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1 Dipartimento di Chimica REGIONE PUGLIA ACCORDO DI PROGRAMMA QUADRO RICERCA SCIENTIFICA II ATTO INTEGRATIVO. RETI DI LABORATORI PUBBLICI DI RICERCA APULIAN FOOD FINGERPRINT: VALORISATION OF APULIAN FOOD PRODUCTS BY MEANS OF NUCLEAR MAGNETIC RESONANCE, MASS SPECTROMETRY AND ELECTRONIC NOSE ANALYSES SCIENTIFIC RESPONSIBLE: PROF. ANTONIO SACCO, DEPARTMENT OF CHEMISTRY, UNIVERSITY OF BARI (Italy). PARTNER: PROF. PIERO MASTRORILLI, DEPARTIMENT DIAC, POLITECNICO DI BARI (Italy).
2 The lab that is being set up will be equipped with the following instrumentation: MHz NMR spectrometer equipped with MAS probe. - High resolution mass spectrometer with ESI and APCI ionization sources. - Electronic nose. - Mass spectrometer of isotopic ratios (IRMS). Here are some research fields that can be investigated using the above equipment. 1) Nuclear Magnetic Resonance (NMR) Within the magnets working at a temperature of 4K, the 700 MHz NMR instrument is the tool using the most intense magnetic field. The salient features of this tool are: 1. Easy to install, in fact: the extension of the magnetic field lines is very short both horizontally and vertically; The magnet is relatively small, unlike the 800 and 900 MHz instruments that require large rooms with ceiling heights over 5 feet. 2. Great dispersion of signals. 3. High sensitivity. In particular, if the instrument is equipped with a cryogenic probe, you can run the proton spectra of samples with concentration of 0.1 µm at within time suitable for 2D and 3D spectroscopy. 4. Great dynamic range. The dynamic range is such that the spectrum of the proton allows, with a scan, to obtain a dynamic range of 0.5 ppm and minimum observable concentration of 0.5 µm. 5. Multinuclear spectroscopy technique with Magic Angle Spinning (MAS). The characteristics listed in paragraphs 2-5, make the 700 MHz NMR spectrometer particularly suitable for the following applications in the research fields below. 2
3 Agri-Food Quali-quantitative compositional studies on liquid food products (oil, wine, fruit juices, milk, etc.). Focus on the minor components that characterize the product investigated. Subsequent statistical analysis, with programs supplied with the instrument, of the NMR data for highlighting any adulteration and characterization of the geographical origin of products. Creation of a database of agri-food products with certain Apulian origin. Agri-Food (MAS probe) Quali-quantitative compositional studies on food products in solid form (flour, bread, mozzarella, meat, fruit, grapes, etc..) Focus on the minor components that characterize the product investigated. Subsequent statistical analysis, with programs supplied with the instrument, of the NMR data for highlighting any adulteration and characterization of the geographical origin of products. Creation of a database of agri-food products with certain Apulian origin. Pharmaceutical Study of the metabolism of drugs in biological fluids. Study of metabolic imbalance induced by drugs in biological fluids. Identification of impurities in synthetic products and finished products ready for commercialization. Pharmaceutical (MAS probe) Study of drug activity by NMR analysis of cell suspensions in vitro. Studies on drug metabolism in cell suspensions in vitro and biopsies. Study of metabolic imbalance induced by drugs in cell suspensions in vitro and biopsies. Biochemical/Pharmaceutical Characterization of the structure of proteins and DNA. Characterization of drug-protein interaction. Materials (MAS probe) Analysis of polymers and composites. Study the interaction between organic and inorganic additives, e.g. for R&D in the field of high performance cements. Study of gel phase systems, e.g. for R&D in the field of high performance cements and of polymers produced in heterogeneous phase and/or in emulsion. Environment Characterization of degradation products due to agricultural products. Characterization of degradation products derived from human activity (waste products). Characterization of degradation products derived by industrial activity. 3
4 2) High Resolution Mass Spectrometry (HRMS) HRMS technique allows to determine the molecular weight of a compound to elucidate the molecular structure, to identify the components of a mixture and to investigate quantitative and mechanistic studies. In the following the food applications are reported even if numerous further applications are possible (food, proteomics, molecular and cellular biology, environmental science, polymer chemistry, pharmacology, archaeology, inorganic analysis, etc...). Agri-Food Characterization of the polyphenolic compounds in foods. Characterization of aromatic extracts used as antioxidants. Characterization of components of complex mixtures such as beer, propolis, oils, etc. Quality control and authenticity tests of various foods. Determination of unwanted substances due to processes and storage (e.g. acryl amide, biogenic amines, heterocyclic aromatic amines, formaldehyde, furan). Determination of biogenic amines. Applications to samples of fresh and aged cheese. Simultaneous determination of heterocyclic aromatic amines in food for babies (meat homogenate). Determination of formaldehyde in various types of food such as, for example, frozen fish. Determination of furan in foods for babies (vegetables and fruit). Simultaneous determination of isoflavonoids in soybeans. Determination of pesticides and not allowed additives. Determination of azo-dyes in samples of tomato and chilli-pepper based sauces. Identification and quantification of compounds present in trace amounts (pesticides, hormones, drug metabolites) in complex matrices. 4
5 3) Isotopic Ratio Mass Spectrometry (IRMS) The IRMS technique allows the measurement of natural isotope ratios ( 13 C/ 12 C, 15 N/ 14 N, 18 O/ 16 O, 35 S/ 34 S, 2 H/ 1 H etc.). Here are only some examples of the different fields of application for IRMS and information that can be obtained from the analyses. Agri-Food: Characterization of food products: 1. by geographic parameters (environmental and climatic factors); 2. on the basis of the diet (animal products). Traceability of food and identification of possible frauds. In particular, the most common adulteration recognizable by isotopic methods include: Fruit juices Fruit juices Honey Natural Vanilla Wine* Wine* Olive oil Cheese addition or dilution with water Addition of cane or corn sugar Addition of cane or corn sugar Addition of artificial vanilla dilution with water addition of beet, cane or corn sugar addition of cheaper oils Supplements of the diet with feed *official methods adopted by the European Union (EU) 5
6 Quality control and authenticity tests of various foods. Determination of the presence of dioxins in contaminated fish or environmental micropollutants. Continuous monitoring of fermentation for the biotechnology industry. Environmental Chemistry: Environmental monitoring Determination of the different kinds of contamination Identification and quantification of environmental biodegradation processes and isotopic fractionation. Study of 13 C and 18 O isotopic exchange between atmosphere and biosphere that achieves metabolic and ecological relationships between the various constituents of the ecosystem. Pharmacology: Authentication and quality control of drugs. Identification of structural biomolecules to determine the sequence of biopolymers. Analysis of 13 C/ 12 C ratio in samples of breath after assumption of suitable substrates, for the diagnosis of various diseases such as infection with Helicobacter pylori. 6
7 4) Sensory Analysis by Electronic Nose The electronic noses are complex systems characterized by a structure similar to that of the human olfactory system: an array of chemical sensors with low selectivity provides a characteristic signature of a chemical mixture which is then classified based on the knowledge gained in an electronic nose training phase. Here are some of the numerous applications of the electronic nose: Agri-Food: Control of fermentation processes, of cooking processes and production quality. Smell release and quality of packaging and containers. Traceability of meat and cheese. Off flavour, ripening and maturing of milk and milk-derivatives. Classification, humidity, microbiological quality and mycotoxin contamination of grain. Freshness of Fish. Control of processes and microbiological quality of meat. Toasting level and quality of grains, mycotoxins in coffee. Freshness of vegetables. Quality and nutritional value of oil. Environmental Monitoring: Analysis of combustible mixtures. Detection of leaks from pipelines. Identification of toxic waste. Water quality. Monitoring of air quality. Monitoring of industrial gas emissions. Identification and control of smell emission. Monitoring of waste water treatment. Everyone interested in using the above mentioned instruments in the future is invited to contact Prof. Antonio Sacco, Department of Chemistry, University of Bari (antonio.sacco@chimica.uniba.it). 7
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