Temporal trends (1953-2012) of toxic and essential elements in red deer antlers from northeastern Poland.

Red deer antlers have a number of advantages that make them a unique material for monitoring trace elements. As antlers are shed and regrown every year, results of toxicological investigations can be applied to a particular region and time. We analyzed the content of four toxic (Pb, Cd, Hg, As) and three essential (Cu, Zn, Fe) trace elements in 254 red deer antler samples spanning between 1953 and 2012. Age of stags did not influence concentrations of analyzed elements in antlers, except for Zn whose level increased with age. The highest concentrations of toxic elements occurred at the beginning of the analyzed period. Levels of Pb, Hg and Zn in antlers decreased over the course of the study. Levels of Cd and As were low and presented a steady trend. Variations in the levels of the analyzed elements in red deer antlers are considered to reflect levels of exposure of animals in their habitat over the sixty-year study period. The range of essential element levels did not indicate any contamination. Environmental conditions in the Mazury Region during the last decades appeared to have improved significantly, as established by declining trends of toxic elements levels in deer antlers.

Total mercury levels in the muscle and liver of livestock and game animals in Poland, 2009-2018.

The bioaccumulation of mercury (Hg) in the food chain may pose a threat to human health. The risk of dietary Hg intake is mostly caused by the consumption of fish and seafood, therefore the knowledge on the exposure from land animal products is limited. In our article, we summarized the results of analyses of Hg in muscle tissue and liver of different livestock and game animals obtained during ten years of official monitoring that was carried out in Poland from 2009 to 2018. The majority of the results in muscle tissue were below the limits of quantification (LOQs). The mean Hg concentrations in muscle tissue ranged from 0.6 to 5.6 µg kg-1 of wet weight and the mean liver Hg concentrations were within the range of 0.8-16.4 µg kg-1 of wet weight, with lowest levels in chickens and highest in wild boars. The results revealed decreasing trends in liver Hg in cattle and cervids over the years, which was congruous with decreasing emission of Hg in Europe. Our results showed that the consumption of meat and liver of livestock and game animals in Poland may be considered to be safe for human health, which was confirmed by the low number of noncompliant samples relative to the applicable legal limits, as well as by estimated dietary exposure.

Development of an Analytical Procedure for the Determination of Multiclass Compounds for Forensic Veterinary Toxicology.

Reported here is a new analytical multiclass method based on QuEChERS technique, which has proven to be effective in diagnosing fatal poisoning cases in animals. This method has been developed for the determination of analytes in liver samples comprising rodenticides, carbamate and organophosphorus pesticides, coccidiostats and mycotoxins. The procedure entails addition of acetonitrile and sodium acetate to 2 g of homogenized liver sample. The mixture was shaken intensively and centrifuged for phase separation, which was followed by an organic phase transfer into a tube containing sorbents (PSA and C18) and magnesium sulfate, then it was centrifuged, the supernatant was filtered and analyzed by liquid chromatography tandem mass spectrometry. A validation of the procedure was performed. Repeatability variation coefficients <15% have been achieved for most of the analyzed substances. Analytical conditions allowed for a successful separation of variety of poisons with the typical screening detection limit at ≤10 µg/kg levels. The method was used to investigate more than 100 animals poisoning incidents and proved that is useful to be used in animal forensic toxicology cases.

The effect of diet enriched with rapeseed meal on endogenous thiouracil contents in urine and milk of cattle.

Thyreostatic compounds, such as thiouracil, are orally active drugs that can be used to increase the weight of cattle before slaughter. Due to potentially teratogenic and carcinogenic effects of their residues on public health, the use of thyreostats in animal production has been banned in the European Union since 1981. Systematic detection of low concentrations of thiouracil in the urine of livestock in many countries is believed to be of endogenous origin due to the use of Brassicaceae plants in the animal diet. Therefore, the purpose of the study was to determine the effects of diets rich in rapeseed meal on formation of thiouracil in urine and milk of dairy cows. For two weeks three cows were subjected to a diet supplemented with rapeseed at 30%, compared to the control cattle diet which contained up to 11% rapeseed. During the experiment, samples of urine and milk were collected and analysed by LC-MS/MS. The increase and decrease of thiouracil concentration in urine samples in different animals was individual and cyclic. The highest concentration of natural thiouracil determined in urine was 3.61 µg l-1. It has been found that endogenous thiouracil exists in two tautomeric forms. A few days of storage of frozen urine samples affected the stability of natural thiouracil, whereas an acidic medium improved the stability of the compound and its isomer, which remained stable even after two months of storage at temperatures below -18°C. Due to the instability of thiouracil, urine samples upon sampling should be delivered to the laboratory as soon as possible or properly preserved. In milk samples, thiouracil was not found above the decision limit of the applied method of 0.63 µg l-1. Preliminary studies have shown that faecal examination for banned thiouracil can be a complementary test for urine samples, and may be helpful in determining the origin of the compound present in urine.

Control of Residues of Thyreostats in Slaughter Animals in Poland in 2011-2017.

INTRODUCTION: In the European Union, the use of thyreostatic drugs for fattening slaughter animals has been banned since 1981 under Council Directive 81/602/EEC. For protection of consumer health against unwanted residues and in compliance with Directive 96/23, each EU country must monitor thyreostats in samples of animal origin. This paper presents the results of research on thyreostatic residues carried out in Poland in 2011-2017. MATERIAL AND METHODS: The material for testing was urine (n = 3,491), drinking water (n = 127), and muscle samples (n = 349) officially collected by Veterinary Sanitary Inspectors in slaughterhouses and farms throughout the country in accordance with the national residue control plan. The samples were examined for the presence of tapazole, thiouracil, methylthiouracil, propylthiouracil, and phenylthiouracil using liquid chromatography tandem mass spectrometry through an accredited method. RESULTS: In four bovine and three porcine urine samples, the permissible thiouracil concentration was exceeded. In one sample of porcine urine, methyl- and propylthiouracil were found. The presence of thiouracil and its derivatives in urine samples is most likely due to feeding animals diet containing cruciferous plants. CONCLUSIONS: The results of research indicate that thyreostats are not used for anabolic purposes in slaughter animals in Poland.

Absence of evidence or evidence of absence? A transfer and depletion study of Sudan I in eggs.

Sudan I is a carcinogenic industrial azo-dye, forbidden for use in food. However, it has been detected in food on several occasions, such as in paprika, used in animal husbandry to enhance egg yolk colour. Therefore, an animal experiment was designed to simulate the transfer of Sudan I to eggs after its unintentional administration to laying hens. A group of laying hens (n=18) received feed contaminated with Sudan I at the raising concentrations: 0.45mg/kg, 4.97mg/kg and 42.1mg/kg. Residues of Sudan I were detected in egg yolks (0.29±0.03µg/kg, mean±SD) only after the administration of the feed contaminated with the dye at the highest concentration. The determined concentrations were much lower than expected based on the compound's lipophilicity. In conclusion, the transfer of Sudan I to eggs was limited and strongly dependent on its concentration in feed.

Multiresidue determination of veterinary medicines in lyophilized egg albumen with subsequent consumer exposure evaluation.

The process of lyophilization causes that the veterinary drugs residues present in egg albumen do not decompose, as it takes place during the process of high-temperature drying. Thus, lyophilized albumen may be a potential source of their residues for consumers. As a consequence, reliable methods for the determination of veterinary medicinal products from egg albumen are needed. The method for the determination of 85 analytes in lyophilized egg albumen was developed and successfully validated. The recoveries were between 84 and 110%, within laboratory repeatability and reproducibility - in the range of 3.29-16.8% and -5.93 to 19.3%. The presence of enrofloxacin and doxycycline was confirmed in real egg albumen samples. The concentrations ranged from 5.65-596µg/kg for doxycycline to 0.89-134µg/kg for enrofloxacin. Nevertheless, the evaluated daily intake and % of the ADI (Acceptable Daily Intake) received by the consumers' were at a toxicologically accepted level.

Can red deer antlers be used as an indicator of environmental and edible tissues' trace element contamination?

Venison is an attractive product for consumers concerned with healthy lifestyle; however, it can contain high levels of toxic elements, and therefore, it is a possible source of hazardous contaminants in human diet. Antlers are suitable bioindicators of environmental metal contamination, and herein, we assessed the ability of trace element levels in antlers to indicate levels in edible soft tissues. We determined the concentrations of lead (Pb), cadmium (Cd), mercury (Hg), arsenic (As), copper (Cu), zinc (Zn), and iron (Fe) in the liver, kidney, muscle, and antlers of 14 free-ranging red deer (Cervus elaphus) from northeastern Poland using atomic absorption spectrometry. We found the highest concentrations of Pb (0.321 ± 0.165 mg/kg), As (0.045 ± 0.074 mg/kg), Zn (105.31 ± 16.33 mg/kg), and Fe (220.92 ± 117.18 mg/kg) in antlers; of Cd (4.974 ± 1.90 mg/kg) and Hg (0.048 ± 0.102 mg/kg) in kidney; and of Cu (7.29 ± 7.02 mg/kg) in the liver. A positive relationship between concentrations in antlers and muscle was found only for Cu (p = 0.001), and it therefore appears that red deer antlers cannot be used as an index for element concentrations in soft tissues. While our results confirm that the Mazury region is little polluted, consumption of red deer offal from this area should be limited according to extant legal limits set for livestock consumption.

Simultaneous Determination of Ten Illegal Azo Dyes in Feed by Ultra-high Performance Liquid Chromatography Tandem Mass Spectrometry.

INTRODUCTION: The paper presents the method of simultaneous determination of 10 illegal azo dyes in feed by ultra-high performance liquid chromatography coupled with tandem mass spectrometry technique. MATERIAL AND METHODS: The dyes were extracted with hexane, evaporated to dryness, and analysed. Separation was achieved in 7 min in a gradient elution using acetonitrile (A) and 0.1% formic acid (B) as a mobile phase. RESULTS: The validation results showed the repeatability of the method, which was evaluated at three levels (50, 500, and 5,000 µg/kg). All the matrix calibration curves for the working ranges were linear (R2 0.9904 to 1.0), the repeatability was between 2.1% and 24%, and recoveries ranged from 77.9% to 120%. The LOD and LOQ were at 1-2 and 5-10 µg/kg for different dyes, respectively. Furthermore, the method was applied in the homogeneity tests of the in-house prepared feed containing Sudan I at the levels of 0.5, 5, and 50 mg/kg. CONCLUSIONS: A sensitive, selective, and fast multiresidue method was successfully developed and validated. Its robustness was confirmed by the analysis of an experimental feed containing Sudan I.

Expression of metallothionein in the liver and kidneys of the red deer (Cervus elaphus L.) from an industrial metal smelting area of Poland.

The metallothionein 1 (MT1) coding sequence of red deer was identified and compared to orthologous sequences from other mammals. Over 90% identity was observed between red deer MT1 amino acid sequence and MT1 sequences of other ruminants. Liver and kidney samples of red deer were collected from the industrial zinc smelting site of Miasteczko Slaskie and from the Masuria Lake District serving as a pollution-free control site. The concentrations of cadmium (Cd), lead (Pb), copper (Cu) and zinc (Zn) were analyzed by the atomic absorption spectrometry technique (AAS). The levels of Cd in the liver of red deer from the metal smelting region was about 8 times higher than for the reference control site. Next, the expression of MT1 mRNA in the liver of red deer was quantified by the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and the expression of MT1/2 protein in the liver and kidneys was analyzed by immunohistochemistry. Positive correlations were found between expression levels for MT1 mRNA and the concentrations of Cu and Zn in liver of red deer, and with the age of animals. Immunohistochemical staining demonstrated the nuclear and cytoplasmatic expression in both liver and kidney tissues, but with no obvious relationship shown for the expression of MT1/2 protein and tissue metal levels. Our results showed that the analysis of MT expression levels in the red deer could not be used independently as a biomarker for identifying exposure to Cd, but could be co-analyzed with tissue metal levels to give better prognosis for environmental exposure to metals.

Influence of enrofloxacin traces in drinking water to doxycycline tissue pharmacokinetics in healthy and infected by Mycoplasma gallisepticum broiler chickens.

Most of antibiotics, administrated in the treatment of poultry diseases are dissolved in drinking water, and it can lead to water supply systems contamination, especially when the regular cleaning is not using. This situation can lead to unconscious administration of low doses of antibiotics to untreated animals. The aim of this study was to clarify the impact of the exposure of enrofloxacin traces (500 µg l(-1)) to doxycycline pharmacokinetics in healthy and experimentally Mycoplasma gallisepticum infected broiler chickens., Two experimental groups, received of enrofloxacin in water and all groups, received 20 mg kg(-1) bw of doxycycline. The compounds concentrations in muscles and livers were determined by LC-MS/MS. The maximum drug tissue concentration (Cmax) of doxycycline was highest in liver obtained from infected chickens which, received enrofloxacin traces (ENR + DC/MG). It was about 40% higher than in healthy chickens from group I which received only doxycycline. It was found that the concentration-time curve AUC(0-t) values in group ENR + DC/MG were almost 75% higher than in the group (DC) and 35% higher than in group (ENR + DC) which also received enrofloxacin traces. The constant exposure of broiler chickens on enrofloxacin traces as well as infection, may significantly influenced on doxycycline tissue pharmacokinetic profile.

Multiresidue method for the simultaneous determination of veterinary medicinal products, feed additives and illegal dyes in eggs using liquid chromatography-tandem mass spectrometry.

A multiclass method was developed for the simultaneous determination of 120 analytes in fresh eggs. The method covers the analytes from the groups of tetracyclines (6), fluoroquinolones (11), sulphonamides (17), nitroimidazoles (9), amphenicols (2), cephalosporins (7), penicillins (8), macrolides (8), benzimidazoles (20), coccidiostats (14), insecticides (3), dyes (12) and others (3). Samples were extracted using 0.1% formic acid in acetonitrile:water (8:2) with the addition of EDTA and cleaned using solid phase extraction with Hybrid SPE cartridges. The chromatographic separation was achieved on C8 column using mobile phase consisting of (A) methanol:acetonitrile (8:2) - (B) 0.1% formic acid in a gradient mode. Validation results according to the Commission Decision 2002/657/EC are as follows: linearity (r⩾0.99), recovery (75-108%), repeatability (CV 1.60-15.9%), reproducibility (CV 2.60-15%), decision limit (CCα 2.25-1156 µg/kg) and detection capability (CCß 2.04-1316 µg/kg). The presented method was used for analysis of 150 real eggs samples taken from monitoring control program.

Application of the yeast-based reporter gene bioassay for the assessment of estrogenic activity in cow's milk from Poland.

Milk contain compounds acting through the estrogen receptor signaling. The still open question whether such estrogens pose a risk for human health, encouraged us to measure the overall estrogenic activity of cow's milk in the in vitro yeast reporter bioassay. First, we assessed the ability of the bioassay to detect estrogens frequently detected in milk. The relative potencies of 16 compounds descended in the order: 17ß-estradiol (17ß-E2), 17α-ethinylestradiol, diethylstilbestrol, dienestrol, 17α-E2, estrone, zearalenone, estriol, equol, genistein, 17ß-E2 glucuronide, bisphenol A, apigenin, daidzein. Flavone, 4-n-nonylphenol and 4-t-octylphenol shown no activity in the bioassay.The estrogenic activities of milk samples without hydrolysis were below the detection limit, whereas in 50% of the deconjugated samples they varied between 0.29 and 0.49 ng EEQ mL(-1). We also compared the estrogenic activity in raw cow's milk collected from rural and industrial locations in Poland. In our pilot study we did not observe statistically significant difference in estrogenic activities in milk collected from the two locations. We found that the daily intake of estrogens with milk may be higher than estrogen levels in human serum. Further studies are warranted to evaluate the significance of milk and dairy as a source of estrogens for humans.

Liquid chromatography-tandem mass spectrometry multiclass method for the determination of antibiotics residues in water samples from water supply systems in food-producing animal farms.

A sensitive liquid-chromatography-electrospray tandem mass spectrometry multiclass method for determination of 45 veterinary compounds belonging to 9 different antibiotic groups, including aminoglicosides (4), ß-lactams (13), diaminopyrimidines (1), fluoroquinolones (10), lincosamides (1), macrolides (5), pleuromutilins (1), sulfonamides (6) and tetracyclines (4), in water from breeding animal watering supply system has been developed. Isolation of the analytes was carried out by solid phase extraction with heptafluorobutyric acid as an ion-pair agent on the Strata-X reversed phase cartridges. All analytes were determined simultaneously in one single run on a C18 column with gradient elution and short analysis time (13 min). Method was validated, average relative recoveries were in the range of 84.3-109.3% with satisfactory precision are repeatability for all compounds are in the range of 4.7-12.2%, within-laboratory reproducibility are in the range of 4.4-13.5% for. The limit of quantitation (LOQ) of the method was in the range of 0.02-10 µg L(-1), depending of analyte. The applicability of the method was tested by determining antimicrobial compounds in real water samples collected from water supply systems in breeding animal farms. The average antibiotics concentration in real water samples were, respectively, in the range of 0.14-1670 µg L(-1).

The effect of environmental mycotoxins on selected ovarian tissue fragments of multiparous female wild boars at the beginning of astronomical winter.

The contamination of plant material with mycotoxins, in particular of the genus Fusarium, is common in the natural environment. Multiparous female wild boars are exposed to feed contaminated with zearalenone (ZEN) and deoxynivalenol throughout the year. The aim of this study was to determine the concentrations of the above mycotoxins in multiparous female wild boars and to describe their effect on the histological structure of the ovaries at the beginning of astronomical winter. Toxicological examinations revealed 0.291 ng/ml of ZEN, 0.406 ng/ml of α-zearalenol (α-ZEL), 0.392 ng/ml ß-zearalenol (ß-ZEL) and an absence of deoxynivalenol (values below the sensitivity of the method) in the blood plasma of multiparous female wild boars. Numerous ovarian follicles at various stages of development, characterized by different degree of damage, were observed. Numerous deformed resting ovarian follicles were noted directly under the epithelium, and signs of follicular atresia and hyalinization were observed. Blood vessels in the medulla of the ovary were dilated, which probably improved the distribution of ZEN in the ovaries. Higher substrate (ZEN) concentrations in the ovaries led to an insignificant increase in the staining intensity of 3ß-HSD and 17ß-HSD clusters. The observed changes could contribute to prolonging the initial stage of late anestrus in multiparous female wild boars.

Determination of carbadox and olaquindox metabolites in swine muscle by liquid chromatography/mass spectrometry.

This paper presents LC-MS/MS method that was developed for the simultaneous determination and confirmation metabolites of carbadox (desoxycarbadox, quinoxaline-2-carboxylic) and olaquindox (3-methylquinoxaline-2-carboxylic acid) residues in pig muscle tissues at concentrations ≤3.0µgkg(-1). Pig muscle tissues were deproteinated with meta-phosphoric acid in methanol and then were extracted with ethyl acetate:dichloromethane (50:50, v/v). The whole extracts were evaporated to dryness in rotary evaporator at 45°C, and dry residues were re-dissolved in 0.5% isopropanol in 1% acetic acid. The LC separation was performed on a C8 column with a gradient system consisting of isopropanol/water/acetic acid and methanol as the mobile phase. Additionally SelexION™ technology to reduce matrix effect was used. The decision limit (CCα) ranged from 1.04µgkg(-1) to 2.11µgkg(-1) and the detection capability (CCß) ranged from 1.46µgkg(-1) to 2.89µgkg(-1). The total recoveries were from 99.8% to 101.2%. The results of validation fulfil the requirement of the confirmatory criteria according to the European Commission Decision 2002/657/EC.

Evaluation of estrogenic activity in animal diets using in vitro assay.

A yeast estrogen bioassay (RIKILT REA) was in-house validated for feed on the 5µg 17ß-estradiol-equivalents per kg level according to EC Decision 2002/657/EC. All the performance characteristics met the criteria as defined in the Decision and the REA is able to detect 17ß-estradiol in animal feed at a low level of 1.15-2µgkg(-1). Subsequently, the developed and validated procedure was applied to determine the estrogenic activity in 24 feed samples intended for food producing animals, pets and laboratory animals. Two batches of rodent diet Murigran and one dog feed have been presented as a suspect, i.e. gave responses above the determined decision limit (CCα) and detection capability (CCß). In assessing the performance of the estrogenic activity in these diets evaluated by comparison with the 17ß-estradiol calibration curve, 17ß-estradiol-equivalence levels of 7.07µg EEQkg(-1) and 9.54µg EEQkg(-1) in two batches of rodent diet and 5.3µg EEQkg(-1) in dog feed have been established. The activities observed in the rodent feed could be explained by chemical analysis, revealing high amounts of genistein, daidzein and trace amounts of zearalenone. In addition, the estrogenic activity in one of rodent feed was above the established CCα, but below the CCß values established and all other samples showed no estrogenic activity with responses below the CCα value, which corresponds to levels below 2µg EEQkg(-1).

Determination of stilbenes and resorcylic acid lactones in bovine, porcine and poultry muscle tissue by liquid chromatography-negative ion electrospray mass spectrometry and QuEChERS for sample preparation.

Liquid chromatography tandem mass spectrometry method was developed and validated to confirm of resorcylic acid lactones: zeranol, taleranol, zearalanone, zearalenone, α and ß-zearalenol and stilbenes in muscle tissue. The compounds were analyzed by LC-MS/MS QTrap 5500 apparatus in negative ionization mode. Chromatographic separation on Poroshell 120-EC C18 (150mm×2.1mm, 2.7µm) column was achieved at 45°C using isocratic elution of mobile phase - methanol/water (65:35, v/v). For the treatment of tissue samples prior to analysis, QuEChERS method was applied based on the extraction of analytes from muscle samples with ethyl acetate, separation of the aqueous and organic phases with application of magnesium sulphate and sodium acetate, the purification of the extract obtained by dispersive SPE with the use of sorbent C18, PSA and magnesium sulphate. The method was validated in accordance with the Commission Decision 2002/657/EC. Good recoveries were obtained (from 83% to 115%) as well as acceptable within-lab reproducibility (<22%). The values of the decision limit CCα and the detection capability CCß for individual compounds are found to be below the recommended concentration set at 1µgkg(-1) and not exceed 0.23µgkg(-1) and 0.39µgkg(-1), respectively. The elaborated method meets the criteria for confirmatory methods and is used in the official control of hormones.

Rapid method for the determination of metamizole residues in bovine muscle by LC-MS/MS.

Metamizole is a pyrazolone non-steroidal anti-inflammatory drug allowed for use in food-producing animals. According to Council Directive 96/23, residues of this drug have to be monitored because of the potential risk to consumers' health. Metamizole is hydrolysed to its marker residue 4-methylaminoantypyrine.This compound is further metabolised to three main metabolites: 4-formylaminoantipyrine, 4-aminoantipyrine and 4-acetylaminoantipyrine. The MRL of 4-methylaminoantipyrine in animal tissues is 100 µg kg(-1). Considering the above points, a method for the detection of four metamizole metabolites in bovine muscles was developed. Analytes were extracted from muscle by a mixture of acetonitrile and sodium acetate buffer. After centrifugation, the supernatant was passed through alumina cartridges, diluted with mobile phase and analysed by using LC-MS/MS. Four metamizole metabolites were separated on a C8 column in 23 min with a gradient of methanol:acetonitrile:ammonium formate solution and analysed by using positive ionisation. Validation of the method indicated a within-laboratory reproducibility in the range of 7-30% and recovery in the range of 45-95%. The method fulfils the criteria for confirmatory methods and, thanks to its labour efficiency, may also be used for screening purposes.

Determination of zeranol, taleranol, zearalanone, α-zearalenol, ß-zearalenol and zearalenone in urine by LC-MS/MS.

An LC-MS/MS method was developed for the sensitive and selective determination of zeranol, taleranol, zearalanone, α-zearalenol, ß-zearalenol and zearalenone in animal urine. Analysis was performed on the free compounds after enzymatic deconjugation. Sample preparation included liquid-liquid extraction followed by solid-phase extraction (SPE) with C18 and NH2 columns. For chromatographic separation of hormones an Inertsil(®) ODS-3 analytical column (150 mm × 2.1 mm, 3 µm) was used. The analytes were determined and identified by LC-MS/MS on a QTRAP5500 instrument with a TurboIon-Spray source operating in negative electrospray ionisation mode. For confirmatory purposes at least two transitions were obtained for each analyte. According to Commission Decision 2002 /657/EC the validation parameters - recovery, repeatability, reproducibility, linearity, specificity, decision limits and detection capabilities - were determined. All parameters were in agreement with 2002/657/EC performance criteria. The apparent recovery ranged from 76.2% to 116.3% for all examined compounds. The repeatability was below 20% and reproducibility did not exceed the limit of 25% for most analytes. Linearity was good for all analytes in the whole range of tested concentrations, as proved by the correlation coefficients greater than 0.99. The decision limits (CCα) ranged from 0.04 to 0.18 µg l(-1) for all analytes whereas the detection capabilities (CCß) ranged from 0.07 to 0.31 µg l(-1), respectively. All CCα and CCß values were below the recommended concentration of 2 µg l(-1). This analytical method will be used in an integrated system for Polish monitoring programmes for the confirmation of violative screened samples.