Paolo Stacchini

Human exposure to lead, cadmium and mercury through fish and seafood product consumption in Italy: a pilot evaluation

The presence of selected toxic heavy metals, such as cadmium (Cd), lead (Pb) and mercury (Hg), was investigated in fish and seafood products, namely, blue mussel, carpet shell clam, European squid, veined squid, deep-water rose shrimp, red mullet, European seabass, gilthead seabream, Atlantic cod, European hake, Atlantic bluefin tuna and swordfish so as to assess their human exposure through diet. Metals were detected by quadrupole inductively coupled plasma mass spectrometry (Q-ICP-MS) and hydride generation atomic absorption spectrometry (Hg-AAS). Measurements of Cd, Pb and Hg were performed by means of analytical methods validated in compliance with UNI CEI EN ISO/IEC 17025 [2005. General requirements for the competence of testing and calibration laboratories. Milano (Italy): UNI Ente Nazionale Italiano di Unificazione]. The exposure assessment was undertaken matching the levels of Cd, Pb and total Hg with consumption data related to fish and seafood products selected for this purpose. In order to establish human health implications, the estimated weekly intakes (EWIs) for Cd, Pb and Hg were compared with the standard tolerable weekly intakes (TWI) for Cd and provisional tolerable weekly intakes (PTWIs) for Pb and Hg stipulated by the European Food Safety Authority (EFSA) and the Food and Agriculture Organization/World Health Organization (FAO/WHO) Joint Expert Committee on Food Additives (JECFA). The found metal concentrations were largely below the maximum levels (MLs) established at the European Union level with the exception of Cd. This metal exceeded the MLs in squid, red mullet, European hake and Atlantic cod. Squid and blue mussel showed the highest Pb concentrations which accounted for 60% and 10% of the MLs, respectively. Highest Hg levels were found in predatory fish. The concentrations of Hg in swordfish, Atlantic bluefin tuna and red mullet accounted for 50%, 30% and 30% of the MLs, respectively. The EWIs for Cd, Pb and Hg related to the consumption of fish and seafood products by the median of the Italian total population accounted for 20%, 1.5% and 10% of the standard TWI for Cd as well as PTWIs for Pb and Hg, respectively. Furthermore, the EWIs estimated using consumption data concerning Italian consumers did not exceed the standard TWI and PTWIs, except for Cd at 95th percentile.

Influence of laboratory homogenization procedures on trace element content of food samples: an ICP-MS study on soft and durum wheat.

Sample contamination as a consequence of abrasion of grinding tools during the homogenization of food materials to be analysed for trace elements was addressed. The possible release of 15 trace elements (Al, As, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Se, Sn, V, Zn) from six different grinding and milling devices, operating either continuously or discontinuously, was evaluated. All the devices were commercially available and were representative of models usually employed in food and agricultural laboratories. Wheat grains belonging to one soft and one durum cultivar were used as test material. The determination of the analyte concentrations in subsamples submitted to the different preparation treatments was performed by quadrupole inductively coupled plasma mass spectrometry (Q-ICPMS). Accordingly, a suitable digestion method was developed and the ArC+ interference affecting Cr determination was evaluated and corrected. Statistical differences with respect to the control were detected for 10 elements (Al, Cd, Co, Cr, Cu, Fe, Mn, Mo, Ni, Pb) and in most cases contamination of the samples was traced back to the composition of the grinding equipment. None of the investigated devices was contamination-free with respect to all of the quantified elements. Abrasion of the grinding tools was higher with durum wheat than with soft wheat as a consequence of their different hardness.

4‐hexylresorcinol as inhibitor of shrimp melanosis: Efficacy and residues studies; evaluation of possible toxic effect in a human intestinal in vitro model (caco‐2); preliminary safety assessment

Studies were performed on the efficacy, residues and in vitro enterocyte toxicity of 4-hexylresorcinol (4-HR), which could be utilized as an inhibitor of shrimp melanosis (black spot). Mediterranean sea shrimp (Parapaeneus longirostris) were treated with solutions of 4-HR in sea-water, at three different concentrations, 25, 50 or 100 mg/kg of shrimp, to test its antioxidative property. As a comparison a group of shrimp was treated with sodium metabisulphite (1 g/kg), while another group was left untreated. 4-HR showed a marked ability to inhibit or slow down melanosis (black spot) in shrimp; the most effective concentration was 100 mg/kg within an optimum period of 7 days but with effects up to the tenth day. During the first 5 days, 4-HR residues in the edible part of the shrimp showed a fast decrease in all three groups, going from initial average values of 20 mg/kg at 0 time, to 0.9 in the group treated at 25 mg/kg; from 42 to 1.8 mg/kg in the group at 50 mg/kg and from 85 to 1.9 mg/kg in the group at 100 mg/kg. In vitro studies on enterocyte-like Caco-2 cells did not indicate any cytotoxic effect up to a concentration of 50 micrograms/ml. Moreover, no inhibition of protein synthesis was observed, which lends further support to the absence of significant damage to the intestinal mucosa induced by 4-HR. The available database on 4-HR pharmacology and toxicology is inadequate to determine even a provisional ADI. There is negative evidence of carcinogenesis and no significant untoward effects were observed in humans when it was used as an anthelmintic. However, it is not possible to determine a NOEL for non-genotoxic effects. 4-HR could become an interesting alternative to the use of sulphites to prevent black spot. However, a more complete database is needed to achieve a regulatory evaluation.

Constituents of aromatic plants: I. Methyleugenol

Methyleugenol (ME) is a natural constituent of the essential oils of a number of plants widely used in foodstuffs as flavouring agents. Its occurrence, routes of intake, and available information on toxicity, kinetics and metabolism are reviewed. In view of the carcinogenic potential of ME, the need to check its presence in food products with effective analytical methods is pointed out.

Cadmium in organs and tissues of horses slaughtered in Italy.

The cadmium content of muscle, liver, kidney and blood samples from 62 horses slaughtered in Italy was investigated. Cadmium was determined by graphite furnace atomic absorption spectrometry (GFAAS) after wet digestion of the samples. The mean and median contents of all samples were (on a fresh weight basis) 75 and 41 μgkg-1 for muscle, 2.46 and 2.10 mgkg-1 for liver, 20.0 and 13.5 mgkg-1 for kidney. The cadmium level in blood samples was always below 6 μgl-1. The cadmium concentrations in muscle, liver and kidney were found to be related to the life span of the specimens and increased with age. Females exhibited higher levels than males, but this difference was significant only in the case of muscle tissue. The geographical origin was recognized as the main factor influencing the cadmium content of the equine specimens analysed. The differences between horses coming from the three main breeding countries considered in this study (Poland, Lithuania, Hungary) were marked and statistically significant. The average intake of cadmium from equine meat was estimated for the general population and for population groups resident in areas with high consumption of this food item. While the cadmium intake from equine meat for the average consumer accounts for about 1% of the total cadmium intake estimated for Italy, in the latter case the enhanced consumption of equine muscle is often accompanied with the consumption of substantial quantities of equine liver and this may lead to high cadmium intakes.

Chromium determination in foods by quadrupole inductively coupled plasma–mass spectrometry with ultrasonic nebulization

Abstract The analytical issues connected with chromium determination in foodstuffs by quadrupole inductively coupled plasma–mass spectrometry (Q-ICP-MS) were addressed, including signal stability, spectral interferences and the use of mathematical correction equations. The analytical performance was compared to that of electrothermal atomisation–atomic absorption spectrometry (ETA-AAS), selected as reference method. Five food certified reference materials (CRMs), including two meat-based CRMs not previously characterized for their Cr content, were included in the study. The use of ultrasonic nebulization (UN) and the adoption of 53Cr as analytical mass allowed precise and accurate results to be obtained by Q-ICP-MS, with lower detection limits than ETA-AAS.

Determination of the banned growth promoter moenomycin A in feed stuffs by liquid chromatography coupled to electrospray ion trap mass spectrometry

Flavomycin complex is an antibiotic banned in the European Union as an additive in feed stuffs. As a consequence, the monitoring programmes for official control within the Community require analysis of feeds for possible illegal use of flavomycin. A method for unambiguous identification and quantification of moenomycin A, the main pharmacologically active component of flamomycin complex, in several feeds by liquid chromatography coupled to electrospray ion trap mass spectrometry (LC/ESI-MS/MS) is herein described for the first time. The method was developed to be used as a confirmative analytical tool for the network of Italian official control laboratories; both the singly and doubly charged molecular ions were observed as precursor ions, from which four product ions were selected for both quantitative analysis and unambiguous identification of moenomycin A. The method was in-house validated for feeds in the concentration range 0.50-30.0 microg/g, according to the Regulation 882/2004/EC requirements. Mean recoveries ranging between 83.9-94.2% and relative standard deviations <23% account for method trueness and repeatability, respectively. Moreover, other analytical performance parameters, i.e. method specificity, ruggedness, the linearity of detector response, the limit of quantification (LOQ), the limit of detection (LOD), and measurement uncertainty were evaluated and reported. The ion trap LC/ESI-MS/MS method is highly selective and reliable; high drug recovery, good reproducibility and an LOQ down to 0.10 microg/g guarantee its applicability for confirmatory purposes in the official control activity in Italy.

Cadmium bioaccumulation in Mediterranean spider crab (Maya squinado): human consumption and health implications for exposure in Italian population.

Cd bioaccumulation pattern was investigated in Mediterranean spider crab (Maya squinado, Herbst, 1788) collected from the northern Adriatic Sea. Specimens were caught in the framework of a monitoring plan in order to quantify the Cd distribution into different organs and tissues of crab. For this purpose, Cd level was studied in appendages, cephalothorax, abdomen as well as gonads. Cd concentrations were found largely below the Maximum Level (ML) established at the European Union (EU) level for muscle from crab appendages (found mean 0.011 mg kg(-1)) and approximately amounted to 2% of the EU ML (0.50 mg kg(-1)). The higher Cd concentrations were found in organs and tissues included in crab body such as abdomen, chephalotorax and gonads with respect to appendages. Chephalotorax showed the highest metal concentration (mean value of 1.19 mg kg(-1)). The possible differences in Cd bioaccumulation rate among crab organs and tissues were also investigated applying a parametric linear regression. A major Cd bioaccumulation rate was revealed in chephalotorax with respect to other analyzed organs and tissues. Furthermore, the evaluation of health risk related to human consumption of the Mediterranean spider crab has been studied for median of total population, median and 95th percentile of consumers of Italy. The observed results highlighted that the consumption of organs and tissues included in crab body such as abdomen, gonads and, in particular, chephalotorax substantially increased the Cd intake reaching also alarming Estimated Weekly Intake (EWI) values especially for median and 95th percentile of Italian consumers.

A basis for estimation of consumption: Literature values for selected food volatiles. Part III∗

Quantitative data on volatile compounds have been reported in 16 food items. No publications reporting quantitative data were found for two of these 16 food products, i.e. avocado and jackfruit. About 550 volatile compounds have been assayed globally in the other 14 food products. Mango and raspberry were the products with the greatest number of volatile compounds; the most representative substances were benzaldehyde, ethyl acetate, limonene, and 2-phenylethanol.

European official control of food: Determination of histamine in fish products by a HPLC-UV-DAD method.

The evaluation of histamine content in fish and fishery products, responsible for scombroid poisoning, is essential to guarantee the safety of food. EU regulation requires validated analytical methods to ensure the verification of compliance with food law in official control activity. To this aim a previous gradient RP-HPLC method with DAD detection was modified and validated, according to international guidelines. The reliability of results was tested by analysing fish reference materials within the participation in European proficiency tests. The method has been used for the analysis of real samples consisting of several fish-based products with considerable differences in matrix composition. This characteristic is of great relevance to be able of apply the method in the field of official control.