MULTIPARAMETRIC SYSTEM FOR WINE MONITORING FERMENTATION
Project number: PN-II-PT-PCCA-2013-4-0297; Contract no. 146/2014
Project Director: Ph.D Petruta-Madalina BUCUR;
Project budget: 1.437.508 lei (1.250.000 lei public budget and 187.508 lei own budget);
Contract period: 39 months (01.07.2014- 30.09.2017);
Contracting Authority: Executive Unit for Higher Education, Research, Development and Innovation Funding- (UEFISCDI);
Contractant: National Institute of Research and Development for Biological Sciences;
Contract number: 146/2014;
Contact: Madalina Bucur
Project Manager: Dr. Petruta Madalina BUCUR
Dr. Petruta Madalina BUCUR (ResearcherID: B-9624-2011) has obtained his Ph.D. in 2007 at University of Bucharest with thesis entitled: Chemical and biochemical sensors for analytical control of some pollutants in environmental sample. Her research interest covers different domains of analytical chemistry and biochemistry such as: enzymatic biosensors, electrochemical sensors, flow injection analysis methods, piezoelectric analysis, enzymatic biosensor development for substrate or inhibitor detection, different methods for enzyme immobilization and stabilization, use of electrochemical mediators. She is author of 17 original papers in peer-reviewed journals and 3 Romanian patents. She was director/responsible of 2 national project: PN-II-RU-TE-0076/2011 and SAFEFOOD, project no. 61030/2007. She has been researcher in 3 European contracts and has extensive experience in national grants (more then 10 projects).
PARTNER 1 (P1)
BABES BOLYAI UNIVERSITY
Responsible P1: Conf. Dr. Delia GLIGOR
Dr. Delia Gligor, is currently conf. PhD. at
the Department of Environmental Analysis and Engineering, Faculty of
Environmental Science and Engineering, Babeș-Bolyai University Cluj-Napoca.
After graduating the Faculty of Chemistry and Chemical Engineering, Physical
Chemistry specialization, has attended the Applied Electrochemistry Master and
the PhD in Electrochemistry (2002). The scientific articles (43 articles in ISI
journals) are published in high impact factor journals (Biosensors &
Bioelectronics, Electrochemical Acta, Electroanalysis, Microchemical Acta,
etc.) and representative for the electrochemistry and analytical chemistry
domain. The research activity is conducted over 17 years in the field of
analytical electrochemistry and was focused on the development and
characterization of new sensors changed with different innovative materials and
their applications in environmental pollution control, clinical laboratory,
biotechnology process control and products quality control. The relevant
contributions are: a book chapter in the prestigious publishing house Nova
Science Publishers, New York, numerous participations in international
conferences in the field and research internships in prestigious laboratories
from foreign universities (Lund University, Sweden, University Henry Poincaré,
Nancy, France and Cannakale Onsekiz Mart University, Turkey). He also
coordinated five national contracts (IDEAS, CEEX ET, CNCSIS AT ANSTI) and one
bilateral contract (
PARTNER 2 (P2)
UNIVERSITY POLITEHNICA OF BUCHAREST
Responsible P2: Conf. Dr. Ion RUSU
Dr. eng. Ion Rusu, Lecturer, University POLITEHNICA of BUCHAREST, Faculty of Electronic, Telecommunication and Information Technology- has obtained his Ph.D. in 2001 at University Politehnica of Bucharest, project: Microwave Circuits and Devices for hyperthermia therapy". His research interest: Electronic devices and microwave circuits, for medical therapies. He is co-author of 31 original papers in conference proceedings or journals and 2 Romanian patent. He has been researcher in over 38 national and international projects/grants and has extensive experience in national grants (director/scientific responsible for 17 research grants). He is co-author of 2 original book, "Biological effects of radiation RF and Microwave" and Absorption of electromagnetic field and human head area ".
PARTNER 3 (P3)
Responsible P3: Dr. eng. Doru Vladimir PUȘCAȘU
Dr. eng. PUȘCAȘU Doru Vladimir , Scientific
researcher I and Scientific Director. He graduated Polytechnic Institute
Traian Vuia from Timisoara in the year 1971 and has as main competences:
elaboration of studies of evaluation the impact on environment, environment
balances and risks evaluations for cement plants and quarries of raw materials,
researches regarding technological processes from cement industry, technical
assistance and technological expertise, upgrading and optimizations of
installations and equipments from building materials industry. He was involved as project director or as
partner in 16 national projects having as thematic the building materials, environment protection and efficient
using of thermal and electrical energy and as partner in international projects
on environment range, out of which we mention NATO/CORINT-
Public budget (lei)
Own Budget (lei)
The objectives of this project are the development of a complete analytical solution for the wine sector consisting of: (1) a multiparametric flow analysis system based on enzymatic biosensors for monitoring of alcoholic and lactic must fermentations (2) mineralogical and geochemical investigations of soil from vineries and (3) detection of heavy metals from soil and winemaking relevant matrices (grapes, must and wine). The European winemaking sector is facing an intense competition from imports of low-cost wines and is under pressure to improve the products quality. Because the winemaking process is an art highly dependent on the producers' experience and skill, fast, cheap and reliable analytical devices and services are vital to obtain an effective control at all stages of the production. The current analysis techniques (FTIR or chromatographic analyses) are long and expensive due to complexity of the sample matrix and too complex to be used in situ by the average winemaker. This multidisciplinary project will develop two enzymatic biosensors for the detection of alcohol (based on oxidases) and respectively lactic acids (based on dehydrogenases). The biosensors may be used independently or integrated in a multiparameter flow analytical system that will quantify the signal using a analog potentiostat. The heavy metal detection is important for two reasons: (1) they are involved in taste and flavor development and (2) to protect the consumers of contaminated products. The soil has a major influence on the grape quality and provides the wine specificity that determine the consumers fidelity for the products obtained from a specific "terroir". The combination of all three analytical systems provides a complete solution adaptable to the specificity of winemaker from various European countries each one with different domestic market and background.
Ø Development of a multiparametric system dedicated to the monitoring of must fermentation (10 December 2014);
Ø Construction of a multiparametric system dedicated to the monitoring of must fermentation - (10 December 2015);
Ø Testing of the performances achived by the multiparametric system for wine monitoring
SHORT PRESENTATION OF THE SCIENTIFIC RESULTS
In this phase CO it was developed a biosensor for alcohol analysis in batch system. The developed biosensor shows satisfactory analytical performance (a good sensibility, selectivity, limit of detection and measurement errors) to analyze the concentration of ethanol in wine samples. Preliminary studies were made to adapt the biosensor developed in a monocanal flow analysis system.P1 have develop new modified electrodes for the oxidation of NADH with low interference demonstrated by tests in synthetic samples.
P2 made studies on the design of analog blocks of the electronic component of biosensors and have made preliminary studies on the assessment of faradaic current supplied to realize biosensor measurement circuit. Simulations performed certify proposed circuit functionality with currents from tens to hundreds of nA. The choice of the connection cables between measuring cell and amplifier instrumentation is very important for electrical performances. For extreme caution regarding noise entering through power supply circuit will be used to power two 9V batteries through a connection that will provide adequate voltage. The entire measurement chain (INA118 and LPF) will be closed in a Faraday cage to shield noise effectively gave foreign sources of disturbance.
Theoretical studies have been conductedby P3 and P1 regarding the geology and soil from two vineyards selected from Stefanesti and Pietroasele viticultural areas. The selected grapes and wine are: Riesling and Cabernet.
Cyclic voltammograms for
Fig. 2. A) Analytical signal obtained with enzymatic biosensor; B) Calibration graph 0.1-12 mM ethanol
Fig.3. The general scheme for potentiostat low pass filter amplifier
In this phase it was developed a flow injection analyses system based on a biosensor for alcohol analysis. The developed system in flow based on alcohol oxidase from Pichia pastoris shows satisfactory analytical performance for the monitoring of alcoholic fermentation of must. Another objective was to develop an enzymatic biosensor based on dehydrogenases for the monitoring of lactic acid in the process of fermentation that present a good analytical performance.
It was made a potentiostat with positive potential and low pass filter using an amplifier. The measured performances are within the simulated results obtained with TINA8, and this confirms a good implementation of the electric scheme.
It was realized a geochemical investigation in two vineyards Stefanesti and Pietroasele: As, Cd, Cr, Co, Cu, Ni, Pb, Se, Zn, Fe, Mn, Ca, Mg, K were analysed in three profile of soil (0-20, 20-40, 40-60 cm).
Fig. 1. Analytical signal obtained in
Fig.2. Potentiostat with low pass filter amplifier.
Fig 3. The concentration in soil of metals: As, Cd, Cr, Co, Cu, Ni, Pb, Se, Zn.
Legend: RS = sample of soil from Stefanesti -Reisling
RP = sample of soil from Pietroasele - Reisling
CS = sample of soil from Stefanesti, -Cabernet
CP = sample of soil from Pietroasele - Cabernet
NV = Normal Value (756/1997)
In this phase it was tested a multiparametric system for alcoholic fermentation of must. It was developed the electrical circuit for an analog potentiostat studied in Phase 2 in order to implement the system for monitoring of the must fermentation by taking in consideration: linearity, stability, low noise, capability to ensure both polarities of voltage Vcell and the possibility to measure both senses of current Icell. Another objectives were to develop a biosensor based on electrode modified with toluidine blue and lactate dehydrogenase and a method based on microelectrodes modified with Prussian Blue in order to obtained microbiosensors based on alcohol oxidase.
It was realized a chromatographic method for the identification and quantification of L and D lactic acid by HPLC and it was made geochemical investigation of soil in two vineyards for Riesling and Cabernet Sauvignon.
In this phase a Control and Processing Circuit was developed and optimized in two variants. In the first variant, based on preliminary experimental results, the 5th order filter was moved from the potentiostat's reaction loop to the CCP output, leading to improvement stability. This filter has led to a significant reduction in the noise at CCP output (of
2.9 times as compared to the non-filter case). The
results of the measurements show a significant signal / noise signal
improvement (approx. 10dB) compared to the case without filter. The developed
circuit was compared to the PG580, highlighting a similar precision in measurement
cell current for the resistor range equivalents of the cell 200 kΩ - 1
MΩ. Comparative experimental
investigations were performed between the proposed circuit and the PGSTAT302N
system, using both a hydrogen peroxide sensor and a laboratory biosensor for
ethanol based on alcool oxidase. Performance achieved with Proposed
1. M.C Radulescu, P. M. Bucur, B. Bucur, G.-L. Radu, Biosensor based on inhibition of monoamine oxidases A and B for detection of β-carbolines, Talanta, 2015,137, pp 94-99, IF=3,54;
2. A. Chira, Bucur Bogdan, T. Galaon, G.-L. Radu, Study of the synthesis and environmental removal of 4,4 '-dipyridine derivatives, Environ Engineering and Management Journal, 2015, 14 (2), pp. 269-275, FI=1,065;
3. R. Bindiu, S. Filipescu, R. Balc, L. Barbu, D.M. Gligor The middle/late Eocene transition in the Eastern Carpathians (Romania) based on foraminifera and calcareous nannofossil assemblages Geological Quarterly, 2016, vol 60, no 1, FI=1,00;
4. M. C. Radulescu, M. P. Bucur, A. Alecu, B. Bucur, G. L. Radu: Electrochemical Determination of Hydrogen Peroxide using a Prussian Blue-Copper Modified Platinum Microelectrode, Analytical Letters, 2016, 49(13), FI=1,088
5. G. Ispas, C. Roba, R. Bălc, M. Coadă, D. Gligor, Assessment of environmental
factors in the Haneș mining area,
6. R. Bălc, T. Tămaș, G. Popiță, G. Vasile, M. Bratu, D.
Gligor, C. Moldovan, Assessment of chemical
elements in soil, grapes and wine
from two representative vineyards
7. B. Bucur, S.I. Hosu, A. Vasilescu : Book chapter "Biosensors for Characterisation of Bioactive Compounds from Wines", Frontiers in Bioactive Compounds, 2016, Vol. 1, 147-189 (Constantin Apetrei) ed. Bentham Science Publishers. ISSN: 2468-6395.
8. Maria-Alexandra Cîmpean, Izabella Crăciunescu, Delia Gligor, Amperometric sensor based on HEMA hydrogels modified with Toluidine Blue for nitrite detection in water samples, Materials Chemistry and Physics, 2017, 200, 233-240 (I.F. = 2.084).
9. Chira A., Bucur B, Radu G.L., Electrodeposited Organic Layers Formed from Aryl Diazonium Salts for Inhibition of Copper Corrosion, Materials, 2017, vol 10, (235), pag. 2-13 (I.F. =2,6)
1. M. C. Radulescu, B. Bucur, M.P. Bucur, G. L. Radu: Biosensor Based on Inhibition of Monoamine Oxidases A and B for Detection of B-Carbolines, ELSEDIMA International Conference, 18 19 September 2014, Cluj-Napoca, Romania (prezentare orala);
2. L. Barbu, D.M. Gligor: Carbon paste electrodes modified with natural zeolite adsorbed with toluidine blue and methylene blue for construction of a newbiosensor for wine control quality ELSEDIMA International Conference, 18 19 September 2014, Cluj-Napoca, Romania;
3. M.C. Radulescu, M.P. Bucur, B. Bucur, G.L. Radu Stable Prussian blue-copper modified (micro)electrodes for sensitive enzymatic biosensors based on oxidases, Seventh international workshop on « Biosensors for food safety and environmental monitoring 19-21 noiembrie 2015, Erfoud, Maroc (prezentare orala);
4. G. Brezeanu, M. Badila, F. Draghici, R. Pascu,
G. Pristavu, F. Craciunoiu, Ion Rusu: High temperature Sensors Based on
Silicon Carbide (SiC) Devices International
5. G. Pristavu, A Vasilica, V. Anghel, R. Pascu, G. Brezeanu, F.
Draghici: High voltage freewheeling diodes in an extended capability led
driving application International
6 R. Pascu, F. Craciunoiu, M. Kusko, M. Mihaila, G. Pristavu, M. Badila,
G. Brezeanu: SiO2/4H-SiC Interface states Reduction by POCl3 post oxidation
annealing, International Semiconductor
7. 1. G.-E. Popița, R. Bălc, G. Vasile, D. Gligor, Metal elements distribution in soil in two vineyards from Romania, Simpozion național cu participare internațională Environment & Progress, 30 octombrie 2015, Cluj Napoca Romania (poster).
8. Gabriela-Emilia Popiță, Ramona Bălc, Tudor Tămaș, Gabriela Vasile, Delia Gligor, C. Moldovan, Trace metal distribution in soils from two Romanian vineyards, European Society for Soil Conservation Conference ESSC 2016, Cluj-Napoca, 15-18 iunie 2016 (poster).
9. Pristavu G, Brezeanu G, Badila M., Draghici F., Pascu F., Craciunoiu F., Rusu I, Pribeanu A, High temperature behavior prediction techniques for non-uniform Ni/SIC Schottky diodes, International Conference on Silicon Carbide and Related Materials 2017 (ICSCRM 2017), 17-22.09.2017, Washington, D.C., SUA
10. Mihaela Ligia Ungureșan, Vlad Mureșan, Delia Gligor, Codruța Varodi,
Adsorption process of phenothiazine solution in dimethylsulfoxide on graphite
C. Patent claim
D. PhD thesis:
Chira Ana successfully sustained her doctoral thesis in 2014.
Updated: September 2017