Interspecific differences in the bioaccumulation of arsenic of three Patagonian top predator fish: Organ distribution and arsenic speciation.

Interspecific differences in arsenic bioaccumulation and organ distribution (muscle, liver, kidney and gills) in three predator fish (creole perch, rainbow trout and brown trout) from a Patagonian lake impacted by volcanic eruptions were studied. Arsenic in fish organs were compared analyzing: 1) temporal (before and after volcanic eruption) and spatial (near and far from the volcano) influence of Puyehue-Cordón Caulle volcanic complex activity on arsenic concentrations; 2) the influence of growth (as total length), organ type and their interactions over arsenic accumulation; and 3) arsenic speciation and total arsenic relationship with carbon to nitrogen ratios (C:N), as a proxy of lipid presence, in fish muscle. In general, total arsenic concentrations in creole perch organs were 2-7 times higher than those recorded in the corresponding organs of salmonids. Arsenic was preferentially accumulated in liver and kidney in the three fish species. The influence of the volcanic activity over arsenic concentrations was more evident in creole perch: organs from creole perch captured closest to the volcano exhibited higher arsenic concentrations. Temporal variations were not so consistent. No clear relationship between arsenic and fish length was observed. Positive and linear relationship between arsenic in all pair of organs was found in creole perch, while rainbow trout showed a quadratic relationship between muscle and the remaining organs, indicating different arsenic assimilation-elimination relationships between organs and fish. The arsenic liver:muscle ratio in the three fish species was greater than 1, suggesting some level of arsenic stress. Arsenobetaine (AB) and dimethylarsinic acid (DMA) were the dominant arsenic species in muscle of these fish, having creole perch 3-4 times higher AB than rainbow trout. A positive relationship between C:N ratio and total arsenic concentrations was found, with higher C:N in creole perchs near the volcano. In terms of food safety, no inorganic arsenic compound were detected, therefore arsenic levels in fish from Lake Nahuel Huapi does not represent any health risk to consumers.

Metals in Feathers of African Penguins (Spheniscus demersus): Considerations for the Welfare and Management of Seabirds Under Human Care.

Bird feathers have been proven to be reliable indicators of metal exposure originating from contaminated food and polluted environments. The concentrations of 15 essential and non-essential metals were investigated in African penguins (Spheniscus demersus) feathers from a Northwestern Italian zoological facility. These birds are exclusively fed with herring from the northeast Atlantic Ocean. Certain elements, such as Hg and Cd, reflected the bioaccumulation phenomena that occur through the marine food chain. The levels of Cr, Mn, and Ni were comparable to those registered in feathers of birds living in polluted areas. These results are important for comparative studies regarding the health, nutrition and welfare of endangered seabirds kept under human care.

PCDD/Fs, DL-PCBs and NDL-PCBs in European catfish from a northern Italian lake: the contribution of an alien species to human exposure.

PCDDs/Fs, DL-PCBs and NDL-PCBs are environmentally persistent substances that have been associated with adverse effects on human health. Humans are mainly exposed to these pollutants through ingestion of contaminated fish and fishery products; the consumption of fatty fish such as European catfish can contribute considerably to the intake of dioxins and PCBs. Samples of fish muscle of the top-of-the-food-chain predator Silurus glanis from the northern Italian Lake Varese were analyzed to detect the levels of 17 PCDD/F and 18 PCB congeners. All samples presented detectable levels of PCDD/Fs and PCBs, with concentrations ranging from 0.001 to 1.310pgg(-1) wet weight (w.w.) for Σ PCDD/Fs, 0.031 to 21.000pgg(-1) w. w. for Σ DL-PCBs, and 0.397 to 117.910ngg(-1) w. w. for Σ NDL-PCBs. One sample exceeded the maximum levels of 6.5pgg(-1) w. w. for the sum of PCDD/Fs and DL-PCBs set by European regulations for fish (1259/2011 EU), while other samples exceeded the action levels of 0.75pgg(-1) w. w. for the sum of PCDD/Fs, and 2pgg(-1) w. w. for the sum of DL-PCBs (277/2012 EU). Consuming contaminated catfish may pose a risk for human health, especially for the subpopulation traditionally accustomed to eating this fish, as well as sensitive individuals, such as children and pregnant women.

Oxidative stress related to chlorpyrifos exposure in rainbow trout: Acute and medium term effects on genetic biomarkers.

Organophosphates (OPs) are derivatives of phosphoric acid widely used in agriculture as pesticides. Chlorpyrifos (CPF) is an OP that is extremely toxic to aquatic organisms. Rainbow trout (Oncorhynchus mykiss) is considered as a sentinel model species for ecotoxicology assessment in freshwater ecosystems. An exposure study was carried out on rainbow trout to investigate genetic responses to CPF-induced oxidative stress by Real-Time PCR, and to determine the accumulation dynamics of CPF and toxic metabolite chlorpyrifos-oxon (CPF-ox) in edible parts, by HPLC-MS/MS. Among the genes considered to be related to oxidative stress, a significant increase in HSP70 mRNA levels was observed in liver samples up to 14 days after CPF exposure (0.05 mg/L). CPF concentrations in muscle samples reach mean values of 285.25 ng/g within 96 hours of exposure, while CPF-ox concentrations were always under the limit of quantification (LOQ) of the applied method. Our findings lead us to consider HSP70 as a suitable genetic marker in rainbow trout for acute and medium-term monitoring of CPF exposure, complementary to analytical determinations.

Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA): emerging contaminants of increasing concern in fish from Lake Varese, Italy.

Perfluoroalkylated substances (PFASs) are highly fluorinated aliphatic compounds with high thermal and chemical stability, used in a range of industrial applications. Extensive screening analyses in biota samples from all over the world have shown the bioaccumulation of PFAS into higher trophic levels in the food chain. Perfluorooctane sulfonic acid (PFOS) and perfluoroctanoic acid (PFOA) are potential reproductive and developmental toxicants and are considered to be emerging endocrine disrupters. Ingestion of fish and other seafood is considered the main source of exposure of these contaminants. Here, we quantified PFOS and PFOA by LC-MS/MS in muscle samples of European perch from Lake Varese, Italy. PFOS was detected in all samples with concentrations of up to 17.2 ng g(-1). Although the reported values were lower than the recommended total daily intake (TDI) proposed by the European Food Safety Authority (EFSA), fish from Lake Varese may be a significant source of dietary PFOS exposure.

Retrospective data analysis of animal poisoning events in Liguria.

Intentional poisoning represents a serious risk to domestic and wild animals, and it can be an environmental and human health issue as well . This paper is a retrospective study, which covers a decade, based on animal poisoning cases and poisoned baits that were submitted for diagnostic examinations to the Veterinary Medical Research Institute for Piedmont, Liguria and the Aosta Valley (IZS-PLVA) in Liguria region. All data were collected through a passive surveillance system introduced in Italy by a decree of the Ministry of Health in January 2009. 43.2% of the animal poisoning cases were confirmed by toxicological analysis, whereas toxic agents were detected in 31.1% of the baits. The most affected animal species were dogs and cats, followed by synanthropic birds,. Only 4% of the total poisoning events analysed involved wild animals and cases of livestock poisoning were minimal. An increased number of cases in January, March, April and August was noticed, but no seasonal trend was detected. The most affected areas were the ones with the highest level of urbanization and population density. The major cause of the poisonings and the most common substances detected in the examined baits were anticoagulants whereas cholinesterase inhibitors, organochlorine pesticides and carbamates were detected in a minor number of cases. This study raises concerns about deliberate animal poisoning in ligurian region and highlights the necessity to fight this phenomenon as it endangers animals, humans and environment.

A novel family with Lamin B1 duplication associated with adult-onset leucoencephalopathy.

BACKGROUND AND AIMS: Duplication of the lamin B1 gene (LMNB1) has recently been described in a rare form of autosomal dominant adult-onset leucoencephalopathy. The aim of the study was to evaluate the presence of LMNB1 gene defects in a series of eight patients with diffuse adult-onset hereditary leucoencephalopathy. METHODS: Clinical features of tested patients included a variable combination of pyramidal, cerebellar, cognitive and autonomic dysfunction. Neuroradiological data (MRI) showed symmetrical and diffuse white-matter lesions in six cases, and multifocal confluent lesions in two. LMNB1 full gene deletion/duplication and point mutations were searched using a TaqMan real-time PCR assay and direct sequencing of all coding exons. RESULTS: One patient carried a 140-190 kb duplication involving the entire LMNB1 gene, the AX748201 transcript and the 3' end of the MARCH3 gene. Clinical and neuroimaging data of this proband and an affected relative overlapped with the features already described in patients with LMNB1 duplication. Lamin B1 expression was found increased in lymphoblasts. No LMNB1 gene defect was identified in the remaining seven probands. CONCLUSIONS: LMNB1 gene duplication appears characteristic of a subset of adult-onset autosomal dominant leucoencephalopathies, sharing autonomic dysfunction at onset, diffuse T2-hyperintensity of supra- and infratentorial white matter, sparing of U-fibres and optic radiations. The variable phenotypes in the remaining cases lacking LMNB1 defects (five with autosomal dominant transmission) suggest that adult-onset leucoencephalopathies are genetically heterogeneous.

Mutations in the lamin B1 gene are not present in multiple sclerosis.

BACKGROUND: Whole gene duplication of the lamin B1 gene (LMNB1), encoding for a protein of the nuclear lamina, causes an adult-onset autosomal dominant leukodystrophy (ADLD). Clinical features of ADLD (onset in adult life, dysautonomic symptoms, followed by pyramidal and cerebellar dysfunctions) partially resemble those of multiple sclerosis (MS), particularly the primary-progressive form. Our aim was to test whether LMNB1 gene mutations were present amongst patients with a diagnosis of MS. METHODS: One hundred eighty-two MS patients were screened for copy number variations of the LMNB1 gene using a qPCR assay. Point mutations in the LMNB1 gene were searched by denaturing high-performance liquid chromatography and direct sequencing in a subgroup of 16 patients with familial MS. RESULTS: No duplication/deletion of the lamin B1 gene was found amongst MS patients, and no point mutation was identified in the familial cases. CONCLUSION: Our work indicates that lamin B1 defects are probably not responsible for signs and symptoms resembling multiple sclerosis.

Determination of amprolium in feed by a liquid chromatography-mass spectrometry method.

As a consequence of the finding of veterinarian drugs in food European Community banned several compounds like coccidiostats as amprolium (APL). This antibiotic has been used as a preventive and clinical anticoccidial drug in chicken. The 2005/187/CE, 2005/925/EC Recommendations ban the use of amprolium as additive in chicken feed. For this reason a rapid and sensitive liquid chromatography-mass spectrometry (LC-MS) method was developed to detect amprolium in chicken feed following the European community proposed technique (1999/27/EC) for sample preparation. Cause the validation is required for the analytical methods used in feed official control, this method was validated according to 2004/882/EC Regulation.

Trace element occurrence in the Pacific oyster Crassostrea gigas from coastal marine ecosystems in Italy.

The Pacific oyster is one of the world's most widespread bivalves and a suitable species for biomonitoring trace elements in marine environments thanks to its bioaccumulation ability. As it is also an edible mollusc, concentrations of harmful elements in its tissues must be monitored. For these purposes, 464 wild individuals were collected from 12 sites along the Italian coasts. The concentration of fourteen trace elements (Al, As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb, Se, Sn, Tl, and Zn) in their tissues was quantified. Among the three heavy metals, cadmium, lead, and mercury, none exceeded the maximum limit for in food set by European Union regulations but Cd in one sample from the Varano Lagoon resulted extremely close to this value. Contamination by Hg of the northern Adriatic and Orbetello Lagoons was also observed. Moreover, there was a positive association between the lagoon's environmental conditions and the bioaccumulation of this element in oysters. Despite the ban instituted 15 years ago on the use of Sn in antifouling paints, this element is still present in several marine environments, as demonstrated in the oysters sampled from harbour areas. Samples collected from harbours also showed very high concentrations of Cu and Zn due to the ability of oysters to accumulate these elements, which have replaced Sn in antifouling paints. Analysis of the samples from most sites indicated a low risk of human exposure to harmful elements through oyster consumption; nonetheless, chemical sanitary controls should focus primarily on Cd, Cu, and Zn.

Heavy metals and essential elements in Italian cereals.

Crops intended for human nutrition and food production containing different essential trace elements, such as copper and zinc, could be contaminated by toxic metals like cadmium and lead. The interrelationship between micronutrients and contaminant trace elements in different cereals was investigated in North-western Italy, where both agricultural and industrial activities are present. Elemental concentrations in sampled cereals were assessed by inductively coupled plasma mass spectrometry (ICP-MS). Rice, oats and barley reached the highest median levels for Al, Cd and Pb content, while corn samples were less contaminated by toxic metals. Regarding essential elements highest median values of Cu and Zn were both found in barley, while Ni median content was higher in oats. Rice had the lowest median levels of essential elements. The correlation study between toxic and essential elements seemed to demonstrate fixed trends in analysed samples, corroborating the importance of a different diet to limit potential adverse effects caused by toxic elements.

Presence of trace metals in aquaculture marine ecosystems of the northwestern Mediterranean Sea (Italy).

Information regarding chemical pollutant levels in farmed fish and shellfish, along with the risks associated with their consumption is still scarce. This study was designed to assess levels of exposure to 21 trace elements in fish (Dicentrarchus labrax), mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) collected from aquaculture marine ecosystems of the northwestern Mediterranean Sea. Metal concentrations showed great variability in the three species; the highest values of the nonessential elements As and Cd were found in oysters while the highest levels of Al, Pb and V were found in mussels. The essential elements Cu, Mn and Zn were highest in oysters, but Fe, Cr, Ni, Se, Co and Mo levels were highest in mussels. Fish had the lowest concentrations for all trace elements, which were at least one order of magnitude lower than in bivalves. The rare earth elements cerium and lanthanum were found at higher levels in mussels than in oysters, but undetectable in fish. The maximum values set by European regulations for Hg, Cd and Pb were never exceeded in the examined samples. However, comparing the estimated human daily intakes (EHDIs) with the suggested tolerable copper and zinc intakes suggested a potential risk for frequent consumers of oysters. Similarly, people who consume high quantities of mussels could be exposed to concentrations of Al that exceed the proposed TWI (tolerable weekly intake).

Comparative liver accumulation of dioxin-like compounds in sheep and cattle: Possible role of AhR-mediated xenobiotic metabolizing enzymes.

PCDDs, PCDFs, and PCBs are persistent organic pollutants (POPs) that accumulate in animal products and may pose serious health problems. Those able to bind the aryl hydrocarbon receptor (AhR), eliciting a plethora of toxic responses, are defined dioxin-like (DL) compounds, while the remainders are called non-DL (NDL). An EFSA opinion has highlighted the tendency of ovine liver to specifically accumulate DL-compounds to a greater extent than any other farmed ruminant species. To examine the possible role in such an accumulation of xenobiotic metabolizing enzymes (XME) involved in DL-compound biotransformation, liver samples were collected from ewes and cows reared in an area known for low dioxin contamination. A related paper reported that sheep livers had about 5-fold higher DL-compound concentrations than cattle livers, while the content of the six marker NDL-PCBs did not differ between species. Specimens from the same animals were subjected to gene expression analysis for AhR, AhR nuclear translocator (ARNT) and AhR-dependent oxidative and conjugative pathways; XME protein expression and activities were also investigated. Both AhR and ARNT mRNA levels were about 2-fold lower in ovine samples and the same occurred for CYP1A1 and CYP1A2, being approximately 3- and 9-fold less expressed in sheep compared to cattle, while CYP1B1 could be detectable in cattle only. The results of the immunoblotting and catalytic activity (most notably EROD) measurements of the CYP1A family enzymes were in line with the gene expression data. By contrast, phase II enzyme expression and activities in sheep were higher (UGT1A) or similar (GSTA1, NQO1) to those recorded in cattle. The overall low expression of CYP1 family enzymes in the sheep is in line with the observed liver accumulation of DL-compounds and is expected to affect the kinetics and the dynamics of other POPs such as many polycyclic aromatic hydrocarbons, as well as of toxins (e.g. aflatoxins) or drugs (e.g. benzimidazole anthelmintics) known to be metabolized by those enzymes.

Dioxins, DL-PCB and NDL-PCB accumulation profiles in livers from sheep and cattle reared in North-western Italy.

Products of animal origin represent the main route of human exposure to dioxins and dioxin-like PCBs (DL-compounds). Recently, concerns have been raised about ovine products, particularly the liver, in which relatively high levels of DL-compounds have been reported. We surveyed ovine and bovine livers in areas with no known sources of dioxin or DL-PCB contamination, in order to assess accumulation patterns for both DL-compounds and non-DL (NDL-) PCBs. None of the ovine and bovine samples exceeded the current Maximum Limits (MLs) for DL-compounds. Liver DL-compound TEQ concentrations were up to 5-fold higher in sheep than in cows. No statistically significant differences in total NDL-PCBs levels were found. The main contributors to TEQ levels were the Penta- and Hexa-chlorinated PCDFs and PCB 126. The results confirm the increased bioaccumulation in ovine liver towards specific DL-compounds even in ewes reared in areas with no known sources of PCDD/Fs or DL-PCBs contamination.

Human exposure to metals due to consumption of fish from an artificial lake basin close to an active mining area in Katanga (D.R. Congo).

The concentrations of 14 essential and nonessential trace elements were determined in fish from Lake Tshangalele, Katanga province, Democratic Republic of Congo. This province has been a place of intensive mining activities for centuries, which have increased in recent years, due to the use of metals such as copper and cobalt for the industries of fast-growing countries. Lake Tshangalele, which receives effluents from metallurgical and mining plants in Likasi, is home to several fish species that are an important part of the diet of the local population, and, therefore, it constitutes a relevant site for documenting the human exposure to metals as a result of a fish diet. The highest concentrations (median levels, dry weight) of cobalt (7.25mgkg(-1)), copper (88.1mgkg(-1)), iron (197.5mgkg(-1)), manganese (65.35mgkg(-1)), zinc (122.9mgkg(-1)) and aluminum (135.4mgkg(-1)) were found in fish collected closest to the copper mining plant, with decreasing concentrations along the lake, up to the dam. In the most contaminated fish samples, values of up to 270.1mgkg(-1) for Al, 173.1mgkg(-1) for Cu, 220.9mgkg(-1) for Zn, 211.0mgkg(-1) for Mn, 324.2mgkg(-1) for Fe, 15.1mgkg(-1) for Co, 4.2mgkg(-1) for Cr, 1.6mgkg(-1) for Cd, 1.9mgkg(-1) for Pb, and 1.8mgkg(-1) for Ni were found. Metal contamination from mining activity resulted in being of great concern because of potential health risks to the local inhabitants due to the consumption of heavily contaminated fish. CAPSULE: High levels of metals, especially cobalt, aluminum, iron, manganese, zinc and cadmium were found in fish from Tshangalele water system.

Mercury and selenium in European catfish (Silurus glanis) from Northern Italian Rivers: can molar ratio be a predictive factor for mercury toxicity in a top predator?

The study of mercury and selenium bioaccumulation in fish is crucially important for evaluating the extent of contamination in freshwater environments, and the possible health risk posed for humans when the antagonistic interactions of these two elements are considered. Several factors affect the risk of mercury intake from fish consumption, including mercury levels, human consumption patterns, and sensitive populations (e.g., pregnant women, foetuses, young children and unknown genetic factors). The protective effects of selenium on mercury toxicity have been extensively publicised in recent years, particularly targeting fish consumers. In this study, mercury (Hg) and selenium (Se) concentrations were determined in the muscle of European catfish (Silurus glanis) collected from North Italian Rivers. Differences in mercury and selenium levels, as a function of size, gender and location were investigated. Hg was strongly related to length, gender and location, while Se levels are not dependent on fish size or location. The mean Se/Hg molar ratio was strongly affected by location, and significantly related to length and age. Selenium was in molar excess of mercury in all sites, with a rank order of mean Se/Hg molar ratio of the Parma River (2.55)>Po River (1.71)>Tanaro River (1.66)>Bormida River (1.36). However, in 37% of analyzed samples, Hg exceeded the maximum level set by 1881/2006/EC and 629/2008/EC in fish muscle. The molar ratio of Se/Hg was <1 only in the presence of significantly high Hg levels (>0.5mg/kg), and therefore the mean molar ratio cannot be considered as a safety criterion in top predator fish.

Human dietary exposure and levels of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), dioxin-like polychlorinated biphenyls (DL-PCBs) and non-dioxin-like polychlorinated biphenyls (NDL-PCBs) in free-range eggs close to a secondary aluminum smelter, Northern Italy.

PCDD/Fs and PCBs are environmentally persistent substances that have been associated with adverse effects on human health. Contamination of soils, animal feed and pastures leads to their bioaccumulation of in food products of animal origin, which are considered the major source of intake of these contaminants in humans. We analyzed eggs from free-range hens, sampled from small farms, located within a distance of 4.5 km from a secondary aluminum smelter in Northern Italy. The concentrations of PCDD/Fs, DL-PCBs and NDL-PCBs were higher in eggs from locations close to the plant, and strongly exceeded the limits set by EU Regulation 1259/2011 (2.5 pg WHO TEQ fat g(-1) for PCDD/Fs, 5.0 pg WHO TEQ g(-1) for PCDD/Fs and DL-PCBs L, 40 ng g(-1) for NDL-PCBs). Without the prompt and immediately applied control strategies, the consumption of contaminated eggs may have posed [corrected] a risk for human health, especially for children (≤9 years) and infants (≤3 years), due to the 2-fold excess of the current exposure limits.

Analysis of mercury and methylmercury concentrations, and selenium:mercury molar ratios for a toxicological assessment of sperm whales (Physeter macrocephalus) in the most recent stranding event along the Adriatic coast (Southern Italy, Mediterranean Sea).

Mass stranding of sperm whales (Physeter macrocephalus) is a rare event in the Mediterranean Sea. In September 2014, a pod of seven sperm whales became stranded along the Adriatic coast of Southern Italy. This is the seventh occurrence of this type since 1555 in this sea basin. Total concentrations of mercury (T-Hg), methylmercury (MeHg) and selenium (Se) were measured from brain, muscle, liver and kidney of three female sperm whales, which died in this event. Analyses showed considerable, age-dependent variations in Hg and Se concentrations in the different organs. The contamination levels of T-Hg in the liver (up to 200 mg kg(-1)) and brain (up to 21 mg kg(-1)) samples were markedly higher than those in the kidney and muscle samples. The liver and brain also showed the highest Se levels. Se:Hg molar ratios ⩾1 were observed in all the organs of the three sperm whales, suggesting that Se could protect the animals from Hg toxicity. The risk of Hg-associated neurotoxicity was assessed by comparing our values to thresholds set for neurotoxicity in mammals, and the role of Se in the detoxification process of T-Hg/MeHg is discussed herein.

Non-dioxin-like polychlorinated biphenyls (NDL-PCBs) in eel, trout, and barbel from the River Roya, Northern Italy.

Non-dioxin-like polychlorinated biphenyls (NDL-PCBs) contribute to the dietary intake of total PCBs. They are consumed via ingestion of contaminated fish and fishery products. Thus, it is important to quantify their levels in aquatic organisms in order to evaluate the risks associated with fish consumption by humans. In 2013, an advisory against consumption of fish from the River Roya (IT) was disseminated after NDL PCBs were found to exceed maximum levels set by EU Regulations. We investigated the presence of NDL PCBs in eel, trout and barbel from the River Roya. We found concentrations in the range of 9.2-27.6 ng g(-)(1)ww in barbel, 9.2-97.0 ng g(-1)ww in trout and 9.0-239.5 ng g(-1)ww in eel. The distribution of congeners among different species did not show significant variations and was characterized by a higher proportion of 153, 138 and 180 congeners, according to their high resistance to metabolic degradation.

Development, validation and application to real samples of a multiresidue LC-MS/MS method for determination of ß2 -agonists and anabolic steroids in bovine hair.

ß(2) -agonists are often abused in cattle breeding because of their effects on animal growth and meat properties. The use of ß(2) -agonists as growth promoters is forbidden in the European Union (Council Directive 96/23/EC classifies them into group A of Annex I), due to their toxicity and carcinogenic properties, as for anabolic steroids, which are often administered in combination with ß(2) -agonists, to promote the storage of proteins and increase muscle size. A unique confirmatory liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the quantitative detection of 13 ß(2) -agonists and anabolic steroids plus the qualitative identification of other three analytes in bovine hair was developed and validated, according to Decision 2002/657/CE. Hair samples were washed with dichloromethane, digested within a NaOH solution and subjected to liquid-liquid extraction. The analysis was performed by high performance liquid chromatography coupled to a triple quadrupole mass spectrometer operating in the selected reaction monitoring mode. The absence of matrix interferents, together with good repeatability of both retention times and relative abundances of diagnostic transitions, allowed the correct identification of all analytes. The quantitative calibrations obtained from spiked blank hair samples proved linear in the range tested. CCα and CCß ranged from 0.5 ng/g to 30 ng/g. Intralaboratory reproducibility (CV%) ranged between 5.0 and 17.7 and trueness between 96% ± 7% and 105% ± 8%. The applicability of the method to real positive samples was demonstrated for both ß(2) -agonists and anabolic steroids. 17α-boldenone was found in most (70%) hair samples obtained from untreated animals, supporting the hypothesis of endogenous production of this steroid.