WO2020024059A1 - Oligosaccharides utilisés comme marqueurs oligomoléculaires - Google Patents

Oligosaccharides utilisés comme marqueurs oligomoléculaires Download PDF

Info

Publication number
WO2020024059A1
WO2020024059A1 PCT/CA2019/051058 CA2019051058W WO2020024059A1 WO 2020024059 A1 WO2020024059 A1 WO 2020024059A1 CA 2019051058 W CA2019051058 W CA 2019051058W WO 2020024059 A1 WO2020024059 A1 WO 2020024059A1
Authority
WO
WIPO (PCT)
Prior art keywords
omt
inulin
oligosaccharides
product
hydrolyzed
Prior art date
Application number
PCT/CA2019/051058
Other languages
English (en)
Inventor
Nicholas Hansen LOW
Jamie Lynn WILLEMS
Original Assignee
University Of Saskatchewan
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by University Of Saskatchewan filed Critical University Of Saskatchewan
Publication of WO2020024059A1 publication Critical patent/WO2020024059A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/73—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/10—Washing or bathing preparations
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
    • C07H3/04—Disaccharides
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H3/00—Compounds containing only hydrogen atoms and saccharide radicals having only carbon, hydrogen, and oxygen atoms
    • C07H3/06—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D7/00—Features of coating compositions, not provided for in group C09D5/00; Processes for incorporating ingredients in coating compositions
    • C09D7/40—Additives
    • C09D7/60—Additives non-macromolecular
    • C09D7/63—Additives non-macromolecular organic
    • C—CHEMISTRY; METALLURGY
    • C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
    • C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
    • C09D7/00—Features of coating compositions, not provided for in group C09D5/00; Processes for incorporating ingredients in coating compositions
    • C09D7/40—Additives
    • C09D7/65—Additives macromolecular
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
    • C—CHEMISTRY; METALLURGY
    • C13—SUGAR INDUSTRY
    • C13K—SACCHARIDES OBTAINED FROM NATURAL SOURCES OR BY HYDROLYSIS OF NATURALLY OCCURRING DISACCHARIDES, OLIGOSACCHARIDES OR POLYSACCHARIDES
    • C13K11/00—Fructose
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/92—Oral administration
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/02—Food

Definitions

  • the present disclosure is in the field of internal tracing systems, in particular internal tracing systems using hydrolyzed inulin oligosaccharides as oligomolecular tags (OMTs).
  • ONTs oligomolecular tags
  • RFID tags are more expensive than barcodes, thereby limiting their application for lower-value products.
  • the major limitations of external traceability systems are that they can be readily removed or manipulated, and that the food or ingredient within the container/package can be substituted in whole or in part without detection.
  • Another proposed internal traceability system is through the addition of oligonucleotides with defined sequences to a product.
  • One of the major advantages of genetic (i.e. DNA) based molecular tags is that a nearly unlimited number of unique sequences can be produced, which would allow producers to change the tag used as required (Paunescu et al, 2016).
  • Oligonucleotides have been added to anti-cancer drugs as a method for product traceability and have also been examined for use in food products, fuels and cosmetics (Puddu et al, 2014. Magnetically recoverable, thermostable, hydrophobic DNA/silica encapsulates and their application as invisible oil tags.
  • Limitations of this approach to food traceability include: whether these oligonucleotides would be considered as food additives, and if so their approval; possible negative reactions from consumers to the addition of oligonucleotides (i.e. foreign DNA) to foods; food labeling issues; stability under food processing conditions; cost; and accuracy/reproducibility.
  • the development of an alternate internal traceability system for foods/food ingredients is long overdue.
  • the present disclosure relates to alternate internal traceability systems for foods/food ingredients.
  • Inulin oligosaccharides are useful as internal traceability tags, for example, because: 1 ) these carbohydrates are not commonly found in the majority of foods; 2) production of a series of different oligosaccharide structures is possible by controlled hydrolysis and blending; 3) inulin is naturally present in some food products (e.g. agave syrup, chicory root); and 4) inulin oligosaccharides are food ingredients used in diabetic and low calorie foods and as such are considered to be safe.
  • the present disclosure describes fractions of inulin oligosaccharides (i.e.
  • OMTs oligomoleculartags
  • the present disclosure describes an OMT for tracing a product, wherein the OMT comprises at least one combined fraction of hydrolyzed inulin, optionally chicory inulin, agave inulin, dahlia tubers inulin, or Jerusalem artichoke inulin, suitably chicory inulin.
  • the at least one combined fraction of hydrolyzed inulin comprises oligosaccharides having the molecular weight between about 300 and about 1 ,800 Da.
  • the at least one combined fraction of hydrolyzed inulin comprises oligosaccharides having a degree of polymerization (DP) of (a) from 5 to 8; (b) from 3 to 7; (c) from 3 to 5; or (d) from 2 to 4.
  • DP degree of polymerization
  • the at least one combined fraction of hydrolyzed inulin comprises oligosaccharides: (a) GF5, GFe, Fs, F6, F7, and Fs;
  • the oligosaccharides are GFs, GF6, Fs, F6, F7, and Fs.
  • the oligosaccharides are GF 4 , GFs, F3, F 4 , Fs, F6, and F7.
  • the oligosaccharides are GF3, GF 4 , F3, F 4 , and Fs.
  • the oligosaccharides are F2, F3, and F 4 .
  • the oligosaccharides are in solid, liquid solution or suspension form, optionally powder form.
  • the product is a food product, a pharmaceutical product, a nutraceutical product, a textile product, a personal care product or an industrial product.
  • the product is hydrophobic.
  • the food product is a whole food or a food ingredient.
  • the whole food is a beverage.
  • the food ingredient is flour or a food additive.
  • the beverage is water, milk, a juice, coffee, tea, or a soft drink.
  • the water is coconut water.
  • the juice is apple, pear, grape, cranberry, grapefruit, tomato, blackberry, raspberry, strawberry, orange, winter melon, pomegranate, aloe vera, clam, sugarcane, kiwifruit, lemon, lime, lychee, cantaloupe, honeydew, papaya, pineapple, guava, prune, beet, carrot, celery, cucumber, dandelion-green, parsley, spinach, tejuino, turnip, watercress or hemp juice.
  • the pharmaceutical product is a biologic or a small molecule.
  • the nutraceutical product is a dietary supplement or a functional food.
  • the person care product is a personal hygiene or a cosmetic product.
  • the industrial product is paint.
  • the OMT is at a concentration of at least about 10 ppm.
  • the OMT is stable for at least 12 months, optionally from about 4.0 °C to about 90 °C.
  • the OMT comprises hydrophobic oligosaccharides.
  • the hydrophobic oligosaccharides comprise acetylated oligosaccharides.
  • a method for producing the OMT described herein comprising hydrolysis of inulin, optionally chicory inulin, agave inulin, dahlia tubers inulin, or Jerusalem artichoke inulin, suitably chicory inulin, by an endo-inulinase, wherein the endo-inulinase is from Arthrobacter, optionally Arthrobacter sp. S37, from Aspergillus, optionally Aspergillus niger, or from Pseudomonas , optionally Pseudomonas mucidolens.
  • the hydrolyzed inulin is separated by size exclusion chromatography.
  • the hydrolyzed inulin is identified by Capillary Gas Chromatography, High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection (HPAE-PAD), High Performance Anion Exchange Chromatography with Pulsed Electrochemical Detection (HPAE-PED), or liquid chromatography-mass spectrometry (LC-MS).
  • HPAE-PAD High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection
  • HPAE-PED High Performance Anion Exchange Chromatography with Pulsed Electrochemical Detection
  • LC-MS liquid chromatography-mass spectrometry
  • a method of tracing a product comprises introducing the OMT described herein into the product, optionally during or after production of the product, optionally before or during packaging of the product.
  • a system of tracing a product comprising the OMT described herein and a detector.
  • FIG. 1 shows HPAE-PAD chromatograms of exemplary oligomolecular tags (OMTs) produced via enzymatic hydrolysis of chicory inulin and size exclusion chromatography.
  • OMT #1 A; fractions 31 -33
  • OMT #2 B; fractions 34-36
  • OMT #3 C; fractions 37-39
  • OMT #4 D; fractions 40-42).
  • F fructose
  • G glucose and the subscripts indicate the number of each residue.
  • FIG. 2 shows HPAE-PAD chromatograms of the natural oligosaccharide profiles of laboratory scale apple juice at select processing stages.
  • D concentrated juice.
  • Fig. 3 shows HPAE-PAD chromatograms of the oligosaccharide profiles of laboratory scale apple juice plus exemplary OMT #2 (fraction 34-36; 40.0 ppm) throughout processing.
  • A mash prior to enzyme treatment
  • B mash after enzyme treatment
  • C filtered juice prior to concentration
  • D concentrated juice.
  • Fig. 4 shows peak areas of exemplary inulin oligosaccharides before and after storage in apple juice for six months.
  • inulin oligosaccharides or“inulooligosaccharides” and the like as used herein refer to inulin that has been hydrolyzed into oligosaccharides, i.e. hydrolyzed inulin oligosaccharides, and also includes hydrolyzed inulin oligosaccharides that have been modified so they are hydrophobic.
  • hydrophilic refers to the property of a substance that has attraction to water. If a hydrophilic molecule is mixed with water the two will tend to form one phase. Hydrophilic molecules tend to be polar molecules.
  • hydrophilic may be used to refer to a food or other materials described herein, where the food or other materials are, for example, water based or water soluble.
  • hydrophobic refers to the property of a substance that lacks attraction to water. If a hydrophobic molecule is mixed with water the two will tend to separate and form two individual phases. Hydrophobic molecules tend to be nonpolar molecules that group together to form micelles rather than be exposed to water.
  • hydrophobic may be used to refer to a food or other materials described herein, where the food or other materials contain hydrophobic molecules, for example, fats and oils.
  • tracing also refers to tracking, monitoring, authenticating, distinguishing, and/or identifying a product.
  • A“fraction” of inulin oligosaccharides comprises any suitable part of inulin oligosaccharides for the tracing a product.
  • the fraction may be obtained by any suitable fractionation methods, for example, size exclusion chromatography.
  • combined fraction refers to a composition comprising two or more inulin oligosaccharides of different structures.
  • the inulin oligosaccharides of different structures have different sizes, masses, and/or chemical compositions.
  • a combined fraction is produced and collected from the methods described herein.
  • inulin oligosaccharides are produced by hydrolyzing inulin using an endo- inulinase.
  • the inulin oligosaccharides are separated by fractionation using size exclusion chromatography procedure, whereby eluent is collected in constant volumes (known as fractions) in different tubes, and the eluent collected from proximal tubes are pooled to provide a“combined fraction”.
  • the letters“F” and“G” as used herein are in reference to components of inulin oligosaccharide fractions refers to the molecular residues fructose and glucose, respectively.
  • the numerical subscript (n) proceeding the letters“F” and“G” refer to the number of residues of each type in a given molecule. Therefore, for example, F6 refers to a hexasaccharide containing 6 fructose residues and GF 4 refers to a pentasaccharide containing 1 glucose and 4 fructose residues.
  • free D-fructose refers to a D-fructose molecule that is not covalently linked to another molecule, such as another carbohydrate molecule.
  • the second component as used herein is different from the other components or first component.
  • A“third” component is different from the other, first, and second components, and further enumerated or“additional” components are similarly different.
  • suitable means that the selection of the particular compound or conditions would depend on the specific synthetic manipulation to be performed, and the identity of the molecule(s) to be transformed, but the selection would be well within the skill of a person trained in the art.
  • an effective amount means an amount of an OMT of the disclosure that is effective to achieve the desired result.
  • an effective amount is an amount that, for example, is sufficient to be detected by a detector described herein, compared to the detection without the OMT.
  • detectable as used herein when referring to detection of carbohydrates, for example, monosaccharides including free D-fructose, or oligosaccharides including F2, F3, F 4, Fs, F6, F7, Fs, GF3, GF 4, GFs, and GF6, means detection within the limits using methods, instruments and systems known in the art, for example, detections/detectors described herein including Capillary Gas Chromatography, High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection (HPAE-PAD), High Performance Anion Exchange Chromatography with Pulsed Electrochemical Detection (HPAE-PED), and liquid chromatography-mass spectrometry (LC-MS).
  • HPAE-PAD High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection
  • HPAE-PED High Performance Anion Exchange Chromatography with Pulsed Electrochemical Detection
  • LC-MS liquid chromatography-mass spectrometry
  • the present disclosure includes an oligomolecular tag (OMT) for tracing a product, wherein the OMT comprises at least one combined fraction of hydrolyzed inulin, optionally chicory inulin, agave inulin, dahlia tubers inulin, or Jerusalem artichoke inulin, suitably chicory inulin.
  • the OMT comprises at least one combined fraction of hydrolyzed chicory inulin.
  • the OMT comprises at least one combined fraction of hydrolyzed agave inulin.
  • the OMT comprises at least one combined fraction of hydrolyzed dahlia tubers inulin.
  • the OMT comprises at least one combined fraction of hydrolyzed Jerusalem artichoke inulin.
  • the combined fraction of hydrolyzed inulin contains oligosaccharides that fall within the range of 300 - 1 ,800 Da.
  • the at least one combined fraction of hydrolyzed inulin comprises oligosaccharides having the molecular weight between about 300 and about 1 ,800 Da.
  • the combined fraction of hydrolyzed inulin can be characterized by a degree of polymerization (DP), which refers to the number of monomeric units in an oligomer molecule.
  • DP degree of polymerization
  • the at least one combined fraction of hydrolyzed inulin have a DP of (a) from 5 to 8; (b) from 3 to 7; (c) from 3 to 5; or (d) from 2 to 4.
  • the combined fraction of hydrolyzed inulin includes oligosaccharides containing various number of fructose residues, either only by themselves (F n ) or further containing a glucose residues (GF n ).
  • the at least one combined fraction of hydrolyzed inulin comprises oligosaccharides:
  • the oligosaccharides are GFs, GF6, Fs, F6, F7, and Fs. In some embodiments, the oligosaccharides are GF 4 , GFs, F3, F 4 , Fs, F6, and F7. In some embodiments, the oligosaccharides are GF3, GF 4 , F3, F 4 , and Fs. In some embodiments, the oligosaccharides are F2, F3, and F 4 . [0068] In some embodiments, the oligosaccharides comprise hydrophobic oligosaccharides. In some embodiments, the hydrophobic oligosaccharides comprise acetylated oligosaccharides.
  • the OMTs described herein are useful in a variety of forms.
  • the OMT comprises hydrolyzed inulin in solid, liquid solution or suspension form, optionally powder form.
  • the OMT comprises hydrolyzed inulin in solid form.
  • the OMT comprises hydrolyzed inulin in liquid form.
  • the OMT comprises hydrolyzed inulin in suspension form.
  • the OMT comprises hydrolyzed inulin in powder form.
  • the solvent for the solution or suspension is water.
  • the OMTs described herein are useful for tracing a variety of products, including agricultural products such as foods, health products such as pharmaceuticals and nutraceuticals, and products related to textile, personal care and industry, all of which can be a hydrophilic or hydrophobic product.
  • the product is a food product, a pharmaceutical product, a nutraceutical product, a textile product, a personal care product or an industrial product.
  • the food product is a whole food or a food ingredient.
  • the food product is a whole food.
  • the food product is a food ingredient.
  • the whole food is a beverage.
  • the food ingredient is flour or a food additive.
  • the beverage is water, milk, a juice, coffee, tea, or a soft drink.
  • the water is coconut water.
  • the juice is apple, pear, grape, cranberry, grapefruit, tomato, blackberry, raspberry, strawberry, orange, winter melon, pomegranate, aloe vera, clam, sugarcane, kiwifruit, lemon, lime, lychee, cantaloupe, honeydew, papaya, pineapple, guava, prune, beet, carrot, celery, cucumber, dandelion-green, parsley, spinach, tejuino, turnip, watercress or hemp juice.
  • the pharmaceutical product is a biologic or a small molecule.
  • the nutraceutical product is a dietary supplement or a functional food.
  • the personal care product is a personal hygiene or a cosmetic product.
  • the industrial product is paint.
  • the OMTs described herein are useful for tracing a product even when the OMTs are in low concentrations.
  • the OMT is at a concentration from at least about 1 , 2, 3, 4, 5, 6, 7, 8, 9, or 10 ppm.
  • the OMT is at a concentration of at most about 100 ppm.
  • the OMT is at a concentration of at least about 10 ppm to at most about 100 ppm.
  • the OMT s described herein are stable and useful for tracing a product at a wide range of temperatures.
  • the OMTs described herein are also stable during ultrahigh temperature (UHT) processing, such as at about 150 °C for up to 15 seconds.
  • the OMTs described herein are also stable during canning processing, such as at about 120 °C for up to 20 minutes.
  • the OMTs described herein are also stable at about 90 °C for at least 3 hours.
  • the OMT is stable at about 150 °C for up to 15 seconds.
  • the OMT is stable at about 120 °C for up to 20 minutes.
  • the OMT is stable at about 90 °C for at least 3 hours. In some embodiments, the OMT is stable for at least 6, 9, 12, 15, 18 or 24 months, optionally from about -80, -40, -20, -10, 0, 4.0 °C to about 25, 30, 40, 50, 60, 70, 80, 90 °C. In some embodiments, the OMT is stable for at least 12 months, from about -80 °C to about 90 °C. In some embodiments, the OMT is stable for at least 12 months, from about 4 °C to about 90 °C. In some embodiments, the OMT is stable for at least 12 months, from about -40 °C to about 50 °C. In some embodiments, the OMT is stable for at least 12 months, from about -20 °C to about 40 °C.
  • the OMTs described herein are produced by controlled hydrolysis using an enzymatic method.
  • OMTs comprising hydrolyzed linulin were produced using endo-inulinases (EC 3.2.1.7) from Arthrobacter sp. S37, which hydrolyzes the glycosidic linkages of inulin to produce oligosaccharides and minimal fructose, with less than 0.01 % (w/w) free D-fructose in the hydrolyzed inulin.
  • endo-inulinases EC 3.2.1.7
  • Arthrobacter sp. S37 which hydrolyzes the glycosidic linkages of inulin to produce oligosaccharides and minimal fructose, with less than 0.01 % (w/w) free D-fructose in the hydrolyzed inulin.
  • endo-inulinases EC 3.2.1.7
  • Arthrobacter sp. S37 which hydrolyzes the glycosidic
  • the present disclosure also includes a method for producing an OMT, comprising:
  • an inulin solution optionally chicory inulin, agave inulin, dahlia tubers inulin, or Jerusalem artichoke inulin, suitably chicory inulin, by adding an endo-inulinase to the solution to produce hydrolyzed inulin, wherein the endo-inulinase is from Arthrobacter, optionally Arthrobacter sp. S37, from Aspergillus, optionally Aspergillus niger, or from Pseudomonas , optionally Pseudomonas mucidolens.
  • the method described herein uses an endo-inulinase that produces less than 0.01 % (w/w) free D-fructose.
  • the endo- inulinase produces less than 0.1 %, 0.09%, 0.08%, 0.07%, 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, or 0.01 % (w/w) free D-fructose.
  • the endo-inulinase produces no detectable level of free D-fructose.
  • the endo-inulinase produces less than 0.1 % (w/w) free D-fructose.
  • the endo-inulinase produces less than 0.01 % (w/w) free D-fructose.
  • the presence of free D-fructose indicates exo-inulinase activity, the presence of which decreases the levels of fructose- containing inulin oligosaccharides.
  • the method for producing an OMT comprises adding chicory inulin, agave inulin, dahlia tubers inulin, or Jerusalem artichoke inulin to a solution and hydrolyzing the inulin by adding an endo-inulinase from Arthrobacter, optionally Arthrobacter sp. S37, from Aspergillus, optionally Aspergillus niger , or from Pseudomonas , optionally Pseudomonas mucidolens.
  • the method of producing an OMT comprises adding chicory inulin to a solution and hydrolyzing the inulin by adding an endo-inulinase from Arthrobacter sp. S37.
  • suitable solvents include water, and buffer solutions containing acetate, citrate and phosphate.
  • Inulin hydrolysis described herein is carried out at between about pH 5.00 and about pH 7.50, for at least 24 hours at 50.0 ⁇ 5.0 °C.
  • the hydrolysis condition is at between about pH 5.00 and about pH 7.50, for at least 24 hours at 50.0 ⁇ 5.0 °C.
  • the hydrolysis condition is at between about pH 5.00 and about pH 7.50, for about 24 hours at 50.0 ⁇ 5.0 °C.
  • the hydrolysis condition is at about pH 5.00 for at least 24 hours at 50.0 ⁇ 5.0 °C.
  • the hydrolysis condition is at about pH 5.00 for about 24 hours at 50.0 ⁇ 5.0 °C.
  • Hydrolyzed inulin oligosaccharides can be further processed to produce hydrophobic hydrolyzed inulin oligosaccharides, such as acetylated hydrolyzed inulin oligosaccharides.
  • Acetylation may be carried out prior to or after hydrolyzed inulin oligosaccharides have been fractionated by size exclusion chromatography.
  • Acetylation takes place at hydroxyl groups of the oligosaccharides such that these hydroxyl groups are esterified to acetate.
  • Acetylation reaction occurs in the presence of an acetylating agent, for example, acetyl chloride or acetic anhydride.
  • Acetylation of hydrolyzed inulin oligosaccharides can be controlled and/or maximized by varying temperatures and duration, in the presence or absence of a solvent and/or a catalyst (e.g . sodium acetate) that are familiar to the skilled person.
  • a solvent and/or a catalyst e.g . sodium acetate
  • at least about 0.1 % to at least about 98%, 99%, 99.9%, or about 100% of available hydroxyl groups of the inulin are acetylated.
  • inulin acetylated to a lesser degree may be used, such as inulin with at least about 0.1 % of available hydroxyl groups acetylated.
  • Acetylated hydrolyzed inulin oligosaccharides can be identified by capillary gas chromatography, for example, with flame ionization detection under appropriate temperature programming conditions known to the skilled person.
  • acetylated inulin is considered to be inulin acetate when the hydroxyl peak of its infra-red spectrum disappears and acetyl peaks appear.
  • Inulin oligosaccharides acetate is hydrophobic and insoluble in water even at elevated temperatures, but is soluble in various organic solvents such as acetone, ethyl acetate, chloroform, and dichloromethane, and in for example, hydrophobic foods and bioproducts.
  • the method for producing OMTs further comprises acetylating the oligosaccharides, thereby providing hydrophobic oligosaccharides. In some embodiments, the method for producing OMTs further comprises acetylating the oligosaccharides, thereby providing acetylated oligosaccharides. In some embodiments, the inulin oligosaccharides comprise hydrophobic inulin oligosaccharides.
  • the inulin oligosaccharides comprise acetylated inulin oligosaccharides.
  • the hydrophobic inulin oligosaccharides comprise acetylated inulin oligosaccharide.
  • hydrophobic hydrolyzed inulin oligosaccharides can be produced using other techniques, such as esterification with other carboxylic acids and/or incorporation of silane and/or siloxane groups.
  • hydrophobic inulin oligosaccharides of the present disclosure for use in tracing hydrophobic product, such as hydrophobic food product, a hydrophobic pharmaceutical product, a hydrophobic nutraceutical product, a hydrophobic textile product, a hydrophobic personal care product or a hydrophobic industrial product described herein.
  • structurally different OMTs are fractionated by size exclusion chromatography.
  • Bio-Gel P-2 Gel with a fractionation range of 100 - 1 ,800 Da is used, and water is selected as the mobile phase as it is inexpensive and food safe.
  • sodium or potassium phosphate is selected as the mobile phase for hydrolyzed inulin.
  • sodium or potassium phosphate is selected as the mobile phase for hydrophobic hydrolyzed inulin.
  • sodium or potassium phosphate is selected as the mobile phase for acetylated hydrolyzed inulin.
  • the hydrolyzed inulin is separated by size exclusion chromatography.
  • a number of methods can be used to accurately identify hydrolyzed inulin comprising oligosaccharides.
  • the present disclosure provides High Performance Anion Exchange Chromatography with Pulsed Amperometric Detection (HPAE-PAD) for the identification of hydrolyzed inulin.
  • HPAE-PAD Pulsed Amperometric Detection
  • the skilled person can readily recognize alternative options that are suitable for identification of hydrolyzed inulin.
  • Capillary Gas Chromatography, High Performance Anion Exchange Chromatography with Pulsed Electrochemical Detection (HPAE-PED) and liquid chromatography-mass spectrometry (LC-MS) are also useful for tracing or identifying hydrolyzed inulin.
  • the hydrolyzed inulin is identified by Capillary Gas Chromatography, HPAE-PAD, HPAE-PED, or LC-MS.
  • the hydrolyzed inulin is identified by Capillary Gas Chromatography.
  • the hydrolyzed inulin is identified by HPAE-PAD.
  • the hydrolyzed inulin is identified by HPAE-PED.
  • the hydrolyzed inulin is identified by LC-MS.
  • the oligosaccharides are identified by Capillary Gas Chromatography, HPAE-PAD, HPAE-PED, or LC-MS.
  • the oligosaccharides are identified by Capillary Gas Chromatography.
  • the oligosaccharides are identified by HPAE-PAD.
  • the oligosaccharides are identified by HPAE-PED.
  • the oligosaccharides are identified by LC-MS.
  • hydrolyzed inulin having molecular weights in the range between 300 and 1 ,800 Da, which were comprised in a combined fraction having a degree of polymerization (DP) of (a) from 5 to 8; (b) from 3 to 7; (c) from 3 to 5; or (d) from 2 to 4.
  • DP degree of polymerization
  • the hydrolyzed inulin comprises oligosaccharides having the molecular weight between about 300 and about 1 ,800 Da.
  • the hydrolyzed inulin is comprised in a combined fraction of hydrolyzed inulin having a degree of polymerization (DP) of (a) from 5 to 8; (b) from 3 to 7; (c) from 3 to 5; or (d) from 2 to 4.
  • DP degree of polymerization
  • the methods described herein produced combined fractions containing oligosaccharides having particular glucose and fructose residues.
  • the combined fraction of hydrolyzed inulin comprises oligosaccharides:
  • the oligosaccharides are GFs, GF6, Fs, F6, F7, and Fs. In some embodiments, the oligosaccharides are GF 4 , GFs, F3, F 4 , Fs, F6, and F7. In some embodiments, the oligosaccharides are GF3, GF 4 , F3, F 4 , and Fs. In some embodiments, the oligosaccharides are F2, F3, and F 4 .
  • the oligosaccharides comprise hydrophobic oligosaccharides.
  • oligosaccharides GFs, GF6, Fs, F6, F7, and Fs comprise hydrophobic oligosaccharides.
  • oligosaccharides GFs, GFs, Fs, F6, F7, and Fs comprise hydrophobic oligosaccharides.
  • oligosaccharides GF 4 , GFs, F3, F 4 , Fs, F6, and F7 comprise hydrophobic oligosaccharides.
  • oligosaccharides GF3, GF 4 , F3, F 4 , and Fs comprise hydrophobic oligosaccharides. In some embodiments, oligosaccharides GFs, GF6, Fs, F6, F7, and Fs comprise acetylated oligosaccharides. In some embodiments, oligosaccharides GFs, GF6, Fs, F6, F7, and Fs comprise acetylated oligosaccharides. In some embodiments, oligosaccharides GF 4 , GFs, F3, F 4 , Fs, F6, and F7 comprise acetylated oligosaccharides. In some embodiments, oligosaccharides GF3, , F3, F 4 , and Fs comprise acetylated oligosaccharides.
  • the OMTs described herein are useful for tracing a product described herein.
  • Introduction of the OMT s into the product can be carried out during the preparation or production of the product or after the product has been produced, and/or before or during packaging.
  • the present disclosure also provides a method of tracing a product comprising introducing an OMT described herein into the product, optionally during or after production of the product, optionally before or during packaging of the product.
  • an OMT is introduced into a product during production of the product.
  • an OMT is introduced into a product after production of the product.
  • an OMT is introduced into a product before packaging of the product.
  • an OMT is introduced into a product after packaging of the product.
  • detection of the OMTs comprises use of a method, instrument or system described herein.
  • the OMTs described herein are useful as part of a system for tracing a product described herein. Accordingly, the present disclosure further provides a system of tracing a product, comprising an OMT described herein and a detector.
  • the detector can be any suitable detector known in the art, for example Capillary Gas Chromatograph, HPAE-PAD, HPAE-PED and LC-MS. In some embodiments, the detector is Capillary Gas Chromatograph, HPAE-PAD, HPAE-PED or LC-MS. In some embodiments, the detector is Capillary Gas Chromatograph. In some embodiments, the detector is HPAE-PAD. In some embodiments, the detector is HPAE-PED. In some embodiments, the detector system is LC-MS.
  • Agar, ammonium sulfate, ampicillin, antifoam A, chloramphenicol, imidazole, inulin (chicory), isopropyl b-D-l-thiogalactopyranoside (IPTG), sodium dihydrogen phosphate (NaH2P0 4 ), sodium hydroxide (NaOH) solution (50% w/w) and Tris-HCI were obtained from Sigma-Aldrich Canada Ltd. (Oakville, ON, Canada).
  • Glycerol, LB broth (Miller, granulated), sodium acetate (NaOAc) and sodium chloride (NaCI) were obtained from Fisher Scientific (Ottawa, ON, Canada).
  • Bio-Gel P-2 gel was obtained from Bio-Rad Laboratories (Hercules, CA, USA). Inulin oligosaccharide powder Orafti® P95 was obtained from Beneo (Mannheim, BW, Germany). The water used throughout this disclosure was obtained from a Milli-QTM water system (Millipore Corp., Milford, MA, USA).
  • a plasmid (pRSET C) containing the gene for an Arthrobacter sp. S37 endo-inulinase was obtained from Dr. S. Kang and stored at -80 °C (Kang et at, 1998. Purification and properties of an endo-inulinase from an Arthrobacter sp. Biotechnology Letters , 20: 983-986). The gene for the endo-inulinase had been modified to contain a lac operon and a His6 tag.
  • the overnight culture was then used to inoculate 20.0 mL of LB broth containing ampicillin (50 pg/mL) and chloramphenicol (35 pg/mL) and incubated at 37.0 ⁇ 2.0 °C until an optical density at 600 nm of approximately 0.5 (OD600 ⁇ 0.5) was reached.
  • Enzyme production was induced by the addition of IPTG to a final concentration of 1.0 mM and the incubation temperature was reduced to 25.0 ⁇ 2.0 °C.
  • Samples of the supernatant were taken after 20.0 and 48.0 hours, and were analyzed by SDS-PAGE using a Mini-Protean II electrophoresis cell (Bio-Rad, Hercules, CA, USA) to confirm the production of endo-inulinase by a band at ⁇ 75 kDa.
  • 50 pL of sample was mixed with a 50 pL b-mercaptoethanol and SDS and heated at 100 °C for 5 minutes prior to being loaded onto a 12% polyacrylamide gel with 1 % SDS. Gels were run for 1.5 hour at a constant voltage (200 V) and stained with Coomassie blue dye.
  • BLUeye prestained protein ladder (GeneDireX, Inc., Sigma-Aldrich Canada Ltd., Oakville, ON, Canada) was used for comparison.
  • the mixture was incubated at 25.0 ⁇ 2.0 °C overnight, and cells were sedimented by centrifugation (Sorvall Instruments Centrifuge, DuPont, Wilmington, DE, USA) at 6,000 rpm for 30 minutes at 4.0 ⁇ 2.0 °C.
  • the cell pellet was re-suspended in 20.0 mM Tris-HCI (pH 7.5), and the cells were disrupted using a French press cell disruptor at 15 kPsi and then centrifuged at 12,000 rpm for 20 minutes at 4 ⁇ 2.0 °C.
  • the supernatant was collected and ammonium sulfate was added to achieve a 20% (w/w) saturation level and the mixture was stored static at 4.0 ⁇ 2.0 °C overnight.
  • the solution was centrifuged at 12,000 rpm for 30.0 minutes at 4.0 ⁇ 2.0 °C and the supernatant was collected. Ammonium sulfate was added to achieve 60% (w/w) saturation and the solution was kept static at 4.0 ⁇ 2.0 °C for 3.5 hours. The solution was then centrifuged at 12,000 rpm for 30.0 minutes at 4.0 ⁇ 2.0 °C and the pellet was re-suspended in 4.0 ml_ of binding buffer (20.0 mM NaH2P0 4 , 500.0 mM NaCI, 5.0 mM imidazole, 10.0% glycerol, pH 7.5) and dialyzed against the binding buffer for 72 hours.
  • the solution was then syringe filtered (cellulose acetate, 0.45 pm pore size; 13 mm diameter; Chromatographic Specialties) and purified by fast protein liquid chromatography (FPLC; BioLogic LP System; Bio-Rad Laboratories; Hercules, CA, USA) employing a HiTrap chelating HP column (GE Healthcare Life Sciences, Mississauga, ON, Canada).
  • FPLC fast protein liquid chromatography
  • the stationary phase was initially equilibrated with 20.0 mL of charging buffer (50.0 mM NiS0 4 , pH 4.0) at a flow rate of 1 .0 mL/min for 20.0 minutes, followed by binding buffer at the same flow rate for 20 min.
  • Inulooligosaccharide Separation Inulooligosaccharides were fractionated employing a Bio-Gel P-2 gel stationary phase with water as the mobile phase.
  • the stationary phase ⁇ 180g was initially hydrated for 16 h at room temperature in water prior to packing into a glass column (120 x 2.5 cm to achieve a bed height of 100 cm).
  • Ten grams of a 3.0% (w/w) aqueous solution of produced inulooligosaccharides was placed onto the column with water elution at a flow rate of 0.6 mL/min with 10 ml_ fractions collected for a period of 13 hours. Individual fractions were monitored for the presence of carbohydrates employing HPAE-PAD.
  • Inulooligosaccharide analysis was carried out using a Dionex ICS 5000 HPLC system (Thermo Fisher Scientific, Waltham, MA, USA) equipped with a Dionex AS autosampler, and an ICS 5000 electrochemical cell with a disposable gold electrode.
  • Analyte separation was accomplished using a Dionex CarboPac PA1 column (4 x 250 mm) in series with a CarboPac PA1 guard column (4 x 50 mm) at room temperature.
  • a linear gradient elution program was used for inulooligosaccharide separation where solvent A was 160 mM NaOH, solvent B was 160 mM NaOH/1.0 M NaOAc and solvent C was 1 .0 M NaOH.
  • Initial conditions were 100.0% A for 1.5 min; linear gradient to 60.0% B at 60.0 min and hold for 1.0 min; 100.0% C at 61.1 min with a hold until 65.0 min; 100.0% A at 65.1 min with a hold until 75.0 min.
  • the flow rate was 1.0 mL/min.
  • the injection volume was 20.0 mI_.
  • the mixture was then allowed to react at room temperature (22 ⁇ 1 °C) with stirring for 90.0 minutes.
  • the enzymes were inactivated by heating in a 90 °C water bath for 10.0 minutes.
  • the juice was extracted by vacuum filtration with VWR Grade 417 filter paper (40 pm pore size; VWR International Incorporated LLC, Mississauga, ON).
  • Juice clarification was achieved through the addition of 3.0 pL of Amylase AG 300 L and 3.8 pL of Pectinex Ultra Clear and the mixture was allowed to react at 50.0 ⁇ 5.0 °C for 90.0 minutes prior to enzyme inactivation.
  • Particles were removed by vacuum filtration through VWR Grade 413 filter paper (5 pm pore size).
  • Juice was concentrated to 70.0 ⁇ 5.0 °Brix using a rotary evaporator. Samples were taken after mash production, after enzyme treatment of the mash, the final juice and concentrate. Laboratory scale apple juice samples were stored at -20 °C until required for further analysis.
  • OMTs Laboratory Scale Apple Juice with Oligomolecular Tags
  • Two of the oligomolecular tags (OMTs) were selected (Fractions 34-36 and 40- 42) and intentionally added at the mashing stage (/.e., after blending but prior to enzyme addition) of laboratory scale apple juice production at two different concentrations (10.0 and 40.0 mg OMTs/100 g mash) to determine if inulooligosaccharides could be utilized as an internal traceability system.
  • the untagged samples at the same stages of laboratory scale apple juice production were used as controls.
  • Non-concentrate samples were syringe filtered into 2.0 ml_ sterilized centrifuge tubes (Fisher Scientific), with one set maintained at room temperature (22.0 ⁇ 2.0 °C) and a second at 4.0 ⁇ 2.0 °C. The same conditions were used for the concentrate samples. Samples were analyzed at the following time periods: 1 day, 2 days, 3 days, 1 week, 2 weeks, 3 weeks, 1 month, 2 months, 3 months, 4 months, 5 months and 6 months by HPAE-PAD. Apple juice samples were diluted to 5.5 ⁇ 0.1 °Brix with water prior to analysis.
  • Oligosaccharides produced from enzymatic inulin hydrolysis were selected as potential internal oligomolecular tags (OMTs) as they, (1 ) can be produced to yield a wide range of structural diversity; (2) have generally recognized as safe (GRAS) status for their addition to foods (FDA, 2014. GRAS notice 000537: Short-chain fructo- oligosaccharides); (3) are stable to human and food glycosidic enzyme activities; (4) offer long term stability under the acid conditions of foods (see results below); and (5) have high water solubility which is useful in the majority of food products.
  • OMTs organic oligomolecular tags
  • inulin and related oligosaccharides in the food industry, with the majority involving their role as a prebiotic/soluble fibre ingredient.
  • Other industrial uses include, lipid replacement in foods such as table spreads, baked goods, dairy products and frozen desserts (Lopez-Molina et ai, 2005. Molecular properties and prebiotic effect of inulin obtained from artichoke ( Cynara scolymus L). Phytochemistry, 66: 1476-1484); and as a low calorie and low glycemic index sweetener (Morris and Morris, 2012.
  • inulin and fructo-oligosaccharide supplementation have also been used as the starting material for ultra-high fructose syrup (UHFS; ⁇ 85 to 97% fructose) production via acid or enzyme hydrolysis (Bajpai and Bajpai, 1991 . Cultivation and utilization of Jerusalem artichoke for ethanol, single cell protein, and high fructose syrup production. Enzyme and Microbial Technology, 13: 359-362; Nakamura et ai, 1995.
  • Oligosaccharides can be produced using either enzymatic or chemical hydrolysis of inulin. Chemical hydrolysis results in poorer inulin oligosaccharide production due to the harsh environmental conditions employed (i.e. HCI [ ⁇ 0.5N]/60 °C/15 min) resulting in the production of high levels of fructose (Avila-Fernandez et al, 201 1 . Production of functional oligosaccharides through limited acid hydrolysis of agave fructans. Food Chemistry, 129: 380-386), which makes the material unsuitable as a source for OMTs.
  • Acid hydrolysis of inulin has a number of additional drawbacks for OMT s production including: final product colour (due to caramelization and Maillard reactions); the use of non-environmentally friendly chemicals; the production of unwanted by-products such as furfurals (e.g. 5- hydroxymethylfurfural [HMF]); requirement of corrosion resistant materials; and the production of unwanted salts during neutralization (Waleckx et al, 201 1. Use of inulinases to improve fermentable carbohydrate recovery during tequila production. Food Chemistry, 124: 1533-1542).
  • inulin oligosaccharides can be synthesized from sucrose by the addition of fructose units using either b-fructofuranosidase (EC 3.2.1.26) or b- fructosyltransferase (EC 2.4.1 .9) (Barthomeuf & Pourrat, 1995. Production of high content fructo-oligosaccharides by an enzymatic system from Penicillium rugulosam. Biotechnology Letters, 17: 91 1 -916), these biochemically controlled reactions result in the production of three oligosaccharides [i.e.
  • Chicory inulin was selected as the substrate as it is a common source of inulin used in the food industry and commercially available with a high degree of purity containing minimal oligosaccharide/monosaccharide concentrations prior to hydrolysis.
  • Other inulin sources were also tested by the present inventors and were found to be useful, which include agave inulin, dahlia tubers inulin, and Jerusalem artichoke inulin.
  • oligosaccharides were collected into the following four combined fractions: #1 , tubes 31 -33 (based on a mobile phase flow rate of 0.6 mL/min and a tube volume of 10.0 mL), contained oligosaccharides with degrees of polymerization (DP) ranging from 5 to 8 (Fig. 1 A); #2, tubes 34-36, had DPs ranging from 3 to 7 (Fig. 1 B); #3, tubes 37-39, had DPs ranging from 3 to 5 (Fig. 1 C); and #4, tubes 40- 42, had DPs ranging from 2 to 4 (Fig. 1 D).
  • DP degrees of polymerization
  • each of the four fractions were analyzed by mass spectrometry and had protonated ions ([M+H] + ) corresponding to the correct molecular weights.
  • combined fraction #4 showed protonated ions at m/z 343.2, 505.4 and 667.6 which agree with the presence of di-, tri- and tetrasaccharides, respectively.
  • Combined fraction #1 was identified to contain GFs (i.e., a hexasaccharide containing one glucose residue and five fructose residues), GFs, Fs, F6, F7, and F8, combined fraction #2 contained GF 4 , GFs, F3, F 4 , Fs, F6, and F7, combined fraction #3 contained GF3, GF 4 , F3, F 4 , and Fs, and combined fraction #4 contained F2, F3, and F 4 (Fig. 1 ). Less than 0.01 % (w/w) of free D-fructose was detected when hydrolyzing inulin with the endo-inulinase described herein.
  • GFs i.e., a hexasaccharide containing one glucose residue and five fructose residues
  • combined fraction #2 contained GF 4 , GFs, F3, F 4 , Fs, F6, and F7
  • combined fraction #3 contained GF3, GF 4 , F3, F 4 , and F
  • OMT #2 i.e., combined fraction #2 consisting of fractions 34-36; Fig. 1 B
  • Apple juice was selected as the model system as it is a commonly consumed and adulterated food product (Brause & Ratterman, 1982. Verification of authenticity of apple juice. Journal of the Association of Official Analytical Chemists, 65: 846-849; Low et ai, 1999. Capillary gas chromatography detection of invert sugar in heated, adulterated, and adulterated and heated apple juice concentrates employing the equilibrium method.
  • Oligosaccharide analysis was conducted on the laboratory scale apple juice containing OMT #2 (40.0 ppm) (Fig. 3A-D) and the blank (Fig. 2A-D) at each stage of processing, specifically, the mash prior to enzyme addition, the mash after enzyme treatment, filtered juice after clarification/prior to concentration and the final juice concentrate.
  • the large off-scale peak at the beginning of the chromatogram ( ⁇ 10.0 minutes) is due to the major carbohydrates/polyol in apple juice, namely fructose, glucose, sucrose and sorbitol (Thavarajah & Low, 2006).
  • the apple mash contained minimal naturally occurring oligosaccharides (retention times >10 minutes; Fig. 2A; Willems & Low, 2016).
  • the added OMTs were readily distinguishable from the naturally occurring apple juice oligosaccharides at this stage (Fig. 3A).
  • After enzyme treatment the formation of oligosaccharides was observed as shown by the appearance of peaks in the chromatograms with retention times >12 minutes (Fig. 2B-D).
  • These oligosaccharides were produced from the hydrolysis of pectin, hemicellulose (/.e., xyloglucan) and starch by the commercial enzymes (Willems & Low, 2016).
  • the carbohydrases used during juice production which include a variety of pectinases, amylases and hemicellulases (Kashyap et a ⁇ , 2001 . Applications of pectinases in the commercial sector: a review. Bioresource Technology, 11, 215-227), have not been reported to have activity on inulin based oligosaccharides. This was supported by the minimal change in oligosaccharide concentration (as measured by peak height; /.e., a detector response variation of 51.0 nC initially to 46.6 nC in the final concentrate).
  • any changes in concentration may be due to harsh processing conditions (/.e., high temperature) and potential loss during steps such as filtration. Therefore, for ease of use and to minimize any potential concentration changes, the present disclosure recommends that the OMTs be added later during production, for example just prior to concentration. This would allow easier addition and more consistent distribution through the addition to a liquid rather than the mash.
  • the same experiment was repeated with a final OMT concentration of 10.0 ppm and the results were similar.
  • OMT #2 (/.e. combined fraction #2) were added to a commercial apple juice concentrate to achieve a final concentration of 40.0 ppm.
  • the commercial juice concentrate was analyzed by HPAE-PAD before and after OMT addition. It was found that the OMTs could be readily differentiated from the oligosaccharides found in commercially produced apple juice, further showing the applicability of these oligosaccharides as markers. The same experiment was repeated with a final OMT concentration of 10.0 ppm and the results were similar.
  • Apple juice was selected for the reasons described above and room temperature and 4.0 °C were selected as they are common storage temperatures forfood and related products. Samples were stored for six months and analyzed by HPAE-PAD for changes in the oligosaccharide content. It was found that at both room temperature and at 4.0 °C that there was no significant change in concentration as determined by peak area after six months of storage for both the single strength and concentrated apple juice samples (Fig. 4).
  • inulin oligosaccharides were produced, purified and analyzed for use as oligomolecular tags (OMTs) for traceability and authenticity.
  • OMTs oligomolecular tags
  • Inulin based OMTs were readily produced from commercially available chicory inulin using an endo-inulinase. Through this process a variety of unique OMTs were produced which could be mixed in various combinations/ratios to produce an even greater assortment of readily distinguishable OMTs for use in traceability and authenticity. Due to their GRAS status these OMTs are useful in both whole foods (e.g ., apple juice) and food ingredients (i.e., flour, food additives) allowing these products to be traced throughout the production chain.
  • whole foods e.g ., apple juice
  • food ingredients i.e., flour, food additives
  • OMTs are useful in high value products such as pharmaceuticals, nutraceuticals, personal care products including personal hygiene and cosmetics, and industrial products (e.g. paints).

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Wood Science & Technology (AREA)
  • Animal Behavior & Ethology (AREA)
  • Genetics & Genomics (AREA)
  • Veterinary Medicine (AREA)
  • Biotechnology (AREA)
  • Public Health (AREA)
  • Dermatology (AREA)
  • Materials Engineering (AREA)
  • Molecular Biology (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Birds (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Polysaccharides And Polysaccharide Derivatives (AREA)
  • Coloring Foods And Improving Nutritive Qualities (AREA)

Abstract

La présente invention concerne des marqueurs oligomoléculaires (OMT) comprenant des oligosaccharides d'inuline hydrolysés utiles pour tracer un produit, ainsi que des procédés de production d'OMT. L'invention concerne également des procédés de traçage d'un produit comprenant un OMT, et des systèmes de traçage d'un produit, les systèmes comprenant un OMT et un détecteur.
PCT/CA2019/051058 2018-08-03 2019-08-01 Oligosaccharides utilisés comme marqueurs oligomoléculaires WO2020024059A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US201862714301P 2018-08-03 2018-08-03
US62/714,301 2018-08-03

Publications (1)

Publication Number Publication Date
WO2020024059A1 true WO2020024059A1 (fr) 2020-02-06

Family

ID=69230445

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CA2019/051058 WO2020024059A1 (fr) 2018-08-03 2019-08-01 Oligosaccharides utilisés comme marqueurs oligomoléculaires

Country Status (1)

Country Link
WO (1) WO2020024059A1 (fr)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20100272652A1 (en) * 2009-03-26 2010-10-28 Seventh Sense Biosystems, Inc. Determination of tracers within subjects
US20110020510A1 (en) * 2008-04-23 2011-01-27 Kim Martha Jozefa Frooninckx Aqueous dispersion of fructan-containing particles, method of preparation and use

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110020510A1 (en) * 2008-04-23 2011-01-27 Kim Martha Jozefa Frooninckx Aqueous dispersion of fructan-containing particles, method of preparation and use
US20100272652A1 (en) * 2009-03-26 2010-10-28 Seventh Sense Biosystems, Inc. Determination of tracers within subjects

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
"Internal Tagging In-depth Research", INFORMA MARKET RESEARCH CO.LTD, June 2010 (2010-06-01), University of Guelph, XP055684747, Retrieved from the Internet <URL:https://www.informaresearch.com/sites/default/files/reports_pdf/U_of_G_Report.pdft.pdf.> *
BORROMEI ET AL.: "Characterization and quantitation of short-chain frutooligosaccharides and inulooligosaccharides in fermented milks by high-performance anion-exchange chromatography with pulsed amperometric detection", JOURNAL OF SEPARATION SCIENCE, vol. 32, no. 21, 2009, pages 3635 - 3642, XP055684750 *
BORRORNEI ET AL.: "Evaluation of Fructooligosaccharides and Inulins as Potentially Health Benefiting Food Ingredients by HPAEC-PED and MALDI-TOF MS", INTERNATIONAL JOURNAL OF ANALYTICAL CHEMISTRY, vol. 2009, 2009, pages 1 - 9, XP055684737 *
RUIZ-MATUTE ET AL.: "Detection of adulterations of honey with high fructose syrups from inulin by GC analysis", J. OF FOOD COMPOSITION & ANALYSIS, vol. 23, no. 3, 2010, pages 273 - 276, XP026990904 *
SINGH ET AL.: "Production of Frutooligosaccharides from Inulin by Endoinulinases and Their Prebiotic Potential", FOOD TECHNOLOGY AND BIOTECHNOLOGY, vol. 48, no. 4, 2010, pages 435 - 450, XP055305545 *
WILLEMS ET AL.: "Authenticity Analysis of Pear Juice Employing Chromatographic Fingerprinting", JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, vol. 62, no. 48, 2014, pages 11737 - 11747, XP055684753 *

Similar Documents

Publication Publication Date Title
BeMiller Carbohydrate analysis
Kapasakalidis et al. Effect of a cellulase treatment on extraction of antioxidant phenols from black currant (Ribes nigrum L.) pomace
Gawkowska et al. Structure-related gelling of pectins and linking with other natural compounds: A review
Harding et al. An introduction to polysaccharide biotechnology
Zykwinska et al. Extraction of green labeled pectins and pectic oligosaccharides from plant byproducts
Larsen et al. Interactions of anthocyanins with pectin and pectin fragments in model solutions
Colodel et al. Optimization of acid-extraction of pectic fraction from grape (Vitis vinifera cv. Chardonnay) pomace, a Winery Waste
Ele-Ekouna et al. Chemical characterization of pectin from green tea (Camellia sinensis)
Diaz et al. Nonenzymatic degradation of citrus pectin and pectate during prolonged heating: effects of pH, temperature, and degree of methyl esterification
Guadalupe et al. Polysaccharide profile and content during the vinification and aging of Tempranillo red wines
Sila et al. Pectins in processed fruits and vegetables: Part II—Structure–function relationships
Hao et al. Advances in preparation of chitooligosaccharides with heterogeneous sequences and their bioactivity
Mrabet et al. Dietary fiber from Tunisian common date cultivars (Phoenix dactylifera L.): Chemical composition, functional properties, and antioxidant capacity
Lenucci et al. Possible use of the carbohydrates present in tomato pomace and in byproducts of the supercritical carbon dioxide lycopene extraction process as biomass for bioethanol production
Orfila et al. Expression of mung bean pectin acetyl esterase in potato tubers: effect on acetylation of cell wall polymers and tuber mechanical properties
Willems et al. Authenticity analysis of pear juice employing chromatographic fingerprinting
Cardelle-Cobas et al. Synthesis of oligosaccharides derived from lactulose and Pectinex Ultra SP-L
Redgwell et al. Cell wall polysaccharides of Chinese Wolfberry (Lycium barbarum): Part 1. Characterisation of soluble and insoluble polymer fractions
Allison et al. Obtaining multiple coproducts from red grape pomace via anthocyanin extraction and biogas production
Mehrländer et al. Structural characterization of oligosaccharides and polysaccharides from apple juices produced by enzymatic pomace liquefaction
Zhang et al. Profiling the water soluble pectin in clear red raspberry (Rubus idaeus L. cv. Heritage) juice: impact of high hydrostatic pressure and high-temperature short-time processing on the pectin properties
Tan et al. Most of the rhamnogalacturonan-I from cultured Arabidopsis cell walls is covalently linked to arabinogalactan-protein
Trindade et al. Biochemical characterization, thermal stability, and partial sequence of a novel exo‐polygalacturonase from the thermophilic fungus Rhizomucor pusillus A13. 36 obtained by submerged cultivation
Santos-Moriano et al. Efficient conversion of chitosan into chitooligosaccharides by a chitosanolytic activity from Bacillus thuringiensis
Mutanda et al. Response surface methodology: synthesis of inulooligosaccharides with an endoinulinase from Aspergillus niger

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 19844204

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 19844204

Country of ref document: EP

Kind code of ref document: A1