METHOD FOR CHARACTERIZING A PRODUCT HAVING ANTIOXIDANT ACTIVITY, AND SYSTEM FOR IMPLEMENTING THIS METHOD

Abstract

The invention relates to a method (100) for characterizing a target product to be characterized having an antioxidant activity, comprising a step (S60) of receiving a signal representative of a measurement of an overall antioxidant activity of the target product to be characterized, the method further comprising the steps of retrieving (S90B) from a database, by means of a computer processing module, (a) identifiers of antioxidant constituents of the product to be characterized, (b) coefficients respectively representative of the antioxidant activities of the constituents in the target product to be characterized, and (c) individual antioxidant activities of the constituents considered individually;and calculate (S90C), by means of the calculation and computer processing module, the concentrations of the antioxidant constituents in the target product to be characterized from the signal representative of an overall antioxidant activity of the target product, the coefficients, and the antioxidant activities. Figure to be published with the abstract: Fig. 2;

Description

Title of the invention: METHOD FOR CHARACTERIZING A PRODUCT EXHIBITING ANTIOXIDANT ACTIVITY, AND SYSTEM FOR IMPLEMENTING THIS METHOD 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 providing it with its antioxidant activity. Prior art

[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 field of health and the food industry, antioxidant molecules and their effects represent an active field of research, these molecules being capable of limiting oxidative stress at the cellular level for an individual while 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, it is considered that taking antioxidants in pharmaceutical, cosmetic, food, or food supplement form is 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] Reference may be made 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, we can cite in particular polyphenols, which include flavonoids and non-flavonoids as well as other phenolic compounds.

[0007] Polyphenols are responsible for the antioxidant power of many drinks, 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] Thus, producers or consumers of these products wish to know more precisely their characteristics concerning their antioxidant activities for prophylactic or curative purposes.

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

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

[0011] Conventional methods of wine analysis consist of 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 counted in weeks, as well as very high costs, which quickly become prohibitive for a producer who would like to monitor the presence of several molecules in his product.

[0012] Alternatively, certain technologies such as PAOT® technology are capable of rapidly and inexpensively 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 in itself allow the composition of any product to be detailed in terms of its antioxidant compounds.

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

[0015] An 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.

[0016] To these ends, the invention relates to a method for characterizing a target product to be characterized having an antioxidant activity, comprising a step of receiving a signal representative of a measurement of an overall antioxidant activity of the target product to be characterized, the method further comprising the steps of recovering from a database, by means of a computer processing module, (a) identifiers of antioxidant constituents of the product to be characterized, (b) coefficients respectively representative of the antioxidant activities of the constituents in the target product to be characterized, and (c) individual antioxidant activities of the constituents considered individually; and calculate, using the calculation and computer processing module (Cale), concentrations of the antioxidant constituents in the target product to be characterized from the signal representative of an overall antioxidant activity of the target product, the coefficients, and the antioxidant activities.

[0017] This method can be implemented using PAOT® technology or any other technology allowing the overall antioxidant activity of a product to be measured.

[0018] A first advantage of this method is that, at the end user level, a single measurement of the overall antioxidant activity of a product makes it possible to deduce all of the concentrations of the constituents of this product which participate in the antioxidant activity of the latter.

[0019] A second advantage is that no particular expertise is necessary for the implementation of the process, the aspects of the process requiring expertise being carried over into the preparatory phases for the determination of the concentrations of antioxidant constituents of the product to be characterized.

[0020] Additional advantages, arising directly from the process, consist of reduced cost, great flexibility and great reactivity 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 by means of an electrochemical measuring device for the total antioxidant activity of a product such as PAOT® technology.

[0021] Thus, the implementation of the method according to the invention, flexible and of modest cost, allows monitoring of the production of products such as wine, fruits and their juices, tea, coffee so as, for example, to optimize their composition with a view to preventing or treating diseases linked to oxidative stress.

[0022] According to other non-limiting characteristics of the invention, taken alone, or in any technically feasible combination:

[0023] - the method may comprise the steps of receiving, by means of the module of computer processing, a signal representative of a type of the target product to be characterized, and to identify a reference product from the database corresponding to the target product to be characterized on the basis of the signal representative of a type of the product to be characterized, the recovery in the database of said identifiers of antioxidant constituents being able to be carried out on the basis of the signal representative of a type of the product to be characterized;

[0024] - the calculation can be carried out in such a way as to consider only anti constituents oxidants whose individual antioxidant activities are multiplied by their concen respective concentrations exceed 5% of the overall antioxidant activity of the reference product or the target product to be characterized;

[0025] - the calculation can be carried out in such a way as to consider only anti constituents oxidants 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

[0026] - the method may comprise the steps of measuring the overall antioxidant activity of the target product by means of a measuring device, and to generate 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 measurement result.

[0027] The invention extends to a system for characterizing a target product to be characterized having antioxidant activity, the system comprising a database and a calculation unit configured to implement the method. BRIEF DESCRIPTION OF THE FIGURES

[0028] Other characteristics and advantages of the invention will emerge from the detailed description of the invention which follows with reference to the appended figures, in which:

[0029] [Fig.l] [Fig.l] illustrates a method according to the invention;

[0030] [Fig.2] [Fig.2] illustrates a system for implementing a method according to the invention; and

[0031] [Fig.3] [Fig.3] represents a table illustrating the implementation of the method of the [Fig.l]. DETAILED DESCRIPTION OF THE INVENTION

[0032] A method according to the invention consists, in a preparation phase, in carrying out on the one hand chemical analyses of products so as to determine their respective compositions in constituents each having an antioxidant action, and on the other hand in determining by measurement the antioxidant activity of each of these constituents considered individually.

[0033] In an application phase, a measurement is carried out of the analysis of the overall antioxidant activity of a target product to be characterized, then the composition of this target product in antioxidant constituents is calculated using the data obtained during the preparation phase and the result of the measurement.

[0034] These two phases and an example of their implementation and associated means are detailed below. Preparation phase

[0035] The preparation phase includes in-depth data collection work which consists first of all in 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 method 100 for characterizing a product having antioxidant activity illustrated by [Fig.l].

[0036] Certain products are known to have antioxidant effects in the context of food consumption, such as wines, beers, teas, coffees or even certain fruit juices, but we can also be interested, for example, in the antioxidant characteristics of biological fluids (blood, urine, etc.) or cosmetic products (creams, lotions, etc.).

[0037] From the point of view of producers, suppliers, or end consumers, it would be interesting to characterize in particular the respective contents of these products in antioxidant constituents.

[0038] In the case of a wine producer, such characterization would allow 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 in one or more specific constituents.

[0039] In a step S20, the respective constituents which give them antioxidant activity are identified and associated with each of the reference products listed during step S10, for example in the form of a table stored in the computer database DB.

[0040] These constituents are designated by Constÿ identifiers indicating a jth constituent of an ith reference product Prod;.

[0041] This step can be carried out by a bibliographic study of the antioxidant constituents already identified.

[0042] Alternatively, the antioxidant activity of the constituents of the identified reference products can be tested using known methods.

[0043] In a step S30-1, the concentrations of the constituents of each of the listed reference products are measured, in a step S30-2, the antioxidant activity of the constituents taken in isolation is measured, and in an optional step S30-3, the overall antioxidant activity of the listed reference products is measured.

[0044] Measurements of antioxidant concentrations and activities can be carried out by conventional methods.

[0045] The measurements of antioxidant activities can be carried out in particular by means of PAOT® technology, which consists, in the case of work on a liquid sample, in measuring the electrochemical changes of an aqueous solution containing a molecule M in the free radical state by means of a working electrode and a reference electrode, following the addition of a sample to be analyzed.

[0046] These changes are explained by variations in the concentrations of the oxidized / reduced forms of M in the presence of antioxidants, the antioxidant activity being obtained by measuring a decrease in the electrochemical potential of the solution and is evaluated according to the following formula: [OO47] antioxidant activity = x 100%

[0048] where EPproductlO is the electrochemical potential at the end of the analysis, i.e. when the potential of the solution stabilizes (automatic stopping of the analysis) and EP-controlO is the electrochemical potential at time 0, the instant of addition of the sample.

[0049] The expression “antioxidant activity” designates both a physicochemical phenomenon and a numerical estimate of this activity which could be reformulated as “level of antioxidant activity”.

[0050] Depending on the products considered, the molecule M used can be of different natures, as determined by the practitioner.

[0051] Reference may be made 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.

[0052] The results of the measurements of steps S30-1 and S30-2 are stored in the database DB, in association with the constituents concerned, data which can for example be visualized in the form of the table represented by [Fig.3] comprising the data corresponding to two products i Prod i, their respective constituents Consty, the concentrations Cy and the respective individual antioxidant activities AAy of each constituent j in a product i, i varying from 1 to 2 for the two products considered, j varying from 1 to 5 for product 1, indicating that 5 constituents are associated with this product 1, j varying from 1 to 3 for product 2, indicating that 3 constituents are associated with this product 1, in this example.

[0053] At this stage, it is optionally possible to introduce at least one criterion for selecting the constituents for each of the products so as to retain in the database BD only the constituents having significant antioxidant activity for the product concerned, during a step S40.

[0054] Alternatively, all constituents can be kept in the database, but only those constituents meeting said selection criterion will be taken into account in subsequent calculations.

[0055] Advantages of this practice are that the database does not need to be exhaustive with respect to the identification of the antioxidant constituents of each of the reference products in order to be usable, which can considerably reduce the amount of work required to supply it with data and make it accessible for the characterization of products about which knowledge is not yet complete.

[0056] Furthermore, a simplification of the data taken into account can allow a simpler implementation of the method and better handling by a non-specialist user.

[0057] 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.

[0058] A second possible selection criterion consists of considering for each reference product only the 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.

[0059] These two criteria require the measurement of the overall antioxidant activity of the reference product considered, which can be recorded in the DB database.

[0060] These criteria can be combined, that is to say that a given constituent of a given product may have to respect one, two, or three of the above criteria to be retained in the database or to be taken into account in subsequent calculations.

[0061] At a step S50, starting from the data obtained, it is possible to define coefficients Coefj indicating the relative contribution of a constituent Constj to the overall antioxidant activity of a reference product Prod; according to equation 2, Coef. = ^ Equation2

[0062] then store them in the DB database, as visible in the table Tab 1.

[0063] The above steps can be carried out gradually, in the laboratory and decentralized ways to gradually enrich the database with listed products for increasingly diverse applications and increasingly precise results.

[0064] The following steps are at the initiative of an end user, who will advantageously use the result of the data collection and calculation work carried out during steps S10 to S50. Application phase

[0065] The application phase consists, for an end user, in measuring the overall antioxidant activity of a target product to be characterized having antioxidant activity and of a type listed in the DB database, 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.

[0066] 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:

[0067] AAGh^product) = 'LkCkAAk Equation 1 where Ck and AAk represent respectively the concentration and the antioxidant activity of a kth constituent of the product.

[0068] Consequently, from (a) the data recorded in the DB database which indicate the concentration and the antioxidant activity of the constituents of a reference sample of a product considered, 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 considered and (c) the overall antioxidant activity of a measurement sample of a target product to be characterized of the same nature as the product considered, it is possible to go back to the composition in antioxidant constituents of the measurement sample of said target product to be characterized.

[0069] 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 AAGiobMes; this measurement is recorded locally in a computer memory Mem by the user, possibly automatically by means of the measuring device used.

[0070] 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.

[0071] At a step S70 following, or preceding, step S60, the user enters into the computer memory Mem the type Typ of the target product, or the nature of this product, of which he has measured the overall antioxidant activity.

[0072] This may be a type of wine, coffee, fruit juice, biological fluid and, more generally, any sample of which antioxidant activity is likely to be measured.

[0073] The user may have at his disposal a display device Disp displaying a list of reference products Prod; 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.

[0074] It is important for the user to choose a reference product 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 return to these concentrations for the target product to be characterized from a single measurement of overall antioxidant activity.

[0075] In this way, it can be considered that the coefficients Coe^ determined in step S50 for a reference product are respectively representative of the antioxidant activities of the constituents Constÿ in the target product to be characterized.

[0076] 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 DB database.

[0077] In a step S80, a signal Sigtyp representative of the type Typ of the target product to be characterized stored in the memory Mem and a signal Sigmes representative of the result of the measurement AAGiObMeS are generated and transmitted by a communication unit Com to a computer calculation and processing unit Cale functionally connected to the database DB.

[0078] The measuring device Mes used by the user, the computer memory Mem, the display device Disp, the input means Inp and the communication unit Com can be integrated within a measuring module Mod, and the computing and processing unit Cale and the database DB can be remote from the measuring module Mod, as illustrated in [Fig.2].

[0079] In particular, the database and the computing and processing unit can be connected to the measurement module Mod via a computer network such as the Internet.

[0080] However, this is only an example configuration, and it is possible to imagine that the database and the computing 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.

[0081] A system Sys for implementing the method 100 can be considered in a form fully integrated in a single device or, alternatively, as a distributed set of devices physically distinct from each other but functionally connected, for example by means of a computer network.

[0082] In any case, the invention is not limited to a particular implementation of the hardware elements used for its implementation,

[0083] At a step S90, thanks to the information on the type of reference product transmitted by the signal Sigtyp, the computing and processing unit Cale identifies (step S90A) a reference product (Prod;) 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 Consfj of this reference product i as well as the associated data, more specifically their respective coefficients Coefy and their individual antioxidant activities AAy, then calculates (step S90C) the concentration q of each of the constituents j of the target product having an antioxidant activity by means of the formula equation 3:

[0084] _ CoefEq. 3 '} “ “

[0085]

[0086]

[0087] It should be noted that the calculation can be carried out by considering only the constituents meeting one or more of the criteria mentioned in step S40. Furthermore, with regard to the second and third selection criteria, the thresholds may 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 just measured by the end user. In a step S100, the computing and processing unit Cale sends back to the communication unit Com the identifiers Consty of the antioxidant constituents and their respective concentrations C deduced from the measurement to store them in the memory. Mem and display them on the display device Disp for the user.

[0088] 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 method according to the invention is quick, simple, and accessible to a non-specialist.

[0089] The general formula of equation 1 is only rigorously exact if each of the antioxidant constituents in product A and their respective antioxidant activities are known.

[0090] However, this general formula will remain usable despite the possible presence of antioxidant constituents which are not known or a voluntary choice to retain in the calculations only the constituents having a 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 which are important and can be effectively studied even if antioxidant constituents are neglected, independently of their relative shares in the overall antioxidant activity of the product.

[0091] Of course, the invention is not limited to the above description, and variant embodiments can be made without departing from the scope of the invention as defined by the claims.

Claims

Claims

1. Method (100) for characterizing a target product to be characterized having an antioxidant activity, comprising a step (S60) of receiving a signal (Sigmes) representative of a measurement of an overall antioxidant activity of the target product to be characterized, the method being characterized in that it further comprises the steps of: - receive (S80), by means of a computer processing unit (Cale), a signal (Sigtyp) representative of a type of the target product to be characterized; - identify (S90A) in a database (DB) a reference product (Prod;) corresponding to the target product to be characterized on the basis of the signal (Sigtyp) representative of a type of the product to be characterized, - retrieve (S90B) in the database (DB) identifiers (Consty) of antioxidant constituents of the target product to be characterized on the basis of the signal (Sigtyp) representative of a type of the product to be characterized; - recover (S90B) from the database (DB), by means of the computer processing unit (Cale), (a) coefficients (Coefy) respectively representative of the antioxidant activities of the antioxidant constituents in the target product to be characterized, and (b) individual antioxidant activities (AAy) of the constituents considered individually; and - calculate (S90C), by means of the computer processing unit (Cale), the concentrations (¢) of the antioxidant constituents (Consty) in the target product to be characterized from the signal (Sigmes) representative of an overall antioxidant activity of the target product, the coefficients (Coefÿ), and the antioxidant activities (AAy) from the equation _ Coef in which Q. represents the concen- I concentrations of antioxidant constituents in the target product, Coef. indicates the relative contributions of the constituents to the overall antioxidant activity of the reference product (Prod;), AAGlohMes 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 Coef . are taken from the equation coef — in which Cj represents the respective concentrations of constituents in the reference product (Prod;).

2. 2. The method according to claim 1, wherein the calculation (S90) is carried out so as to consider only antioxidant constituents whose individual antioxidant activities (AAy) multiplied by their respective concentrations (Cy) exceed 5% of the overall antioxidant activity of the reference product or the target product to be characterized.

3. 3. The method according to claim 1, in which the calculation (S90) is carried out so 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 the target product to be characterized.

4. The method according to any one 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 (S 80): - the signal (Sigtyp) 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) representative of an overall antioxidant activity of the target product on the basis of a result of the measurement (S60).

5. 5. System (Sys) for characterizing a target product to be characterized having antioxidant activity, the system comprising a database (DB) and a calculation unit (Cale) configured to implement the method according to any one of claims 1 to 4.

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