Mette Erecius Poulsen

Adverse effects on sexual development in rat offspring after low dose exposure to a mixture of endocrine disrupting pesticides.

The present study investigated whether a mixture of low doses of five environmentally relevant endocrine disrupting pesticides, epoxiconazole, mancozeb, prochloraz, tebuconazole and procymidone, would cause adverse developmental toxicity effects in rats. In rat dams, a significant increase in gestation length was seen, while in male offspring increased nipple retention and increased incidence and severity of genital malformations were observed. Severe mixture effects on gestation length, nipple retention and genital malformations were seen at dose levels where the individual pesticides caused no or smaller effects when given alone. Generally, the mixture effect predictions based on dose-additivity were in good agreement with the observed effects. The results indicate that there is a need for modification of risk assessment procedures for pesticides, in order to take account of the mixture effects and cumulative intake, because of the potentially serious impact of mixed exposure on development and reproduction in humans.

Results from the monitoring of pesticide residues in fruit and vegetables on the Danish market, 1998?99

To check compliance with the maximum residue levels in foods and to monitor the residue levels to enable an evaluation of the exposure of the Danish population to pesticides, a monitoring programme for pesticides residues in fruit and vegetables was performed. Sampling plans were designed based on previous findings and on consumption data. Samples (n = 4404) of mainly fresh conventionally and organically grown fruit and vegetables were collected at wholesalers and importers and at food processing companies. Of the samples, 3% were frozen products and 5% organically grown. Of the samples, 34% were of Danish origin, with 66% from other countries. Two accredited laboratories performed analyses of the samples using capillary gas chromatography, liquid chromatography and a spectrophotometric method. Reporting levels were set at the lowest calibration level or at the limit of determination. Residues were found in 60% of the samples of fruit but in only 18% of the vegetable samples. Residues above the maximum residue levels were found in 6% of all samples of fruit and in 2% of the vegetable samples.

Probabilistic assessment of the cumulative dietary exposure of the population of Denmark to endocrine disrupting pesticides

The four pesticides epoxiconazole, prochloraz, procymidone and tebuconazole, are commonly used pesticides, all suspected of acting as endocrine disrupters. In the present study, we assessed the acute cumulative dietary exposure to the women of child bearing age and the general population of Denmark to these pesticides from the intake of fruit and vegetables. The assessment was carried out using the probabilistic approach combined with the relative potency factor (RPF) approach. Residue data for prochloraz, procymidone, and tebuconazole were obtained from the Danish monitoring programme 2006-2009, while residue data for epoxiconazole were obtained from the Swedish monitoring programme carried out in the period 2007-2009. Food consumption data were obtained from the Danish nationwide dietary survey conducted in 2000-2002. Relative potency factors for the four pesticides were obtained from rat studies. Prochloraz was used as the index compound. All four pesticides increased nipple retention in male offspring, and epoxiconazole, prochloraz, and tebuconazole also increased the gestation period in pregnant rat dams. For women of childbearing age, the high-end cumulative exposure (99.9th percentile) was calculated to 9% of the Adjusted Reference Value (ARV) for the effect on nipple retention and to 1% of the ARV for the effect on increased gestation period.

Survey of pesticide residues in table grapes: Determination of processing factors, intake and risk assessment

The differences in residue pattern between Italy and South Africa, the main exporters of table grapes to the Danish market, were investigated. The results showed no major differences with respect to the number of samples with residues, with residues being found in 54–78% of the samples. Exceedances of the European Union maximum residue limit (MRL) were found in five samples from Italy. A number of samples were rinsed to study the possible reduction of residues. For copper, iprodione, procymidone and dithiocarbamates a significant effect of rinsing was found (20–49% reduction of residues). However, no significant effect was found for organophosphorus pesticides and pyrethroids, whereas the number of samples with residues of benzilates, phenylamids and triazoles was insufficient to demonstrate any significant effects. An intake calculation showed that the average intake from Italian grapes was 3.9 µg day−1 for pesticides and 21 µg day−1 for copper. Correspondingly, the intakes from South African grapes were 2.6 and 5.7 µg day−1, respectively. When the total exposure of pesticides from grapes were related to acceptable daily intake, expressed as the sum of Hazard Quotients, the exposure were approximately 0.5% for Italian samples and 1% for South African samples.

Deposition of lindane in Denmark

Abstract The deposition of α- and γ-HCH (Lindane) was measured at four different stations in 1990–1992. γ-HCH was found in peak concentrations of 27–122 ng/l in spring and early summer, while the mean concentrations from the rest of the year were about 7 ng/l. The levels of α-HCH were about 2 ng/l without any detectable seasonal variation. The deposited amounts of α- and γ-HCH are probably caused by long-range transport from European countries south and west of Denmark.

Development of an LC-MS/MS method for the determination of pesticides and patulin in apples

A method for the simultaneous determination of 33 pesticides or degradation products together with patulin in apples by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed. The method involved homogenization of the apples, extraction with ammonium acetate–acetic acid solution in methanol–water by ultrasonication, filtration, and determination by LC-MS/MS. The repeatability and within-laboratory reproducibility for the three spiking levels 0.02, 0.04 and 0.2 mg kg−1 were between 4% and 35%. In general, the repeatability and reproducibility were about 10–20%. The limits of quantification (LOQs) were between 0.01 and 0.14 mg kg−1. The method was used on incurred samples from parts of the ISAFRUIT project financed by the European Commission under the 6th Framework Programme. Samples were analysed at four different stages: after harvest, after storage (controlled), after a water bath, and after 28 days at room temperature. Pesticide residues were found at all stages, but no significant differences in the concentration were seen between the stages analysed. The concentration decreased significantly only for tolylfluanid after storage at room temperature for 28 days when only 0–6% of the original amount of tolylfluanid remained in the apples. No patulin was found in the apples stored for 28 days at room temperature and no growth of Penicillium expansum was observed on these apples. However, when the apples were inoculated with a spore suspension of P. expansum, high concentrations of patulin were found.

Estimation of the uncertainty in a multiresidue method for the determination of pesticide residues in fruit and vegetables.

The estimation of uncertainty of an analytical result has become important in analytical chemistry. It is especially difficult to determine uncertainties for multiresidue methods, e.g. for pesticides in fruit and vegetables, as the varieties of pesticide/commodity combinations are many. In the present study, recommendations from the International Organisation for Standardisations (ISO) Guide to the Expression of Uncertainty and the EURACHEM/CITAC guide Quantifying Uncertainty in Analytical Measurements were followed to estimate the expanded uncertainties for 153 pesticides in fruit and vegetables. Data from in-house validation were used in the estimation of the uncertainty. No significant difference in the relative standard deviation for reproducibility (RSDR) were found between the different concentration levels at concentration levels exceeding 2.5 times the detection limit. Therefore, it was possible to pool the RSDR within a single matrix. However, a difference in RSDR between matrices was seen, thus the poorest RSDR of the investigated matrices was chosen for the uncertainty estimation. The expanded uncertainties ranged from 7 to 78% with an average of 32% and median of 32%. Furthermore, only RSDR contributed to the uncertainty estimation.

Cumulative dietary exposure of the population of Denmark to pesticides.

We used the Hazard Index (HI) method to carry out a cumulative risk assessment after chronic dietary exposure to all monitored pesticides in fruit, vegetables and cereals for various consumer groups in Denmark. Residue data for all the pesticides were obtained from the Danish monitoring programme during the period 2004-2011. Food consumption data were obtained from DANSDA (the DAnish National Survey of Diet and physical Activity) for the period 2005-2008. The calculations were made using three different models to cope with residues below the limit of reporting (LOR). We concluded that a model that included processing factors and set non-detects to ½ LOR, but limited the correction (Model 3), gave the most realistic exposure estimate. With Model 3 the HI was calculated to be 0.44 for children and 0.18 for adults, indicating that there is no risk of adverse health effects following chronic cumulative exposure to the pesticides found in fruit, vegetables and cereals on the Danish market. The HI was below 1 even for consumers who eat more than 550 g of fruit and vegetables per day, corresponding to 1/3 of the population. Choosing Danish-produced commodities whenever possible could reduce the HI by a factor of 2.

Clean-up of cereal extracts for gas chromatography-tandem quadrupole mass spectrometry pesticide residues analysis using primary secondary amine and C18.

The level of co-extracted matrix in wheat and oat extracts obtained by the QuEChERS method (EN 15662) is high and the occurrence of free fatty acids generates a major matrix peak in TIC chromatograms (rt. 13-22min). Matrix can compromise the analytical performance in pesticide analysis using GC-MS/MS. In order to reduce the amount and the effects of matrix we tested the effect of using six different amounts of primary secondary amine (PSA) (0, 25, 50, 100, 150 and 200mg/ml extract) with and without the addition of six different amounts of C18 (0, 25, 50, 100, 150 and 200mg/ml extract) in the dispersive solid phase extraction (dSPE) procedure. dSPE clean-up using 25mg/ml extract significantly reduced the major matrix peak observed for wheat extracts. Higher amounts of PSA reduced the analytical response for iprodione and malathion. For oat extract 50-150mg PSA/ml extract was needed to obtain equally low intensity of the matrix peak. For oat the analytical responses of the target pesticides generally increased with increasing amount of PSA. C18 had no significant effect on the intensity of the major matrix peaks and even resulted in lower analytical responses for several of the target pesticides. Based on the present study it is concluded that the optimal dSPE clean-up procedure employs 25mg PSA/ml extract for wheat and 150mg PSA/ml extract for oat.

Influence of different disease control pesticide strategies on multiple pesticide residue levels in apple

Summary Seven pesticide application strategies were investigated to control apple scab (Venturia inaequalis) and powdery mildew (Podosphaera leucotricha) and, at the same time, fulfil the new quality standards implemented by some German retailers. These demand that pesticide residues should be below 80% of European Maximum Residue Levels (EU MRL) and that the number of residues present at levels above 0.01 mg kg−1 should be limited to a maximum of four. The strategies fulfilled the requirement to use combinations of different active substances in order to prevent the emergence of resistance to pesticides. The trials were conducted at two sites in Switzerland, in 2007, and all strategies and applications were in accordance with actual practice. Four replicates of apple samples from each strategy were then analysed for pesticide residues. The incidence of infection with apple scab and powdery mildew were monitored during the season in order to evaluate the efficacy of the different strategies. The efficacies of the different strategies against apple scab and powdery mildew were between 84% and 100% successful. In general, the level of pesticide residues found correlated with application rate and time, and no measured residue level exceeded the EU–MRL. The numbers of residues present at > 0.01 mg kg−1 were between two and five.