Dioxins, dioxin-like PCBs and non-dioxin-like PCBs in foodstuffs: occurrence and dietary intake in The Netherlands.

Data on occurrence of dioxins (polychlorinated dibenzo-p-dioxins [PCDDs] and dibenzofurans [PCDFs]), dioxin-like PCBs (polychlorinated non-ortho and mono-ortho biphenyls) and non-dioxin-like PCBs (as represented by the so-called indicator-PCBs: congeners 28, 52, 101, 118, 138, 153 and 180) in food products consumed in The Netherlands that were collected in measurement programs carried out during 1998 and 1999, and combined with food consumption data to assess the dietary intake of these persistent food contaminants. The estimated median life-long-averaged intake of the sum of dioxins and dioxin-like PCBs in the population is 1.2 pg WHO-TEQ (toxic equivalents) per kg body weight (bw) per day, while the estimated median life-long-averaged intake of indicator-PCBs is 5.6 ng per kg bw per day. The contribution of different food groups to the total intake of both dioxins + dioxin-like PCBs and non-dioxin-like PCBs is fairly uniformly distributed over the foods consumed: meat products (23% and 27%, respectively), dairy products (27% and 17%, respectively), fish (16% and 26%, respectively), eggs (4% and 5%, respectively), vegetable products (13% and 7%, respectively), and industrial oils and fats (17% and 18%, respectively). Compared with earlier intake estimations the present estimation shows a continued reduction in the intake of dioxins as well as PCBs. This reduction is related to the decrease in the concentration of these substances in the majority of foodstuffs. Nevertheless, a small part of the population still has a rather high life-long averaged intake: 8% of the population is exposed to intake levels above the tolerable weekly intake for dioxins and dioxin-like PCBs of 14 pg WHO-TEQ per kg bw per week, as recently derived by the Scientific Committee on Food of the European Commission. For the non-dioxin-like PCBs an internationally accepted maximum intake level is still lacking. However, to provide risk managers with a health-based guideline to prevent health effects of exposure to non-dioxin-like PCBs, the (international) derivation of a tolerable daily intake is recommended. Monitoring the dietary intake of PCBs is just as important as monitoring the intake of dioxins and dioxin-like PCBs, and attempts to decrease the exposure to both compound classes need continuous attention.

Fast sample preparation involving MASE and coupled column normal phase liquid chromatography for the rapid trace analysis of dioxins in air-dust samples from fire catastrophe emissions.

A new approach has been developed and tested for the urgent analysis of dioxins in samples of air-dust filters originating from catastrophe emissions. The procedure consists of a fast extraction of the sample with microwave solvent extraction (MASE) and acetone as solvent followed by a fast cleanup of the extract with normal phase coupled column liquid chromatography (LC/LC). The multi-dimensional LC/LC system employs a 50 mm x4.6 mm i.d. column packed with 3mum silica and a 150 mm x4.6 mm i.d. column packed with 5mum PYE as the first and second analytical column, respectively. Iso-hexane is used on both columns to perform cleanup and dichloromethane to perform efficient back-flush elution of the compounds from the second column. The obtained polarity-based separation in the first dimension and molecular-structure based separation in the second dimension provides a fast and powerful cleanup. Validation was done by analysing samples of homemade RIVM air-dust with aged residues (n=8, spiking level about 15pgmg(-1) per compound) of dioxins/furans and samples of reference Urban Dust SRM 1649a (n=4) with both the new approach and the existing conventional procedure and were instrumentally analyzed with capillary gas chromatography and high resolution mass spectrometric detection (GC/HRMS). In comparison to the existing conventional procedure, the new approach reduces sample processing from several days to several hours per sample. As regards the aged-residue air-dust samples, the new method shows a good accuracy, precision and high selectivity providing a performance in good agreement with the existing procedure. In SRM air-dust, the concentration of a few compounds obtained by the new method was below (10-50%) the certified value.

Levels of certain metals, organochlorine pesticides and dioxins in cord blood, maternal blood, human milk and some commonly used nutrients in the surroundings of the Aral Sea (Karakalpakstan, Republic of Uzbekistan).

UNLABELLED: Since the 1960s a massive decline in the volume of the Aral Sea has occurred as a result of the diversion of the supplying rivers to cotton irrigation schemes. The contaminated sediment of the former seabed has been disseminated over the surrounding area by strong winds. This deterioration of the ecosystem has created a hazardous situation for the health of approximately 3.5 million people. This pilot study was undertaken to assist Karakalpak health workers in investigating the degree of exposure to metals, persistent organochlorine pesticides (OCPs) and dioxins during the most vulnerable prenatal and postnatal period and to provide guidelines for future research. All subjects came from areas located within 200 km of the southern border of the Aral Sea. Blood was obtained from 18 pregnant women and 28 newborns (cord blood) to determine the levels of metals, lead, cadmium, zinc and selenium and the OCPs alpha-, beta- and gamma-hexachlorocyclohexane (HCH; lindane is the product name of gamma-hexachlorocyclohexane), hexachlorobenzene (HCB), and isomers and metabolites of dichlorodiphenyltrichloroethane (DDT), i.e. op-DDT, pp-DDT, pp-DDE and pp-TDE. Levels of metals were also determined in blood from 28 non-pregnant women. In addition, levels of 17 dioxins were determined in 41 human milk samples and 1batch of formula. Information about possible dietary sources of the fat-soluble OCPs and dioxins was obtained from a pooled sample of butter and from seven cottonseed oil samples. A mass-selective detector coupled to a gas chromatograph equipped with a large volume injector was used to analyse the selected OCPs, whereas the dioxins were determined by gas chromatography with high-resolution mass spectrometry. The levels of metals in cord and maternal blood were consistent with concentrations observed in European countries. Only three women (7%) had lead levels greater than 100 ppb. The most notable pollutants in maternal and cord blood were the OCPs, notably HCB, beta-HCH, pp-DDE and the most toxic dioxin, 2,3,7,8-TCDD, when calculated as TEQs. A similar pattern was observed in human milk: beta-HCH and pp-DDE levels of more than 1,000 ng g(-1) fat were found in 68 and 43% of the subjects, respectively. Levels of 2,3,7,8-TCDD were six times higher than those observed in Western Europe. Traces of pp-DDE were detected in the batch of formula milk. Contaminated animal fat, but not cottonseed oil, is the most likely dietary source of OCPs and dioxins. CONCLUSION: Further epidemiological research is needed to elucidate the health implications of these pollutants on perinatal and maternal health, including lactation. More importantly, an investigation should be initiated to identify the emission sources of persistent organic pollutants in Karakalpakstan and adjacent regions.

Comparison of various liquid chromatographic methods involving UV and atmospheric pressure chemical ionization mass spectrometric detection for the efficient trace analysis of phenylurea herbicides in various types of water samples.

The performance of mass spectrometric (MS) detection and UV detection in combination with reversed-phase liquid chromatography without and with the use of coupled column RPLC (LC-LC) has been compared for the trace analysis of phenylurea herbicides in environmental waters. The selected samples of this comparative study originated from an inter-laboratory study. For both detection modes, a 50 mm x 4.6 mm I.D. column and a 100 mm x 4.6 mm I.D. column packed with 3 microm C18 were used as the first (C-1) and second (C-2) column, respectively. Atmospheric pressure chemical ionization mass spectrometry was performed on a magnetic sector instrument. The LC-LC-MS analysis was carried out on-line by means of direct large volume (11.7 ml) injection (LVI). The performance of both on-line (LVI, 4 ml of sample) and off-line LC-LC-UV (244 nm) analysis was investigated. The latter procedure consisted of a solid-phase extraction (SPE) of 250 ml of water sample on a 500 mg C18 cartridge. The comparative study showed that LC-LC-MS is more selective then LC-LC-UV and, in most cases, more sensitive. The LVI-LC-LC-MS approach combines direct quantification and confirmation of most of the analytes down to a level of 0.01 microg/l in water samples in less then 30 min. As regards LC-LC-UV, the off-line method appeared to be a more viable approach in comparison with the on-line procedure. This method allows the screening of phenylurea's in various types of water samples down to a level of at least 0.05 microg/l. On-line analysis with LVI provided marginal sensitivity (limits of detection of about 0.1 microg/l) and selectivity was sometimes less in case of surface water samples. Both the on-line LVI-LC-LC-MS method and the LC-LC-UV method using off-line SPE were validated by analysing a series of real-life reference samples. These samples were part of an inter-laboratory test and contained residues of herbicides ranging from 0.02 to 0.8 microg/l. Beside good correlation between the methods the data agreed very well with the true values of the samples.

Determination of linuron and related compounds in soil by microwave-assisted solvent extraction and reversed-phase liquid chromatography with UV detection.

The combination of microwave-assisted solvent extraction (MASE) and reversed-phase liquid chromatography (RPLC) with UV detection has been investigated for the efficient determination of phenylurea herbicides in soils involving the single-residue method (SRM) approach (linuron) and the multi-residue method (MRM) approach (monuron, monolinuron, isoproturon, metobromuron, diuron and linuron). Critical parameters of MASE, viz, extraction temperature, water content and extraction solvent were varied in order to optimise recoveries of the analytes while simultaneously minimising co-extraction of soil interferences. The optimised extraction procedure was applied to different types of soil with an organic carbon content of 0.4-16.7%. Besides freshly spiked soil samples, method validation included the analysis of samples with aged residues. A comparative study between the applicability of RPLC-UV without and with the use of column switching for the processing of uncleaned extracts, was carried out. For some of the tested analyte/matrix combinations the one-column approach (LC mode) is feasible. In comparison to LC, coupled-column LC (LC-LC mode) provides high selectivity in single-residue analysis (linuron) and, although less pronounced in multi-residue analysis (all six phenylurea herbicides), the clean-up performance of LC-LC improves both time of analysis and sample throughput. In the MRM approach the developed procedure involving MASE and LC-LC-UV provided acceptable recoveries (range, 80-120%) and RSDs (<12%) at levels of 10 microg/kg (n=9) and 50 microg/kg (n=7), respectively, for most analyte/matrix combinations. Recoveries from aged residue samples spiked at a level of 100 microg/kg (n=7) ranged, depending of the analyte/soil type combination, from 41-113% with RSDs ranging from 1-35%. In the SRM approach the developed LC-LC procedure was applied for the determination of linuron in 28 sandy soil samples collected in a field study. Linuron could be determined in soil with a limit of quantitation of 10 microg/kg.

Potential of a simple mechanical extraction system for the rapid extraction of pesticides from foodstuffs. Application to the determination of organochlorine pesticides in lettuce.

The efficiency of a simple mechanical extraction system as applied in veterinary drug analysis has been tested in the field of pesticide residue analysis. As a first application, the system was used for the extraction of organochlorine pesticides from vegetables. The convenience of this simple extraction system consists of performing mechanical extraction in disposable polyethylene-based extraction bags, reducing considerably manual operation and cross-contamination. For the tested compounds and matrix (lettuce), recoveries of the 10 organochlorine pesticides performed at two spiking levels (n = 4 each level and compound) ranging between 0.06 and 3.3 mg/kg were between 60 and 80%, with most standard deviations <5%. The extraction method appeared to be simple and fast with a great potential for the analysis of many pesticide-matrix combinations.

Semi-permeable surface analytical reversed-phase column for the improved trace analysis of acidic pesticides in water with coupled-column reversed-phase liquid chromatography with UV detection. Determination of bromoxynil and bentazone in surface water.

The coupled-column (LC-LC) configuration consisting of a 3 microm C18 column (50 x 4.6 mm I.D.) as the first column and a 5 microm C18 semi-permeable-surface (SPS) column (150 x 4.6 mm I.D.) as the second column appeared to be successful for the screening of acidic pesticides in surface water samples. In comparison to LC-LC employing two C18 columns, the combination of C18/SPS-C18 significantly decreased the baseline deviation caused by the hump of the co-extracted humic substances when using UV detection (217 nm). The developed LC-LC procedure allowed the simultaneous determination of the target analytes bentazone and bromoxynil in uncleaned extracts of surface water samples to a level of 0.05 microg/l in less than 15 min. In combination with a simple solid-phase extraction step (200 ml of water on a 500 mg C18-bonded silica) the analytical procedure provides a high sample throughput. During a period of about five months more than 200 ditch-water samples originating from agricultural locations were analyzed with the developed procedure. Validation of the method was performed by randomly analyzing recoveries of water samples spiked at levels of 0.1 microg/l (n=10), 0.5 microg/l (n=7) and 2.5 microg/l (n=4). Weighted regression of the recovery data showed that the method provides overall recoveries of 95 and 100% for bentazone and bromoxynil, respectively, with corresponding intra-laboratory reproducibilities of 10 and 11%, respectively. Confirmation of the analytes in part of the samples extracts was carried out with GC-negative ion chemical ionization MS involving a derivatization step with bis(trifluoromethyl)benzyl bromide. No false negatives or positives were observed.

Rapid determination of glyphosate in cereal samples by means of pre-column derivatisation with 9-fluorenylmethyl chloroformate and coupled-column liquid chromatography with fluorescence detection.

A rapid procedure for the determination of glyphosate in cereals has been developed. Convenient sample pretreatment is carried out by (i) a overnight standing extraction of 1.0 g homogenized sample with 20 ml of water, (ii) centrifugation of the samples, (iii) a passing of 2.5 ml of the clear layer through a 100 mg C18 solid-phase extraction cartridge and (iv) collection of the last 1.5 ml of the eluent into a calibrated tube. For the instrumental analysis, the efficient approach developed earlier for environmental water samples [J.V. Sancho, F. Hernández, F.J. LUpez, E.A. Hogendoorn, E. Dijkman, P. van Zoonen, J. Chromatogr. A, 737 (1996) 75] was successfully adopted for the determination of glyphosate in the obtained cereal extracts. The procedure includes a 15 min derivatisation step of the analyte with 9-fluorenylmethyl chloroformate and a 16 times dilution step prior to instrumental analysis employing coupled-column LC with fluorescence detection. The developed procedure has a sample throughput of more than 25 samples per day and a limit of quantification of 0.5 mg/kg. The method was validated by analyzing freshly spiked cereal samples and samples with aged residues at levels between 1.0 and 10 mg/kg. The overall recovery of the freshly spiked samples was 86% (n = 10) with a repeatability of 6.5% and a reproducibility of 9.5%. For samples with aged residues recoveries performed at different time intervals (range 80-150 days) did not differ significantly; the overall recovery (n = 10) was 74% with a repeatability and reproducibility of 14 and 20%, respectively.

Screening and analysis of polar pesticides in environmental monitoring programmes by coupled-column liquid chromatography and gas chromatography-mass spectrometry.

Screening and analysis of polar pesticides based on coupled-column reversed-phase liquid chromatography (LC-LC) and GC- or LC-MS is a powerful tool in the execution of environmental monitoring programmes. This paper presents a unified approach utilising LC-LC screening followed by GC-MS confirmation. As polar pesticides are not generally amenable to GC a widely applicable derivation technique is used. The results demonstrate that the proposed LC and MS techniques are capable of analysing a wide range of polar pesticides down to levels of 0.1 microgram/l (EU limit for drinking water). LC switching techniques for group analysis or individual compounds rely on the reversed-phase retention and the UV detectability of the pesticides in combination with the choice of the LC columns. Fast miniaturised derivatization prior to GC-MS forms an integral part in the proposed strategy. In order to avoid extraction losses, derivation in the aqueous sample, preferably with electrophoric reagents with enhanced sensitivity in GC-NICI-MS are employed where possible. In this communication, method development and validation fitting in the strategy are evaluated and the results of the combined approach are discussed.

Determination of organochlorine compounds in fatty matrices. Application of rapid off-line normal-phase liquid chromatographic clean-up.

A clean-up method for organochlorine compounds in fatty samples based on normal-phase liquid chromatography (NPLC) is described. To this end, an existing clean-up procedure which uses column switching for the separation of organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) from the fat matrix was simplified to a single silica LC column procedure. The use of an LC column packed with 3 microns silica enables complete fat/OCP separation in a total fraction volume of 12 ml, and results in a fully automated clean-up procedure that takes only 32 min per sample. The method showed average recoveries of 80-110% in the concentration range of 1-510 micrograms/kg, with relative standard deviations of less than 10%. Limits of determination were in the range of 0.5-50 micrograms/kg. The simplified approach has shown its potential for a variety of samples, such as milk, pork fat, animal feed and cod liver oil, showing its general applicability to fatty samples.

Automated solid-phase extraction coupled to gas chromatography with electron-capture detection: a combination of extraction and clean-up of pyrethroids in the analysis of surface water.

The combination of automated solid-phase extraction (SPE) and large-volume introduction gas chromatography electron-capture detection (GC-ECD) is used for the determination of synthetic pyrethroids in surface and drinking water. The selectivity that is needed for the use of GC-ECD of environmental samples is achieved by addition of a polarity modifier to the sample. In this study, sufficient methanol was added to 19 ml of sample to obtain a level of 30% of modifier prior to SPE. The cartridge is washed with one additional ml of 30% methanol in water. After drying, the cartridge is eluted with toluene. GC-ECD analysis is performed by on-column introduction of 100 microliters of the toluene extract. The whole procedure was automated by using an automated solid-phase extraction apparatus. Limits of detection (LODs) for the pyrethroids in surface water are in the range of 1 to 8 ng/1 (signal-to-noise ratio = 3); even lower limits of detection are possible for drinking water.

Analysis of organic micropollutants in the lipid fraction of foodstuffs.

An overview is given of current techniques for the analysis of organic micropollutants that accumulate in the fatty fraction of foodstuffs, such as pesticides, polychlorinated biphenyls, polycyclic aromatic hydrocarbons, polychlorinated dibenzodioxins and polychlorinated dibenzofurans. Isolation and clean-up are considered to be of great importance in the field of residue analysis. In general, problems are related to the low levels of the individual compounds at which they usually occur and the complexity of extraction and clean-up procedures for isolating and separating analytes from matrix components and other contaminants. Therefore, special attention is focused on sample pretreatment and on coupled chromatographic techniques, showing developments towards multi-residue methods, miniaturization and automation of analytical procedures. Coupling of chromatographic techniques with spectroscopic techniques is also considered as an important tool for identification and confirmation purposes.

Determination of misonidazole and desmethylmisonidazole in plasma by high-performance liquid chromatography with reductive electrochemical detection.

A reversed-phase high-performance liquid chromatographic method with reductive mode electrochemical detection was developed for the determination of misonidazole and desmethylmisonidazole in plasma. A thin-layer amperometric detector with glassy carbon working electrode was used to detect the nitroimidazoles at a potential of -0.60 V. The calibration curves were linear. The within-day and day-to-day coefficients of variation were below 3% for plasma misonidazole concentrations of 6-60 mg/l and 1-15 mg/l for desmethylmisonidazole. Electrochemical detection limits were between 2 and 4 pg, which is about 10-20 times lower than that obtained by detection at 323 nm. Limits of quantitation of the nitroimidazoles in plasma were in the order of 1-2 micrograms/l. Under the described conditions other nitroimidazoles and nitro compounds can also be detected with ultimate sensitivity by reductive mode electrochemical detection.