Institutul Național de Cercetare-Dezvoltare pentru Tehnologii Criogenice și Izotopice Ramnicu Valcea



Financing contract no.: 146PD/2020

Project No.: PN-III-P1-1.1-PD-2019-0325

Project implementation period: 01.09.2020-30.08.2022



The proposal is based on a multi-disciplinary approach focused to develop new comprehensive and accessible mapping techniques of the apple-juice-cider-vinegar technological process in terms of stable isotope and multi-element patterns by representing it in a statistical model used to track biogeochemical processes, as scientific basis for investigating complex foodstuff.

The project objective is to provide new tools to determine strong unchangeable bonds in the iso-chemical state of the fruit to the based products and to offer the first elements from which reconstruction of this very important relationship between climate, region, technology and chemistry can be derived from simple equations.

Specifically, the proposal will provide a better characterization and understanding of those processes and markers that may differentiate the chemical and isotopic pattern of apples from different cultivars and of its derived fermentation products and will develop an advanced statistical model for data evaluation and interpretation, able to ensure a correct evaluation and interpretation regarding the apple products authenticity.

Besides highlighting the opportunities offered by the existing research infrastructure, these services will bring important information feedback required to promote uniform guidelines for apple products standardization and for their more effective control on the market facilitating thus the cooperation of research entities with stakeholders from the food industry sector.

Essential data will be obtained in order to achieve objective such: advanced investigations on authenticity and traceability indicators based on isotopic and polyphenolic fingerprint and quantitative and qualitative analysis of apple/apple products; and chemometrics methods for providing an integrated statistical approach for classification and distinguishing apples fruits and derived products according to the origin.

Project Team

Project leader: PhD. Chem. Oana Romina BOTORAN ;
National Research and Development Institute for Cryogenics and Isotopic Technologies – ICSI Rm. Valcea; ICSI Analytics Department

Mentor: Prof. PhD. Nicoleta STĂNCIUC ;
Dunărea de Jos University of Galați, Faculty of Food Science and Engineering, Department of Food Science, Food Engineering and Applied Biotechnology, Integrated Center of Research, Expertise and Technological Transfer in the Food Industry (BioAliment – TehnIA)


General objectives
A complex map of the apple-juice-cider-vinegar technological process based on the isotopic, compositional and bioactive compounds fingerprint, according to botanical and geographical origin, using chemometric methods and mathematical models will be performed. Will be performed a highly proactive approach based on a comprehensive data base of isotopic and compositional parameters that can be used both to detect adulterated products as soon as they appear on the market and even to dissuade the defrauders before they begin. Using a multi-variables approach and given the leading project target, the objectives to be achieved by the proposed study are to:

O1. Investigating the key elements needed to develop new analytical tools for certification and authentication of the apple-juice-cider-vinegar chain and set the strategy of selecting, collecting and preparing the appropriate test samples for further analytical investigations; set of a referential product database;

O2. Generate the specific isotopic (e.g. 13C, 2H, 15N, 18O), compositional (mineral content, organic acids, sugars, amino acids) and bioactive compounds (polyphenols, including phenolic acids, anthocyanins, flavonoids, etc.) fingerprint of apple and apple-based products; develop and optimise separation procedures and analytical methods to improve the quantification of marker compounds;

O3. Assess bioactive compounds (polyphenols, including phenolic acids, anthocyanins, flavonoids, etc) and antioxidant activities of apple/apple products, to evaluate the functionality of these products and establish biochemical indicators (e.g. polyphenoloxidase, peroxidase) in order to advance the knowledge concerning the impact of enzymatic browning on the quality of apples and derivative products;

O4. Develop appropriate mathematical models for metabolic flux analysis for apples based-products mapping using multivariate statistical methods based on constructed isotopic fingerprint of fruit juice products from individual isotope contents (e.g. 13C, 2H, 15N, 18O) in different apple components (e.g. pulp, sugars and proteins) and phytochemical and biochemical data using dedicated analytical techniques;

O5. Provide an integrated statistical approach to enable the correct evaluation and interpretation of isotopic data and promote accessible, more sensitive and cost-effective procedures based on applications of fingerprinting techniques in detecting fraudulent fruit juices.

SMART objectives


Expected results


  • Database with raw material chain profile and isotopic signature of authentic and artificial fruit-based products;
  • Validation of advanced methods for apple products composition using specific techniques (e.g. chromatography, spectrophotometry, and spectrometry) in combination with proper multivariate statistical analysis methods;
  • Development and implementation of new integrated statistical approach using mathematical models for metabolic flux analysis, stable isotope complex mixing models and Bayesian models to realize a correct evaluation and interpretation;
  • Database for traceability on the apple-juice-cider-vinegar chain.


Total budget: 239.350 RON
Budget allocation by stages (in RON):
Stage 1: Investigating the key elements needed to develop new analytical tools for certification and authentication of the apple-juice-cider-vinegar chain. Set the strategy of selecting, collecting and preparing test samples for further analytical investigations and designing the referential database.
Total budget: 55.000
• Stage 2: Generate the specific isotopic, compositional and bioactive compounds fingerprint of apple and apple-based products.
Total budget: 110.000
• Stage 3: Development and validation of a information system and a database with origin markers for the apple-juice-cider-vinegar chain.
Total budget: 74.350

Project progress: public results & dissemination

Public results

  • Stage 1/2020
  • Stage 2/2021
  • Stage 3/2022


Manuscripts published in per review ISI and BDI journals:

  • Silviu Laurentiu BADEA, Roxana Elena IONETE, Diana COSTINEL, Constantin NECHITA, Mihai BOTU, Oana Romina BOTORAN, Changes of carbon-isotope ratios in soil organic matter relative to parent vegetation and site specificity, acceptat spre publicare in revista Notulae Botanicae Horti Agrobotanici Cluj-Napoca, Volume 48, Issue 4, 2020 (10.15835/nbha48412138), IF/2019 = 1.168 (Q2).

Scientific manifestations:

  • European Biotechnology Congress 2020, September 24-26, 2020:, Roxana Elena IONETE, Diana COSTINEL, Nicoleta STANCIUC, Oana Romina BOTORAN, Stable isotopes signatures in fruits as tool to inferred climate variations – prezentare poster.


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