Evaluation and validation of two different chromatographic methods (HPLC and LC-MS/MS) for the determination and confirmation of ochratoxin A in pig tissues

This study was undertaken to compare two different analytical methods for the determination and confirmation of ochratoxin A (OTA) in blood serum, kidney and liver of pigs. Sample clean-up was based on liquid-liquid phase extraction. The detection of OTA was accomplished with high-performance liquid chromatography (HPLC) combined either with fluorescence detection (FL) or electro spray ionization (ESI+) tandem mass spectrometry (MS–MS). Comparative method evaluation was based on the investigation of 90 samples of blood serum, kidney and liver per animal originating from different regions of Serbia. The analytical results are discussed in view of the respective method validation data and the corresponding experimental protocols. In general, analytical data obtained with (LC–MS–MS) liquid chromatography electro spray tandem mass spectro metry detection offered comparable good results at the sub-ppb concentration level. The results indicate that the liquid chromatography electro spray tandem mass spectrometric (LC-MS/MS) method was more specific and sensitive for the analysis and confirmation of ochratoxin A in pig tissues then high pressure liquid chromatography (HPLC) method after methylation of OTA.     

Isomeric analysis of PCDD/PCDF in waste incinerator fly ash by GC-MS/MS

The development of an analytical method for the analysis of PCDD/PCDF in flyash using a bench analytical system comprising of a gas chromatograph fitted with an ion trap detector operated in the tandem MS mode is described. The optimum settings for the most important MS/MS parameters are given, including those for the less reported mono- to tri-chlorinated dioxin and furan congener groups. Flyash samples from three waste incineration plants representing a decommissioned 1970s plant design, an upgraded and still operating plant originally designed in the 1970s, and a modern 1990s design operating plant have been analysed for PCDD/PCDF. The flyash samples were analysed for PCDD/PCDF using the methods developed and the total PCDD/PCDF, I-TEQ values and isomeric profiles are reported. The flyash from the older decommissioned incinerator had very significantly higher concentrations of PCDD/PCDF compared to the modern incinerator flyash.     

EPA SITE Demonstration of the International Waste Technologies/Geo-Con In Situ Stabilization/Solidification Process

This paper presents an EPA evaluation of the first field demonstration of an in situ stabilization/solidification process for contaminated soil under the EPA Superfund Innovative Technology Evaluation (SITE) program. Demonstration of this process was a joint effort of two vendors: ? International Waste Technologies (IWT) of Wichita, Kansas, who provided the treatment process, specifically the proprietary additive called HWT-20, and

? Geo-Con, Inc., of Pittsburgh, Pennsylvania, who provided both engineering and hardware for the in situ soil treatment.

The field demonstration took place in April, 1988 at a site in Hialeah, Florida, contaminated mainly with polychlorinated biphenyls (PCBs). EPA tested the soil before and after treatment and the EPA evaluation of this process is based on results from this testing. A year later, in April 1989, EPA tested again the treated soil and results of that testing were compared to those of the demonstration. Results of the EPA evaluation of the IWT process, the Geo-Con performance, and treatment costs are discussed separately.     

Inclusion of 13C12-labelled mono-, di-, and tri-chlorinated dibenzo-p-dioxin and dibenzofuran standards in US EPA methods 0023A/8290

13C12-Labelled mono-, di-, and tri-chlorinated dibenzo-p-dioxin (CDD) and chlorinated dibenzofuran (CDF) standards have been tested for their applicability to standard EPA sampling and analytical Methods 0023A/8290. These methods target for analysis only the tetra- through octa-CDD/CDF homologues. Extension of the isotope dilution method to include those lower chlorinated homologues is important toward obtaining reliable species concentration data on the complete, mono- to octa-chlorinated homologue profile. These data will improve our ability to model poly-CDD/CDF concentrations through understanding mechanisms of poly-CDD/CDF formation, chlorination, and dechlorination.     

Application of GC-MS/MS for the analysis of PCDD/Fs in sewage effluents

The application of high resolution gas chromatography in combination with tandem mass spectrometry in an ion trap was tested to substitute the expensive high resolution mass spectrometry in analysing polychlorinated dibenzo-p-dioxins and furans in sewage effluents. In tandem mass spectrometry, a set of parameters has to be optimised in order to attain the required sensitivity. To the best of our knowledge, this is the first time a method development for analysing PCDD/Fs with GC-MS/MS in an ion trap is described in this detail. Nine parameters are varied, including isolation window, collision induced dissociation (CID) amplitude, CID time, acquisition mass range, broadband amplitude, CID bandwidth, modulation range, filament current and ion trap temperature. This technique can be adapted to other analytes. By this optimisation, limits of detection of 0.01-0.05 ng/l are obtained. With its selectivity and sensitivity, tandem mass spectrometry is a powerful tool for the determination of PCDD/Fs in water samples. 55 sewage effluent samples from Germany were analysed.     

Determination of polycyclic aromatic hydrocarbons and substitutes (Nitro-, Oxy-PAHs) in urban soil and airborne particulate by GC-MS and NCI-MS/MS

A method involving high resolution gas chromatography combined with ion trap (HRGC-MS/MS) and negative chemical ionisation (NCI) was developed for the determination of nitrated or oxygenated polycyclic aromatic hydrocarbons (nitro-PAHs, oxy-PAHs) and other electrophilic substitutes in soil samples. Efficient clean-up was achieved by a combination of methods for the determination of PAHs in soil and nitro-PAHs in aerosol using solid phase extraction (SPE) and semipreparative high performance liquid chromatography (HPLC). In samples of surface soil from the city of Basle (Switzerland), nitro-PAHs (mainly 3-nitrofluoranthene and 1-nitropyrene) were found in concentrations between 30 and 800 ng/kg dry weight. Oxy-PAHs and parent PAHs revealed 10²-10⁴-fold higher levels. Nitro-PAHs which are up to 10⁵ times more mutagenic seem to be less persistent in soil than the parent forms, although their entire mutagenic potential has to be estimated as being on the same order of magnitude. In urban air particulate matter, the amounts of nitro-PAHs (2–62 pg/m³) were 10–100 times lower than oxyPAHs and parent PAHs which were both found in a similar range. 3-nitrobenzanthrone, a recently described suspected human carcinogen has not yet been detected. Using multivariate statistical analysis, it was possible to elucidate similarities or special characteristics of substances in a given matrix reflecting their chemical properties or specific emission sources.     

Determination of acidic herbicides in cereals by QuEChERS extraction and LC/MS/MS

A simple and rapid method has been studied for the determination of acidic herbicides (2,4-D, Dichlorprop, Dichlorprop-p, Fluazifop, Fluroxypyr, MCPA, Mecoprop and Mecoprop-p) on cereals (rye). The method involves an alkaline hydrolysis with sodium hydroxide in order to release covalently bound compounds, prior to QuEChERS extraction, followed by neutralization and analysis via liquid chromatography-double mass spectrometry LC/MS/MS. The performance of the method either with or without alkaline hydrolysis was studied in terms of recovery rates and limit of quantification (LOQ). In either case, recoveries were determined at four spiking levels (0.02 mg/kg, 0.05 mg/kg, 0.1 mg/kg and 0.5 mg/kg) with 5 replicates for each level. Mean recoveries ranged from 90 to 120 %, whereas relative standard deviations (RSD %) proved to be less than 20 %. Quantitative analysis was carried out by the internal standard (Nicarbazin) and the LC/MS/MS analysis was performed in electrospray ionisation (ESI) negative mode using a Zorbax XCB Eclipse column. The developed method was applied to the analysis of several cereals commercially available like as rye flour, oat meal, oat flakes and dehusked oat. Residue levels were found below the limit of quantification (LOQ) of the method. The method has been tested in EU Proficiency Tests for cereals with good results.     

Application of a LC–MS/MS method for multi-mycotoxin analysis in infant formula and milk-based products for young children commercialized in Southern Brazil

ABSTRACT

An analytical method based on liquid chromatography–tandem mass spectrometry (LC–MS/MS) was validated and applied for the analysis of aflatoxin M₁ (AFM₁), ochratoxin A (OTA) and deoxynivalenol (DON) in infant formula and milk-based products for young children commercialized in Brazil. A total of 38 samples were evaluated, including 12 infant formula, 14 follow-on formula and 12 samples of milk-based products. AFM₁ was detected in 12 (32%) samples, and seven (18%) samples contained AFM₁ levels above the method limit of quantification in a concentration range between 0.013 and 0.067 ng mL^?1 (0.026 ± 0.019). Two samples of milk-based products exceeded the maximum level (ML) fixed by the European Union for AFM₁ in baby foods, however, all samples were in agreement with the levels established by the Brazilian regulation. OTA and DON were not detected in any of the analyzed samples.     

Analysis of dioxins and furans in environmental samples by GC-ion-trap MS/MS

The work aims to characterise PCDDs/Fs in environmental matrices by high resolution gas chromatography coupled to low resolution quadrupole ion-trap mass spectrometry (GC-QITMS/MS). In particular, the study was mainly focused in different environmental samples such as sewage sludge, soils, and sediments and in combustion residues as fly ashes. Previous experiences have already detected and quantified PCDDs/PCDFs in all matrices studied. Moreover, isomer-specific analysis of 2,3,7,8-PCDDs/Fs compounds corroborated good correlation between GC-QITMS/MS and a well-established technique such as high resolution mass spectrometry (HRMS). Finally, QC measures included the participation in intercalibration exercises. Successful results were achieved using GC-QITMS/MS in the analysis of fly ashes, soils and sediment materials. In general, GC-QITMS/MS constitutes an interesting alternative for routine analysis of dioxins in such as matrices.     

Analytical method using gas chromatography and ion trap tandem mass spectrometry for the determination of S-triazines and their metabolites in the atmosphere

Gas chromatography-ion trap detector (GC-ITD) was used to detect atmospheric triazines and their degradation products in the gaseous and particulate phases. Because triazines and their metabolites are expected to be present at very low concentrations and enclosed in the complex atmospheric matrix, the analytical method used was both highly selective and sensitive. These two properties were obtained by associating chromatography with ion trap tandem mass spectrometry (GC-ITD (MS/MS)). To develop this method, a comparison between the two ionization modes (electron impact and positive-chemical-ionisation) in single-MS was first conducted to choose the parent ions of the five target analytes, i.e. atrazine, desethylatrazine, deisopropylatrazine, terbuthylazine and desethylterbuthylazine. Then, a MS/MS method was optimised by parameters such as the radio frequency storage level and the collision-induced dissociation excitation voltage. Finally, a last step enabled the development of a calibrating program based on the quantification of daughter ions. With this analytical procedure, the detection limits varied between 0.8 and 15 pg m(-3) depending on the compounds under study. This method was tested with success for four atmospheric samples collected in Strasbourg (France) in which four of the five target compounds were detected.     

Rapid PCDD/PCDF screening method for fly ash with ion trap MS/MS

A rapid screening method for the polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/PCDF) in fly ash has been developed. The screening is done in two steps: (a) the extraction by the accelerated solvent extraction and (b) the quantitative measurement by ion trap tandem mass spectrometer (MS/MS). Anhydrous sodium sulfate was added to the fly ash containing activated carbon in order to increase the extraction efficiency. The selectivity of the ion trap MS/MS was confirmed with 13C-labelled internal standard. Then the results of the screening method were compared with those obtained by the conventional analytical method using high-resolution mass spectrometer (HRMS).     

Development of an analytical strategy based on liquid chromatography-high resolution mass spectrometry for measuring perfluorinated compounds in human breast milk: application to the generation of preliminary data regarding perinatal exposure in France

Perfluorinated compounds (PFCs) are man-made chemicals for which endocrine disrupting properties and related possible side effects on human health have been reported, particularly in the case of an exposure during the early stages of development, (notably the perinatal period). Existing analytical methods dedicated to PFCs monitoring in food and/or human fluids are currently based on liquid chromatography coupled to tandem mass spectrometry, and were recently demonstrated to present some limitations in terms of sensitivity and/or specificity. An alternative strategy dedicated to the analysis of fourteen PFCs in human breast milk was proposed, based on an effective sample preparation followed by a liquid chromatography coupled to high resolution mass spectrometry measurement (LC-HRMS). This methodology confirmed the high interest for HRMS after negative ionization for such halogenated substances, and finally permitted to reach detection limits around the pg mL(-1) range with an outstanding signal specificity compared to LC-MS/MS. The proposed method was applied to a first set of 30 breast milk samples from French women. The main PFCs detected in all these samples were PFOS and PFOA with respective median values of 74 (range from 24 to 171) and 57 (range from 18 to 102) pg mL(-1), respectively. These exposure data appeared in the same range as other reported values for European countries.     

Polyfluoroalkyl phosphate esters and perfluoroalkyl carboxylic acids in target food samples and packaging—method development and screening

Polyfluoroalkyl phosphate mono-, di-, and tri-esters (mono-, di-, and triPAPs) are used to water- and grease-proof food packaging materials, and these chemicals are known precursors to perfluoroalkyl carboxylic acids (PFCAs). Existing analytical methods for PAPs lack sample clean-up steps in the sample preparation. In the present study, a method based on ultra performance liquid chromatography coupled to tandem mass spectrometry (UPLC/MS/MS) was developed and optimized for the analysis of mono-, di-, and triPAPs, including a clean-up step for the raw extracts. The method was applied to food samples and their PAP-containing packaging materials. The optimized UPLC/MS/MS method enabled the separation and identification of a total of 4 monoPAPs, 16 diPAPs, and 7 triPAPs in the technical mixture Zonyl®-RP. For sample clean-up, weak anion exchange solid phase extraction columns were tested. PAPs standard solutions spiked onto the columns were separated into a fraction containing neutral compounds (triPAPs) and a fraction with ionic compounds (mono- and diPAPs) with recoveries between 72–110 %. Method limits of quantification for food samples were in the sub to low picogram per gram range. For quantitative analysis of PAPs, compound-specific labeled internal standards showed to be essential as sorption and matrix effects were observed. Mono-, di-, and/or triPAPs were detected in all food packaging materials obtained from the Swedish market. Up to nine diPAPs were detected in the food samples, with the 6:2/6:2 and 6:2/8:2 diPAPs as the dominant compounds. DiPAP concentrations in the food samples ranged from 0.9 to 36 pg/g, which was comparable to individual PFCA concentrations in the same samples. Consumption of food packed in PAP-containing materials could be an indirect source of human exposure to PFCAs.     

Solid-state UV–MALDI–MS assay of transition metal dithiocarbamate fungicides

The determination of transition metal containing dithiocarbamate fungicides represents a challenging aspect of analytical object. They have a low stability, low solubility and stabilize versatile coordination monomers, dimers, disulfides and/or S-oxidized derivatives. Their diverse biological activities and agricultural implementation encompass plant prevention and crop protection against a variety of plants containing fungi and diseases of 400 pathogens and 70 cultures. Nonetheless, those dithiocarbamates (DTCs) are banned for agricultural use in Europe or have expiration at years 2016–2017 because of their highly toxic degradation products and/or metabolites, in particular ethylene thiourea; they found large-scale implementations in materials research and medicine. Despite the broad interdisciplinary of DTC application, due to the above reasons, they have received little attention in the rapidly growing field of analytical chemistry, and in particular, the analytical mass spectrometry. Therefore, the study reported on qualitative, quantitative and structural analysis of ten DTCs (1–10), using the matrix assisted laser desorption/ionization (UV–MALDI)–Orbitrap–mass spectrometry (MS) contributed considerably to the implementation of the method for environmental and foodstuffs monitoring. Its ultrahigh resolving power and capacity for direct solid-state analysis, at limited number of sample pretreatment steps, at concentration levels of analytes of up to femtogram per gram resulted to achievement of a highly precise analytical information for these non-trivial objects. The presented fully validated method and technique is based on the successful ionization of DTCs embedded in three novel organic salts (M₁–M₃). In this regard, the reported MS and the single-crystal X-ray diffraction data as well as the quantum chemical one are able to correlate the molecular structures in condense and in the gas phase. Despite the novelty of the fundamental methodological character of the research reported, the promising metrology contributed to the applied aspect of the UV–MALDI–MS as a robust analytical method for environmental and foodstuffs monitoring, which is tested on two commercially available crop protecting products such as Mancozeb® and Antracol®, respectively.     

Adsorption/thermal desorption-GC/MS for the analysis of pesticides in the atmosphere

An analytical methodology using Automatic Thermal Desorption (ATD) and GC/MS was developed for the determination of the pesticides alachlor, atrazine, captan, formothion, lindane and phosalone in atmospheric samples. This methodology was developed to evaluate the atmospheric contamination by pesticides during treatments and by post-application. Atmospheric samples were collected by using (4 i.d. x 89 mm) stainless steel sampling tubes containing 125 mg of adsorbents at a flow rate of 80 ml min-1. Different types of adsorbents were tested for their ability to efficiently trap pesticides under study: Tenax TA, Carbopack Y, Carbopack B, Carbotrap, Carboxen, Chromosorb 106 and XAD-4. Results of experiment show that Tenax gives the better results for all the pesticides used but the use of the thermal-desorption method, especially for pesticides with low volatility and/or poor thermal stability presents some difficulties. This method was validated by the analysis of the contamination of atmosphere, through volatilization by post-application processes, of atrazine in a parcel of 1 ha.     

Canister-Based Method for Monitoring Toxic VOCs in Ambient Air

The availability of reliable, accurate and precise monitoring methods for toxic volatile organic compounds (VOCs) is a primary need for state and local agencies addressing daily monitoring requirements related to odor complaints, fugitive emissions, and trend monitoring. The canister-based monitoring method for VOCs is a viable and widely used approach that is based on research and evaluation performed over the past several years. This activity has involved the testing of sample stability of VOCs in canisters and the design of time-integrative samplers. The development of procedures for analysis of samples in canisters, including the procedure for VOC preconcentration from whole air, the treatment of water vapor in the sample, and the selection of an appropriate analytical finish has been accomplished. The canister-based method was initially summarized in the EPA Compendium of Methods for the Determination of Toxic Organic Compounds in Ambient Air as Method TO-14. Modifications and refinements are being added to Method TO-14 in order to obtain a Statement of Work for the Superfund Contract Laboratory Program for Air. This paper discusses the developments leading to the current status of the canisterbased method and provides a critique of the method using results obtained in EPA monitoring networks.     

Indoor Emissions from Conversion Varnishes

ABSTRACT

Conversion varnishes are two-component, acid-catalyzed varnishes that are commonly used to finish cabinets. They are valued for their water and stain resistance, as well as their appearance. They have been found, however, to contribute to indoor emissions of organic compounds. For this project, three commercially available conversion varnish systems were selected. A U.S. Environmental Protection Agency (EPA) Method 24 analysis was performed to determine total volatile content, and a sodium sulfite titration method was used to determine uncombined (free) formaldehyde content of the varnish components. The resin component was also analyzed by gas chromatography/mass spectroscopy (GC/MS) (EPA Method 311 with an MS detector) to identify individual organic compounds. Dynamic small chamber tests were then performed to identify and quantify emissions following application to coupons of typical kitchen cabinet wood substrates, during both curing and aging. Because conversion varnishes cure by chemical reaction, the compounds emitted during curing and aging are not necessarily the same as those in the formulation. Results of small chamber tests showed that the amount of formaldehyde emitted from these coatings was 2.3–8.1 times the amount of free formaldehyde applied in the coatings. A long-term test showed a formaldehyde emission rate of 0.17 mg/m²/hr after 115 days.     

An enzyme-linked immunosorbent assay for the determination of dioxins in contaminated sediment and soil samples

A 96-microwell enzyme-linked immunosorbent assay (ELISA) method was evaluated to determine PCDDs/PCDFs in sediment and soil samples from an EPA Superfund site. Samples were prepared and analyzed by both the ELISA and a gas chromatography/high resolution mass spectrometry (GC/HRMS) method. Comparable method precision, accuracy, and detection level (8 ng kg(-1)) were achieved by the ELISA method with respect to GC/HRMS. However, the extraction and cleanup method developed for the ELISA requires refinement for the soil type that yielded a waxy residue after sample processing. Four types of statistical analyses (Pearson correlation coefficient, paired t-test, nonparametric tests, and McNemar's test of association) were performed to determine whether the two methods produced statistically different results. The log-transformed ELISA-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin values and log-transformed GC/HRMS-derived TEQ values were significantly correlated (r=0.79) at the 0.05 level. The median difference in values between ELISA and GC/HRMS was not significant at the 0.05 level. Low false negative and false positive rates (<10%) were observed for the ELISA when compared to the GC/HRMS at 1,000 ng TEQ kg(-1). The findings suggest that immunochemical technology could be a complementary monitoring tool for determining concentrations at the 1,000 ng TEQ kg(-1) action level for contaminated sediment and soil. The ELISA could also be used in an analytical triage approach to screen and rank samples prior to instrumental analysis.     

Modeling Evidence of Incloud Transformation Of Sulfur Dioxide to Sulfate

Methylene chloride (CH₂Cl₂) is presently under study by the U.S. EPA and other agencies to determine if the compound presents an unreasonable risk to human health and to determine if regulatory action is needed to reduce the risk. This paper describes one portion of the study which required the development and validation of a method of sampling and analysis for source emission measurements.

Prior to source sampling, laboratory experiments were conducted to determine the best sample container in which to collect an integrated sample. It was found that CH₂Cl₂ remained stable in Tedlar bags for at least four weeks. The analytical method selected was gas chromatography with flame ionization detection (GC/FID). During the field portion of the study, both manufacturer and user emission sources of CH₂Cl₂ were tested. Multiple sets of simultaneous quadruplicate bag samples were collected to determine the precision of both sampling and analysis in the field. All samples were analyzed at the test site after collection and then returned to Research Triangle Park. Samples were subsequently reanalyzed in the laboratories of Radian and the U.S. EPA using three GC/FID instruments and two types of GC columns. The range of concentrations from the sources was 100 ppm to 27,000 ppm CH₂Cl₂. A statistical analysis of samples collected simultaneously showed no difference in the samples, proving good precision In both sampling and analysis. Some of the sample bags returned from the test sites developed leaks indicating that immediate on-site analysis is best. A comparison of results obtained in the field and the two laboratories showed that interand intra-laboratory precision was within 10 percent.     

Development and applications of a mass spectrometry/mass spectrometry method for the quantification of polychlorinated dibenzo-p-dioxins and dibenzofurans

Mass spectrometry/mass spectrometry (MS/MS) has been demonstrated to be a valuable technique in the analysis of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs, PCDFs). In this study, we demonstrate both the value and limitations of high-resolution mass spectrometry (HRMS) and MS/MS in the analysis of PCDDs and PCDFs. We have found that in certain cases, MS/MS can solve analytical problems that cannot be solved by high-resolution mass spectrometry and we demonstrate the reliability and reproducibility of MS/MS in the analysis of PCDDs and PCDFs in environmental matrices.