Method for marking materials for authentication and / or traceability of said materials

Abstract

The present invention relates to a method for marking a material, for example for the purpose of authenticating and / or tracing the material, in particular the batch of origin. The method comprises the steps of: providing a marking composition selected from molecular tags; and incorporating the marking composition into the material. Also according to the present invention, the marking composition comprises particles, preferably those that remain intact after incorporation into the material, comprising: a uniform porous matrix comprising a plurality of pores; an information support added to the uniform porous matrix, the information support consisting of a polymer comprising an information sequence; and particle surfaces having a coating comprising at least one fluidizing agent suitable for promoting uniform dispersion of the particles within the material.

Description

[Technical Field]

[0001] The present invention relates to the technical field of marking materials with molecular tags, for example for the purpose of authentication and / or traceability of said materials. [Background technology]

[0002] Marking methods have two major applications that give a competitive advantage to a material or finish: - authentication to protect against counterfeiting, among other causes of commercial loss; - Traceability, as brands and retailers are increasingly under pressure from consumers to ensure transparency covering their entire supply and production chains It is positioned in relation to.

[0003] Typical marking methods are based on traditional barcodes, QR Codes (Quick Response Codes), or even RFID (Radio Frequency Identification) chips with rapid reading systems as well as authentication and batch tracking systems located on or within the product packaging.

[0004] However, these "surface" tags are not applicable to some specific materials nor to situations where the code should not be visible to the naked eye.

[0005] Molecular tags address these shortcomings through their nanoscale footprint and difficulty in counterfeiting.

[0006] In effect, a "molecular tag" is a constituent element of the molecular composition of the material in which it is incorporated; it is an integral part of the material mass of interest.

[0007] Molecular tags are thus true molecular markers that allow the evolution of a material to be traced back to a batch over time.

[0008] This authentication technology also adds value to materials by collecting data from tagged materials and tracking tags embedded in finished products.

[0009] Molecular tags therefore offer an innovative solution for physically marking, tracing and authenticating materials from the producer to the retailer of products made from these materials, thus ensuring transparency throughout the entire supply chain (linear or circular).

[0010] However, current molecular tags are not entirely satisfactory.

[0011] For example, WO2013143014 discloses the encapsulation of informative DNA in non-porous silica nanoparticles suitable for invisible and secure marking or information that is part of a product. Specifically, the DNA is packed between a silica core and a silica shell.

[0012] However, since silica nanoparticles have non-negligible solubility in water at basic pH values ​​(above pH 9), the silica shell may be unstable in alkaline materials, and the dispersion of colloidal silica nanoparticles will be affected.

[0013] Specifically, DNA can also be protected by a nanoparticle outer layer composed of covalently linked siloxanes.

[0014] However, to release the DNA, the siloxane covalent bond must be broken with hydrofluoric acid, an acid that is relatively dangerous to handle.

[0015] Thus, there is a need for molecular tags that can be well dispersed within a target material mass, preferably as a uniform dispersion, without modifying its properties.

[0016] Moreover, preferably, the information support must be secured in the material and then retrieved along with the associated information sequence.

[0017] The process of extracting information from products crafted from target materials must be simple and safe enough to be transferred to the analytical laboratory. Summary of the Invention [Means for solving the problem]

[0018] To remedy the above-mentioned drawbacks of the state of the art, the present invention proposes a method for marking materials, for example with the aim of authenticating and / or traceability of said materials, and in particular of the batches of origin.

[0019] This marking method is as follows: - providing a marking composition selected from molecular tags; - incorporating said marking composition into said material; Includes.

[0020] Also according to the present invention, the marking composition comprises particles, advantageously those which remain intact after incorporation into the material, and which are: a uniform porous matrix comprising a plurality of pores; - an information support added to said homogeneous porous matrix; - the particle surface having a coating comprising at least one fluidizing agent suitable for promoting uniform dispersion of said particles within said material; Includes.

[0021] Preferably, the combination of information support, uniform porous matrix and fluidizing agent is chosen to provide self-association, thus facilitating dissociation, since no chemically reactive species are required to form covalent bonds.

[0022] Such a marking method is particularly interesting because the particles of the marking composition are particularly well dispersed within the target material mass without modifying its properties. This optimized dispersion is particularly interesting for increasing the sensitivity when recovering molecular tags in the material. Furthermore, the fluidizing agent advantageously improves the stability of the information support and the uniform porous matrix.

[0023] Moreover, the information support is advantageously well protected in the material and then recovered together with the associated information sequence.

[0024] Preferably, the information support is stabilized by association with the porous matrix and fluidizing agent, thereby providing protection within the material, which can then be retrieved along with the associated information sequence.

[0025] The use of a uniformly porous matrix has obvious advantages since it is possible to load significantly more information support than for particles of comparable composition and size.

[0026] The addition of information support to the porous material and surface modification with a flow agent ensures the stability of the encoded data and its applicability to a range of materials.

[0027] Finally, preferably, in another advantageous aspect, the particles are stable in the material but degradable when exposed to the environment, with the degradation products advantageously resulting in molecules that are environmentally friendly and safe with respect to human health.

[0028] Other non-limiting and advantageous properties of the process according to the invention, taken individually or in all technically possible combinations, are: - the homogeneous porous matrix is ​​composed of at least 90% w / w of a salt, preferentially an inorganic salt, consisting of an ionic assembly of positively charged cations and negatively charged anions or consisting of a metal oxide, and said homogeneous porous matrix is ​​insoluble in said material. The particles can be degraded by metal chelating agents or by specific pH conditions (as acidic or alkaline solutions), preferably mild acidic conditions, more preferably a pH between 2 and 5, more preferably aqueous solutions of citric acid, acetic acid, or hydrochloric acid. The particles have a size in the range of 200 nm to 100 μm, preferably 500 nm to 10 μm, more preferably 750 nm to 3 μm. The particles have a pore size in the range of 2 nm to 500 nm, preferably in the range of 2 to 200 nm, more preferably in the range of 10 nm to 100 nm. The information support is contained within the pores and / or absorbed onto the surface of the homogeneous porous matrix. The information sequence of said information support results from a code conversion of an ordered bit system, the latter resulting from the encoding of a raw data set. the information support is chosen from among nucleic acids and nucleic acid analogues, preferably DNA, more preferably single-stranded or double-stranded DNA, the information support advantageously consisting of nucleic acid fragments, preferably having a size of less than 500 bp, preferably in the range of 50 to 250 bp; - the information support is associated with, preferably adsorbed and / or contained in, said homogeneous porous matrix; The superplasticizer is selected from surface treatment agents for inorganic fillers (see Katz, Harry S. et Mileski, JV (eds.). Handbook of fillers for plastics. Springer Science & Business Media, 1987, an overview of inorganic fillers includes precipitated calcium carbonate, barite, kaolin, talc, silica, carbon black, etc.), which include saturated fatty acids, unsaturated fatty acids, phospholipids, lipid mixtures, long alkyl chain surfactants, amphiphilic polymers, resin acids, waxes, hydrophilic polymers, peptides and proteins, polysaccharides, or other inorganic salts. - fluidizing agents that improve dispersion in hydrophobic materials, preferably amphiphilic molecules or surfaces, such as hydrophilic saturated fatty acids, advantageously with a carbon chain length of C15 to C20, such as palmitic acid and stearic acid, unsaturated fatty acids, advantageously with a carbon chain length of C15 to C20, such as oleic acid and linoleic acid, phospholipids, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, lipid mixtures, preferably biosourced or biodegradable, such as lecithin, lipophilic moieties The organic amphiphilic molecules carrying the hydroxyl group are selected from among phosphonates, glycols, alcohols, phenates, sulfonates, salicylates, succinic anhydrides, waxes such as beeswax, resin acids such as cycloaliphatic carboxylic acids and abietic acid, as well as their salts, esters, and ethers, bifunctional polymers with hydrophobic moieties such as Eudragit®, or some filler coupling agents such as titanates (LICA12®) or zirconates (NZ12®). the superplasticizer is selected from among superplasticizers that improve the dispersion in hydrophilic materials, such as hydrophilic polymers, such as polycarboxylic acids, condensed phosphoric acids, polyacrylates, polyamines, polyacrylamides, polyaminoacyl, polystyrene, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), polymaleic acid (PMA), polyepoxysuccinic acid (PESA), polysaccharides (dextran, alginate, chitosan, chondroitin, cellulose, pectin, carboxymethyl inulin, etc.), peptides (such as polyaspartic acid, polyglutamic acid), proteins, lignin, preferably biosourced or biodegradable polymers, other inorganic salts (such as calcium phosphate, hydroxyapaptite). The material is selected from liquid, semi-solid or solid materials, such as plastics, thermosets, varnishes, rubbers, paints, oils, lubricants, medicines, cosmetics and food. The incorporation step consists of incorporating the marking composition into the material during the manufacturing process of said material.

[0029] The present invention also relates to a marking composition for marking a material, for example intended for the authentication and / or traceability of said material, in particular of the batch of origin, in which said marking composition comprises particles, advantageously those which remain intact after incorporation into said material, and which are: a uniform porous matrix comprising a plurality of pores; - an information support added to the homogeneous porous matrix, the information support consisting of a polymer containing an information sequence; - the particle surface having a coating comprising at least one fluidizing agent suitable for promoting uniform dispersion of said particles within said material; Includes.

[0030] The present invention also relates to materials comprising the marking compositions of the present invention.

[0031] The materials are as follows: - hydrophobic materials (e.g. plastics, oils, waxes, varnishes, lubricants) or - Hydrophilic materials (e.g. ink, paint, water-based thermosets, milk, cement) It is possible.

[0032] The different features, variations and embodiments of the invention can be associated with one another in various combinations, unless they are mutually incompatible or mutually exclusive. [Brief explanation of the drawings]

[0033] [Figure 1]Optical fluorescence microscopy (×100) photographs of molecular tags obtained with a Nikon H600L fluorescence microscope are provided: 10 mg / g of green fluorescent Atto488-labeled double-stranded 70 bp DNA (Supporting Information) and CaCO3 porous microparticles (vehicle) loaded with 0.3% w / w sodium polyacrylate (flow agent). A / Optical micrograph (differential interference contrast) of a 6 μm molecular tag. B / Green fluorescence micrograph of the same 6 μm molecular tag. The green fluorescence is due to Atto488-labeled DNA. C / Optical micrograph (differential interference contrast) of a 2.5 μm molecular tag. D / Green fluorescence micrograph of the same 2.5 μm molecular tag. [Figure 2] Figure 1 provides a diagram depicting the modification of physicochemical properties of molecular tags as a function of the presence or absence of fluidizing agent. Particle size (size in μm) and polydispersity measurements (volume frequency in %) in water obtained with a Malvern Mastersizer 3000 equipped with a Hydro2000S modular system. A: 2.5 μm homogeneously porous calcium carbonate vehicle alone. B: 2.5 μm hydrophilic molecular tags well dispersed in water (fluidizing agent: sodium polyacrylate 0.5% w / w). [Figure 3] Figure 1 shows the protective effect of vehicle + fluidizing agent combinations on the chemical reactivity of DNA to reactive genotoxic molecules such as isothiocyanates (FITC: fluorescein isothiocyanate or RITC: rhodamine isothiocyanate). Left: Plot of relative DNA fluorescein fluorescence (fluorescence in arbitrary units) after 48 h of reaction with FITC. 1: water, 2: 80 bp DNA without FITC (negative control), 3: 80 bp DNA + FITC (positive control), 4: 80 bp DNA + FITC loaded in vehicle at 10 mg / g, 5: 80 bp DNA + FITC loaded at 3.5 mg / g in vehicle associated with the fluidizing agent sodium polyacrylate (0.5% w / w). Right: 4% low-melting agarose electrophoresis gel of the same DNA fragments. A / fluorescein fluorescence shown; B / BET-stained DNA fragments shown. DNA is protected from isothiocyanates only when loaded into a fluidizing agent-associated vehicle. CaCO3 vehicle alone is not sufficient to protect DNA. [Figure 4] A 4% agarose plate is provided to track the electrophoresis of DNA fragments generated after PCR of DNA extracted from thermoplastic. The thermoplastic is marked with a hydrophobic molecular tag (a homogeneous porous CaCO3 vehicle loaded with 10 mg / g of 70 bp DNA and covered with 2% w / w stearic acid). The 70 bp DNA information support extracted from the solidified molded polymer is amplified with 2 × 35 bp extension primers (2 × [20 bp primer for DNA amplification + 15 bp for integration into the plasmid]), resulting in the amplification of a 100 bp DNA oligomer. 1: Negative control (untreated water), 2: Negative control (untreated digestion buffer), 3: Negative control (extraction polymer PURASORB® PDLG7507), 4: Negative control (extraction polymer PURASORB® PDLG5010), 5: Extraction polymer PURASORB® PDLG7507 marked with 2 ppm molecular tag, 6: Extraction polymer PURASORB® PDLG5010 marked with 1 ppm molecular tag, 7: Positive control (PCR of 1 ng DNA), L: Untreated reference TriDye™ Ultra Low Range DNA Ladder from NEB. DETAILED DESCRIPTION OF THE INVENTION

[0034] Various other features of the present invention will emerge from the following description.

[0035] The present invention relates to a method for marking materials, for example for the purpose of authenticating and / or traceability of said materials, especially of originating batches, by means of a marking composition.

[0036] As used herein, "material" or "object" refers to a material that is marked (or subsequently marked) with a marking composition that includes at least a molecular tag.

[0037] The material can be an element, component, part, or substance that constitutes or can be made into something.

[0038] The marking compositions are particularly useful in verification methods that allow differentiation and / or identification of materials.

[0039] Therefore, the verification method is suitable for distinguishing genuine material from counterfeit material.

[0040] Additionally, verification methods are suitable for identifying materials (eg, by batch type, manufacturing location, manufacturing time).

[0041] Generally, the marking method of the present invention provides the marked material with a competitive advantage, namely: - authentication to indicate the origin of the material and, in particular, to protect against counterfeiting; and - Traceability, enabling manufacturers and trade partners, as well as consumers and authorities, to identify, authenticate and track goods throughout the supply chain; The aim is to grant

[0042] According to the present invention, the marking method comprises the following steps: - providing a marking composition selected from molecular tags; - incorporating said marking composition into said material; Includes.

[0043] According to the invention, starting from a material containing a marking composition, a recovery step can be performed on said material so as to recover the information support.

[0044] Preferably, said recovering step is effected to extract said information support on a product elaborated from the material.

[0045] As used herein, a "molecular tag" advantageously relates to a compound suitable for the identification and / or authentication of a material of interest.

[0046] A "molecular tag" is a constituent element of the molecular composition of the material in which it is incorporated. It is an integral part of the material mass of interest.

[0047] In other words, the present invention also relates to the use of said marking composition for marking materials, for example for the purpose of authentication and / or traceability of said materials, especially of originating batches.

[0048] General Remarks on Marking Compositions In the present invention, the marking composition comprises at least three components: a uniform porous matrix comprising a plurality of pores; - an information support added to the homogeneous porous matrix, the information support consisting of a polymer containing an information sequence; - the particle surface having a coating comprising at least one fluidizing agent suitable for promoting uniform dispersion of said particles within said material; The particle comprises a molecular tag comprising a combination of:

[0049] Typically and preferably, the combination of information support, uniform porous matrix and fluidizing agent is chosen to provide self-association, thus facilitating dissociation, since no chemically reactive species are required to form covalent bonds.

[0050] Specifically, "self-association" refers to the ability of chemical entities to naturally and spontaneously interact with one another without the need for the formation of covalent chemical bonds or the presence of specific chemical reactants. It is a phenomenon in which chemical entities interact with one another due to intermolecular forces such as van der Waals forces, hydrogen bonding, ionic interactions, etc., without forming permanent chemical bonds.

[0051] This self-association can occur when molecules or chemical compounds possess functional groups or properties that cause them to be attracted to one another, and these interactions can be reversible, allowing the participating entities to dissociate at a later time without the need for complex chemical reactions.

[0052] In the present invention, the selected components, i.e., the information support, the uniform porous matrix, and the fluidizing agent, are designed to spontaneously interact with each other without the need for reactive species to form covalent bonds, which is useful for retrieving the information support because such interactions are not permanent, simplifying the subsequent dissociation process.

[0053] Preferably, the marking composition comprises a "matrix flow agent" couple adapted to the information support and target materials.

[0054] As used herein, the term "particle" or "particulate" refers to the smallest entity in a marking composition that can be identified as a particulate by its appearance.

[0055] Advantageously, the particles have a size ranging from 200 nm to 100 μm, preferably from 500 nm to 10 μm, more preferably from 750 nm to 3 μm.

[0056] Preferably, the size is within any combination selected from the following values: 0.5 μm, 1 μm, 1.5 μm, 2 μm, 2.5 μm, 3 μm, 3.5 μm, 4 μm, 4.5 μm, 5 μm, 5.5 μm, 6 μm, 6.5 μm, 7 μm, 7.5 μm, 8 μm, 8.5 μm, 9 μm, 9.5 μm, 10 μm.

[0057] The term "size" advantageously means the physical characteristic size of a particle linked to a measurement method (or dimensional analysis by suitable techniques), for example by laser diffraction or sieving or image analysis.

[0058] The size of a solid particle is advantageously determined by its "equivalent spherical diameter", or in other words, its volume diameter (d v ) corresponds to the "equivalent spherical diameter" of

[0059] The particle size, and in particular the particle size distribution, of such particles is advantageously determined by the D, also called "median size" or "median diameter" 50 or d 50 It is defined by the value of

[0060] D 50 represents the particle size at which 50% of the volume (or mass) has a smaller (or larger) particle size, in other words, D 50 is the diameter corresponding to 50% of the cumulative frequency of the number, mass, or volume.

[0061] Thus, as used herein, "median size" in relation to a particle advantageously means the 50th percentile particle diameter (volume median particle diameter D50) in its volume size distribution (e.g., as measured by a particle size analyzer based on laser diffraction / scattering spectroscopy).

[0062] Particle size distribution can be measured by laser diffraction using a Mastersizer 2000 or 3000 equipped with a Hydro2000S system.

[0063] Generally, the particles (preferably in combination with a fluidizing agent) advantageously form a protective agent for the information support.

[0064] As used herein, the term "protective agent" refers to a molecule capable of preventing information support damage caused by environmental factors such as UV light, ionizing radiation, genotoxic chemical agents, or by naturally occurring agents such as reactive oxygen species, reactive nitrogen species, reactive carbonyl species, lipid peroxidation products, alkylating agents, etc.

[0065] The particles of the marking composition advantageously remain intact after incorporation into the target material.

[0066] As used herein, the term "remain intact" means that the particles are contained in the subject material with the information support remaining attached / associated with the uniform porous matrix.

[0067] Also, the term "remain intact" means that at least a portion of the particles are encapsulated and the homogeneous porous matrix is ​​fractionated or divided due to the marking method, but the information support remains attached / associated with the homogeneous porous matrix.

[0068] Generally, the concentration of information support in the particles is advantageously between 0.1 and 500 mg, preferably between 10 and 500 mg, of information support per gram of homogeneous porous matrix.

[0069] Preferably, the concentration is: 10 mg g -1 , 50 mg g -1 , 100 mg g -1 , 150 mg g -1 , 200 mg g -1 , 250 mg g -1 , 300 mg g -1 , 350mg·g -1 , 400mg·g -1 , 450mg·g -1 , 500mg·g -1 The range is any combination selected from the following:

[0070] Uniformly porous matrix The homogeneous porous matrix advantageously forms a "vehicle", ie a compound that can be loaded with information support and allows its incorporation into the target material.

[0071] The homogeneous porous matrix advantageously aims to carry the information support by enclosing the latter.

[0072] As used herein, "homogeneous" or "homogenous" means that the matrix lacks any sequential layers (eg, lacks a core-shell).

[0073] As used herein, "porous" means that the matrix is ​​a solid that contains pores, i.e., cavities, channels, or gaps whose depth is greater than their width. The term "porous" also means that the matrix is ​​capable of adsorbing the information support described herein.

[0074] Generally, the term "porosity" as used herein also refers to the following, in accordance with IUPAC recommendations (Rouquerol, 1994): - in mesoporous particles, pores with a free diameter in the range of 2 to 50 nm, or - pores with a free diameter greater than 50 nm in macroporous particles; means.

[0075] The "free diameter" or "pore size" of a pore refers to the pore diameter measured in the dry state using measurement techniques such as quantitative nitrogen sorption isotherms and analytical methods such as density functional theory (DFT) and the Barrett-Joyner-Halenda (BJH) method, which are used to extract pore size distributions from experimental isotherms based on the Kelvin model of pore filling.

[0076] For the assessment of larger pores, mercury intrusion (pores sized from 3.2 nm to over 400 μm) and capillary flow porometry (pores sized from 13 nm to 500 μm) can be used.

[0077] Preferably, the homogeneous porous matrix of the present invention comprises: - About 1~1500m 2 / g maximum surface area, and / or - pore sizes in the range of 2 to 500 nm, preferably 2 to 200 nm, more preferably 10 nm to 100 nm; It has.

[0078] Preferably, the maximum surface area is one of the following values: 1, 10, 100, 200, 300, 400, 500, 600, 700, 800, 900, 1000, 1100, 1200, 1300, 1400, 1500 m 2 / g.

[0079] Preferably, the pore size is within any combination selected from the following values: 2 nm, 10 nm, 20 nm, 30 nm, 40 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm.

[0080] The "surface area" is advantageously obtained by applying the Brunauer-Emmett-Teller (BET) method, which derives surface area from physical sorption isotherm data. According to ISO 9277-1995, specific surface area is measured using nitrogen measurements via the BET method.

[0081] Porosity can also be visualized in transmission electron microscopy (TEM) images. TEM images taken in bright-field imaging mode show contrast variations between light and dark regions within a single particle, with the light areas representing areas where electrons do not interact significantly with the material. These areas correspond to pores in the material.

[0082] More preferentially, the size of the pores can be adapted to the size of the information support.

[0083] In a preferred embodiment, the homogeneous porous matrix contains at least 90% w / w of salts, preferentially: - an ionic assembly of positively charged cations and negatively charged anions, or - metal oxides, for example, ZrO2, It is composed of inorganic salts consisting of:

[0084] As used herein, a mineral compound (salt, metal oxide) is considered "insoluble" if less than 1 gram of it is completely soluble in 1 L of the target solvent. A table of water-insoluble salts can be found in JH Perry, Techniques de l'Ingenieur "Solubilite dans l'eau des composes mineraux", Chemical Engineers Handbook, Ref: K590 v1 (1992) https: / / doi.org / 10.51257 / a-v1-k590. For insoluble compounds, solubility is given in milligrams of anhydrous substance per liter of solvent (typically water) as a function of temperature. For example, only 11.2 mg of CaCO3 is soluble in 1 L of water at 18°C.

[0085] The anion is advantageously a carbonate, phosphate, sulfate, borate, silicate (e.g., CO 2- , PO4 3- , SO4 2- , BO3 3- , SiO3 2- etc.), or a combination thereof.

[0086] The cations are advantageously selected from calcium, magnesium, barium, aluminum, zinc, strontium, manganese (e.g., Ca 2+ , Mg 2+ , Ba 2+ , Al 3+ , Zn 2+ , Sr 2+ , Mn 2+ etc.), or a combination thereof.

[0087] Preferably, the particles consist primarily (preferably greater than 90% w / w) of calcium carbonate, calcium phosphate, barium sulfate, barium phosphate, barium carbonate, borosilicate, calcium silicate, manganese carbonate, or combinations thereof.

[0088] The calcium carbonate is preferably mesoporous calcium carbonate, i.e. the chemical element CaCO3,XH2O crystallized in the form of particles exhibiting the structural particularity of being organized in a network of channels of variable pore size.

[0089] The calcium carbonate porous particles may be, for example: - Volodkin DV,Petrov AI Prevot M,Sukhorukov GB.Matrix polyelectrolyte microcapsules:new system for macromolecule encapsulation.Langmuir.2004 Apr 13;20(8):3398-406.doi:10.1021 / la036177z.PMID:15875874, - Volodkin DV,Larionova NI,Sukhorukov GB.Protein encapsulation via porous CaCO3microparticles templating.Biomacromolecules.2004 Sep-Oct;5(5):1962-72.doi:10.1021 / bm049669e.PMID:15360312, - Anna Vikulina,Joseph Webster,Denis Voronin,Evgenii Ivanov,Rawil Fakhrullin,Vladimir Vinokurov,Dmitry Volodkin,Mesoporous additive-free vaterite CaCO3crystals of untypical sizes:From submicron to Giant,Materials & Design,Volume 197,2021,109220,https: / / doi.org / 10.1016 / j.matdes.2020.109220, - Ferreira AM, Vikulina AS, Volodkin D. CaCO3crystals as versatile carriers for controlled delivery of antimicrobials.J Control Release.2020 Dec 10;328:470-489.doi:10.1016 / j.jconrel.2020.08.061.Epub 2020 Sep 5.PMID:32896611, is disclosed in.

[0090] The homogeneous porous matrix can also be chosen from among biogenic and biosourced homogeneous porous matrices.

[0091] Such a homogeneous porous matrix is, for example, selected from among coccoliths.

[0092] Coccoliths are produced by microalgae, e.g., Emiliania huxleyi, which have favorable properties (e.g., size of 3 μm, pore diameter of 4 nm, and pore size of 19 nm). 2 .g -1 It exhibits a specific surface area of ​​.

[0093] Coccoliths include, for example: - Jakob, I., Chairopoulou, MA, Vucak, M., Posten, C., & Teipel, U. (2017).Biogenic calcite particles from microalgae-Coccoliths as a potential raw material.Engineering in life sciences,17(6),605-612.DOI:10.1002 / elsc.201600183,PMID:28701909, or - Lomora, M., Shumate, D., Rahman, AA, & Pandit, A. (2019). Therapeutic applications of phytoplankton, with an emphasis on diatoms and coccolithophores. Advanced Therapeutics, 2(2), 1800099; https: / / doi.org / 10.1002 / adtp.201800099, is disclosed in.

[0094] The homogeneous porous matrix is ​​advantageously insoluble in said material (the matrix is ​​incapable of forming such a solution in said material).

[0095] The homogeneous porous matrix advantageously integrates other organic and / or inorganic chemical elements.

[0096] Said organic and / or inorganic chemical elements are advantageously capable of influencing the particle properties, such as its size, its structure and its properties.

[0097] The organic and / or inorganic chemical entities are chosen, for example, from short polymers, fatty acids, short organic molecules such as ethylene glycol, or other inorganic salts.

[0098] The particles are advantageously inert in terms of information support stability, for example: metal chelators, such as EDTA (ethylenediaminetetraacetic acid), EGTA (ethylene glycol-bis(β-aminoethyl ether)-N,N,N',N'-tetraacetic acid), NTA (N,N-bis(carboxymethyl)glycine), DTPA (diethylenetriaminepentaacetic acid), or natural and biodegradable L-glutamic acid diacetate (GLDA), and / or specific pH conditions (as an acidic aqueous solution), advantageously for a short exposure time (preferentially less than 1 hour), - mild acidic conditions combined with chelating agents, It is decomposed by:

[0099] Citric acid, a natural molecule derived from lemons, is both a calcium chelator and a weak acid, so the particles are easily and rapidly soluble in citric acid solutions.

[0100] As used herein, the term "decompose" or "decomposition" refers to dissolving the homogeneous porous matrix (in the form of soluble salts or gases), advantageously using a metal chelator and / or acidic pH conditions to release the information support.

[0101] According to IUPAC (Muller, P. "Glossary of terms used in physical organic chemistry" (IUPAC Recommendations 1994, Pure and Applied Chemistry, vol. 66, no. 5, 1994, pp. 1077-1184. https: / / doi.org / 10.1351 / pac199466051077), "chelation" refers to the formation or existence of two or more separate coordinate bonds between a polydentate (multiple-bonded) ligand and a single central atom.

[0102] Such ligands are usually organic compounds and are called chelant, chelator, chelating agent, or sequestering agent.

[0103] Metal chelators are chemical compounds that react with metal ions to form stable, water-soluble metal complexes. These agents reorganize the chemical composition of the metal, improving its general stability and likelihood of binding to other substances.

[0104] Preferably, the particles are degradable by mild acidic conditions, ie, a pH of 2 to 5, for example, an aqueous solution of citric acid, acetic acid, or hydrochloric acid.

[0105] Preferably, the pH is within any combination selected from the following values: 2, 3, 4, 5.

[0106] At a pH below 5, DNA is susceptible to depurination (i.e., loss of purine bases from the DNA), so chelating agents are preferred.

[0107] Acidic conditions are preferably used with very short extraction times (preferentially less than 1 hour) to accelerate the degradation process induced by the chelating agent.

[0108] Thus, advantageously, particle decomposition does not require the handling of acids (eg, hydrofluoric acid, nitric acid, chlorosulfuric acid, perchloric acid, triflic acid, fluoroantimonic acid) that are significantly harmful to human health.

[0109] For example, calcium carbonate porous particles may be: - 100mM EDTA solution, pH 5, or - an aqueous solution of acetic acid at pH 3, - Ca 2+ and CO3 2- a 1% (w / v) aqueous solution of citric acid at pH 2, which results in immediate particle dissolution from ionic dissociation; It can be dissolved using

[0110] Generally speaking, LDHs (layered double hydroxides) are excluded from the present invention as they do not form a uniform porous matrix.

[0111] In fact, LDHs are not porous particles. The layered structure of LDHs creates interlayer spaces between the layers, and these interlayer spaces can contain anions and water molecules. However, these spaces are not pores within the meaning of the present invention.

[0112] Information Support An information support is added to the uniform porous matrix, and the information support comprises a polymer containing an information sequence.

[0113] The information support can be loaded onto and / or into the homogeneous porous matrix by adsorption or co-precipitation.

[0114] In other words, the information support is advantageously contained within the pores and / or on the surface of a uniform porous matrix.

[0115] Or, in other words, the information support is advantageously contained at least in part or entirely within the pores of the uniform porous matrix.

[0116] The information sequence of said information support advantageously results from a code conversion of an ordered bit system, the latter resulting from an encoding of a raw data set.

[0117] As used herein, "transcoding" advantageously means the use of a code to represent information to ensure its integrity (error detection and correction) and to guarantee its security (encryption / ciphering), and therefore this code preferably has an error correction system.

[0118] Encryption methods include, for example: - Erlich et al.,DNA Fountain enables a robust and efficient storage architecture,Science 355,950-954(2017), - Cheng Kai Lim, Saurabh Nirantar, Wen Shan Yew, Chueh Loo Poh, Novel Modalities in DNA Data Storage,Trends in Biotechnology,Volume 39,Issue 10,2021,Pages 990-1003.https: / / doi.org / 10.1016 / j.tibtech.2020.12.008, or - Ceze, L., Nivala, J. & Strauss, K.Molecular digital data storage using DNA.Nat Rev Genet 20,456-466(2019).https: / / doi.org / 10.1038 / s41576-019-0125-3, will be disclosed.

[0119] As used herein, the term "encryption method" refers to a process that converts data into an ordered binary code, which is then converted into a chemical structure that becomes an information carrier.

[0120] Nucleic acids such as DNA, including nucleic acid analogs, can be used to store digital information by designing sequences of nucleotide bases that encode the zeros and ones of the digital information. There are a variety of techniques and encoding schemes known to those skilled in the art for representing digital information using nucleotide bases.

[0121] Advantages of using nucleic acids and / or analogs rather than other storage media to store digital information include information density and longevity.

[0122] During sequencing and synthesis of nucleic acids and / or analogs, substitution, insertion, and deletion errors occur, and data must be protected against these errors. Additionally, environmental factors such as reactive oxygen species and UV radiation can alter nucleic acid sequences. Therefore, to offset these drawbacks, encoding and error correction codes, such as Huffman codes and Reed-Solomon codes, known to those skilled in the art, can be used.

[0123] According to the encryption method, the ordered binary code data is thus converted into a nucleic acid sequence transform, resulting in an information sequence.

[0124] Finally, the encrypted data can be deciphered by determining the nucleic acid base sequence of the information sequence.

[0125] As used herein, the term "encrypted data" advantageously refers to each element of data (e.g., text, image, file) that can be encrypted or encoded in an information sequence, and that can be deciphered from the information support recovered from the product, and that can be decoded or decrypted according to a selected method, where the decoded or decrypted data can be used to verify the nature of the material, or to authenticate the material, or to eliminate counterfeit items.

[0126] Therefore, in a preferred embodiment, the information support is selected from among nucleic acids and nucleic acid analogues, preferably DNA, more preferably single-stranded or double-stranded DNA.

[0127] As used herein, the term "nucleic acid" refers to an oligomer made up of nucleotides, which can be deoxyribonucleotides or ribonucleotides. The compound can be naturally occurring or synthetically produced deoxyribonucleotides or ribonucleotides. Synthetically produced nucleic acids can be naturally occurring sequences or non-naturally occurring sequences. The terms "ribonucleic acid" and "RNA" refer to an oligomer made up of ribonucleotides. The terms "deoxyribonucleic acid" and "DNA" refer to an oligomer made up of deoxyribonucleotides.

[0128] The terms "nucleic acid," "polynucleotide," "target polynucleotide," and "target nucleic acid" are used interchangeably.

[0129] As used herein, the term "nucleic acid analog" refers to a nucleic acid that can contain one or more analogs (e.g., modified backbones, sugars, or nucleobases). Some non-limiting examples of analogs include the following: 5-bromouracil, peptide nucleic acids, xenonucleic acids, morpholino compounds, locked nucleic acids, diol nucleic acids, threose nucleic acids, dideoxynucleotides, cordycepin, 7-deaza-GTP, fluorophores (e.g., rhodamine or fluorescein linked to a sugar), thiol-containing nucleotides, biotin-linked nucleotides, fluorescent base analogs, CpG islands, methyl-7-guanosine, methylated nucleotides, inosine, thiouridine, pseudouridine, dihydrouridine, stevioside, and tetanoside.

[0130] The information support preferably consists of nucleic acid fragments, preferably having a size of less than 500 bp, preferably in the range of 50-250 bp.

[0131] Advances in sequencing, the primary form of information retrieval, or "reading," are making DNA data storage more feasible. The primary methods for reading DNA include sequencing by synthesis (SBS), coupled with third-generation sequencing methods that involve single-molecule sequencing via nanopores or enzymatic well-based reactions.

[0132] Preferably, the informative sequence comprises an identifiable sequence, also referred to as a "detectable sequence", e.g., a nucleic acid sequence or nucleic acid analog sequence that can be detected and sequenced by specific techniques (e.g., hybridization, PCR techniques, sequencing, capture, electrochemical detection).

[0133] Superplasticizer As specified above, the particles have a particle surface with a coating comprising at least one fluidizing agent suitable for promoting uniform (or homogeneous) dispersion of said particles within said material. In the present invention, "uniform dispersion" also encompasses "homogeneous dispersion" or "random dispersion", advantageously without agglomerations of particles.

[0134] "Uniformly dispersed" advantageously means that the particles are uniformly, or at least approximately uniformly, spaced or distributed within the material.

[0135] As used herein, a dispersion is defined as a system in which discrete particles of one material are dispersed in a continuous phase of another material. The two phases can be the same or different states of matter.

[0136] Dispersions are classified in several different ways, including how large the particles are in relation to the particles of the continuous phase, whether or not settling occurs and Brownian motion is present.

[0137] Generally, dispersions of particles large enough to settle are called suspensions, and those of smaller particles are called colloids and solutions. For IUPAC definitions, see "Terminology of polymers and polymerization processes in dispersed systems (IUPAC Recommendations 2011)". Pure and Applied Chemistry. 83(12):2229-2259, and Compendium of Polymer Terminology and Nomenclature (IUPAC Recommendations 2008) (2nd ed.). RSC Publ. p. 464.

[0138] As used herein, the essence of coating advantageously consists in the surface modification of said particles, i.e. the modification of the physicochemical properties of the surface through the application of chemical substances, preferably a surface coating to form surface treated particles.

[0139] The surface coating is advantageously an adhesive layer, ie a coverage of the surface of said particles by said fluidizing agent.

[0140] As used herein, fluidizing agent refers to a microparticle surface treatment selected to modify the handling characteristics of the particles in the target material.

[0141] In the context of the present invention, the term "fluidizing agent" advantageously relates to a functionalizing agent that modifies the physicochemical properties of the particles to improve their incorporation into the target material.

[0142] The "flow agent" is advantageously selected from among a crystallization inhibitor, a dispersant, a surface treatment agent, an agent that enhances the flowability of the microparticle dispersion, or a combination thereof.

[0143] The at least one fluidizing agent is advantageously chosen directly depending on the nature of the target material, which is, inter alia: - Hydrophobic materials (oils, plastics, waxes, hydrophobic polymers), or - aqueous media or hydrophilic materials (paints, thermosetting prepolymers, inks, milk, cement), It is possible.

[0144] For example, to incorporate molecular tags into a hydrophobic plastic such as PVC (polyvinyl chloride), the fluidizing agent is selected from among fatty acids, such as stearic acid, to improve the dispersibility of the particles.

[0145] Conversely, to incorporate molecular tags into thermosetting prepolymers made up of aqueous latex solutions, fluidizers that improve the dispersibility of particles in this type of material are chosen from among sodium polyacrylates.

[0146] Preferably, fluidizing agents that form covalent layers (which make the particles incapable of being broken down using chelating agents such as organosilanes or siloxanes) are excluded from the present invention.

[0147] When modifying the surface properties of the particles, the fluidizing agent advantageously acts on at least one particle property, therefore, in other words: - changes in the surface charge (zeta potential) that modify the stability behavior and / or the interaction with other components of the material, - changes in the aggregation / settling properties of the particles (alone in a solvent or in interaction with other particles of said material), - if the target material is a liquid, a change in particle dispersibility, where as used herein particle dispersibility corresponds to the ease of dispersing a powder in a liquid to achieve spatial uniformity and a desired particle size (see ISO / TS22107:2021); - If the target material is solid, a change in the uniformity of the particle dispersion (no particle agglomeration), It is possible to measure.

[0148] According to the present invention, the physicochemical properties of the molecular tags can be adapted to the material to be marked as follows. - hydrophilic molecular tags can be generated using hydrophilic fluidizing agents, preferably for incorporation into hydrophilic materials, or Hydrophobic molecular tags can be generated using hydrophobic fluidizing agents, preferably for incorporation into hydrophobic materials.

[0149] Advantageously, the fluidizing agent imparts several properties to the particle, thus improving its dispersibility in the solvent or environment.

[0150] The zeta potential of particles typically ranges from -100 to +100 mV. The magnitude of the zeta potential is a predictor of the colloidal stability of a solution. Default zeta potential analytical limits are -150 mV to +150 mV, respectively. See Tech Note: Zeta potential quality report for the Zetasizer Nano.

[0151] For example, the zeta potential of porous calcium carbonate microparticles can be changed from a negative to a positive zeta potential by coating them with a positive polymer.

[0152] The zeta potential of CaCO porous microparticles is determined to be -12.2 mV in water at a fixed pH. Adsorption of poly(allylamine hydrochloride) (PAH, Mw ∼70 kDa) fluidizer onto the surface of the particles is sufficient to achieve a zeta potential value of +10 mV (see Volodkin DV et al. Matrix polyelectrolyte microcapsules: a new system for macromolecule encapsulation. Langmuir. 2004 Apr 13;20(8):3398-406. doi:10.1021 / la036177z. PMID:15875874).

[0153] In another example, surface modification of nanosized precipitated calcium carbonate nanoparticles (nano-PCC) with a fluidizing agent (e.g., alginate, pectin, acrylamide (A-PAM), or nanofibrillated cellulose (NFC)) resulted in a charge reversal, expressed as a change in the sign of the zeta potential from positive to negative.

[0154] The addition of NFC to nano-PCC dispersions caused a charge reversal, shifting the zeta potential from approximately +10 mV to −20 mV in fixed pH experiments.

[0155] The stability of the saturated dispersions was determined by measuring the change in turbidity, ie the light transmitted through the sample, with a Turbiscan Ma2000 instrument.

[0156] Without modification, nanoPCC dispersions were unstable, observed as a rapid increase in light transmission due to settling of flocculated particles (more than 40% in just a few minutes). Pectin- and NFC-modified nanoPCC dispersions were stable and did not show any change in turbidity as a function of time.

[0157] The stability of nano-PCC particles was also improved with A-PAM and alginate; no phase separation was detected, although a slight increase in light transmission (Δ turbidity about 8%) was observed.

[0158] Conversely, the addition of 1 mL of 1% hydrophobizing agent, e.g., alkenyl succinic anhydride (ASA), to a dispersion of unmodified nano-PCC particles caused aggregation of the formed mixture, and phase separation was observed as a rapid increase in Δ turbidity (%), which reached 80% in 120 min.

[0159] Such superplasticizers, ASA, carrying branched iso-alkenyl chains (C14-C22), are widely used, inter alia, in the surface sizing of paper, paperboard, and cardboard, as well as in the hydrophobization of cellulose fibers. They destabilize particles in aqueous media but enhance their dispersibility in organic solvents and hydrophobic materials such as plastics. (See Nypeloe T et al., "Tailoring Surface Properties of Paper Using Nanosized Precipitated Calcium Carbonate Particles." ACS Appl. Mater. Interfaces 2011, 3, 9, 3725-3731. https: / / doi.org / 10.1021 / am200913t).

[0160] The effect of a fluidizing agent on particles can be measured by a number of techniques well known in the prior art depending on the target parameters.

[0161] In relation to surface charge, the electrophoretic mobility of the studied samples can be advantageously measured using a ZetaSizer Nano ZS (Malvern, UK) operating at a wavelength of 633 nm. This instrument measures the electrophoretic mobility of particles and converts it to zeta potential (ZP) using the von Smolkovskiy equation. Results are expressed as the average of at least three independent measurements.

[0162] Regarding the hydrodynamic radius, the particle mean hydrodynamic diameter Dh of the particles is advantageously determined by DLS using a Zetasizer 3000 (Malvern, UK) equipped with a 10 mW He-Ne laser (633 nm) as the light source. Measurements are performed at a scattering angle of 90°, and the reported results are the average of three independent measurements.

[0163] Regarding flocculation / settling, optical analyzers are advantageously used to study suspension behavior and determine settling speed (Turbiscan MA 2000, Formulaction, Toulouse, France). This instrument detects and measures the changes in concentrated and opaque suspensions. Turbiscan performs step-by-step vertical scanning of the entire sample with a pulsed near-infrared light source (λ=850 nm), converting the macroscopic aspects of the mixture into two graphics (see Mengual O, Meunier G, Cayre I, Puech K, Snabre P. TURBISCAN MA 2000: multiple light scattering measurement for concentrated emulsion and suspension instability analysis. Talanta. 1999 Sep 13; 50(2): 445-56. doi: 10.1016 / s0039-9140(99)00129-0. PMID: 18967735). Transmission and backscattering data were obtained from the respective detectors; the transmission detector receives light traveling across the sample (at 0° from the incident beam), while the backscattering detector receives light backscattered by the sample at 135° from the incident beam. The stability of the dispersion was determined by measuring the change in turbidity, i.e., the light transmitted through the sample. The Turbiscan provides backscattered light flux (Δturbidity in % relative to an external standard) as a function of sample height. Turbiscan data can report the change in transmitted light measured from the middle of the test tube over time. Turbidity measurements were started immediately after dispersion preparation.

[0164] Turbiscan is also used to determine particle dispersion in liquids. The stability of particle dispersions is assessed by Turbiscan Stability Index (TSI) values.

[0165] With regard to examining the uniformity of particle dispersion in a solid material, the particles can be directly observed using scanning electron microscopy (SEM) or atomic force microscopy (AFM).

[0166] With regard to recrystallization properties, the stabilization of unstable porous vehicles is advantageously directly observable by microscopic techniques such as optical microscopy or scanning electron microscopy, depending on their size.

[0167] Regarding the surface area of ​​microparticles, it is standard practice to apply the Brunauer-Emmett-Teller (BET) method, which derives the surface area from physical sorption isotherm data. In accordance with ISO 9277-1995, the specific surface area was measured using nitrogen measurements via the BET method.

[0168] Regarding the surface energy, the contact angle is advantageously measured by a video optical contact angle tester (CA; OCA25, Eastern-Dataphy, Germany).

[0169] In practice, the fluidizing agent advantageously comprises a wide variety of molecules, including coatings comprising organic molecules such as fatty acids, polymers, resins, etc., and / or coatings comprising inorganic salts.

[0170] In a preferred embodiment, the at least one fluidizing agent is selected from among surface treatment agents for inorganic fillers according to their hydrophilic or hydrophobic properties.

[0171] Fluidizers that impart hydrophobicity to particles and thereby improve their dispersibility in hydrophobic solvents or environments are generally amphiphilic molecules or surfactants, such as: saturated fatty acids, advantageously those with a carbon chain length of C15 to C20, such as palmitic acid and stearic acid; unsaturated fatty acids, advantageously those with a carbon chain length of C15 to C20, such as oleic acid and linoleic acid; phospholipids, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, lipid mixtures, preferably biosourced or biodegradable, such as lecithin; organic amphiphilic molecules carrying a lipophilic moiety, such as phosphonates, glycols, alcohols, phenates, sulfonates, salicylates, succinic anhydrides, - Waxes, such as beeswax, shellac, vegetable waxes, resin acids, such as cycloaliphatic carboxylic acids and abietic acid, as well as their salts, esters and ethers; bifunctional polymers with hydrophobic moieties, such as Eudragit®, or some filler coupling agents, such as titanates (LICA12®) or zirconates (NZ12®), is.

[0172] Fluidizing agents that improve or impart dispersibility of particles in hydrophilic materials include the following: - hydrophilic polymers, such as polycarboxylic acids, condensed phosphates, polyacrylates, polyamines, polyacrylamides, polyaminoacyl, polystyrene, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), polymaleic acid (PMA), polyepoxysuccinic acid (PESA), polysaccharides (dextran, alginate, chitosan, chondroitin, cellulose, pectin, carboxymethyl inulin, etc.), peptides (e.g. polyaspartic acid, polyglutamic acid), proteins, lignin, preferably biosourced or biodegradable polymers (see Vroman I et al. Biodegradable Polymers. Materials (Basel). 2009;2(2):307-344. Published 2009 Apr 1. doi:10.3390 / ma2020307); other inorganic salts, such as calcium phosphate, hydroxyapaptite, Examples include:

[0173] As used herein, in accordance with IUPAC Recommendations 2003, a "biodegradable polymer" is a polymer that is susceptible to degradation due to biological activity, with degradation being accompanied by a decrease in its molar mass.

[0174] For example, the particles may be treated or coated with a hydrophobizing agent, such as an aliphatic carboxylic acid, or with a surfactant.

[0175] Suitable aliphatic acids are, for example, C15 to C28 fatty acids such as stearic acid, palmitic acid, myristic acid, lauric acid, or mixtures thereof.

[0176] The particles can also be treated or coated with hydrophilic polymers to make them cationic or anionic, for example with polyacrylate or polyaspartic acid (anionic) or polyarginine, polylysine (cationic).

[0177] Synthesis of particles of marking composition As shown in the examples, the information support and the fluidizing agent are advantageously simultaneously incorporated from solution onto a homogeneous porous matrix.

[0178] For example, the information support and fluidizing agent are advantageously simultaneously incorporated from solution onto a preformed homogeneous porous matrix by direct adsorption of macromolecules (physisorption).

[0179] In an alternative embodiment, depending on the nature of the "flow agent," the information support and flow agent can also be added sequentially to the porous vehicle.

[0180] For example, the support and flow agent can be added sequentially by direct adsorption in the case of a fatty acid such as stearic acid.

[0181] Generally, the combination of information support, uniform porous matrix and fluidizing agent is advantageously self-associating and therefore does not require chemically reactive species to form covalent bonds, facilitating their dissociation.

[0182] material The present invention also relates to a material comprising the marking composition according to the present invention.

[0183] In general, the material is advantageously chosen from liquid, semi-solid or solid materials, such as plastics, thermosets, varnishes, rubbers, lubricants, paints, oils, medicines, cosmetics and foods.

[0184] The material can be any solid traceable item, whether in storage or in transport, such as an electronic device, an item of clothing, paper, fiber, or fabric, or any other commercial item or cash or value.

[0185] Additionally, the commercial items marked with the molecular tags can be liquids such as inks, oils, dyes, sprays, and the like.

[0186] As a commercial item, the product can be a commodity item such as paper, metal, wood, plastic, rubber, powder, and the like.

[0187] Additionally, the material may be a pharmaceutical or food product.

[0188] For example, the material is a viscous aqueous prepolymer that can be irreversibly cured by heating to form a thermoset (see page 898 of "Curing Thermoset Polymers," IUPAC, 2004, 76, 889. (Definitions of terms relating to reactions of polymers and to functional polymeric materials (IUPAC Recommendations 2003))).

[0189] In contrast to this, the target material can be a thermoplastic material, which, in contrast to thermosetting polymers, is transformed without a chemical reaction. No thermal degradation is exhibited and the molecular structure of the polymer is not modified during the transformation. Thermoplastics are advantageously defined in "ISO 7792-1:2012, Plastics - Thermoplastic polyester (TP) molding and extrusion materials - Part 1: Designation system and basis for specifications."

[0190] In a preferred embodiment, the incorporation step advantageously consists of incorporating the marking composition into the material during the manufacturing process of said material.

[0191] In other words, the present invention also relates to a process for making a material, comprising the step of incorporating said marking composition into said material during its making process.

[0192] Preferably, the material is a generation batch. As used herein, "generation batch" advantageously relates to a manufacturing process in which material is made in a specified batch or amount over a period of time.

[0193] The batch is advantageously run through a series of steps in a large scale manufacturing process to produce the final desired material.

[0194] Generally, the concentration of the marking composition is advantageously from 0.1, preferably from 0.5 to 150 mg of marking composition per liter or kg of material (0.1 to 150 ppm, 1 ppm=1 mg / kg).

[0195] Preferably, the concentration is within any combination selected from the following values: 0.1, 0.5, 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150 mg of marking material per liter or kg of material.

[0196] Generally, the particles of the marking composition are advantageously intact in the target material.

[0197] Extracting and reading information support The material is advantageously processed to extract particles and then extract information support from said extracted particles.

[0198] The information sequence is advantageously extracted from the material via an "information sequence extraction method", ie a process for obtaining the information sequence from the material.

[0199] The particles can then be treated with the previously disclosed decomposition conditions (metal chelators or acidic pH conditions) to specifically release the information support.

[0200] For example, a calcium carbonate homogeneous porous matrix can be treated with an acidic solution (pH 2, preferably citric acid) or an EDTA solution, or a combination thereof, to release the information support.

[0201] For this reason, in general, the extraction method is advantageously carried out, for example, by dissolving the homogeneous porous matrix in an aqueous solution of the complexing agent, followed by extraction and purification steps to recover the information support.

[0202] To extract information support from molecular tags containing hydrophobic fluidizers embedded in hydrophobic materials, extraction is advantageously performed by combining a hydrophobic solvent (e.g., toluene, chloroform, dichloromethane, ethyl acetate) in combination with an aqueous chelator digestion solution. The information support may be in an aqueous fragmented state.

[0203] Once the vehicle is dissolved, the extracted information support can be purified and amplified.

[0204] Finally, the reading step corresponds to sequencing the information support and decoding the information sequence via the selected encoding method.

[0205] If the marked material is solid, the particles are extracted by first dissolving or melting the material in a suitable solvent. The choice of solvent is highly dependent on the product material. Suitable solvents can be easily identified by those skilled in the art and include organic solvents as well as aqueous (acidic or basic) solvents.

[0206] Generally, the particles are advantageously dissolved using an aqueous chelating agent solution or an aqueous acidic solution.

[0207] Preferably, the organic solvent containing the particles can be directly contacted with the dissolution solution and subsequently extracted (containing the information support in the aqueous phase). Alternatively, an intermediate step can be performed to separate the microparticles from the organic solvent, for example, by centrifugation or filtration.

[0208] Various other modifications of the invention may be made within the scope of the appended claims. [Example]

[0209] Example 1: Synthesis of size-controlled mesoporous vaterite microparticles (vehicles) In a typical experiment, 50 mL of 0.33 M Na2CO3 solution was rapidly poured into an equal volume of 50 mL of 0.33 M CaCl2 solution at 20 °C, and after 30 seconds of vigorous controlled agitation (650 rpm) with a digital electronic overhead stirrer, the white precipitate was filtered off on a Satorlon Polyamide filter (pore size 0.2 μm).

[0210] The precipitate was thoroughly washed twice with pure water to remove excess ions, and then washed twice with 100% ethanol to improve the dispersion properties.

[0211] The resulting powder was dried for approximately 12 hours at 55° C. This procedure results in fairly uniform spherical CaCO microparticles with an average diameter in the 6 μm range.

[0212] The size of the microparticles can be modified by changing several parameters such as salt concentration, agitation speed, or temperature.

[0213] In another typical experiment, 50 mL of 0.6 M Na2CO3 solution was rapidly poured into an equal volume of 50 mL of 0.6 M CaCl2 solution at 20° C. After 30 seconds of vigorous, controlled agitation (650 rpm) with a digital electronic overhead stirrer, the white precipitate was filtered off on a Satorlon Polyamide filter (pore size 0.2 μm).

[0214] Applying the same filtration and drying procedure results in fairly uniform spherical CaCO3 microparticles with an average diameter of 2.5 μm.

[0215] Example 2: Generation and synthesis of information-supporting DNA sequences Short barcodes encoding DNA sequences (typically 10 bp to 210 bp) were computer-generated and contained digital information composed of alphanumeric characters.

[0216] For example, digital information can be encoded in nucleobases (barcode coding sequences) via the Reed-Solomon error-correcting code algorithm. This format allows for the storage of digital information in polynucleotide molecules.

[0217] To amplify the DNA sequence, 2 x 20 bp orthogonal (non-interacting) polymerase chain reaction primer binding regions are generated on either side of this barcode coding sequence. The association of the barcode coding sequence + 2 x 20 bp primer binding regions on each side provides the informative supporting sequence.

[0218] In a specific embodiment aimed at stabilizing DNA molecules, a DNA strand complementary to the information-supporting sequence carrying the digital information can be generated.

[0219] Finally, chemically synthesize double-stranded DNA information support (ranging from 50 bp to 250 bp).

[0220] Example 3: Loading of information support and hydrophilic fluidizer (surface functionalizing agent), synthesis of hydrophilic molecular tag The double-stranded nucleotide information support and fluidizer are incorporated by direct adsorption of macromolecules from solution onto preformed CaCO3 microspheres (physisorption).

[0221] In a typical experiment, DNA at a concentration of 15 μM and 0.5% (w / w) sodium polyacrylate salt (fluidizing agent) based on the mass of the CaCO3 microparticles are dissolved in loading buffer (KCl 0.5N, pH 6.5).

[0222] 50 mg of CaCO3 microparticles were then resuspended in a 2 ml Eppendorf tube containing 500 μL of loading buffer, vortexed vigorously, and incubated under agitation for at least 4 hours.

[0223] The zeta potential of the microparticles ranged from positive to excessive Ca at pH 8.0–9.0. 2+ It is important to note that the pH of the loading buffer is chosen to favor the interaction of the particles with negatively charged macromolecules, since the charge at pH 6.5 changed to a negative value that can be explained by the pH change. Thus, at pH 6.5, the microparticles have a positive global net charge.

[0224] After incubation, the microparticles were centrifuged to remove the loading buffer and washed once with clean KCl 0.5N pH 6.5 buffer (5 min) to remove non-adsorbed macromolecules. The microparticles were washed twice with 70% ethanol solution and dried at 55°C in a hood.

[0225] DNA loading can be determined indirectly by measuring the disappearance of DNA absorbance (260 nm) in the loading and washing buffers.

[0226] In a typical experiment, 6 μm molecular tags are loaded at concentrations ranging from 0.1 to 2 μg of DNA per mg of inorganic vehicle.

[0227] In another typical experiment, 2.5 μm molecular tags are loaded at concentrations ranging from 2 to 15 μg of DNA per mg of inorganic vehicle, depending on the DNA loading solution concentration.

[0228] Example 4: Synthesis of hydrophobic molecular tags by sequential loading of information support followed by hydrophobic fluidizer In a typical experiment, DNA is dissolved in loading buffer (KCl 0.5N, pH 6.5) at a concentration of 15 μM.

[0229] 50 mg of CaCO3 microparticles were then resuspended in a 2 ml Eppendorf tube containing 500 μL of loading buffer, vortexed vigorously, and incubated under agitation for at least 4 hours.

[0230] After incubation, the microparticles were centrifuged to remove the loading buffer and washed once with clean KCl 0.5N pH 6.5 buffer (5 min) to remove non-adsorbed macromolecules. The microparticles were washed twice with 70% ethanol solution and dried at 55°C in a hood.

[0231] DNA loading can be determined indirectly by measuring the disappearance of DNA absorbance (260 nm) in the loading and washing buffers.

[0232] In a typical experiment, 6 μm molecular tags are loaded at concentrations ranging from 0.5 to 10 μg of DNA per mg of inorganic vehicle.

[0233] In another typical experiment, 2.5 μm molecular tags are loaded at concentrations ranging from 2 to 15 μg of DNA per mg of inorganic vehicle, depending on the DNA loading solution concentration.

[0234] Stearic acid fluidizer is dissolved at 1 mg / mL in an ethanol / water solution (2 / 1, v / v) at 60°C.

[0235] Once dried, the DNA-loaded microparticles are mixed with a portion of stearic acid solution, the amount of stearic acid added being 2% (w / w), for example, 1 mg for 50 mg of DNA-loaded microparticles.

[0236] Once mixed with the stearic acid solution, the microparticles are incubated at 58° C. for 4 hours under vigorous agitation in a sealed tube to prevent evaporation of the ethanol and water.

[0237] After incubation, the microparticles were centrifuged to remove the stearic acid loading buffer and washed once with clean ethanol / water 2 / 1 (v / v) (5 min) to remove non-adsorbed molecules. The microparticles were washed twice with 70% ethanol solution and dried at 55 °C in a hood.

[0238] During this process, it can be observed that the colored fluorescent DNA remains well confined within the microparticles, and no DNA is released.

[0239] Example 5: Microparticle size analysis The size of the microparticles can be measured directly in DIC (differential interference contrast) or fluorescence microscopy photographs. Photographs were taken with a Nikon H600L fluorescence microscope (Japan) equipped with an ANDOR 5.5 Neo camera, a Lumencor Spectra X Light Engine Computer Controllable source, and fluorescence filters (DAPI, FITC, RITC) using NIS Elements software version 4.40—see Figure 1.

[0240] Manually measured average sizes of 200 microparticles give sizes of 6.28±0.59 μm for microparticles made at 0.3 M salt concentration and 2.32±0.32 μm for microparticles made at 0.6 M salt concentration, respectively.

[0241] Example 6: Effect of fluidizer on the miscibility, polydispersity, and aggregation tendency of microparticles in aqueous media (Figure 2) Measurement of the hydrodynamic volume and diameter of microparticles diluted in distilled water can be performed using a Malvern Mastersizer3000 (particle size distribution analyzer) particle sizer equipped with a Hydro2000S system for measuring the size of microparticles diluted in a continuous water stream.

[0242] First, we must point out that the hydrophobic microparticles tested (stearic acid fluidizer), whether empty or loaded with DNA, are not compatible with these measurements because they are not miscible with water; in fact, they float on the water.

[0243] On the other hand, this type of measurement focuses on the dispersibility of the hydrophilic fluidizer sodium polyacrylate.

[0244] Microparticles without a fluidizer (calcium carbonate vehicle alone) tend to form aggregates of various sizes (6.5 μm or 90 μm for 2.5 μm particles), but the addition of 0.5% w / w hydrophilic sodium polyacrylate fluidizer suppresses aggregate formation and produces a single population of non-aggregated microparticles with a hydrodynamic diameter of 2.96 μm.

[0245] The effect of a fluidizer (0.5% w / w sodium polyacrylate) on the sedimentation properties of the vehicle is also confirmed by Turbiscan measurements at 10 mg / mL in water. The bare vehicle settles in water in 1 hour, while the hydrophilic fluidizer-modified vehicle settles much more slowly (4 hours). Conversely, the vehicle modified with a hydrophobic fluidizer (stearic acid 2% w / w) is no longer miscible with water and tends to float on the water surface.

[0246] Example 7: Direct release and amplification of nucleotide information supports from microparticles One particular advantage of calcium carbonate microparticles is that they can be easily degraded and removed by acidic pH (<5) and / or the addition of chelating agents such as EDTA.

[0247] Thus, in a typical experiment, the microparticles are degraded with a "degradation buffer" (EDTA 100 mM, pH 5).

[0248] The disintegration of the particles is almost instantaneous as evidenced by the appearance of CO2 bubbles from the dissociation of the particles.

[0249] The milky solution becomes clear when all the vehicle is dissolved. If the particles are in a basic buffer (pH > 8), the pH of the solution must first be lowered to below 7 before adding the chelating agent. Higher concentrations of chelating agent solutions can be used (500 mM EDTA or 200 mM EGTA, pH 5 solutions). Alternatively, a relatively large amount of vehicle can be used to dissolve the chelating agent and Ca. 2+ The moles of chelating agent must be adjusted to achieve equimolar concentrations between the ions.

[0250] After 5 minutes of vigorous agitation, the informative nucleotide support can be easily purified using a commercially available DNA purification kit (NEB PCR Cleanup or Macherey-Nagel NucleoSpin Gel and PCR Cleanup) and then amplified by PCR.

[0251] Another option for the release of information support is to use a 1% citric acid (w / v) solution at pH 2. To dissolve calcium carbonate (CaCO) using citric acid (C6H8O7), the following chemical formula is used: 3CaCO3+2C6H8O7→Ca3(C6H5O7)2+3CO2+3H2O According to the formula, there needs to be enough acid to react with calcium carbonate and convert it to calcium citrate (Ca(CHO)), carbon dioxide (CO), and water (HO).

[0252] The resulting calcium citrate (Ca3(C6H5O7)2) is a non-toxic calcium salt of citric acid that is used as a food additive (E333).

[0253] This purification step involves the addition of Mg 2+ It is essential to eliminate EDTA or EGTA or citrate chelating agents which inhibit the PCR reaction by chelating ions.

[0254] Finally, to extract the digital information, one simply has to sequence the amplified DNA fragments on a polynucleotide sequencer.

[0255] For hydrophobic molecular tags, such as stearic acid-coated vehicles, an equal volume of chloroform is added to the digestion buffer and vehicle. After vigorous agitation and demixing of the two immiscible solvents (chloroform and aqueous digestion solution), the information support is present in the aqueous phase, ready for analysis.

[0256] Example 8: Demonstration of DNA-protective effect of a combination of homogeneous porous vehicles and fluidizing agents against chemically reactive genotoxic small molecules (isothiocyanates) - Figure 3 First, to investigate the protective effect against chemical reactivity in an aqueous environment, we defined double-stranded DNA (70-mer or 80-mer) modified at the 5' end to carry a free amine functional group (amine-DNA). Indeed, primary amines are highly reactive toward isothiocyanates, which are molecules that can react with the 5' amine or the amine functional groups of the nitrogenous bases adenine and guanine. Next, to quantify molecular reactivity, we selected fluorescent isothiocyanates (FITC: fluorescein isothiocyanate and RITC: rhodamine isothiocyanate). Therefore, if DNA is not well protected, it can become fluorescent. In a specific example, three different samples of amine-DNA were subjected to reaction with 100 equivalents of FITC or RITC for 48 hours. The first sample was amine-DNA alone, the second sample was amine-DNA loaded into a 2.5 μm uniformly porous calcium carbonate vehicle (10 mg amine-DNA loaded per gram of vehicle), and the third sample was amine-DNA loaded into a 2.5 μm uniformly porous calcium carbonate vehicle (3.5 mg amine-DNA loaded per gram of vehicle) and functionalized with 0.5% (w / w) hydrophilic sodium polyacrylate fluidifier. Briefly, the amount of DNA was adjusted to 25 μg in 100 μL sodium carbonate / bicarbonate buffer, pH 10. Then, 100 equivalents of RITC or FITC diluted in 10 μL DMF was added, shaken vigorously, and incubated for 48 hours under agitation, away from light. The reaction was stopped by adding 100 μL Tris HCl buffer, 500 mM, pH 8 (quencher). Next, adjust the pH of the solution to 5 with dilute HCl solution. Finally, dilute the sample to 500 μL with "dissolution buffer" (EDTA 100 mM pH 5). The 500 μL sample solution should be clear, indicating that the microparticles are well dissolved; otherwise, add a small amount of acetic acid or dilute HCl. Once the particles are dissolved, purify the 500 μL DNA sample on a PD MiniTRAP G25 desalting column (Cytiva) equilibrated with water pH 7, according to the manufacturer's instructions.The purified DNA is then lyophilized and reconcentrated in 100 mM Tris HCl buffer, pH 8. The concentration of each sample is adjusted to the same concentration and then analyzed by electrophoresis in a 4% agarose electrophoresis gel for 45 minutes using a Fisherbrand™ Horizontal Mini Gel Electrophoresis System. Fluorescein and rhodamine fluorescence can be observed before BET staining using a BioRad Gel Doc XR+ imager and image lab software. Once the gel is made, the DNA can be repurified using a Macherey Nagel (NucleoSpin Gel and PCR Clean-up) purification kit, readjusted to the same concentration, and fluorescence can be measured using an EnSpire Multimode Plate Reader (Perkin) microplate reader. All experiments performed yield the same results: DNA alone and DNA loaded into a homogeneous porous calcium carbonate vehicle react similarly to isothiocyanates (RITC or FITC). Thus, a homogeneous porous vehicle alone does not protect DNA from this type of small reactive molecule. In contrast, when 0.5% (w / w) sodium polyacrylate fluidizer is added, more than 80% of the DNA is protected and does not react with the fluorescent molecule. It is the association of three entities, a synergistic tryptic (support + vehicle + fluidizer), that results in molecular tags that protect more than 80% of the informative DNA.

[0257] Example 9: Incorporation of molecular tags into thermosetting polymers The base formulation for the thermoset was kindly provided by an industrial partner. Due to proprietary rights, the detailed composition cannot be disclosed, but it is important to note that the pH of this material is basic (pH > 9). In a typical experiment, preloaded 2.5 μm hydrophilic molecular tags (a homogeneous porous calcium carbonate vehicle loaded with 70 bp double-stranded DNA at 10 mg / g and functionalized with 0.5% (w / w) hydrophilic sodium polyacrylate fluidizer) are directly diluted into an alkaline aqueous thermosetting prepolymer at a selected dilution factor (e.g., 50 mg of molecular tags per L of prepolymer). The aqueous mixture is then vigorously agitated to favor microparticle dispersion and homogenization.

[0258] Curing of the thermosetting prepolymer containing the molecular tag (polymerization and cross-linking of the prepolymer) can then be achieved by heat treatment (100° C. for 3 hours) to result in the formation of a "tagged" thermoset.

[0259] The resulting polymerized, insoluble thermoset is solid. Typically, 250 μl of heated thermosetting prepolymer will give 50 mg of solid thermoset.

[0260] Example 10: Extraction and amplification from cured thermoset The polymerized thermoset can be treated to extract the informational nucleotide support.

[0261] Briefly, 50 mg of solid is ground into micrometric particles in a cryogenic grinder (CryoMill, RETSH) in order to favor contact between the "disintegration buffer" (EDTA 100 mM, pH 5) and the molecular tags.

[0262] For the same reason, solid thermoset micrometric particles (50 mg) are "pre-swollen" in 600 μL of chloroform (or toluene) for 1 hour.

[0263] Then, 200 μL of "degradation buffer" is added, and the sample is vigorously agitated for at least 1 hour to degrade the calcium carbonate vehicle. To enhance DNA purification yields, for highly diluted DNA samples, poly(A) from Sigma is added to the "degradation buffer" at 1 μg / mL during extraction. Poly(A) is used as a carrier for quantitative precipitation or purification of DNA and RNA.

[0264] After decantation, in the case of chloroform, the nucleotide information support is in the upper aqueous phase, while the hydrophobic components that may perturb the PCR reaction are in the lower phase.

[0265] In order to eliminate EDTA, which may also inhibit the PCR reaction, the aqueous phase containing the information support is purified with a classical DNA purification kit (NEB PCR Clean up) before the PCR reaction allowing the information to be amplified.

[0266] The purified DNA can then be sequenced.

[0267] Example 11: Incorporation of molecular tags into biodegradable thermoplastics (e.g., polylactic acid (PLA) or polycaprolactone (PCL) or DL-lactide and glycolide copolymer (PDLG) or L-lactide and ε-caprolactone copolymer (PLC)) GMP-grade thermoplastic polymers and copolymers are purchased from Corbion Purac®. In a typical experiment, PDLG7507 or PDLG5010 (a 75:25 molar ratio DL-lactide and glycolide copolymer) granules were completely dissolved in chloroform at a concentration of 10 wt% (10 g of copolymer diluted in 100 mL of chloroform) by stirring at 40°C for 60 minutes. 1 mg of preloaded hydrophobic molecular tag (10 μg of informative DNA per mg of 2.5 μM CaCO3 vehicle, 2% w / w stearic acid fluidifier) ​​was diluted in 1 mL of ethanol, vortexed, and then directly incorporated into the 10% w / v copolymer / chloroform solution under vigorous mixing (10 minutes). The resulting mixture was cast into a clean Teflon® Petri dish, and the chloroform was allowed to evaporate at ambient temperature and pressure under a chemical hood.

[0268] The resulting solid film was finally dried in an oven at 60°C for 48 h.

[0269] Initially, the final concentration of molecular tags was 100 ppm (100 mg in 1 kg) and the concentration of informative DNA support (10 μg in 10 g of copolymer) corresponded to 1 ppm.

[0270] To see if this would work at higher dilutions, we tested concentrations of 2 ppm, 1 ppm, and 0.1 ppm in the molecular tag (20, 10, and 1 ppb of DNA relative to the target material, respectively).

[0271] We also compared hydrophilic molecular tags (fluidizer: sodium polyacrylate) versus hydrophobic molecular tags (fluidizer: stearic acid). Microscopic observations within molded thermoplastic polymers showed that hydrophilic labels tended to form small clusters of 3-5 microparticles, while hydrophobic labels were well dispersed (no aggregates at all).

[0272] Example 12: Extraction and Amplification of Information Support from Thermoplastics - Figure 4 Solid thermoplastics (dry polymers or copolymers) can be treated to extract the informational nucleotide support. Briefly, 100 mg of the solid was cut and placed in a vial. 500 μL of chloroform was then added to the vial, and the polymer was resolubilized under vigorous agitation. Subsequently, 500 μL of "dissolution buffer" (100 mM EDTA, pH 5, supplemented with 1 μg of polyA carrier) was added, and the vial was stirred for 1 hour. After decantation, the nucleotide information support was in the upper aqueous phase; 300 μL of the aqueous phase was then removed using a sterile syringe, and the DNA was subsequently purified using a Macherey-Nagel NucleoSpin Gel and PCR Clean-up Kit.

[0273] PCR was performed for 25 cycles (98°C for 10 s, 52°C for 30 s, 72°C for 10 s) on an Applied Biosystems® Veriti® Thermal Cycler, 96 wells. Briefly, 5 μL of purified DNA sample was added to 25 μL of Phusion® High-Fidelity PCR Master Mix along with HF buffer (New England Biolabs), 15 μL of PCR-grade water (Roche), and 5 μL primer mix (forward and reverse 35 bp primers, 10 μM each).

[0274] Once amplified, DNA fragments are visualized by electrophoresis in a 4% agarose gel for 45 minutes using a Fisherbrand® Horizontal Mini Gel Electrophoresis System. DNA fragments are revealed by BET staining using a BioRad Gel Doc XR+ imager and image lab software.

[0275] All experiments worked regardless of the type of thermoplastic tested (PLA, PLC, PDLG, and PCL). It should be noted that a negative control was performed using unmarked thermoplastic to ensure that the extracted DNA was not due to laboratory contamination. The purified DNA could then be sequenced.

[0276] Example 13: Incorporation of molecular tags into commercial nail polish and extraction of information support In a typical experiment, preloaded 2.5 μm hydrophilic molecular tags (uniformly porous calcium carbonate vehicle loaded with 70 bp double-stranded DNA at 10 mg / g and functionalized with 0.5% (w / w) hydrophilic sodium polyacrylate fluidizer) are diluted at a selected dilution factor (e.g., 1% w / w) between identical unloaded vehicle microparticles. This mixture is then directly incorporated into 5 mL of commercially available nail polish ("Go Green"® by Yves Rocher) at a selected dilution factor (e.g., 5 to 0.1 ppm relative to the nail polish) and vigorously agitated to favor molecular tag dispersion. The nail polish is then applied to ceramic spheres (MP Biomedicals® 1 / 4 in. Ceramic Spheres) and dried overnight in an oven. Specifically, after drying (evaporation of the nail polish solvent, such as ethyl acetate), the dry mass represents 30% of the deposited mass (50 mg) of varnish.

[0277] Next, for extraction, the colored ceramic spheres were added directly to ethyl acetate solvent (400 μL) and agitated to cause discoloration of the ceramic spheres. 400 μL of "decomposition buffer" (100 mM EDTA, pH 5 supplemented with 1 μg of poly(A)) was then added, and the sample was vigorously agitated for at least 1 hour to decompose the calcium carbonate vehicle.

[0278] After decantation, facilitated by rapid centrifugation (10,000 rpm for 1 minute), the nucleotide information support is in the lower aqueous phase, while the hydrophobic components that may disrupt the PCR reaction are in the organic ethyl acetate upper phase. Finally, the information nucleotide support is purified from the aqueous lower phase using a classical DNA purification kit (NEB PCR Clean-up) before the PCR reaction, allowing the information to be amplified.

[0279] This example was reproduced and confirmed using molecular tags diluted to 0.1 ppm (1 ppb DNA).

[0280] Example 14: DNA sequencing and digital data reading The amplified and purified polynucleotide information support can be sequenced on a polynucleotide sequencer.

[0281] Finally, the alphanumeric data can be read and re-extracted from the nucleotide sequence by decrypting the digital information with the computational algorithm selected in Example 2.

Claims

1. 1. A method for marking materials, for example for the purpose of authenticating and / or traceability of said materials, in particular of batches of origin, said marking method comprising: - providing a marking composition selected from molecular tags; - incorporating said marking composition into said material; Including, The marking composition comprises particles, advantageously those that remain intact after incorporation into the material, and which particles comprise: a uniform porous matrix containing a plurality of pores; - an information support added to said homogeneous porous matrix, said information support consisting of a polymer containing an information sequence; the particle surface having a coating comprising at least one fluidizing agent suitable for promoting the uniform dispersion of said particles within said material; A marking method comprising:

2. 2. The marking method according to claim 1, characterized in that the uniform porous matrix is ​​composed of at least 90% w / w of a salt, preferentially an inorganic salt, consisting of an ionic assembly of positively charged cations and negatively charged anions or consisting of a metal oxide, and the uniform porous matrix is ​​insoluble in the material.

3. 3. The marking method according to claim 1 or 2, characterized in that the particles are decomposed by a metal chelating agent or specific pH conditions, preferably mild acidic conditions, more preferably a pH between 2 and 5, more preferably an aqueous solution of citric acid, acetic acid, or hydrochloric acid.

4. Marking method according to any one of claims 1 to 3, characterized in that the particles have a size in the range of 200 nm to 100 μm, preferably 500 nm to 10 μm, more preferably 750 nm to 3 μm.

5. 5. A marking method according to any one of claims 1 to 4, characterized in that the particles have a pore size in the range of 2 nm to 500 nm, preferably 2 to 200 nm, more preferably 10 nm to 100 nm.

6. 6. A marking method according to any one of claims 1 to 5, characterized in that the information support is contained within the pores and / or absorbed onto the surface of the homogeneous porous matrix.

7. Marking method according to any one of claims 1 to 6, characterized in that the information sequence of the information support results from a code conversion of an ordered bit system, the latter resulting from the encoding of a raw data set.

8. Marking process according to any one of claims 1 to 7, characterized in that the information support is chosen among nucleic acids and nucleic acid analogues, preferably DNA, more preferably single-stranded or double-stranded DNA.

9. 9. A marking method according to claim 8, characterized in that the information support consists of nucleic acid fragments, preferably having a size of less than 500 bp, preferably in the range of 50 to 250 bp.

10. Marking method according to any one of claims 1 to 9, characterized in that the information support is associated with, preferably adsorbed on and / or contained in, the homogeneous porous matrix.

11. 11. The marking method according to any one of claims 1 to 10, characterized in that the fluidizing agent is selected from saturated fatty acids, unsaturated fatty acids, phospholipids, lipid mixtures, organic amphiphilic molecules carrying lipophilic moieties, amphiphilic polymers, resin acids, waxes, hydrophilic polymers, peptides and proteins, polysaccharides, surface treatment agents for inorganic fillers, including inorganic salts, more preferably said fluidizing agent is selected from fluidizing agents that improve the dispersibility of hydrophobic or hydrophilic materials.

12. 12. The marking method according to any one of claims 1 to 11, characterized in that the material is selected from the group consisting of liquid, semi-solid or solid materials, such as plastics, thermosets, varnishes, rubbers, paints, oils, lubricants, medicines, cosmetics and food.

13. A marking method according to any one of claims 1 to 12, characterized in that the step of incorporating comprises incorporating the marking composition into the material during the manufacturing process of the material.

14. A marking composition for marking a material, for example for the purpose of authenticating and / or traceability of said material, in particular of a batch of origin, said marking composition comprising particles, advantageously remaining intact after incorporation into said material, said particles comprising: a uniform porous matrix containing a plurality of pores; - an information support added to said homogeneous porous matrix, said information support consisting of a polymer containing an information sequence; the particle surface having a coating comprising at least one fluidizing agent suitable for promoting the uniform dispersion of said particles within said material; A marking composition comprising:

15. A material comprising the marking composition according to claim 14 or the marking composition obtained by the marking method according to any one of claims 1 to 13.

Images