Ambrogina Albergamo

Heavy metals in aromatic spices by inductively coupled plasma-mass spectrometry

ABSTRACT Objective of this study was to determine the content of Cd, Hg, As and Pb in common spices traded in the Italian market, using inductively coupled plasma-mass spectrometry (ICP-MS). The results were compared with the maximum limits established by the national Legislative Decree (LD) no. 107 implementing the Council Directive 88/388/EEC and by international organisations, such as Food and Agriculture Organization (FAO) and World Health Organization (WHO). Food safety for spices was assessed considering the tolerable weekly intake (TWI) and the provisional tolerable weekly intake (PTWI), respectively, for Cd and Hg and the 95% lower confidence limit of the benchmark dose of 1% extra risk (BMDL01) for As and Pb. Investigated elements in all samples were within the maximum limits as set by the national and international normative institutions. Nevertheless, the heavy metal content of some spices exceeded the PTWI, TWI and BMDL01, which needs attention when considering consumer’s health.

Metabolite and mineral profiling of “Violetto di Niscemi” and “Spinoso di Menfi” globe artichokes by 1H-NMR and ICP-MS

Abstract Globe artichoke has been long considered a nutraceutical food for its valuable content of bioactive compounds. However, beside a well-known polyphenol profile, poor information is available about its metabolite and mineral composition. The aim of this study was to investigate edible parts of Sicilian artichokes, ‘Spinoso di Menfi’ and ‘Violetto di Niscemi’, by 1H NMR and ICP-MS for elucidating these compositional aspects. Although bracts and hearts of both artichokes shared a very similar metabolite pattern, ‘Spinoso di Menfi’ showed a higher number of metabolites, such as amino acids and polyphenols, than ‘Violetto di Niscemi’. ‘Spinoso di Menfi’ was also marked by higher levels of macro- and microelements when compared to ‘Violetto di Niscemi’. Also, artichoke heart demonstrated to accumulate higher mineral levels than bracts. 1H NMR and ICP-MS successfully profiled metabolites and metals in such plant food, partially covering the lack of literature data about ‘Spinoso di Menfi’ and ‘Violetto di Niscemi’ artichokes.

Nano Liquid Chromatography Directly Coupled to Electron Ionization Mass Spectrometry for Free Fatty Acid Elucidation in Mussel

Recently the miniaturization of liquid chromatography (LC) systems and progresses in mass spectrometry instrumentation have enabled direct introduction of the effluent coming from a nanoLC column into the high-vacuum region of an electron ionization source. In the present research, a nanoLC system was directly coupled to an electron ionization mass spectrometer (EI-MS) without any interface or modification of the ion source. The advantage with respect to atmospheric pressure ionization techniques, normally coupled with LC, is major identification power because of a more extensive and reproducible fragmentation pattern, without any matrix effect or mobile-phase interference. In particular, a nanoLC/EI-MS method was developed for elucidation of the free fatty acid profile in mussel samples, avoiding a previous derivatization step, required when gas chromatographic analysis is involved. A total of 20 fatty acids were reliably identified through the comparison with commercial libraries. A quantitative determination was also carried out by using the response factors approach along with the internal standard method, allowing for quantification of 14 fatty acids. Among them, palmitic acid resulted the most abundant, followed by ω6 arachidonic acid. The quantitative data were compared with those obtained by a well-established technique, such as gas chromatography with flame ionization detection (GC-FID). Both nanoLC/EI-MS and GC-FID methods were validated and similar results were obtained in terms of limit of detection and quantification, resulting in the picomole range, and sensitivity as well was not significantly different, as demonstrated by comparing the slope values of the calibration curves (p < 0.05, from a t-test).

Sample preparation techniques coupled to advanced chromatographic methods for marine organisms investigation

OBJECTIVE of this work was to develop suitable extraction methodologies for the isolation of lipids from fish, mussels and clams from the Mediterranean sea, and their successive analysis by means of advanced chromatographic instrumentation. More specifically, three different sample preparation methodologies were adopted: Folchs, Bligh & Dyers and maceration. The lipidic extracts, after application of two different methylation procedures, were subjected to monodimensional and comprehensive two-dimensional GC analyses, in order to compare the fingerprints of samples derived from different extraction and transesterification methodologies. Triacylglycerols (TAGs) were analyzed by an off-line combination of silver-ion liquid chromatography with non-aqueous reversed phase liquid chromatography. In both LC and GC analyses, mass spectrometric detectors were used, which greatly supported the identification procedure. In particular, with respect to HPLC, mass spectrometry with atmospheric pressure chemical ionization in positive mode was applied.

Free fatty acid profiling of marine sentinels by nanoLC-EI-MS for the assessment of environmental pollution effects

The present work aims to elucidate the free fatty acid (FFA) profile of the mussel Mytilus galloprovincialis caged in an anthropogenically impacted area and in a reference site through an innovative and validated analytical approach for the assessment of biological alterations induced by marine pollution. The FFA pattern is involved in the regulation of different cellular pathways and differs with respect to metabolic stimuli. To this purpose, the lipid fraction of mussels coming from both sampling areas was extracted and the FFA fractions were isolated and purified by a solid phase extraction; then, nano-scale liquid chromatography coupled to electron ionization mass spectrometry (nanoLC-EI-MS) was employed for the characterization of the two samples. A total of 19 and 17 FFAs were reliably identified in the mussels coming from the reference and polluted site, respectively. Significant qualitative and quantitative differences found in saturated, monounsaturated and polyunsaturated species may be exploited as typical pollution biomarkers (e.g. alteration of the fatty acid biosynthetic system and lipotoxicity) and explain adverse and compromising effects (e.g. oxidative stress and inflammatory processes) related to environmental pollution.

Monitoring of neonicotinoid pesticides in beekeeping

Abstract The decline of pollinating species is correlated to the extensive use of neonicotinoids against pest insects for crop protection. In this study, the concentrations of neonicotinoid insecticides were determined in honeybees, honeycomb and honey samples, collected in Spring 2015 (blooming period) from different areas in Sicily (IT), to carry out an evaluation of bees products’ safety and an overview of neonicotinoid contamination in beekeeping. The results obtained showed only the presence of clothianidin in bee samples and these concentrations don’t represent a risk for bees’ vitality and safety. The absence of residue in all honey samples, instead, showed the quality of bee products.

Anti-angiogenic activity and phytochemical screening of fruit fractions from Vitex agnus castus

Abstract Although the antitumour activity of Vitex agnus castus fruits has been already addressed, no work has yet assessed their anti-angiogenic potential. To this purpose, several extractive fractions of such fruits were tested on zebrafish embrios by EAP assay, so that only the bioactive fractions could be subsequently tested on the chick chorioallantoic membrane by CAM assay. Bioactive fractions were also phytochemically screened to identify those bioactive compounds responsible for anti-angiogenic activity. A marked inhibition of vessel formation was detected only in zebrafish embryos treated with chloroform or ethyl acetate fractions. Considering CAM assay, chloroform fraction induced a strong reduction of microvasculature and haemoglobin content; while lower anti-angiogenic effects of the ethyl acetate fraction were determined. Phytochemical analyses confirmed the presence of several bioactive anti-angiogenic compounds. Overall, obtained preliminary results highlighted a potential anti-angiogenic activity of V. agnus castus fruits.

Chemical characterization of a variety of cold-pressed gourmet oils available on the Brazilian market

Different specialty extra virgin oils, produced by cold-pressing fruits/nuts (olive, pequi, palm, avocado, coconut, macadamia and Brazil nut) and seeds (grapeseed and canola), and retailed in the Brazilian region of Minas Gerais, were chemically characterized. Specifically, for each type of oil, the fatty acid composition was elucidated by GC-FID, the contents of selected polyphenols and squalene were determined respectively by UHPLC-MS and UHPLC-PDA, whereas minerals were explored by means of ICP-MS. Olive oil was confirmed to have the highest MUFA content due to a valuable level of oleic acid, while oils from grapeseed, Brazil nut and canola were marked by nutritionally important PUFA levels. The highest SFA content found in coconut oil was mainly due to the high levels of lauric acid, known for its advantageous HDL-raising effects. As for polyphenols, gourmet oils from palm, coconut and canola showed higher levels of phenolic acids (e.g. p-hydroxybenzoic, ferulic, syringic, acids) than olive oil, which was though characterized by peculiar antioxidants, such as tyrosol and hydroxytyrosol. Also, olive oil had the highest amount of squalene, followed by the oil from Brazil nut. Finally, all the investigated oils had very low levels (order of μg/kg) of pro-oxidant elements, such as Cu, Fe and Mn. Overall, these findings may fill the gaps still present in literature on certain compositional aspects of commercially available gourmet oils.

Phytochemical screening by LC-MS and LC-PDA of ethanolic extracts from the fruits of Kigelia africana (Lam.) Benth

Abstract Kigelia africana is a tree native to Africa, with a local employment in numerous fields, ranging from traditional medicine to cosmetics and religious rituals. Parts of the plant generally used are stem bark, fruits, roots and leaves. The fruits, which have a singular ‘sausage’ shape, are widely exploited by local folk, in particular for applications/products involving genito-urinary apparatus of both human genders. The scope of this work was to make a consistent chemical investigation on this plant species, in order to clarify and increase the information at present available in literature. To this aim, ethanolic extracts of K. africana fruits were analysed by high-performance liquid chromatography with photodiode array (HPLC-PDA) and electrospray-mass spectrometry (HPLC-ESI-MS) detection, revealing the presence of polyphenols and iridoids. The two detection systems used along with standard co-injection and comparison with previous reports, led to the identification and quantification of six phenolic compounds and three iridoids.

Effects of fasting and refeeding on the digestive tract of zebrafish (Danio rerio) fed with Spirulina (Arthrospira platensis), a high protein feed source

Abstract In the present work, morphological and molecular effects of short-term feed deprivation and refeeding with Spirulina (Arthrospira platensis) on zebrafish digestive tract were determined. Once elucidated the proximate composition of Spirulina feed, immunohistochemical and western blot analyses of peptide transporter (PepT1) and cholecystokinin (CCK8) were carried out in the gastrointestinal tract of zebrafish, previously morphologically investigated. Two and five fasting days caused not only morphostructural alterations, but also the downregulation of PepT1 and CCK8 proteins. Conversely, the recovery of normal morphological conditions, along with an increased PepT1 and CCK8 expression, were observed after refeeding with Spirulina. The increase of PepT1 expression in zebrafish may be responsible for the enhanced CCK8 secretion, so that both proteins may contribute to an improved digestion process during refeeding. These observations could be supported not only by compensatory mechanisms induced by fasting and refeeding but also by an higher protein quality of Spirulina-based diet.