PN-II-PT-PCCA-2013-4-0297

 

 

 

 

 

                                                             

 

MULTIPARAMETRIC SYSTEM FOR WINE MONITORING FERMENTATION

 

 

 

PROJECT PRESENTATION

 

 

Project number: PN-II-PT-PCCA-2013-4-0297; Contract no. 146/2014

 

Project Director: Ph.D Petruta-Madalina BUCUR;

 

Project category: PCCA;

 

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

 

 

 

PARTNERS

 

 

 

PROJECT COORDINATOR

 

NATIONAL INSTITUTE OF RESEARCH AND DEVELOPMENT FOR BIOLOGICAL SCIENCES- Bioanalysis Center

     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

 Faculty of Environmental Science and Engineering

     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 (Turkey).

 

PARTNER 2 (P2)

UNIVERSITY POLITEHNICA OF BUCHAREST

Faculty of Electronics, Telecommunications and Information Technology

     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)

CEPROCIM S.A.

     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-STI program - Software system for the control of pollution dynamics generated by the cement plants. He is member of Engineers Association from Romania – AGIR and he represented Romania in the Technical Working Group from EIPPCB Sevilla. Since 2012 he is correspondent of the Academy of Technical Sciences from Romania.  Also he was member in the Scientific Committee of the “First Nanostructured Multifunctional Materials National Conference (NMM07)”, with international participation, 2007, and of International Conference “International Proficiency Testing Conference” (PT- Conf) in 2007, 2009, 2011 and president of the Conference on Science and Engineering of Oxide Materials – CONSILOX (2004, 2008, 2012).

 

 

FINANCIAL RESOURCES

 

 

YEAR

Public budget (lei)

Own Budget (lei)

TOTAL(lei)

2014

85646

12854

98500

2015

822274

123340

945614

2016

460038

55312

515350

2017

346087

41607

387694

TOTAL (lei)

1250000

187508

1437508

 

ABSTRACT

 

 

 

 

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.

 

PHASES

 

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 fermentation (30 June 2016).

 

 

 

SHORT PRESENTATION OF THE SCIENTIFIC RESULTS

 

Phase 1/2014

 

                  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.

 

 

Fig.1. Cyclic voltammograms for EPC-Z (A); EPC-Z-AT (B) și EPC-Z-AM (C) electrodes.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 


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

 

 

 

               Scientific Report of Phase 1.pdf

 

Phase 2/2015

 

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 FIA with the enzymatic biosensor based on alcohol oxidase for must sample obtained from white grapes.

 

C:\Users\mu\Pictures\Slide Shows\20151113_145702.jpg

 

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)

 

 

Scientific Report of Phase 2.pdf

 

Phase 3/2016

 

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.

C:\Users\Andrei\Desktop\Cablaj.jpg 

 

Scientific report of Phase 3

 

Phase 4/2017

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 CPC - response time, output current (0.25 - 1.55 μA) and sensitivity (~ 35 mA / M) - are comparable to those of the professional device, but at a significantly reduced cost. It was observed a good correlation between the results obtained with the potentiostat produced and efficient commercial equipment. The development of a biosensor for lactic acid has also been finalized and used to analyze real wine samples. A HPLC methods with UV or MS detection have been developed and optimized for the detection of acids from real and synthetic samples of wine.

 

 

 

Final report

DISSEMINATION

 

A. Articles:

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, Alba County, Romania, Chemie der Erde – Geochemistry, 2016, submitted.

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 in Romania, Journal of Soil and Water Conservation,  2016, submitted.

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)

 

B. Conferences:

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 Semiconductor Conference-CAS 2015, 12-14 octombrie 2015. Sinaia, Romania (prezentare orala);

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 Semiconductor Conference-CAS 2015, Sinaia, Romania, 12-14 octombrie 2015 (prezentare orala);

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 Conference-CAS 2015, Sinaia, Romania, 12-14 octombrie 2015 (prezentare orala);

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 electrodes, CEEC-TAC4, 4th Central and Eastern Europe Conference on Thermal Analysis and Calorimetry, Chișinău, Moldova, 28-31 august 2017 (poster).

 

C. Patent claim

A00610/01.09.2017: I. Rusu, G. Brezeanu, G. Pristavu, F. Draghici, A. Enache, B. Bucur, M.P. Bucur, G.L. Radu: Potentiostat de zgomot redus pentru masuratori amperometrice in solutii agitate magnetic.

 

D. PhD thesis:

Chira Ana successfully sustained her doctoral thesis in 2014.

 

Updated: September 2017