{MALDI}-{TOF} Mass Spectrometry Applications for Food Fraud Detection

Chemical analysis of food products relating to the detection of the most common frauds is a complex task due to the complexity of the matrices and the unknown nature of most processes. Moreover, frauds are becoming more and more sophisticated, making the development of reliable, rapid, cost-effective new analytical methods for food control even more pressing. Over the years, MALDI-TOF MS has demonstrated the potential to meet this need, also due to a series of undeniable intrinsic advantages including ease of use, fast data collection, and capability to obtain valuable information even from complex samples subjected to simple pre-treatment procedures. These features have been conveniently exploited in the field of food frauds in several matrices, including milk and dairy products, oils, fish and seafood, meat, fruit, vegetables, and a few other categories. The present review provides a comprehensive overview of the existing MALDI-based applications for food quality assessment and detection of adulterations

Recent Applications of Solid Phase Microextraction Coupled to Liquid Chromatography

Solid phase microextraction (SPME) is one of the most popular sample preparation methods which can be applied to organic compounds allowing the simultaneous extraction and pre-concentration of analytes from the sample matrix. It is based on the partitioning of the analyte between the extracting phase, generally immobilized on a fiber substrate, and the matrix (water, air, etc.), and has numerous advantages such as rapidity, simplicity, low cost, ease of use and automation, and absence of toxic solvents. Fiber SPME has been widely used in combination with various analytical instrumentation even if most of the work has been done coupling the extraction technique with gas and liquid chromatography (GC and LC). This manuscript presents an overview of the recent works (from 2010 to date) of solid phase microextraction coupled to liquid chromatography (SPME-LC) relevant to analytical applications performed using commercially available fibers or lab-made fibers already developed in previous papers, and to improved instrumental systems and approaches

Measurement of squalene in olive oil by fractional crystallization or headspace solid phase microextraction coupled with gas chromatography

Squalene is the most abundant component in the unsaponifiable fraction of olive oil with strong antioxidant properties. Its concentration in olive oils varies between 0.2 and 16.2 g/kg depending on the cultivar(s) used. The propose of this work was to determine the effectiveness of two different extraction methods for squalene determination by gas chromatography (GC) coupled to a flame ionization detector (FID) or to mass spectrometry (MS). In a first approach, oil samples were dissolved in methanol/acetone mixture 7:3 (v/v) and triglycerides separated by fractional crystallization at −20°C. The organic layer was removed, reduced to dryness and the residue reconstituted in n-heptane (containing squalane as external standard) and analyzed by GC-FID. A headspace (HS) solid phase microextraction (SPME) GC-MS method has been also developed in order to have an environmentally friendly (i.e. solventless) extraction procedure. The linear range investigated with both methods was 1.0-10 g/kg. Within-day and between days precision values, expressed as RSD%, were 4 and 7% (GC-FID), and 3 and 6% (GC-MS), respectively. The limit of detection (LOD) at a signal-to-noise (S/N) ratio of 3 were 0.019 (GC-FID) and 0.003 (GC-MS) g/kg; the limit of quantification (LOQ) calculated at S/N = 10 were 0.063 (GC-FID) and 0.008 (GC-MS) g/kg, well below the typical squalene concentration levels found in olive oils. The obtained percentage recoveries were 70 ± 2 (GC-FID) and 98 ± 3 (GC-MS), and were not concentration dependent. The potential of the method has been demonstrated by the analysis of several different olive oil samples produced from different cultivars and different locations

Antibacterial activities of peptides from the water-soluble extracts of Italian cheese varieties.

Water-soluble extracts of 9 Italian cheese varieties that differed mainly for type of cheese milk, starter, technology, and time of ripening were fractionated by reversed-phase fast protein liquid chromatography, and the antimicrobial activity of each fraction was first assayed toward Lactobacillus sakei A15 by well-diffusion assay. Active fractions were further analyzed by HPLC coupled to electrospray ionization-ion trap mass spectrometry, and peptide sequences were identified by comparison with a proteomic database. Parmigiano Reggiano, Fossa, and Gorgonzola water-soluble extracts did not show antibacterial peptides. Fractions of Pecorino Romano, Canestrato Pugliese, Crescenza, and Caprino del Piemonte contained a mixture of peptides with a high degree of homology. Pasta filata cheeses (Caciocavallo and Mozzarella) also had antibacterial peptides. Peptides showed high levels of homology with N-terminal, C-terminal, or whole fragments of well known antimicrobial or multifunctional peptides reported in the literature: alphaS1-casokinin (e.g., sheep alphaS1-casein (CN) f22-30 of Pecorino Romano and cow alphaS1-CN f24-33 of Canestrato Pugliese); isracidin (e.g., sheep alphaS1-CN f10-21 of Pecorino Romano); kappacin and casoplatelin (e.g., cow kappa-CN f106-115 of Canestrato Pugliese and Crescenza); and beta-casomorphin-11 (e.g., goat beta-CN f60-68 of Caprino del Piemonte). As shown by the broth microdilution technique, most of the water-soluble fractions had a large spectrum of inhibition (minimal inhibitory concentration of 20 to 200 microg/mL) toward gram-positive and gram-negative bacterial species, including potentially pathogenic bacteria of clinical interest. Cheeses manufactured from different types of cheese milk (cow, sheep, and goat) have the potential to generate similar peptides with antimicrobial activity

Development, Optimization, and Comparison of Different Sample Pre-Treatments for Simultaneous Determination of Vitamin E and Vitamin K in Vegetables

The absence of vitamin E from the diet can lead to cardiovascular disease, cancer, cataracts, and premature aging. Vitamin K deficiency can lead to bleeding disorders. These fat-soluble vitamins are important nutritional factors that can be determined in different methods in vegetables. In this work, the simultaneous determination of α-tocopherol, α-tocopheryl acetate, phylloquinone, and menaquinone-4 by gas chromatography–mass spectrometry (GC–MS) has been optimized using both direct injection and solid phase microextraction (SPME). Three different sample pre-treatment approaches based on: (A) solid–liquid–liquid–liquid extraction (SLE–LLE), (B) SLE, and (C) SPME were then applied to extract the target analytes from vegetables samples using menaquinone as internal standard. All the procedures allowed the determination of the target analytes in onion, carrot, celery, and curly kale samples. Similar results were obtained with the three different approaches, even if the one based on SPME offers the best performance, together with a reduced use of solvent, time consumption, and experimental complexity, which makes it the preferable option for industrial applications

Analytical investigations on lindane bioremediation capability of the demosponge Hymeniacidon perlevis

Lindane is an organochlorine pesticide that has been widely used to treat agricultural pests. It is of particular concern because of its toxicity, persistence and tendency to bioaccumulate in terrestrial and aquatic ecosystems. In this context, we investigated the ability of the demosponge Hymeniacidon perlevis to bioremediate lindane polluted seawater during in vitro experimentation. Lindane was extracted by solid-phase micro-extraction (SPME) and determined by gas chromatography–mass spectrometry (GC–MS). Furthermore, we assessed the role exerted in lindane degradation by bacteria isolated from the sponge. Sponges showed low mortality in experimental conditions (lindane concentration 1 μg/L) and were able to remove about 50% of the lindane content from seawater in 48 h. Bacteria isolated from sponges showed a remarkable remediating capacity (up to 97% of lindane removed after 8-days). A lindane metabolite was identified, 1,3,4,5,6-pentachloro-cyclohexene. The results obtained are a prelude to the development of future strategies for the in situ bioremediation of this pollutan

Bovine and soybean milk bioactive compounds: Effects on inflammatory response of human intestinal Caco-2 cells

In this study the effects of commercial bovine and soybean milks and their bioactive compounds, namely genistein, daidzein and equol, on the inflammatory responses induced by lipopolysaccharide (LPS) treatment of human intestinal Caco-2 cells were examined, in terms of nitric oxide (NO) release and inducible nitric oxide synthetase (iNOS) expression. Both milks and their bioactive compounds significantly inhibited, dose-dependently, the expression of iNOS mRNA and protein, resulting in a decreased NO production. The NF-κB activation in LPS-stimulated intestinal cells was also examined. In all cases we observed that cell pre-treatment before LPS activation inhibited the IkB phosphorylation. Accordingly, quantification of bioactive compounds by solid phase microextraction coupled with liquid chromatography has shown that they were absorbed, metabolized and released by Caco-2 cells in culture media. In conclusion, we demonstrated that milks and compounds tested are able to reduce LPS-induced inflammatory responses from intestinal cells, interfering with NF-kB dependent molecular mechanisms

Determination of Commercial Animal and Vegetable Milks’ Lipid Profile and Its Correlation with Cell Viability and Antioxidant Activity on Human Intestinal Caco-2 Cells

none9openantonella aresta; Stefania De Santis; Alessia Carocci; Alexia Barbarossa; Andrea Ragusa; Nicoletta De Vietro; Maria Lisa Clodoveo; Filomena Corbo; Carlo ZAMBONINAresta, Antonella; De Santis, Stefania; Carocci, Alessia; Barbarossa, Alexia; Ragusa, Andrea; De Vietro, Nicoletta; Lisa Clodoveo, Maria; Corbo, Filomena; Zambonin, Carl

MALDI-TOF/MS Analysis of Extracellular Vesicles Released by Cancer Cells

The direct shedding of extracellular vesicles (EVs) from the plasma membrane is a recognized fundamental method for the intercellular transfer of properties in both physiological and pathological conditions. EVs are classified according to origin, biogenesis, size, content, surface markers, and/or functional properties, and contain various bioactive molecules depending on the physiological state and the type of the cells of origin including lipids, nucleic acids, and proteins. The presence of tumor-derived EVs in body fluids such as blood, ascites, urine, and saliva, together with the important role played in the tumor microenvironment where they intervene at different levels from oncogenesis to metastasis, make EVs a priority target for cancer studies. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) can play a leading role in the analysis and characterization of EVs and their load due to its intrinsic advantages such as high throughput, low sample consumption, speed, the cost-effectiveness of the analysis, and the ease of use. This work reviews the main MALDI-TOF applications for the analysis and characterization of extracellular vesicles in the tumor field