Francesco Siano

Determination of estragole, safrole and eugenol methyl ether in food products

Abstract A rapid and effective procedure for the extraction and determination of 4-allylanisole (estragole), 4-allyl-1,3-benzodioxole (safrole) and 4-allyl-1,2-dimetoxybenzene (eugenol methyl ether) in food products was developed. These compounds were isolated from food matrices by employing a simultaneous distillation–extraction (SDE) technique with a Likens and Nickerson apparatus and using dichloromethane as extraction solvent. The method was validated by conducting recovery studies on fortified food products at several concentrations. Recoveries ranged between 94 and 105%. The analyses were performed by capillary gas-chromatography, the compounds identified by retention times and confirmed by GC/MS. The detection limits, determined both in standard solutions and in foods, were 10, 5 and 8 ng/ml for estragole, safrole and eugenol methyl ether, respectively and the calibration curves showed a good linearity for all the three compounds in the concentration range 0.5–25 ppm, with correlation coefficients ranging between 0.996 and 1.000. In a number of successive analyses, the estragole peak area repeatability (RSD) was 0.20 ng/ml, whereas for safrole resulted 0.33 ng/ml and for eugenol methyl ether 0.45 ng/ml. The estragole, safrole and eugenol methyl ether levels were measured in 42 samples, for a total of six product types, including “Pesto” sauce, tomato sauce containing basil, “Cola tasting” beverages, Bologna sausage (polony), Vienna sausage (wurstel) and fresh basil. The levels ranged from below the detection limit up to 19.30 mg/kg for estragole in “Pesto” sauce.

Extraction and characterization of vegetable oils from cherry seed by different extraction processes

The aim of this work was to compare the content and composition of oil from cherry seeds using supercritical fluid extraction (SFE) using carbon dioxide as solvent, and Soxhlet extraction using diethyl ether. The compositions of free fatty acids and phytosterols, achieved with two extraction methods, were analyzed. Gas chromatographic analysis of unsaponifiables cherry seed oil samples, obtained with SFE and Soxhlet extraction, permitted to identify in particular two sterols, the s-sitosterol and campesterol. Brassicasterol or stigmasterol were not present in the extracts. The percentages of monitored components changed in function of extraction used procedure, allowing to assert that, with SFE, higher yield of phytosterols was provided. Preliminary data showed that there were significant differences also in the fatty acid composition of cherry seed oil obtained with the two extraction procedures (by means supercritical carbon dioxide or solvent). With the Soxhlet extraction, 48.6% of saturated fatty acids, 17.74% of unsaturated acids, 32.75% of polyunsaturated are obtained. On the contrary, with SFE decreased the percentage of saturated fatty acids (38.43%) in favor of larger amounts of unsaturated (19.93%) and polyunsaturated (36.04%). The supercritical fluid extraction procedure proved so effective in getting an oil of superior quality, without the possibility of degradation phenomena due to the solvent or heat. In both extraction procedures the cherry seed oil contained high percentages of arachidic acid (C20:0) and linoleic acid (C18:2, ?-6). An interesting result was also the presence of nervonic acid in remarkable amount, in all examined samples. Nervonic acid is a long-chain monounsaturated fatty acid (C24:1, ?-9), of marine origin, precursor of the neuronal cell membrane glycolipids, with a key role in the modulation of ion channels and membrane receptors, and therefore widely used as a neurotrophic factor in food supplements and nutraceuticals.

Betaines and related ammonium compounds in chestnut (Castanea sativa Mill.)

Chestnut fruits, being poor of simple sugars and consisting mainly of fibers and starch, are among the constituents of Mediterranean diet. While numerous studies report on content of proteins and amino acids in chestnut, no one has appeared so far on betaines, an important class of nitrogen compounds ubiquitous in plants for their protective action in response to abiotic stress. In this study, we analyzed by HPLC-ESI-tandem mass spectrometry, in fruits and flours of varieties of chestnut cultivated in Italy, the composition of betaines and ammonium compounds intermediates of their biosynthesis. Besides the parent amino acids, the compounds quantified were choline, glycerophosphocholine, phosphocholine, glycine betaine, N-methylproline, proline betaine (stachydrine), β-alanine betaine, 4-guanidinobutyric acid, trigonelline, N,N,N-trimethyllysine. Interestingly, some uncommon derivatives of pipecolic acid, such as N-methylpipecolic acid, 4-hydroxypipecolic acid and 4-hydroxy-N-methylpipecolic acid were identified for the first time in chestnut samples and characterized by MS(n) tandem mass spectrometry.

Potential Anticancer Effects of Polyphenols from Chestnut Shell Extracts: Modulation of Cell Growth, and Cytokinomic and Metabolomic Profiles

In this study, a hydroalcoholic chestnut shell extract was characterized and tested on six different human cell lines. Gallic, ellagic, and syringic acids were the most abundant non-condensed compounds in the chestnut extract, as determined by high performance liquid chromatography (HPLC). Tannins were mainly represented by condensed monomeric units of epigallocatechin and catechin/epicatechin. After 48 h of treatment, only the human hepatoblastoma HepG2 cells reached an inhibition corresponding to IC50 with an increase of apoptosis and mitochondrial depolarization. The cytokinome evaluation before and after treatment revealed that the vascular endothelial growth factor (VEGF) and the tumor necrosis factor (TNF)-α decreased after the treatment, suggesting a potential anti-angiogenic and anti-inflammatory effect of this extract. Moreover, the metabolome evaluation by 1H-NMR evidenced that the polyphenols extracted from chestnut shell (PECS) treatment affected the levels of some amino acids and other metabolites. Overall, these data highlight the effects of biomolecules on cell proliferation, apoptosis, cell cycle and mitochondrial depolarization, and on cytokinomics and metabolomics profiles.

Content of micronutrients, mineral and trace elements in some Mediterranean spontaneous edible herbs.

BackgroundThe analysis of mineral elements composition was determined in three wild edible herbs (Cichorium intybus L., Sonchus asper L. and Borago officinalis) collected in seven different sampling sites which were characterized by different pollution grade. The detection of mineral elements (Ca, K, Mg and Na), micronutrients (Cu, Fe, Li, Mn and Zn) and heavy metals (As, Cd, Hg, Ni and Pb) was performed.ResultsThe results obtained show that in most cases a direct relationship appeared between the amount of elements and the sampling sites. The highest concentrations of heavy metals were found in samples grown in polluted soils. These evaluations showed that contaminants in plants may reflect the environmental state in which they develop.ConclusionThe examined species are a good source of mineral elements and micronutrients, making them particularly adapt to integrate a well-balanced diet. The accumulation of heavy metals showed that contaminants in plants may reflect the environmental state in which they develop. Results showed high concentrations of heavy metals in samples taken in locations characterized by high human activity and in some samples from the local market, of which no one knows the collection area.

Comparative analysis of C-glycosidic flavonoids from Prosopis spp. and Ceratonia siliqua seed germ flour

Seed germ of South American algarrobo (Prosopis species) and European carob (Ceratonia siliqua) contains nutritionally interesting proteins, lipids and phenolics. Using reversed phase-HPLC-diode array detector and nanoflow-HPLC coupled to tandem mass spectrometry (MS/MS), we comparatively characterized and semi-quantified flavonoids from germ of three Argentinean algarrobo (Prosopis alba, Prosopis nigra and Prosopis ruscifolia) and one European carob species. The patterns of glycosylated flavonoids were very similar each other, confirming the taxonomic parentage of the species and supporting their functional similarity on a molecular basis, in view of the use of seed germ flour (SGF) for food applications. The predominant phenolic compounds were apigenin 6,8-C-di-glycoside isomers, namely isoschaftoside and schaftoside, accounting for 3.22-5.18 and 0.41-0.72 mg/g SGF, respectively. C. siliqua germ contained relatively high amounts of further glycosilated derivatives of (iso)schaftoside, which occurred at a lower abundance in Prosopis. Apigenin 6,8-C-di-glycosides have been described as potent α-glucosidase inhibitors, suggesting that food preparations obtained with Prosopis spp. and C. siliqua SGF might contribute to modulate the digestion of carbohydrates in humans. CHEMICAL COMPOUNDS Isoschaftoside (PubChem CID: 13644661); Schaftoside (PubChem CID: 442658); Vicenin-2 (PubChem CID: 442664); Isovitexin (PubChem CID: 162350).

Integrated Analytical Methods to Characterize Lipids from Prosopis spp. and Ceratonia siliqua Seed Germ Flour

Flour from seed germ of European carob (Ceratonia siliqua) and South American algarrobo (Prosopis spp.) is a potential ingredient for health-promoting baked products. Herein, lipids from germ of three Argentinean Prosopis (P. alba, P. nigra, and P. ruscifolia) and one European carob species were characterized in detail, exploiting an array of up-to-date analytical techniques. Total lipids ranged from 7.1 to 8.1% (w/w). Linoleic acid (C18:2, ω-6) predominated the GC flame ionization detector profiles of fatty acid in all samples (43.3–50.6%). Prosopis spp. contained 6–7% of C20–C24 fatty acids and, consistently, C56–C60 triacylglycerols, as detected by MALDI-TOF mass spectrometry (MS), which were practically missing in C. siliqua germ. Phospholipids were isolated by hydroxyapatite chromatography, characterized by MALDI-TOF MS and grossly quantified by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. Because of a relatively high content (9.5–11.8%, v/v), phospholipids might increase the antioxidant potential and improve the baking performances of flour fortified with carob and algarrobo seed germ.