBPs have been identified as human carcinogens and are thought to cause allergic asthma. Therefore, the elimination of DBPs is one major aim of pool water treatment. Using membrane filtration as an alternative treatment technology, DBPs can be removed more efficiently than with conventional treatment. In this study membrane filtration and genotoxicity testing were applied for the characterization of pool water constituents and for the identification of the necessary molecular weight cut off of the membrane for an efficient elimination. Two-step membrane filtration revealed that most of the DBPs (as adsorbable organically bound halogen, AOX) were present in the molecular weight fraction below 1000 g/mol. The fraction below 200 g/mol contained more than 30% of the AOX. The distribution of the dissolved organic carbon (DOC) across the fractions was similar to that of the AOX. The genotoxicity was found to be strongest in the low-molecular weight fraction. Thus, considerable DBP removal by membrane treatment requires membranes with low-molecular weight cut offs down to 200 g/mol. The comprehensive elimination of the genotoxic compounds requires further treatment steps.

Drowning in disinfection byproducts? Assessing swimming pool water.

Disinfection is mandatory for swimming pools: public pools are usually disinfected by gaseous chlorine or sodium hypochlorite and cartridge filters; home pools typically use stabilized chlorine. These methods produce a variety of disinfection byproducts (DBPs), such as trihalomethanes (THMs), which are regulated carcinogenic DBPs in drinking water that have been detected in the blood and breath of swimmers and of nonswimmers at indoor pools. Also produced are halogenated acetic acids (HAAs) and haloketones, which irritate the eyes, skin, and mucous membranes; trichloramine, which is linked with swimming-pool-associated asthma; and halogenated derivatives of UV sun screens, some of which show endocrine effects. Precursors of DBPs include human body substances, chemicals used in cosmetics and sun screens, and natural organic matter. Analytical research has focused also on the identification of an additional portion of unknown DBPs using gas chromatography (GC)/mass spectrometry (MS) and liquid chromatography (LC)/MS/MS with derivatization. Children swimmers have an increased risk of developing asthma and infections of the respiratory tract and ear. A 1.6-2.0-fold increased risk for bladder cancer has been associated with swimming or showering/bathing with chlorinated water. Bladder cancer risk from THM exposure (all routes combined) was greatest among those with the GSTT1-1 gene. This suggests a mechanism involving distribution of THMs to the bladder by dermal/inhalation exposure and activation there by GSTT1-1 to mutagens. DBPs may be reduced by engineering and behavioral means, such as applying new oxidation and filtration methods, reducing bromide and iodide in the source water, increasing air circulation in indoor pools, and assuring the cleanliness of swimmers. The positive health effects gained by swimming can be increased by reducing the potential adverse health risks.