Chromatographic Methods for the Determination of Glyphosate in Cereals Together with a Discussion of Its Occurrence, Accumulation, Fate, Degradation, and Regulatory Status.

The European Union's recent decision to renew the authorization for the use of glyphosate until 15 December 2033 has stimulated scientific discussion all around the world regarding its toxicity or otherwise for humans. Glyphosate is a chemical of which millions of tons have been used in the last 50 years worldwide to dry out weeds in cultivated fields and greenhouses and on roadsides. Concern has been raised in many areas about its possible presence in the food chain and its consequent adverse effects on health. Both aspects that argue in favor of toxicity and those that instead may indicate limited toxicity of glyphosate are discussed here. The widespread debate that has been generated requires further investigations and field measurements to understand glyphosate's fate once dispersed in the environment and its concentration in the food chain. Hence, there is a need for validated analytical methods that are available to analysts in the field. In the present review, methods for the analytical determination of glyphosate and its main metabolite, AMPA, are discussed, with a specific focus on chromatographic techniques applied to cereal products. The experimental procedures are explained in detail, including the cleanup, derivatization, and instrumental conditions, to give the laboratories involved enough information to proceed with the implementation of this line of analysis. The prevalent chromatographic methods used are LC-MS/MS, GC-MS/SIM, and GC-MS/MS, but sufficient indications are also given to those laboratories that wish to use the better performing high-resolution MS or the simpler HPLC-FLD, HPLC-UV, GC-NPD, and GC-FPD techniques for screening purposes. The concentrations of glyphosate from the literature measured in wheat, corn, barley, rye, oats, soybean, and cereal-based foods are reported, together with its regulatory status in various parts of the world and its accumulation mechanism. As for its accumulation in cereals, the available data show that glyphosate tends to accumulate more in wholemeal flours than in refined ones, that its concentration in the product strictly depends on the treatment period (the closer it is to the time of harvesting, the higher the concentration), and that in cold climates, the herbicide tends to persist in the soil for a long time.

Organochlorines as contaminants in butter and other shortenings: occurrence and exposure assessment.

In the present work, experimental measurements were carried out on dietary fats purchased in stores or bought online and produced in different countries of the world. Shortenings of animal origin (lard, butter) or vegetable-made (margarine, coconut oil, hydrogenated palm/soybean oil) were selected. The concentrations of organochlorines (OC pesticides and PCBs) were measured to ascertain whether the level of these contaminants in food has decreased since they were banned and to assess the today consumer exposure to such xenobiotics. Analyses were carried out by GC-ECD and GC-MS according to the United Nations Environment Program on persistent organic pollutants. Organochlorines were detected in 85% of the samples analysed. The observed levels were not high (maximum value 5.54 ± 2.33 ng g-1) and point out a slow decline over the last years. Butter from Europe was the most studied matrix showing HCB and 4,4'-DDE as the main organochlorines that were constantly detected above the limit of quantitation. This can be explained if the past use of the two pesticides is considered. Consumption data from the EFSA database were combined with the HCB and 4,4'-DDE concentrations measured in the present research to obtain the Estimated Daily Intakes (EDIs) of the two contaminants from butter. The comparison with the Tolerable Daily Intakes (TDIs) set by World Health Organization indicates that the observed EDIs are far below the TDIs, so allowing to conclude that the risk deriving from the intake of the investigated organochlorines through butter is acceptably low for European consumers. Results relating to some non-Aroclor PCBs detected in shortenings, such as PCB 47, which have recently been discovered to be released into the environment are also reported.

Authenticity assessment of dairy products by capillary electrophoresis.

Milk and derivatives are a very important part in the diet of the world population. Products from goat, buffalo, and sheep species have a greater economic value than the cow ones, therefore, authenticity frauds by improperly adding cow's milk occur frequently: dairy products are among the seven more attractive foods for adulteration. Milk from each of the above-cited animal species has its own definite profile of whey proteins (variants of α-lactalbumin and ß-lactoglobulin) and its definite profile of caseins (variants of αS1 -, αS2 -, ß-, and κ-casein). Such proteins can be usefully exploited as markers of authenticity by using capillary electrophoresis which is the technique of choice for the analysis of proteins. Due to the multiple adjustable parameters that are unknown to other analytical techniques, capillary electrophoresis is able to detect frauds in milk mixtures and cheese with little use of solvents, fast analysis time, and ease of operation. This makes it attractive and competitive for routine checks that are very important to fight the adulteration market. Advantages and limitations are discussed.

Quality of Fish-Oil-Based Dietary Supplements Available on the Italian Market: A Preliminary Study.

The global market of food supplements is growing rapidly with a large turnover. Fish oil supplements represent a significant part of this turnover as they are believed to have important health benefits. Conversely, there are few papers in the literature about the quality control of fish oil capsules. As prior studies illustrate, a perfect agreement with the label is rarely found, and in some isolated cases, large amounts of soybean oil are also detected, indicating a true adulteration rather than a non-compliance with the label. None of the available studies refer to the Italian market, which ranks first in Europe in the consumption of food supplements. In this present communication, a quality control of fish-oil-based supplements from the Italian market was carried out for the first time. With minor deviations, all results showed substantial agreement with the label. However, the most important conclusion from this research is that compliance with the label is not enough to judge a product of good quality. The analysis of the overall fatty acid composition showed that some supplements have a high level of saturated fatty acids, and therefore they did not undergo a proper purification process. This may represent a safety issue since the purification process also allows the removal of toxic contaminants.

Non-dioxin-like PCBs: a survey on fishery and aquaculture from the Mediterranean area.

A sampling campaign from 21 sites in Italy was conducted: 15 species from fishery and three species from aquaculture, for a total of 40 determinations, were considered. A careful sample preparation preceded the instrumental analysis that was carried out by means of GC-ECD and GC-MS. Good laboratory practice was achieved by the participation in proficiency tests, by the use of certified reference materials and by applying other directives recommended by international organisations. Concentrations measured in this work were compared with a TDI proposed by some international bodies: for a person weighing 70 kg one-third of the samples from fishery, when consumed, lead to exceed this TDI if the average fish daily consumption per capita is considered. Based on the data obtained here some hypotheses on environmental spreading and influence of PCBs on human health are made. Some suggestions about the preparation of fish for consumption are also given.

Fish contamination by polychlorobiphenyls: the mass spectrometric ortho effect in a new and easy gas chromatography-mass spectrometry method for the analysis of the seven indicators. The case of Bluefin Tuna.

A simple instrumental procedure was developed to carry out the not simple analysis of PCBs in fish samples. PCBs with the same degree of chlorination (the isomers) are expected to be totally indistinguishable among them by all existing detectors and by all existing mass spectrometers, and there is no apparent solution in those frequent cases where two isomers chromatographically coelute. Generally such coelutions are solved by means of multidimensional GC, but it is a complex technique impractical for most laboratories. The present research focuses on the seven important "indicator PCBs" by using the mass spectrometer in an innovative way. The "mass spectrometric ortho effect" was usefully exploited in addressing coelutions between isomers. Other new important observations on mass spectra were decisive in solving the apparent problem arising from coelutions between higher chlorinated PCBs with the lower chlorinated ones when low-resolution MS is used. With the proposed procedure, the seven indicators are analyzed in a simple way and with a degree of accuracy never observed with the conventional gas chromatography. The method was applied to some Bluefin Tuna fish samples of big size suspected to have not negligible levels of PCBs due to the high position of this species in the food chain. The supposition was partly confirmed. On the basis of the results here obtained, the recently introduced EU Regulation on six of the seven indicators shows one critical point: in the present paper, an amendment to the Regulation is proposed. A number of important validation measures are reported.

Organochlorine pesticide residues: an extensive monitoring of Italian fishery and aquaculture.

A sampling campaign from 21 sites in Italy was carried out to investigate the presence of organochlorine pesticide residues in different fish species. Samples came from marine fishery and either from sea- or freshwater aquaculture. Fish feed used in some fish farms were also analyzed. Pesticides studied belong to Persistent Organic Pollutants widely used in the past such as DDT, chlordane, heptachlor, and others. To ensure good quality results and proper data validation the main existing guidelines in the field were applied. The instrumental technique was a Dual column-Dual detector Gas Chromatography (GC-ECD and Ion Trap GC-MS) which allowed that complementary data on the same sample were acquired. Results for fishery showed a wide range of concentrations depending from the area and species examined. DDT, the major OC pesticide detected, varied from 0.02 to 130.03 ng g(-1) edible portion. As regards the products of aquaculture we observed slightly lower average levels of pollutants in a more narrow range of concentration: this is probably due to fish feed used as shown by some measures performed in the present study. Organochlorine pesticide residues were detected in all samples examined but they were generally well below the existing tolerance or action levels. Also the estimated daily intakes are well below than those recommended by WHO. This is a good indication about OCPs in the areas investigated but some further considerations on fish safety must be taken into account. An example on how fishes may act as bioindicators is reported.

Analysis of fatty acids in 12 Mediterranean fish species: advantages and limitations of a new GC-FID/GC-MS based technique.

When fatty acids in fish are analyzed, results in percentage form (profile analysis) are mostly reported. However, the much more useful results expressed as mg/100 g (absolute analysis) is the main information required. Absolute methods based on calibration curves are of good accuracy but with a high degree of complexity if applied to a great number of analytes. Procedures based on the sequence profile analysis-total FA determination-absolute analysis may be suitable for routine use, but suffer from a number of uncertainties that have never been really resolved. These uncertainties are mainly related to the profile analysis. In fact, most profile analyses reported in the literature disagree about the number and type of fatty acids monitored as well as about the total percentage to assign to their sum so leading to possible inaccuracies; in addition the instrumental response factor for all FAME (fatty acid methyl esters) is often considered as a constant, but this is not exactly true. In this work, a set of 24 fatty acids was selected and studied on 12 fish species in the Mediterranean area (variable in lipid content and month of sampling): in our results, and in these species, this set constitutes, on average, 90 ± 3 % of the total fatty acid content. Moreover the error derived from the assumption of a unique response factor was investigated. Two different detection techniques (GC-FID and GC-MS) together with two capillary columns (different in length and polarity) were used in order to acquire complementary data on the same sample. With the protocol here proposed absolute analyses on the 12 cited species are easily achievable by the total FA determination procedure. The accuracy of this approach is good in general, but in some cases (DHA for example) is lower than the accuracy of calibration-based methods. The differences were evaluated on a case by case basis.

New trends in the seafood market. Sutchi catfish (Pangasius hypophthalmus) fillets from Vietnam: Nutritional quality and safety aspects.

Sutchi catfish (Pangasius hypophthalmus) produced in the freshwater basins of Vietnam, available on the Italian market as frozen or thawed fillets, were studied for their nutritional quality and safety aspects. Proximate composition, mineral content, fatty acid profile, unsaponifiable components of the lipid fraction and drip loss during thawing at 5°C were determined on the fillets. Fillets were characterised by high moisture levels (80-85%) and low protein (12.6-15.6%) and lipid (1.1-3.0%) contents. Total lipids were characterised by low cholesterol levels (21-39mg/100g), high percentages of saturated fatty acids (41.1-47.8% of total fatty acid) and low percentages of polyunsaturated fatty acids (12.5-18.8% of total fatty acids), which were mainly represented by linoleic acid (44-59% of total polyunsaturated fatty acids). The mineral composition was characterised by a high sodium content (222-594mg/100g), probably partially due to the sodium tripolyphosphate (E 451) used to retain moisture. As regards safety aspects, the quality of the samples analysed was good, with low residue levels of mercury, organochlorine pesticides and polychlorinated biphenyls.