METHOD FOR CHARACTERIZING A PRODUCT WITH ANTIOXIDANT ACTION AND SYSTEM FOR CARRYING OUT THIS METHOD

Description

technical field

[0001] The field of the invention is that of food products, food supplements or pharmaceuticals exhibiting antioxidant activity, and more specifically that of determining the composition of such a product in molecules giving it its antioxidant activity. Previous technique

[0002] An antioxidant molecule is a molecule that slows down or prevents the oxidation of other chemical substances upon contact with it.

[0003] In the health and food industry, antioxidant molecules and their effects represent an active field of research, these molecules being able to limit oxidative stress at the cellular level for an individual, whereas this oxidative stress can become pathological when it is excessive, for example when species such as ROS or RNS (respectively Reactive Oxygen Species and Reactive Nitrogen Species in English terminology) are produced in excess by the body.

[0004] In this context, the intake of antioxidants in pharmaceutical, cosmetic, food, or dietary supplement form is considered likely to limit the damage linked to excess ROS and NRS and thus prevent the occurrence of various diseases such as neurodegenerative disorders (Parkinson's disease, Alzheimer's disease, cerebral ischemia), cardiovascular diseases (atherosclerosis and hypertension), diabetes mellitus, respiratory diseases (asthma), development of cataracts and cancers (colorectal, prostate, breast, lung, bladder cancers).

[0005] We can refer to the publication by Qader et al., “Natural Nrf2 Activators from Juices, Wines, Coffee, and Cocoa”, Beverages 2020, 6(4), 68.

[0006] Among the antioxidant molecules, polyphenols, which include flavonoids and non-flavonoids as well as other phenolic compounds, are particularly noteworthy.

[0007] Polyphenols are responsible for the antioxidant power of many beverages, with, to give some examples of this large family of compounds, chlorogenic acid for coffee, quercetin, chlorogenic acid and resveratrol for wine, catechins and phenolic acids for beer, quercetin and chlorogenic acid for tea or quercetin, chlorogenic acid and 4-ethyl catechol for fruit juices.

[0008] Therefore, producers or consumers of these products wish to know more precisely their characteristics regarding their antioxidant activities for prophylactic or curative purposes.

[0009] However, a single product, a red wine for example, can contain several dozen antioxidant phenolic molecules (anthocyanins, dihydroflavonols, flavanoles, flavanones, flavonoids, tyrosol etc...) each participating in the overall antioxidant activity of the wine in question.

[0010] In addition, the variety and method of growing the grapes, the winemaking method, and the duration and conditions of storage (temperature, exposure to light, humidity, etc.) all influence the composition of the wine.

[0011] Conventional wine analysis methods involve measuring one or more specific molecules, which requires a level of skill and specialization that few laboratories possess, analysis times that can in practice be measured in weeks, and very high costs that quickly become prohibitive for a producer who would like to monitor the presence of several molecules in their product.

[0012] Alternatively, some technologies such as PAOT® technology are capable of rapidly and cost-effectively measuring the total antioxidant activity of a product by electrochemical methods, as detailed in patent application FR 3 089 011.

[0013] However, this is a global measurement on a given product and this type of measurement does not by itself allow for a detailed breakdown of the composition of any product in its antioxidant compounds.

[0014] We therefore see that there is an unmet need for a method of determining the antioxidant compounds of any product that is fast, cheap and accessible to a large number of users without being limited to a few qualified professionals.

[0015] The publication by Erica Sharpe et al., "Metal oxide based multisensor array and portable database for field analysis of antioxidants", Sensors and Actuators B: Chemical, Volume 193, 2014 presents a method for detecting polyphenols and characterizing samples containing an antioxidant. Description of the invention

[0016] One object of the invention is to provide a method capable of leading to the determination of the concentrations of compounds exhibiting antioxidant activity, or antioxidant compounds, in any product exhibiting antioxidant activity.

[0017] For these purposes, the invention relates to a method for characterizing a target product to be characterized exhibiting antioxidant activity, comprising a step of receiving a signal representative of a measurement of the overall antioxidant activity of the target product to be characterized, the method being characterized in that it further comprises the steps of receiving, by means of a computer processing unit, a signal representative of a type of the target product to be characterized; identifying in a database a reference product corresponding to the target product to be characterized on the basis of the signal representative of a type of the product to be characterized; retrieving from the database identifiers of antioxidant constituents of the target product to be characterized on the basis of the signal representative of a type of the product to be characterized;retrieve from the database, using the computer processing unit, (a) coefficients respectively representing the antioxidant activities of the antioxidant constituents in the target product to be characterized, and (b) individual antioxidant activities of the constituents considered individually; and calculate, using the computer processing unit, the concentrations of the antioxidant constituents in the target product to be characterized from the signal representing an overall antioxidant activity of the target product, the coefficients, and the antioxidant activities from the equation; C ′ j = Coef j × AA GlobMes AA j in which C '< j represents the concentrations of antioxidant constituents in the target product, Coefficient j indicates the relative contributions of the constituents to the overall antioxidant activity of the reference product, AA GlobMes represents the overall antioxidant activity of the target product and AA jrepresents the individual antioxidant activities of the constituents considered individually, and the relative contributions of the constituents to the antioxidant activity indicated by Coefficient j are derived from the equation Coef j = C j ∑ j C j in which C j represents the respective concentrations of the constituents in the reference product (Prod i).

[0018] This process can be implemented using PAOT® technology or any other technology that allows for the measurement of the overall antioxidant activity of a product.

[0019] A first advantage of this process is that, at the end user level, a single measurement of the overall antioxidant activity of a product makes it possible to deduce all the concentrations of the constituents of that product which participate in the antioxidant activity of the latter.

[0020] A second advantage is that no special expertise is required to implement the process, as the aspects of the process requiring expertise are deferred to the preparatory phases for determining the concentrations of antioxidant constituents of the product to be characterized.

[0021] Additional advantages, arising directly from the process, include reduced cost, high flexibility and high responsiveness of the process, since it only requires the user to prepare a single sample for a single measurement which can be carried out very simply using an electrochemical measurement device of the total antioxidant activity of a product such as PAOT® technology.

[0022] Thus, the implementation of the process according to the invention, which is flexible and of modest cost, allows for monitoring the production of products such as wine, fruits and their juices, tea, and coffee in order, for example, to optimize their composition with a view to preventing or treating diseases related to oxidative stress.

[0023] According to other non-limiting features of the invention, taken alone, or according to any technically feasible combination: The calculation may be carried out in such a way as to consider only antioxidant constituents whose individual antioxidant activities multiplied by their respective concentrations exceed 5% of the overall antioxidant activity of the reference product or target product to be characterized; the calculation may be carried out in such a way as to consider only antioxidant constituents whose sum of antioxidant activities weighted by their respective concentrations exceeds 80% of the overall antioxidant activity of the reference product or target product to be characterized; and the process may include the steps of measuring the overall antioxidant activity of the target product by means of a measuring device, and of generating the signal representative of a type of the target product to be characterized on the basis of information stored in a computer memory and the signal representative of an overall antioxidant activity of the target product on the basis of a result of the measurement.

[0024] The invention extends to a system for characterizing a target product to be characterized exhibiting antioxidant activity, the system comprising a database and a computing unit configured to implement the process. BRIEF DESCRIPTION OF THE FIGURES

[0025] Other features and advantages of the invention will become apparent from the detailed description of the invention that follows, with reference to the accompanying figures, in which: [ Fig.1 ] There [ Fig.1 ] illustrates a method according to the invention; [ Fig. 2 ] There [ Fig. 2 ] illustrates a system for implementing a process according to the invention; and [ Fig.3 ] There [ Fig.3 ] represents a table illustrating the implementation of the process of the [ Fig.1 ]. DETAILED DESCRIPTION OF THE INVENTION

[0026] A process according to the invention consists, in a preparation phase, of carrying out on the one hand chemical analyses of products in order to determine their respective compositions in constituents each having an antioxidant action, and on the other hand of determining by measurement the antioxidant activity of each of these constituents considered individually.

[0027] In an application phase, we carry out a measurement of the analysis of the overall antioxidant activity of a target product to be characterized, then we trace back to the constitution of this target product in antioxidant constituents using the data obtained during the preparation phase and the result of the measurement.

[0028] These two phases and an example of their implementation and associated means are detailed below. Preparation phase

[0029] The preparation phase includes background data collection work, which consists first of all of identifying and listing, in a computer database DB, reference products having antioxidant properties due to their content of antioxidant constituents, during a step S10 of a process 100 for characterizing a product exhibiting antioxidant activity illustrated by the [ Fig.1 ].

[0030] We know of certain products that have antioxidant effects when consumed in food, such as wines, beers, teas, coffees or certain fruit juices, but we can also be interested in, for example, the antioxidant characteristics of biological fluids (blood, urine...) or cosmetic products (creams, lotions...).

[0031] From the perspectives of producers, suppliers, or end consumers, it would be interesting to characterize in particular the respective content of these products in antioxidant constituents.

[0032] In the case of a wine producer, such characterization would allow for more precise monitoring of the influence of wine production parameters, ranging from the type of grape and its cultivation method to the winemaking process and storage conditions on its content of one or more specific constituents.

[0033] At step S20, the respective constituents that give each of the reference products listed during step S10 are identified and associated with them, for example in the form of a table stored in the computer database DB.

[0034] These constituents are designated by identifiers Const ij indicating a jth constituent of an ith reference product Prod i.

[0035] This step can be carried out by a literature review of already identified antioxidant constituents.

[0036] Alternatively, the antioxidant activity of the constituents of the identified reference products can be tested using known methods.

[0037] At step S30-1, the concentrations of the constituents of each of the listed reference products are measured, at step S30-2, the antioxidant activity of the constituents taken in isolation is measured, and at an optional step S30-3, the overall antioxidant activity of the listed reference products is measured.

[0038] Measurements of antioxidant concentrations and activities can be carried out using conventional methods.

[0039] Antioxidant activity measurements can be carried out in particular using PAOT® technology, which, in the case of work on a liquid sample, consists of measuring the electrochemical changes of an aqueous solution containing a molecule M in the free radical state using a working electrode and a reference electrode, following the addition of a sample to be analyzed.

[0040] These changes are explained by variations in the concentrations of oxidized / reduced forms of M in the presence of antioxidants, antioxidant activity being obtained by measuring a decrease in the electrochemical potential of the solution and is evaluated according to the following formula: activité antioxydante = EPproduct 10 − EPcontrol 0 EPcontrol 0 × 100 % where EPproduct10 is the electrochemical potential at the end of the analysis, i.e. when the potential of the solution stabilizes (automatic stop of the analysis) and EP-control0 is the electrochemical potential at time 0, the instant of the addition of the sample.

[0041] The expression "antioxidant activity" refers both to a physico-chemical phenomenon and to a numerical estimate of this activity, which could be reformulated as "level of antioxidant activity".

[0042] Depending on the products considered, the molecule M used may be of different natures, as determined by the practitioner.

[0043] We can refer to the publication by Joël Pincemail et al., “PAOT-Liquid® Technology: An Easy Electrochemical Method for Evaluating Antioxidant Capacity of Wines”, Diseases 2019 Jan 21;7(1):10.

[0044] The measurement results from steps S30-1 and S30-2 are stored in the database DB, in association with the relevant constituents; this data can, for example, be visualized in the form of the table represented by the [ Fig.3] including the data corresponding to two products i Prod i, their respective constituents Const ij, the concentrations C ij and the respective individual antioxidant activities AA ij of each constituent j in a product i, i ranging from 1 to 2 for the two products considered, j ranging from 1 to 5 for product 1, indicating that 5 constituents are associated with this product 1, j ranging from 1 to 3 for product 2, indicating that 3 constituents are associated with this product 1, in this example.

[0045] At this stage, it is optional to introduce at least one criterion for selecting constituents for each of the products so as to retain in the database only those constituents with significant antioxidant activity for the product concerned, during an S40 step.

[0046] Alternatively, all components can be kept in the database, but subsequent calculations will only consider those components that meet the said selection criterion.

[0047] Advantages of this practice include the fact that the database does not need to be exhaustive in identifying the antioxidant constituents of each of the reference products to be usable, which can significantly reduce the amount of work needed to populate it with data and make it accessible for characterizing products about which knowledge is not yet complete.

[0048] Furthermore, simplifying the data taken into account can allow for simpler implementation of the process and better handling by a non-specialist user.

[0049] A first possible selection criterion consists of considering for each reference product only the antioxidant constituents whose antioxidant activities, weighted by their respective concentrations, exceed a threshold defined by the user, for example 5%, preferably 10%, of the overall antioxidant activity of the product considered.

[0050] A second possible selection criterion consists of considering for each reference product only those antioxidant constituents whose sum of antioxidant activities weighted by their respective concentrations, i.e. the product of the antioxidant activity by the concentration for each of the constituents considered, exceeds 80%, preferably 90%, of the overall antioxidant activity of the product considered.

[0051] These two criteria require the measurement of the overall antioxidant activity of the reference product in question, which can be recorded in the DB database.

[0052] These criteria can be combined, meaning that a given constituent of a given product may have to meet one, two, or three of the above criteria to be included in the database or to be taken into account in subsequent calculations.

[0053] At step S50, based on the data obtained, we can define coefficients Coef j indicating the relative contribution of a constituent Const j to the overall antioxidant activity of a reference product Prod i according to equation 2, Coef j = C j ∑ j C j then store them in the database DB, as seen in Table Tab 1.

[0054] The above steps can be carried out gradually, in the laboratory and in a decentralized manner so as to gradually enrich the database of listed products for increasingly diverse applications and increasingly precise results.

[0055] The following steps are initiated by an end user, who will make good use of the results of the data collection and calculation work carried out during steps S10 to S50. Application phase

[0056] The application phase consists, for an end user, of measuring the overall antioxidant activity of a target product to be characterized having antioxidant activity and of a type listed in the database DB, with a view to determining, from this single measurement and the information stored in the database, the concentrations of the antioxidant constituents of this target product.

[0057] Such a determination is made possible by the fact that the overall antioxidant activity of any product represents the sum of the antioxidant activities of each of its components weighted by the concentration of the component constituting the product, as represented by the general formula Equation 1: AA Glob produit = ∑ k C k AA k Or C k And AA k represent respectively the concentration and antioxidant activity of a kth constituent of the product.

[0058] Consequently, from (a) the data recorded in the DB database which indicate the concentration and antioxidant activity of the constituents of a reference sample of a product under consideration, each constituent being considered individually, (b) the relationship given above between overall antioxidant activity and individual antioxidant activity of each of the constituents of the product under consideration and (c) the overall antioxidant activity of a measurement sample of a target product to be characterized of the same nature as the product under consideration, it is possible to trace back to the antioxidant constituent composition of the measurement sample of said target product to be characterized.

[0059] Thus, a first step S60 to be carried out by the end user consists of measuring the overall antioxidant activity of a target product to be characterized, which gives a result AA GlobMes; this measurement is recorded locally in a computer memory Mem by the user, possibly automatically by means of the measuring device used.

[0060] The measurement method used to determine the overall antioxidant activity of a target product is preferably the same as that used to determine the antioxidant activities of the constituents of the reference products during the preparatory phase.

[0061] At a step S70 following, or preceding, step S60, the user enters into computer memory Mem the type Typ of the target product, or the nature of this product, whose overall antioxidant activity he has measured.

[0062] This could be a type of wine, coffee, fruit juice, biological fluid and, more generally, any sample in which antioxidant activity is likely to be measured.

[0063] The user may have at his disposal a display device Disp displaying a list of reference products Prod i mentioned in the database DB, and may choose from this list a reference product corresponding to the target product by input means Inp according to conventional computer methods, so as to associate with the result of the measurement carried out the type of product on which the measurement was made within the computer memory Mem.

[0064] It is important for the user to choose a reference product that is as close as possible to the target product in terms of chemical composition: in a given reference product, the nature and relative concentrations of the antioxidant constituents are largely constant, and this is what makes it possible to reliably trace back to these concentrations for the target product to be characterized from a single measurement of overall antioxidant activity.

[0065] In this way, we can consider that the coefficients Coef ij determined in step S50 for a reference product are respectively representative of the antioxidant activities of the constituents Const ij in the target product to be characterized.

[0066] The user may, for example, be a winemaker wishing to characterize a wine of a given grape variety from a certain year by comparison with reference data obtained for the same wine from a previous year and recorded in the database DB.

[0067] At an S80 step, a Sig typ signal representing the type Typ of the target product to be characterized stored in the Mem memory and a Sig mes signal representing the result of the AA GlobMes measurement are generated and transmitted by a Com communication unit to a Calc computer calculation and processing unit functionally linked to the DB database.

[0068] The user-operated measurement device Mes, the computer memory Mem, the display device Disp, the input means Inp, and the communication unit Com can be integrated within a Measurement Module Mod, and the computing and processing unit Calc and the database DB can be remote from the Measurement Module Mod, as illustrated by the [ Fig. 2 ].

[0069] In particular, the database and the computer computing and processing unit can be connected to the Mod measurement module via a computer network such as the internet.

[0070] However, this is only an example configuration, and one can imagine that the database and the computer calculation and processing unit are also integrated within the measurement module Mod, or that the measurement is carried out completely independently of any integrated system and that its result is entered manually into the computer memory Mem.

[0071] A process implementation system 100 can be considered as a fully integrated form in a single piece of equipment or, alternatively, as a distributed set of physically separate but functionally connected pieces of equipment, for example by means of a computer network.

[0072] In any case, the invention is not limited to a particular implementation of the hardware elements used for its implementation,

[0073] At step S90, using the information on the type of reference product transmitted by the signal Sig typ, the Calc computing and processing unit identifies (step S90A) a reference product (Prod i) from the database (DB) corresponding to the product to be characterized and retrieves (step S90B) from the database DB the identifiers of the constituents j Const ij of this reference product i as well as the associated data, more specifically their respective coefficients Coef ij and their individual antioxidant activities AA ij, then calculates (step S90C) the concentrations C '< j of each of the constituents j of the target product having antioxidant activity by means of the formula equation 3: C ′ j = Coef j × AA GlobMes AA j

[0074] It should be noted that the calculation can be carried out by considering only the constituents that meet one or more of the criteria mentioned in step S40.

[0075] Furthermore, with regard to the second and third selection criteria, the thresholds can be calculated as percentages of the total antioxidant activity of the reference product or, alternatively, as percentages of the total antioxidant activity of the target product as just measured by the end user.

[0076] At step S100, the Calc computing and processing unit sends the Const ij identifiers of the antioxidant constituents and their respective concentrations back to the Com communication unit. C '< j deduced from the measurement to store them in the Mem memory and display them on the Disp display device for the user.

[0077] We see that, unlike a classic approach, which requires a heavy investment in terms of time spent, cost and contact to be made with specialists, the process according to the invention is fast, simple, and accessible to a non-specialist.

[0078] The general formula of equation 1 is strictly accurate only if each of the antioxidant constituents in product A and their respective antioxidant activities are known.

[0079] However, this general formula will remain usable despite the possible presence of antioxidant constituents that are not known or a voluntary choice to retain in the calculations only the constituents with significant antioxidant activity, either because the known constituents allow a good approximation (one can refer to the criteria of step S40), or because it is the variations in the characteristics of a product that are important and can be effectively studied even if antioxidant constituents are neglected, regardless of their relative shares in the overall antioxidant activity of the product.

[0080] Of course the invention is not limited to the above description, and alternative embodiments can be made without departing from the scope of the invention as defined by the claims.

Claims

1. Method (100) for characterizing a target product to be characterized that exhibits antioxidant activity, comprising a step (S60) of receiving a signal (Sigmes) which is representative of a measurement of the overall antioxidant activity of the target product to be characterized, the method being characterized in that it further comprises the steps of: - receiving (S80), by means of a computer processing unit (Calc), a signal (Sigtyp) which is representative of a type of the target product to be characterized; - identifying (S90A), in a database (DB), a reference product (Prodi) corresponding to the target product to be characterized on the basis of the signal (Sigtyp) which is representative of a type of the product to be characterized, - retrieving (S90B), in the database (DB), identifiers (Constij) of antioxidant constituents of the target product to be characterized on the basis of the signal (Sigtyp) which is representative of a type of the product to be characterized; - retrieving (S90B), in the database (DB) by means of the computer processing unit (Calc), (a) coefficients (Coefij) which are representative, respectively, of the antioxidant activities of the antioxidant constituents in the target product to be characterized, and (b) individual antioxidant activities (AAij) of the constituents considered individually; and - calculating (S90C), by means of the computer processing unit (Calc), the concentrations (C'j) of the antioxidant constituents (Constij) in the target product to be characterized from the signal (Sigmes) which is representative of the overall antioxidant activity of the target product, from the coefficients (Coefij) and from the antioxidant activities (AAij) using the equation C ′ j = Coef j × AA GlobMes AA j , where C'j represents the concentrations of the antioxidant constituents in the target product, Coefj indicates the relative contributions of the constituents to the overall antioxidant activity of the reference product (Prodi), AAGlobMes represents the overall antioxidant activity of the target product, and AAj represents the individual antioxidant activities of the constituents considered individually, and the relative contributions of the constituents to the antioxidant activity, indicated by Coefj are derived from the equation Coef j = C j ∑ j C j , where Cj represents the respective concentrations of the constituents in the reference product (Prodi).

2. Method according to claim 1, in which the calculation (S90) is performed considering only the antioxidant constituents of which the individual antioxidant activities (AAij), multiplied by their respective concentrations (Cij), exceed 5% of the overall antioxidant activity of the reference product or the target product to be characterized.

3. Method according to claim 1, in which the calculation (S90) is performed considering only the antioxidant constituents of which the sum of antioxidant activities, weighted by their respective concentrations, exceeds 80% of the overall antioxidant activity of the reference product or the target product to be characterized.

4. Method according to any of the preceding claims, further comprising the steps of: - measuring (S60) the overall antioxidant activity of the target product by means of a measuring device (Mes); and - generating (S80): - the signal (Sigtyp) which is representative of a type of the target product to be characterized on the basis of information stored in a computer memory (Mem); and - the signal (Sigmes) which is representative of the overall antioxidant activity of the target product on the basis of a result of the measurement (S60).

5. System (Sys) for characterizing a target product to be characterized that exhibits antioxidant activity, the system comprising a database (DB) and a computing unit (Calc), which are configured to implement the method according to any of claims 1 to 4.