Flavour and Metabolites Lab

LABORATORY FOR SAMPLE PREPARATION

NIR spectroscopy

Near infra-red spectroscopy is a spectroscopic method with which the near infra-red range of the electromagnetic spectrum (78 to 2500 nm) is used. It is used in the agricultural sciences as well as medicine, pharmaceutics and atmospheric chemistry. Here it is used for the non-destructive analysis of apples, namely for determining the total sugar and starch content in order to predict the ripening of the apple.

Analytical scale

Inside is an electromagnet which generates a force able to compensate for the weight of the sample being measured, with the result of the measurement being achieved by measuring the force required to reach a state of equilibrium.

Vibration mill

Samples of apples or wood can be ground into dust with this device. 

Distilled water

This water is free of impurities, mineral salts, micro-organisms and dissolved gases. It is used in the laboratory as a solvent or chemical reagent.

Sample processing

ph. Laimburg Research Centre/ivo corrà

Rotary evaporator

: this device is used to reduce (concentrate) solutions by evaporating the solvent. 

Drying oven

Liquid chromatograph coupled with time-of-flight mass spectrometer (LC-TOF)

: with liquid chromatography, the compounds that need to be analysed are split in a separation column with the help of chemical solvents and then analysed with the high resolution mass spectrometer.

Liquid chromatograph coupled with DAD-FLD

: with this device, the compounds that need to be analysed are split in a separation column with the help of chemical solvents and then analysed using a diode array detector (DAD) or fluorescence detector (FLD).

Liquid chromatograph coupled with triple quadropole mass spectrometer

: individual polyphenols, vitamins and amino acids in foodstuffs and plant extracts can be quantified using this device.

Liquid chromatograph coupled with triple quadropole mass spectrometer

: individual polyphenols, vitamins and amino acids in foodstuffs and plant extracts can be quantified using this device.

UV/VIS spectrophotometer

UV/VIS spectroscopy uses the electromagnetic waves of ultraviolet (UV) and visible (VIS) light. This method is used to analyse the total content of anthocyanins (plant pigments), polyphenols, tannins, the antioxidative capacity (antioxidants) and the total chlorophyll content of food and vegetable samples.

Gas chromatograph coupled with triple quadropole mass spectrometer (GC-TQMS)

: this device is used to analyse the aromas in wine, apples, apple juice and strawberries or fatty acids in milk and cheese.

Gas chromatography coupled with flame ionisation detector (GC-FID)

: with gas chromatography, the molecules being analysed are separated in a capillary column by increasing the temperature (50-250 ºC) and then analysed with the flame ionisation detector (FID).

Gas chromatography coupled with time-of-flight mass spectrometer (GC-TOF)

: with gas chromatography, the molecules being analysed are separated in a capillary column by increasing the temperature (50-250 ºC) and then analysed with a high resolution mass spectrometer. Volatile substances such as apple and wine aromas can be quantified using this technique.

Ion chromatograph (IC)

Using ion chromatography, substances can be separated by virtue of their charge. Individual sugars and organic acids in apples, wine grapes and wine can be analysed using this method.

Gas chromatograph coupled with quadropole mass spectrometer (GC-QMS)

: with gas chromatography, the molecules being analysed are separated in a capillary column by increasing the temperature (50-250 ºC) and then analysed using a quadropole mass spectrometer (MS). Volatile substances such as aromas in apples and wines as well as esterified fatty acids in milk products can be analysed using this technique.

Freeze dryer

See point 1 Device used for carefully drying the product. This equipment does not contain refrigeration chambers. 

Freeze dryer

Freeze drying (lyophilisation) is a method by which the product is carefully dried - the ice crystals sublimate directly into a gaseous state without an interim liquid phase occurring. Freeze drying is used, for example, in the manufacture of instant drink powders, fruit for muesli mixtures, herbs and spices. In food analysis, it is used to remove the water from fruit and other foodstuffs with a high water content prior to analysis. This freeze dryer also contains refrigeration chambers.

Freezer room

Refrigerators and freezers for storing the samples

Polyphenols

Natural chemical compounds occurring in plants. These secondary plant substances constitute bioactive substances such as colourings, flavourings or tannins and are supposed to protect the plants from predators or attract insects for pollination through their colour. Many of these polyphenols are considered to be beneficial to health; some, among other things, have an anti-inflammatory effect or help to prevent cancer. Here we are interested in the polyphenols in apples.  

Metabolomics

: investigates the metabolic characteristics of a cell and/or of a tissue or organism as well as the metabolites involved. Here, the processes involved in the infection of a vine with downy mildew, which is caused by the fungus plasmopara viticola, are investigated.  

Project PinotBlanc

Grapes of the variety Pinot Blanc, one of the leading varieties in South Tyrol, are distinguished by a relatively high acid content and a fresh, fruity aroma. However, the typical aromas are suffering from global warming in South Tyrol. The project is looking into whether moving to higher and cooler growing locations would be a possible way of ensuring the quality of the Pinot Blanc. In the Laboratory of Flavours and Metabolites, the aroma profiles of Pinot Blanc wines from mountain and valley locations have been chemically characterised. 

Solvent room

Various solvents are securely stored here. 

Sample collection

To apply to conduct testing, samples must be submitted with the relevant documents. More information:  http://www.laimburg.it/it/servizi/formulari.asp

Centrifuge

Multi-rotator

: due to the fact that it can rotate in different directions and is able to combine movements according to requirements, this device enhances the efficiency of sample preparation.

Thermomixer

: this device is used to carry out reactions and extractions at high temperatures.

Ultrasound bath

: this device is used to carry out chemical reactions and extractions at high temperatures.

Hotplate

: for heating up the samples.

Extractor

: used in the preparation phase to dissolve (extract) the compounds from the samples. 

pH meter

Measuring device for determining and displaying the pH value of a solution.

Mobile laboratory hood

Laboratory hoods are used with chemical applications to protect the laboratory environment and the operator. They protect individuals from inhaling toxic vapours and reduce the risk of fire and explosion.

Centrifuge

Centrifuges are used to separate the liquid phase of solid substances and particles.

Vortex

The plate vibrates in different directions. This enables solvents and reagents to be mixed until homogeneous. 

Extraktor

: used in the preparation phase to dissolve (extract) the compounds from the samples.

Dewar flask for storing liquid nitrogen

DROMYTAL

The spotted wing drosophila (Drosophila suzukii) is a pest that was introduced from Asia. In the DROMYTAL project, researchers from Laimburg Research Centre and the Free University of Bolzano have developed an innovative strategy for combating the pest. The method is based on a yeast lure, with which an insecticide is mixed (attract & kill). In the Laboratory of Flavours and Metabolites, the metabolites of the bioactive yeast cultures are tested to identify those cultures that stimulate feeding and which as a result are best suited to attracting the spotted wing drosophila.

VOCs

VOCs (volatile organic compounds) make up the aroma of a foodstuff. Each apple variety features a different aroma profile, i.e. a complex mixture of countless VOCs that is typical for each one. In this study, the VOCs in different apple varieties were identified. 

LABORATORY OF FLAVOURS AND METABOLITES

The Laboratory of Flavours and Metabolites researches and conducts experiments in the fields of food quality and plant health. Using modern analytical methods, naturally occurring ingredients in agricultural products (apples, apple juice, grapes, wine, cheese, milk) and plant organs (leaves, roots, wood) are determined and quantified in order to test their quality, characteristics and purity. 

Liquid chromatography

: method for separating chemical substances. These substances can be identified and quantified with the help of standards (i.e. their precise concentration in a sample can be determined). 

ANALYSIS LABORATORY

Gas chromatography

Gas chromatography is a method for analysing mixtures that are separated into individual chemical compounds. The method can only be used for volatile components in a temperature range between room temperature and approx. 350 ºC (boiling range up to 400 ºC).  

Isotope analysis

To guarantee the quality of foodstuffs and to protect producers and consumers, it is important to be able to verify the origin of agricultural produce. In the Isotope Analysis project, Laimburg Research Centre, Eco-Research and the Free University of Bolzano are developing a method of verifying origin, which is based on the analysis of isotopes of the element strontium.

DIC PILOT PLANT

The Laboratory of Nuclear Magnetic Resonance

The Laboratory of NMR Spectroscopy deals with the qualitative and quantitative characterisation of small molecules and proteins in foodstuffs and plants with the help of a methodology that uses high-resolution nuclear magnetic resonance (NMR). NMR spectroscopy is a non-invasive and non-destructive laboratory analysis technology that enables us to obtain a fingerprint of molecules that are of interest and thus to characterise their structure and molecular dynamics. This technology can be used to determine the authenticity, typicity and origin of local agricultural and food products as well as to carry out structural, kinetic and functional studies of biological molecules and identify new, unknown molecules. 

NMR sample loading device

After preparation, the NMR test tube is placed on the NMR sample loader with the sample being examined. Both NMR devices are equipped with an automatic sample loading tool. This enables serial measurements to be carried out in a sequence programmed by the operator. The 400 MHz device shown here allows up to 64 samples to be analysed, whilst the device depicted on the right (600 MHz NMR) is able to load up to 30 samples one after another. The possibility of measuring several samples sequentially is especially useful when examining the metabolomics of agricultural and food products, i.e. their different ingredients.   The analysis of several samples thus enables a statistical analysis that is useful for identifying the typicity of the product and its origin.ph: NOI Techpark/ivocorrà

600MHz NMR spectrometer

Like the 400 MHz NMR spectrometer, the 600 MHz spectrometer is also equipped with a "broadband” probe that allows one- and two-dimensional spectra and a range of interesting nuclei to be recorded for investigating the chemical structure of unknown molecules. However, the 600 MHz spectrometer enables the recording of spectra with higher resolution and is therefore better suited for analysing more complex matrices or examining macromolecules with a high molecular weight, such as proteins or nucleic acids.

Drying module

The final step in the process consists in removing the residual moisture collected during the texturing phase. This evaporation takes place in the drying module, where the apples are brought to 50 degrees under a vacuum. In this way, the moisture can be completely removed in around two hours. The plant in the photograph is a prototype.

Automatic filling machine

Automatically calculates the exact filling weight of each pouch.

Manual packaging machine

400MHz NMR Spectrometer        

The NMR spectrometer is an essential tool for determining the structure of molecules and their movements and interactions. An NMR spectrometer consists of a superconductive coil that generates a permanent magnetic field and is immersed in a jacket of liquid helium, which ensures superconductivity at temperatures close to absolute zero (~ -273° C). A second outer jacket containing liquid nitrogen enables the helium to be constantly cooled. The gases are injected into the jackets via a turret on the magnetic head. To reduce the evaporation of the liquid helium, liquid nitrogen is used to minimise the thermal gradients in the Dewar. In the centre of the spectrometer is the bore, a cylindrical container that holds the static field and the probe.  The magnetic field generated by the NMR machine makes it possible to align the nuclei of the molecules being examined (as though they were microscopic magnetic needles) and to explore the properties of the individual atoms by generating spectra.         The 400 MHz NMR spectrometer is equipped with a “broadband” probe that allows one and two-dimensional spectra and various interesting nuclei to be recorded with which the chemical structure of unknown molecules can be discovered.ph. NOI Techpark/ivo corrà

Cylinder with liquid nitrogen

It is necessary to add liquid nitrogen to both spectrometers weekly in order to prevent the evaporation of the liquid helium and thus preserve its superconductivity.  It is added in compliance with all necessary safety standards. 

Cooling of NMR samples

The NMR sample cooler, which is installed on the 600MHz spectrometer, makes it possible to store the samples in the repository (i.e. before or after measuring) at the desired temperature. This is especially important in the case of NMR samples with which a highly volatile solvent is used (e.g. chloroform). 

NMR samples  

The sample that is to be examined (from small molecules to proteins, from simple to complex matrices) is placed in a 5mm tube with a length of approx. 18 cm and then hermetically sealed with the red cap that can be seen in the photograph. The research conducted in the NMR Laboratory at Laimburg Research Centre allows both liquid (water-soluble compounds or organic solvents such as chloroform) and semi-solid samples to be analysed.ph: NOI Techpark/ivo corrà

NMR console

Contains all of the electronics needed for the proper functioning of the NMR machine. For example, the NMR console is responsible for generating the electromagnetic radiation which then interacts with the sample molecules in the magnet.

Low temperature dryer

The apple slices are placed in a low temperature dryer, in which they are dried over a relatively long period but at low temperatures until there is a residual moisture level of only around 10 to 15%. This creates a plastic structure which can be deformed in the subsequent texturing process. 

Module for the drying process

This is the module for the drying process which differs fundamentally from all other existing drying systems, in that it consists of a steam generator, a vacuum container and a texturing reactor. Hot steam is fed into the inside of the reactor for a few seconds and followed by the creation of an instantaneous vacuum. This vacuum allows the instant evaporation of the moisture accumulated in the tissue, reducing the pressure and temperature and bringing the product below the glass transition temperature. As a result, the structure obtained during the treatment is stabilised. 

How does instant controlled pressure drop technology (DIC) work?

The usual methods of manufacturing dry food products result in a collapse of the structure. Consequently, the product loses porosity, uneven layers form and it becomes rubbery. In contrast, the innovation of the DIC technology is that the product is textured by a sudden drop in pressure from overpressure to vacuum. The treatment causes the tissue to partially expand with the original structure being restored. This produces a porous and more permeable material that is crisper and more brittle. The DIC plant consists of three sections: In the pre-drying module, the product achieves an appropriate moisture level and a rubbery-plastic consistency. In the texturing module, overpressure is generated using a jet of hot steam that softens the structure of the product. Next there is an instantaneous pressure drop, which causes the tissue to expand. Finally, in the third and final module, the textured product is almost completely dehydrated. The resulting dried end product exhibits some improvements in regard to consistency, rehydratability and appearance, such as the colour.