EP4688876A1

EP4688876A1 - Formation of amidated pectin from high methoxyl pectin

Info

Publication number: EP4688876A1
Application number: EP24720400.1A
Authority: EP
Inventors: Mario Alberto DELGADO HERNANDEZ; Hannes LUDEMANN
Original assignee: Cargill Inc
Current assignee: Cargill Inc
Priority date: 2023-04-06
Filing date: 2024-04-01
Publication date: 2026-02-11
Also published as: CN121013869A; WO2024211217A1

Abstract

The present invention relates to an amidated pectin, and a method of making the amidated pectin. In particular, the present invention relates to an amidated pectin formed from a wet precipitated high methoxyl pectin. The present invention further relates to a method for producing a low methoxyl amidated pectin, the method includes the following step: adding a wet precipitated high methoxyl pectin to a suspension that includes one or more alcohols and ammonia to form a low methoxyl amidated pectin.

Description

FORMATION OF AMIDATED PECTIN FROM HIGH METHOXYL PECTIN

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of European Patent Application No. 23167014.2, filed April 6, 2023, which is incorporated by reference herein in its entirely.

FIELD OF THE INVENTION

[0002] The present invention relates to an amidated pectin, and a method of making the amidated pectin. In particular, the present invention relates to an amidated pectin formed from a wet precipitated high methoxyl pectin.

BACKGROUND OF THE INVENTION

[0003] Pectin is found naturally in plant cell walls. Although pectin can be extracted from a large number of plants or fruits, the main industrial or conventional sources include citrus peel and apple pomace. The main use of pectin is as a gelling agent, thickening agent, viscosifying agent and/or as a stabiliser in food. As a result, pectin is commonly used in jams, jellies, acid milk beverages and dessert bases as a food additive. Pectin can also be used as dietary' fibers and as an effective food emulsifier. Pectin also has potential uses in other fields such as medicine as a carrier for controlled drugs or bioactive release and in many other nonfood applications.

[0004] Pectin is a polysaccharide comprising a chain of galacturonic acid units which are linked by a-1,4 glycosidic bonds. The galacturonic acid chain is partly esterified as methyl esters. Though the galacturonic acid chain is the backbone of pectin, other chemicals such as acety l and methyl ester groups can be incorporated into the chain. These groups can be important for distinguishing between different pectin types. A review of pectin properties and characteristics can be found in D. Gawkowska et al. “Structure-Related Gelling of Pectins and Linking with Other Natural Compounds: A Review” Polymers (Basel), 2018 Jul; 10(7): 762, the disclosure of which is hereby incorporated by reference. [0005] Pectin can be split into two categories by its degree of methylation, which is the percentage of carboxyl groups which have undergone esterification with methanol. The degree of methylation is of great importance for the use of pectins as gelling agents and depending on the degree of methylation the pectin is classified as either a high methoxyl pectin (HM pectin) or a low methoxyl pectin (LM pectin). If more than 50 percent of the carboxyl groups are methylated then the pectin is a HM pectin, and if less than that degree of methylation has occurred then the pectin is a LM pectin.

[0006] HM pectin is the most commonly used pectin and can be split into two categories: rapid set HM pectin and slow set HM pectin. Rapid set pectin will set faster and at a higher temperature than slow set pectin, and thus is preferable in the production of food and/or beverage products which typically require a certain mouthfeel or the suspension of various particulate matter such as whole or portioned fruits, nuts and the like. Slow set pectin is preferable for food and/or beverage products which do not require the presence of suspended particulate matter. HM pectins can form gels with sugar and acid, forming what is known in the art as low water activity gels or sugar acid pectin gels. Slow set pectin can take from 1 to 2 days to set into a gel. These gels have a 3-dimensional network of pectin molecules in which the solvent (usually water), optionally with the co-solutes of sugar and acid are immobilised resulting in a system that can resist deformation.

[0007] LM pectins differ from HM pectins in that the LM pectins only require the presence of a divalent metal ion to initiate the reaction that causes the pectin to set into a gel. The most commonly used divalent metal ion is calcium. In general, the gel forming abi li ty of LM pectins increases with a decreasing degree of methylation.

[0008] Recently, LM pectins have been amidated to form LM amidated pectin. LM ami dated pectin is a modified form of LM pectin in which some of the galacturonic acid units are converted by ammonia into carboxylic amides. In traditional methods of producing LM amidated pectin, a HM pectin needs to be first extracted from its natural source. The HM pectin is converted into LM pectin, which undergoes amidation to form the LM amidated pectin. The conversion of the HM pectin into LM amidated pectin follows the reaction scheme:

RCOOCH3 + NH4OH => RCONH2 + CH3OH +H2O [0009] Traditional methods of producing LM amidated pectin from a natural pectin in citrus fruits are set out below.

Traditional method 1: A traditional method of producing a LM amidated pectin from a natural pectin sourced from citrus fruits, wherein the traditional method includes a dry amidation step at a temperature of from 10 to 14 °C.

[00010] Traditional method 1 relates to an example of producing LM amidated pectin using traditional methods. In traditional method 1, HM pectin is first extracted from citrus peel and then dried. The extracted and dried HM pectin then undergoes amidation at a temperature of from 10 to 14 °C to form the LM amidated pectin.

[00011] FIG. 1 depicts a flow diagram for traditional method 1.

[00012] Step 1: Extraction of HM pectin from citrus peel

[00013] First, citrus peel is mixed in acidic, hot water to extract the HM pectin in aqueous form from the citrus peel. The temperature and pH of the acidic, hot water is from 68 to 72°C, the pH is from pH 1.50 to 1.70 and the residence time is from 8 to 12 hours.

[00014] The HM pectin in aqueous form is clarified by using centrifugation, filtration or other conventional separation technologies known in the art to obtain a clarified HM pectin in aqueous form.

[00015] The clarified HM pectin in aqueous form is then concentrated according to any technique known in the art and an appropriate amount of an alcohol is added to form an alcohol/water mix. The amount of alcohol added depends on the amount of alcohol required to cause the clarified HM extract to precipitate out from its aqueous form as a precipitate containing the HM pectin.

[00016] The precipitate containing the HM pectin is separated from the alcohol/w ater mix by mechanical separation, wherein any mechanical separation known in the art can be used. The HM pectin obtained from the separation is then washed three times with alcohol at 60 weight % of the total weight of the alcohol to form a washed HM pectin. [00017] The washed HM pectin is dried and ground until a homogeneous particle size distribution is achieved to form a dry HM pectin. Generally, the dry HM pectin has a moisture content of 10 weight % of the total weight of the dry HM pectin. The dry HM pectin can be blended with sugar to form a standardised HM pectin.

[00018] Step 2: Amidation of dry HM pectin

[00019] The dry HM pectin is placed in a suspension which comprises alcohol and ammonia. The alcohol is present at from 60 to 70 weight % of the total weight of the suspension. The ammonia is present at 2.8 to 3.5 weight % of the total weight of the suspension. The suspension is maintained at a temperature of from 10 to 14 °C. The dry HM pectin is in the suspension for from 5 to 8 hours. After from 5 to 8 hours have passed, the dry HM pectin undergoes amidation to form LM amidated pectin.

[00020] The LM amidated pectin is filtered out of the suspension by any filtration technique known in the art. The alcohol and ammonia pass to a recovery step.

[00021] The LM amidated pectin is washed with alcohol at 60 weight % of the total weight of the alcohol to form a washed LM amidated pectin.

[00022] The pH of the washed LM amidated pectin is adjusted until the pH lies from 4.0 to 4.5. The pH is adjusted with acid, such as nitric acid. The result is a pH adjusted LM amidated pectin.

[00023] The pH adjusted LM amidated pectin is washed with alcohol at 60 weight % of the total weight of the alcohol three times to form a second-w ashed LM amidated pectin.

[00024] The second-washed LM amidated pectin undergoes a drying process to form a dried LM amidated pectin with a moisture content of 10 weight % of the total w eight of the dried LM amidated pectin. Any dry ing processes known in the art can be used. [00025] The dried LM amidated pectin is then ready for quality analysis and further modification. Further modification can include standardisation with sugar to make a commercial product.

[00026] Typically, the alcohol used in traditional method 1 is isopropyl alcohol.

[00027] Traditional method 2: A traditional method of producing, a LM amidated pectin from a natural pectin sourced from citrus fruits, wherein the traditional method includes a calcium-removal step and a dry amidation step at a temperature of from 12 to 14 °C.

[00028] Traditional method 2 relates to an example of producing LM amidated pectin using traditional methods. In traditional method 2, HM pectin is firstly extracted from citrus peel and then dried. The extracted and dried HM pectin then undergoes amidation at a temperature of from 12 to 14 °C to form the LM amidated pectin.

[00029] FIG. 2 depicts a flow diagram for traditional method 2.

[00030] Step 1: Extraction of HM pectin from a citrus peel

[00031] Firstly, citrus peel is mixed in acidic, hot water to extract the HM pectin in aqueous form from the citrus peel. The temperature of the acidic, hot water is from 70 to 80°C, the Ph is from Ph 1.90 to 2. 10 and the residence time is from 3 to 3.5 hours.

[00032] The HM pectin in aqueous form is clarified by using centrifugation, filtration or other conventional separation technologies known in the art to obtain a clarified HM pectin in aqueous form.

[00033] Calcium present in the clarified HM pectin in aqueous form is removed using ion exchange with a cationic resin to form a de-calcified HM pectin in aqueous form.

[00034] The de-calcified HM pectin in aqueous form is concentrated according to any technique known in the art to form a concentrated HM pectin in aqueous form. [00035] The degree of esterification (DE) of the concentrated HM pectin in aqueous form is modified by adding an acidic solution to the concentrated HM pectin in aqueous form.

Typically, the acid used is nitric acid, sulfuric acid and/or hydrochloric acid. The degree of esterification (DE) is modified at a temperature of from 30 to 35°C, an acidity of from 0.80 to 1.0 % by mass and a residence time of from 72 to 96 hours. The resultant product is a DE modified HM pectin in aqueous form and the DE modified HM pectin in aqueous form has a degree of esterification (DE) of from 54 to 58 %.

[00036] An appropriate amount of alcohol is added to the DE modified HM pectin in aqueous form to form an alcohol/water mix. The amount of alcohol added depends on the amount of alcohol required to cause the DE modified HM pectin to precipitate out from its aqueous form as a precipitate containing the HM pectin.

[00037] The precipitate containing the HM pectin is separated from the alcohol/water mix by mechanical separation, wherein any mechanical separation technique known in the art can be used.

[00038] The precipitate containing the HM pectin is then w ashed three times with alcohol at 60 weight % of the total weight of the alcohol to form a washed HM pectin.

[00039] The washed HM pectin is dried and ground until a homogeneous particle size distribution is achieved to form a dry HM pectin. Generally, the dry HM pectin has a moisture content of 10 weight % of the total weight of the dry HM pectin. The dry' HM pectin can be blended with sugar to form a standardised HM pectin.

[00040] Step 2: Amidation of dry HM pectin

[00041] The dry HM pectin is placed in a suspension which comprises alcohol and ammonia. The alcohol is present at from 60 to 70 weight % of the total weight of the suspension. The ammonia is present at 2.8 to 3.5 weight % of the total weight of the suspension. The suspension is maintained at a temperature of from 12 to 14 °C. The dry HM pectin is in the suspension for from 5 to 8 hours. After 5 to 8 hours have passed, the dry HM pectin undergoes amidation to form LM amidated pectin. [00042] The LM amidated pectin is filtered out of the suspension. The alcohol and ammonia pass to a recovery step.

[00043] The LM amidated pectin is washed with alcohol at 60 weight % of the total weight of the alcohol to form a washed LM amidated pectin.

[00044] The pH of the washed LM amidated pectin is adjusted until the pH is from 4.0 to 4.5. The pH is adjusted with acid, such as nitric acid. The result is a pH adjusted LM amidated pectin.

[00045] The pH adjusted LM amidated pectin is washed with alcohol at 60 weight % of the total weight of the alcohol three times to form a second-washed LM amidated pectin.

[00046] The second-washed LM amidated pectin undergoes a drying process to form a dried LM amidated pectin with a moisture content of 10 weight % of the total weight of the dried LM amidated pectin. Any drying processes known in the art can be used.

[00047] The LM amidated pectin is then ready for quality⁷ analysis and further modification. Further modification can include standardisation with sugar to make a commercial product.

[00048] Typically, the alcohol used in traditional method 2 is isopropyl alcohol.

[00049] Traditional method 3: A traditional method of producing, a LM amidated pectin from a natural pectin sourced from citrus fruits, wherein the traditional method includes a calcium-removal step and a dry amidation step at a temperature of from 7 to 8 °C.

[00050] Traditional method 3 relates to an example of producing LM amidated pectin using traditional methods. In traditional method 3, HM pectin is firstly extracted from citrus peel and dried. The extracted and dried HM pectin then undergoes amidation at a temperature of from 7 to 8 °C. to form the LM amidated pectin.

[00051] FIG. 2 depicts a flow diagram for traditional method 3. [00052] Step 1: Extraction of IM pectin from a citrus peel

[00053] Firstly, citrus peel is mixed in acidic, hot water to extract the HM pectin in aqueous form from the citrus peel. The temperature of the acidic, hot water is from 70 to 80 °C, the pH is from pH 1.90 to 2. 10 and the residence time is from 3 to 3.5 hours.

[00054] The HM pectin in aqueous form is clarified by using centrifugation, filtration or other conventional separation technologies known in the art to obtain a clarified HM pectin in aqueous form.

[00055] Calcium present in the clarified HM pectin in aqueous form is removed using ion exchange with a cationic resin to form a de-calcified HM pectin in aqueous form.

[00056] The de-calcified HM pectin in aqueous form is concentrated according to any technique known in the art to form a concentrated HM pectin in aqueous form.

[00057] The degree of esterification (DE) of the concentrated HM pectin in aqueous form is modified by adding an acidic solution to the concentrated HM pectin in aqueous form.

Typically, the acid used is nitric acid, sulfuric acid and/or hydrochloric acid. The degree of esterification (DE) is modified at a temperature of from 30 to 35°C, an acidity of from 0.80 to 1 .0 % by mass and a residence time of from 72 to 96 hours. The resultant product is DE modified HM pectin in aqueous form and has a degree of esterification (DE) of from 54 to 58 %.

[00058] An appropriate amount of alcohol is added to the DE modified HM pectin in aqueous form to form an alcohol/water mix. The amount of alcohol added depends on the amount of alcohol required to cause the HM pectin to precipitate out from its aqueous form as a precipitate containing the HM pectin.

[00059] The precipitate containing the HM pectin is separated from the alcohol/water mix be mechanical separation, wherein any mechanical separation technique known in the art can be used.

[00060] The precipitate containing the HM pectin is then washed three times with alcohol at 60 weight % of the total weight of the alcohol to form a washed HM pectin. [00061] The washed HM pectin is dried and ground until a homogeneous particle size distribution is achieved to form a dry HM pectin. Generally, the dry HM pectin has a moisture content of 10 weight % of the total weight of the dry HM pectin. The dry HM pectin can be blended with sugar to form a standardised HM pectin.

[00062] Step 2: Amidation of dry HM pectin

[00063] The dry HM pectin is placed in a suspension which comprises alcohol and ammonia. The alcohol is present at from 60 to 70 weight % of the total weight of the suspension. The ammonia is present at 4.0 to 5.0 weight % of the total weight of the suspension. The suspension is maintained at a temperature of from 7 to 8 °C. The dry HM pectin is in the suspension for from 3 to 5 hours. After 3 to 5 hours have passed, the dry HM pectin undergoes amidation to form LM amidated pectin.

[00064] The LM amidated pectin is then filtered out of the suspension. The alcohol and ammonia pass to a recover}⁷ step.

[00065] The LM amidated pectin is washed with alcohol at 60 weight % of the total weight of the alcohol to form a washed LM amidated pectin.

[00066] The pH of the washed LM amidated pectin is adjusted until the pH lies from 4.0 to 4.5. The pH is adjusted with acid, such as nitric acid. The result is a pH adjusted LM amidated pectin.

[00067] The pH adjusted LM amidated pectin is washed with alcohol at 60 weight % of the total weight of the alcohol three times to form a second washed LM amidated pectin.

[00068] The second washed LM amidated pectin undergoes a drying process to form a dried LM amidated pectin with a moisture content of 10 weight % of the total weight of the dried LM amidated pectin. Any dry ing processes known in the art can be used. [00069] The dried LM amidated pectin is then ready for quality analysis and further modification. Further modification can include standardisation with sugar to make a commercial product.

[00070] Typically, the alcohol used in traditional method 3 is isopropyl alcohol.

[00071] Other methods of forming LM amidated pectin are set out in US2480710A and

WO98/58968A1.

[00072] The product of the well-known methods may suffer from glycosidic breakdown and aggregation during amidation, which leads to the amidated pectin having a changed conformation due to degradation caused during production.

[00073] There is therefore a need for an improved LM amidated pectin wherein the improved LM amidated pectin has the same or similar properties to LM pectin.

[00074] There is therefore a need for an improved method of making a LM amidated pectin, wherein the method does not cause the HM pectin to suffer from glycosidic breakdown and aggregation during the production of the LM amidated pectin.

SUMMARY OF THE INVENTION

[00075] The present invention relates to an amidated pectin, and a method of making the amidated pectin. In particular, the present invention relates to an amidated pectin formed from a wet precipitated high methoxyl pectin.

[00076] Representative features of the present invention are set out in the following clauses, which stand alone or may be combined, in any combination, with one or more features disclosed in the text and/or figures of the specification.

[00077] The present invention is as set out in the following clauses:

1. A method for producing a low methoxyl amidated pectin, the method comprising the following step: adding a wet precipitated high methoxyl pectin to a suspension comprising one or more alcohols and ammonia to form a low methoxyl amidated pectin.

2. The method of clause 1, wherein the low methoxyl amidated pectin has a degree of methylation of below 50 %.

3. The method of clause 1 or clause 2. wherein the wet precipitated high methoxyl pectin has a degree of methylation of above 50 %.

4. The method of any one of clauses 1 to 3, wherein the wet precipitated high methoxyl pectin has an intrinsic viscosity of from 500 to 650 ml/g. or, from 510 to 640 ml/g, or. from 525 to 615 ml/g.

5. The method of any one of clauses 1 to 4, wherein the low methoxyl amidated pectin has an intrinsic viscosity’ of from 450 to 650 ml/g, or, from 475 to 625 ml/g, or, from 488 to 600 ml/g.

6. The method of any one of clauses 1 to 5, wherein the wet precipitated high methoxyl pectin is in the suspension with the one or more alcohols and ammonia at a temperature of from 1 to 12 °C, or. from 2 to 10 °C, or, from 3 to 8 °C, or, from 4 to 6 °C.

7. The method of any one of clauses 1 to 6, wherein the wet precipitated high methoxyl pectin is in the suspension with the one or more alcohols and ammonia for a reaction time of from 1 to 8 hours, or, from 2 to 7 hours, or from 2.5 to 6.5 hours, or. from 3 to 6 hours.

8. The method of any one of clauses 1 to 7, wherein the w et precipitated high methoxyl pectin has a dry substance content of from 20 to 80 w eight %, or, from 30 to 70 weight %, or, from 40 to 60 weight %, or. from 45 to 55 weight %, or, 50 weight % of the total weight of the wet precipitated high methoxyl pectin.

9. The method of any one of clauses 1 to 8, wherein the ammonia is present in the suspension at from 1 to 10 weight %, or, from 2 to 8 weight %, or, from 3 to 7 w eight %, or, from 4 to 6 weight % of the total weight of the suspension. 10. The method of any one of clauses 1 to 9, wherein the one or more alcohols is present in the suspension at from 50 to 80 weight %, or, from 55 to 75 weight %. or, at 60 to 70 weight % of the total weight of the suspension.

11. The method of any one of clauses 1 to 10, wherein the one or more alcohols are any one of tert-amyl alcohol, benzyl alcohol, 1 ,4-butanediol, 1, 2, 4-butanetriol, butanol. 1-butanol, 2- butanol. tert-butyl alcohol, diethylene glycol, ethanol, ethylene glycol. 2-ethylhexanol, furfuryl alcohol, glycerol, iso-butanol, isopropyl alcohol, methanol, 2-(2-methoxyethoxyl)ethanol, 2- methy 1-1 -butanol, 2-methyl-l -pentanol, 3-methyl-2-butanol, neopentyl alcohol, 2-pentanol, 1,3- propanediol, propan-l-ol, propylene glycol, propylene glycol methyl ether, isopropanol, or, any combination thereof.

12. The method of any one of clauses 1 to 11, further comprising the step of: fdtering the low methoxyl amidated pectin from the suspension to form a fdtrate comprising the one or more alcohols and ammonia and a residue comprising the low methoxyl amidated pectin, wherein the step of filtering the low methoxyl amidated pectin from the suspension stops the reaction between the high methoxyl pectin, one or more alcohols and ammonia.

13. The method of clause 12, further comprising the step of: neutralising the residue comprising the low methoxyl amidated pectin with acidulate alcohol until the residue comprising the low methoxyl amidated pectin has a pH of from 3.0 to 5.0, or, 4.0 to 5.0, or, 4.0 to 4.5.

14. The method of clause 13, wherein the acidulate alcohol comprises a mineral acid; optionally, wherein the mineral acid is nitric acid; optionally, wherein the acidulate alcohol further comprises any one of tert-amyl alcohol, benzyl alcohol , 1 ,4-butanediol, 1, 2. 4-butanetriol, butanol, 1-butanol, 2-butanoL tert-butyl alcohol, diethylene glycol, ethanol, ethylene glycol, 2-ethylhexanol, furfuryl alcohol, glycerol, isobutanol, isopropyl alcohol, methanol, 2-(2-methoxyethoxyl)ethanol, 2-methyl-l -butanol, 2- methyl-1 -pentanol, 3-methyl-2-butanol, neopentyl alcohol. 2-pentanol, 1,3-propanediol. propan- l-ol. propylene glycol, propylene glycol methyl ether, isopropanol, or. any combination thereof. 15. The method of any one of clauses 12 to 14, further comprising removing or extracting water and/or alcohol from the residue comprising the low methoxyl amidated pectin.

16. The method of clause 15, wherein removal or extraction of water and/or alcohol from the residue comprising the low methoxyl amidated pectin is carried out by pressing.

17. The method of clause 16, wherein the pressing step is performed until the low methoxyl amidated pectin has a dry substance content of from 30 to 70 weight %, or, from 40 to 60 weight %, or, from 45 to 55 weight %, or, 50 weight % of the total weight of the low methoxyl amidated pectin.

18. The method of any one of clauses 1 to 17, further comprising a drying step wherein water and/or alcohol is removed from the low methoxyl amidated pectin.

19. The method of clause 18, wherein the drying step is performed until the low methoxyl amidated pectin has a moisture content of from 1 to 30 weight %, or from 2 to 20 weight &. or from 7.5 to 15.0 weight 5, or from 9 to 11 weight %, or, 10 weight % of the total weight of the low methoxyl amidated pectin.

20. The method of any one of clauses 1 to 19, further comprising the step of: grinding the low methoxyl amidated pectin; and/or, blending the low methoxyl amidated pectin with sugar.

21. The method of any one of clauses 1 to 20, wherein the wet precipitated high methoxyl pectin is produced via the following method: mixing an aqueous citrus peel in hot, acidic water until a high methoxyl pectin is extracted from the citrus peel; centrifuging, filtering and/or concentrating the high methoxyl pectin; adding an acidic solution to the high methoxyl pectin to modify the degree of esterification (DE) of the high methoxyl pectin; adding alcohol to the high methoxyl pectin until a precipitated high methoxyl pectin is formed; and separating the precipitated high methoxyl pectin from the alcohol to form a wet precipitated high methoxyl pectin. 22. The method of clause 21, wherein the citrus peel is sourced from lemons, mandarin oranges, grapefruit, sweet orange, pomelo, clementines, limes, kumquat, bitter oranges, blood oranges, satsumas, yuzus, tangelos, or, a combination thereof.

23. The method of clause 21 or clause 22, wherein the step of mixing the aqueous citrus peel in hot acidic water until high methoxyl pectin is extracted from the citrus is conducted at a temperature of from 55 to 100 °C, or, 60 to 90 °C, or, from 65 to 85 °C, or, 70 to 80 °C.

24. The method of any one of clauses 21 to 23, wherein the step of mixing the aqueous citrus peel in hot acidic water until high methoxyl pectin is extracted from the citrus is conducted at a pH of from 1.5 to 2.5, or, from 1.75 to 2.25, or, from 1.9 to 2.10.

25. The method of any one of clauses 21 to 24, wherein the step of mixing the aqueous citrus peel in hot acidic water until high methoxyl pectin is extracted from the citrus is conducted for from 2 to 4.5 hours, or, from 2.5 to 4 hours, or. from 3 to 3.5 hours.

26. The method of any one of clauses 21 to 25, wherein the hot, acidic w ater comprises any one of nitric acid, phosphoric acid, sulfuric acid, citric acid, malic acid, succinic acid, adipic acid, gluconic acid, tartaric acid, fumaric acid, or, a combination thereof

27. The method of any one of clauses 21 to 26, further comprising the step of removing calcium present in the high methoxyl pectin by ion exchange.

28. The method of any one of clauses 21 to 27, wherein the high methoxyl pectin is concentrated, until the HM pectin is present in w ater at a concentration of from 15 to 40 g/1, preferably from 20 to 37 g/1, more preferably from 25 to 35 g/1, most preferably from 27 to 30 g/1-

29. The method any one of clauses 21 to 28, wherein the step of adding an acidic solution to the high methoxyl pectin to modify the degree of esterification (DE) of the high methoxyl pectin is conducted until a degree of esterification of from 40 to 75 %, or, from 45 to 70 %, or, from 50 to 65 %. or, from 54 to 58 %. 30. The method of any one of clauses 21 to 29, wherein the step of adding an acidic solution to the high methoxyl pectin to modify the degree of esterification (DE) content of the high methoxyl pectin is conducted at a temperature of from 20 to 45 °C, or, from 25 to 40 °C, or, from 30 to 35 °C.

31. The method of any one of clauses 21 to 30, wherein the step of adding an acidic solution to the high methoxyl pectin to modify the degree of esterification (DE) of the high methoxyl pectin is conducted until the acid content is at from 0.5 to 1.5 weight %, or, from 0.6 to 1.25 weight %, or, from 0.7 to 1.1 weight % of the total weight of the acid and high methoxyl pectin together.

32. The method of any one of clauses 21 to 31, wherein the step of adding an acidic solution to the high methoxyl pectin to modify the degree of esterification (DE) of the high methoxyl pectin is conducted for 50 to 110 hours, or from 60 to 105 hours, or, from 70 to 100 hours, or, from 72 to 96 hours.

33. The method of any one of clauses 21 to 32, wherein the step of adding an acidic solution to the high methoxyl pectin to modify the degree of esterification (DE) of the high methoxyl pectin is conducted with any one of the following acids: nitric acid, sulfuric acid, hydrochloric acid, phosphoric acid, citric acid, malic acid, succinic acid, adipic acid, gluconic acid, tartaric acid, fumaric acid, or, a combination thereof; optionally, wherein the acid is present in water.

34. The method of any one of clauses 21 to 33, further comprising removing or extracting water from the wet precipitated high methoxyl pectin.

35. The method of clause 34, wherein removal or extraction of water from the wet precipitated high methoxyl pectin is carried out by pressing, optionally, wherein the wet precipitated high methoxyl pectin is pressed until the wet precipitated high methoxyl pectin has a dry substance of from 30 to 70 weight %, or, from 40 to 60 weight %, or, 50 weight % of the total weight of the wet precipitated high methoxyl pectin.

36. A method of forming a dairy product, optionally for use in milk, condensed milk, dried milk, milk-drinks, cream, butter, yoghurt, cheese, custard, cottage cheese, cream cheese, curd, frozen dairy products such as frozen custard, frozen yogurt and ice-cream, gelato, powdered milk, evaporated milk, sour cream, soured milk, whey, whey protein or whipped cream, wherein the method comprises the steps of including the product of any one of clauses 1 to 35 in a dairy product.

37. A low methoxyl amidated pectin obtained, or obtainable by, the method of any one of clauses 1 to 35.

38. The amidated low methoxyl pectin of clause 37 for use in daily' products, optionally, for use in milk, condensed milk, dried milk, milk-drinks, cream, butter, yoghurt, cheese, custard, cottage cheese, cream cheese, curd, frozen dairy products such as frozen custard, frozen yogurt and ice-cream, gelato, powdered milk, evaporated milk, sour cream, soured milk, whey, wheyprotein or whipped cream.

BRIEF DESCRIPTION OF THE DRAWINGS

[00078] FIG. 1 is a flow diagram of a traditional method of producing LM amidated pectin from dry HM pectin.

[00079] FIG. 2 is a flow diagram of a traditional method of producing LM amidated pectin from dry HM pectin.

[00080] FIG. 3 is a flow diagram of the method according to the present invention for producing a LM amidated pectin from a wet precipitated HM pectin.

[00081] FIG. 4 is a graph showing the intrinsic viscosity differences between the HM pectin and the same pectin after amidation for LM amidated pectins formed via the traditional dry amidation process and LM amidated pectin formed via the method of the present invention.

DETAILED DESCRIPTION

[00082] Embodiments of the invention are described below with reference to the accompanying drawings. The accompanying drawings illustrate various embodiments of systems, methods, and embodiments of various other aspects of the disclosure. Any person with ordinary skills in the art will appreciate that the illustrated element boundaries (e.g.. boxes, groups of boxes, or other shapes) in the figures represent one example of the boundaries. It may be that in some examples one element may be designed as multiple elements or that multiple elements may be designed as one element. In some examples, an element shown as an internal component of one element may be implemented as an external component in another and vice versa. Furthermore, elements may not be drawn to scale. Non-limiting and non-exhaustive descriptions are described with reference to the following drawings. The components in the figures are not necessarily to scale, emphasis instead being placed upon illustrating principles.

[00083] Embodiments of the present disclosure will be described more fully hereinafter with reference to the accompanying drawings in which like numerals represent like elements throughout the several figures, and in which example embodiments are shown. Embodiments of the claims may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

[00084] The words "comprising,” “having,” “containing,” and “including,” and other forms thereof, are intended to be equivalent in meaning and be open ended in that an item or items following any one of these words is not meant to be an exhaustive listing of such item or items, or meant to be limited to only the listed item or items. It must also be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural references unless the context clearly dictates otherwise. Although any sy stems and methods similar or equivalent to those described herein can be used in the practice or testing of embodiments of the present disclosure, the preferred systems and methods are now described.

[00085] Some of the terms used to describe the present invention are set out below:

[00086] “Amidation” refers to the modification of a pectin to include at least one amide group (-NH2).

[00087] “Degree of methylation” refers to the percentage of carboxyl groups esterified with methanol. If more than fifty percent of the carboxyl groups are methylated the pectin is called high methoxyl pectins (HM), and if less than that degree of methylation has occurred then the pectin is called low methoxyl pectins (LM).

[00088] ‘"High methoxyl pectin” or “HM pectin” refers to a pectin extracted from the peels of citrus fruit, wherein a degree of the original carboxyl groups present in the pectin have undergone esterification with methanol. High methoxyl pectins have a degree of methylation of above 50 %.

[00089] “Low methoxyl pectin” or “LM pectin” refers to a pectin extracted from the peels of citrus fruit, wherein a degree of the original carboxyl groups present in the pectin have undergone esterification with methanol. Low methoxyl pectins have a degree of methylation of below 50 %.

[00090] “Low methoxyl amidated pectin” or “LM amidated pectin” or “LMA pectin” refers to a modified form of LM pectin in which some of the galacturonic acid residues are converted by ammonia into amide groups.

[00091] “Mix” or “mixture” refers to a substance containing two or more difference components.

[00092] “Wet precipitated” refers to a substance which has a moisture content of from 20 to 80 weight %, or, from 30 to 70 weight %, or, from 40 to 60 weight %, or, from 45 to 55 weight %, or, 50 weight % of the total weight of the substance. Preferably, the moisture content is 50 weight % of the total weight of the substance. The substance can be HM pectin with a moisture content of 50 weight % of the total weight of the HM pectin.

[00093] A method of producing, a LM amidated pectin from a natural pectin sourced from citrus fruits, wherein the method includes a wet precipitated amidation step.

[00094] According to a first aspect of the present disclosure there is provided a method for producing LM amidated pectin from a natural pectin sourced from citrus fruits. In particular, the method requires amidation of a wet precipitated HM pectin.

[00095] Advantageously, amidation of a wet precipitated HM pectin results in a LM amidated pectin that has not undergone the same degree of glycosidic breakdown during amidation as traditional amidation of pectins which require amidation of the HM pectin to occur whilst the HM pectin is dry. Advantageously, the resultant LM amidated pectin has similar physical properties, such as intrinsic viscosity, as the HM pectin prior to amidation. [00096] To obtain the LM ami dated pectin, wet precipitated HM pectin is firstly extracted from a natural source of pectin, such as citrus fruits. The wet precipitated HM pectin then undergoes amidation to form the LM amidated pectin.

[00097] FIG. 3 depicts a flow diagram for a method according to the first aspect of the present disclosure.

[00098] Step 1: Extraction of HM pectin from a natural source of pectin

[00099] The HM pectin is extracted from a natural source of pectin. The natural source of pectin can be fruits such as apples, pears, plums, guavas, quince, gooseberries and citrus fruits. Examples of citrus fruits include, but are not limited to, lemons, mandarin oranges, grapefruit, sweet orange, pomelo, clementines, limes, kumquat, bitter oranges, blood oranges, satsumas, yuzus, tangelos, or, a combination thereof. Preferably, the HM pectin is extracted from the peel of citrus fruits.

[000100] The HM pectin is extracted from a natural source by placing the natural source in acidic, hot water.

[000101] Optionally, the temperature of the acidic, hot water is from 55 to 100 °C. preferably 60 to 90 °C, more preferably from 65 to 85 °C, most preferably 70 to 80 °C.

[000102] Optionally, the pH of the acidic, hot water is from 1.5 to 2.5, preferably from 1.75 to 2.25, most preferably from 1.9 to 2.10.

[000103] Optionally, the pH of the acidic, hot water is controlled by adding an acid to hot water. The acid can be any one or more of nitric acid, phosphoric acid, sulfuric acid, citric acid, malic acid, succinic acid, adipic acid, gluconic acid, tartaric acid, fumaric acid, or, a combination thereof. Preferably, the acid is nitric acid.

[000104] Optionally, the residence time of the natural source in the acidic, hot water is from 2 to 4.5 hours, preferably from 2.5 to 4 hours, most preferably from 3 to 3.5 hours. [000105] Preferably, the acidic, hot water is at a temperature of from 70 to 80°C with a pH of from 1.90 to 2. 10 and the natural source has a residence time in the acidic, hot water of from 3 to 3.5 hours.

[000106] The HM pectin is extracted in an aqueous form. The HM pectin in aqueous form can be clarified using centrifugation and filtration.

[000107] Optionally, the HM pectin in aqueous form is clarified with centrifugation and then filtration.

[000108] Once centrifugation and filtration are complete, a filtered aqueous form containing the HM pectin is formed.

[000109] Calcium present in the filtered aqueous form containing the HM pectin can be removed by using ion exchange. Optionally, a cationic resin is used to perform the calcium removal.

[000110] The product of the calcium removal is a de-calcified aqueous form containing the HM pectin. Optionally , the HM pectin in the de-calcified aqueous form containing the HM pectin comprises from 600 ppm of calcium or less, preferably from 550 ppm of calcium or less, most preferably from 500 ppm of calcium or less.

[000111] The de-calcified aqueous form containing the HM pectin can be concentrated by evaporation to form a concentrated HM pectin in aqueous form. Any methods known in the art for concentrating a liquid can be used.

[000112] Optionally, the concentrated HM pectin in aqueous form comprises the HM pectin at from 15 to 40 g/1, preferably from 20 to 37 g/1, more preferably from 25 to 35 g/1, most preferably from 27 to 30 g/1.

[000113] The degree of esterification (DE) of the concentrated HM pectin in aqueous form can be modified by adding an acidic solution to the concentrated HM pectin in aqueous form. The resultant product is a DE modified HM pectin in aqueous form. [000114] Optionally, the degree of esterification (DE) can be modified by adding acid to the concentrated HM pectin in aqueous form. Optionally, the acid is one or more of nitric acid, phosphoric acid, sulfuric acid, citric acid, malic acid, succinic acid, adipic acid, gluconic acid, tartaric acid, fumaric acid, or, a combination thereof Preferably, the acid is nitric acid.

Optionally, the acidic solution comprises the acid in water. Optionally, the acid is at from 0.05 to 0. 15 weight % of the total volume of the acidic solution and concentrated HM pectin in aqueous form, or more preferably, from 0.06 to 0. 12 weight % of the total volume of the acidic solution and concentrated HM pectin in aqueous form, or most preferably, from 0.7 to 1. 1 weight % of the total volume of the acidic solution and concentrated HM pectin in aqueous form.

[000115] Optionally, the temperature of the step for modifying the degree of esterification (DE) is from 20 to 45 °C, preferably from 25 to 40 °C, most preferably from 30 to 35 °C.

[000116] Optionally, the residence time of the concentrated HM pectin in aqueous form in the acidic, hot water is from 50 to 1 10 hours, preferably from 60 to 105 hours, more preferably from 70 to 100 hours, most preferably from 72 to 96 hours.

[000117] Optionally, the resultant product has a degree of esterification (DE) of from 40 to 75 %, preferably from 45 to 70 %, more preferably from 50 to 65 %, most preferably from 54 to 58 %.

[000118] Preferably, modification of the degree of esterification (DE) of the concentrated HM pectin in aqueous form requires a temperature of from 30 to 35°C, an acid content of from 0.70 to 1.10 weight % of the total weight of the mixture, the residence time of the concentrated HM pectin in aqueous form in the acidic, hot water is from 72 to 96 hours and the resultant product has a degree of esterification (DE) of from 54 to 58 %.

[000119] An appropriate amount of alcohol is added to the DE modified HM pectin in aqueous form to form an alcohol/water mix. The amount of alcohol added depends on the amount of alcohol required to cause the HM pectin to precipitate out from its aqueous form as a precipitate containing the HM pectin. [000120] Optionally, the alcohol used is of isopropanol, tert-amyl alcohol, benzyl alcohol , 1,4-butanediol, 1, 2. 4-butanetriol, butanol, 1 -butanol, 2-butanol. tert-butyl alcohol, di ethylene glycol, ethanol, ethylene glycol, 2-ethylhexanol, furfuryl alcohol, glycerol, iso-butanol, isopropyl alcohol, methanol, 2-(2-methoxyethoxyl)ethanol, 2 -methyl- 1 -butanol, 2-methyl-l- pentanol, 3-methyl-2-butanol, neopentyl alcohol, 2-pentanol, 1,3-propanediol, propan-l-ol, propylene glycol, propylene glycol methyl ether, or, any combination thereof. Preferably, the alcohol is isopropyl alcohol.

[000121] The precipitate containing HM pectin can be further washed to remove any impurities present. Optionally, the precipitate containing HM pectin is further washed with alcohol to form a washed HM pectin.

[000122] Optionally, the alcohol used is isopropanol, tert-amyl alcohol, benzyl alcohol , 1,4-butanediol, 1, 2, 4-butanetriol, butanol, 1 -butanol, 2-butanol, tert-butyl alcohol, di ethylene glycol, ethanol, ethylene glycol, 2-ethylhexanol, furfuryl alcohol, glycerol, iso-butanol, isopropyl alcohol, methanol, 2-(2-methoxy ethoxy l)ethanol, 2 -methyl- 1 -butanol. 2-methyl-l- pentanol, 3-methyl-2 -butanol, neopenty l alcohol, 2-pentanol, 1,3-propanediol, propan-l-ol, propylene glycol, propylene glycol methyl ether, or, any combination thereof. Preferably, the alcohol is Isopropyl alcohol.

[000123] The washed HM pectin can be pressed to form a wet precipitated HM pectin. The wet precipitated HM pectin has a dry substance of from 20 to 80 weight %, preferably 30 to 70 weight %, more preferably from 40 to 60 weight %, even more preferably from 45 to 55 weight %. most preferably 50 weight % of the total weight of the wet precipitated HM pectin is achieved. Preferably, the wet precipitated HM pectin has a dry substance of 50 weight % of its total weight.

[000124] Optionally, the wet precipitated HM pectin is a slow set pectin.

[000125] Optionally, the wet precipitated HM pectin can be further dried to form a dry HM pectin. Optionally, the dry' HM pectin is dried until it has a moisture content of from 5 to 15 weight %, preferably from 7.5 to 12.5 weight %, most preferably 10 weight % of the total weight of the dry HM pectin. The dried HM pectin can then be grinded until the dry HM pectin has a homogeneous particle size distribution, and/or blended with sugar to form a standardised HM pectin.

[000126] Step 2: Amidation of wet precipitated HM pectin

[000127] The wet precipitated HM pectin can then be ami dated by being placed in a suspension with alcohol and ammonia to form a LM amidated pectin.

[000128] Optionally, the alcohol is any one or more of tert-amyl alcohol, benzyl alcohol, 1 ,4-butanediol, 1, 2. 4-butanetriol, butanol, 1 -butanol, 2-butanol. tert-butyl alcohol, di ethylene glycol, ethanol, ethylene glycol, 2-ethylhexanol, furfuryl alcohol, glycerol, iso-butanol, isopropyl alcohol, methanol, 2-(2-methoxyethoxyl)ethanol, 2-methyl-l -butanol, 2-methyl-l- pentanol, 3-methyl-2 -butanol, neopentyl alcohol, 2-pentanol, 1,3-propanediol, propan- l-ol, propylene glycol, propylene glycol methyl ether, isopropanol, or, any combination thereof. Preferably, the alcohol is Isopropyl alcohol.

[000129] Preferably, the alcohol is present at from 60 to 70 weight % of the total weight of the suspension, the ammonia is present at from 4 to 6 weight % of the total weight of the suspension, the temperature is from 4 to 6 °C and the reaction mixture is maintained from 3 to 6 hours.

[000130] Optionally, the alcohol can be present at from 50 to 80 weight %, preferably from 55 to 75 weight %, most preferably at 60 to 70 weight % of the total weight of the suspension. Preferably, the alcohol is present at from 60 to 70 weight % of the total weight of the suspension.

[000131] Optionally, the ammonia is present at from 1 to 10 weight %, preferably from 2 to 8 weight %, more preferably from 3 to 7 weight %, most preferably from 4 to 6 weight % of the total weight of the suspension. Preferably, the ammonia is present at from 4 to 6 weight % of the total weight of the suspension.

[000132] Optionally, the reaction mixture containing the wet precipitated HM pectin, alcohol and ammonia is at a temperature of from 1 to 12 °C, preferably from 2 to 10 °C, more preferably from 3 to 8 °C. most preferably from 4 to 6 °C. Preferably, the temperature is from 4 to 6 °C.

[000133] Optionally, the reaction mixture is maintained for from 1 to 8 hours, preferably from 2 to 7 hours, even more preferably from 2.5 to 6.5 hours, most preferably from 3 to 6 hours. Preferably, the reaction mixture is maintained from 3 to 6 hours.

[000134] To stop the reaction from occurring in the reaction mixture, the LM amidated pectin can be filtered from the alcohol and ammonia with filtration to produce a filtrate comprising the alcohol and a residue comprising the LM amidated pectin.

[000135] Optionally, the filtrate comprising the alcohol and ammonia can pass to a recovery step.

[000136] Optionally, the residue comprising the LM amidated pectin can be neutralised to stop further reaction from occurring to form a neutralised LM amidated pectin.

[000137] Optionally, the reaction is neutralised with acidulated alcohol. Optionally, the acidulate alcohol comprises a mineral acid. Optionally, the mineral acid is nitric acid.

Optionally, the acidulate alcohol further comprises any one of tert-amyl alcohol, benzyl alcohol , 1 ,4-butanediol, 1, 2, 4-butanetriol, butanol, 1 -butanol, 2-butanol, tert-butyl alcohol, diethylene glycol, ethanol, ethylene glycol, 2-ethylhexanol, furfuryl alcohol, glycerol, iso-butanol, isopropyl alcohol, methanol, 2-(2-methoxyethoxyl)ethanol, 2-methyl-l -butanol, 2-methyl-l- pentanol, 3-methyl-2 -butanol, neopentyl alcohol. 2-pentanol, 1,3-propanediol. propan-l-ol, propylene glycol, propylene glycol methyl ether, isopropanol, or, any combination thereof. Preferably, the alcohol is isopropyl alcohol.

[000138] Optionally, the residue comprising the LM amidated pectin is neutralised with acidulate alcohol until the pH of the neutralised LM amidated pectin is from 3.0 to 5.0, preferably from 4.0 to 5.0, most preferably from 4.0 to 4.5. Preferably, the residue comprising the LM amidated pectin is neutralised with acidulate alcohol comprising nitric acid and isopropyl alcohol until the neutralised LM amidated pectin has a pH of from 4.0 to 4.5.

[000139] The neutralised LM amidated pectin can be washed with alcohol. [000140] Optionally, the alcohol is any one of tert-amyl alcohol, benzyl alcohol , 1,4- butanediol, 1, 2, 4-butanetriol, butanol, 1 -butanol, 2-butanol, tert-butyl alcohol, diethylene glycol, ethanol, ethylene glycol, 2-ethylhexanol, furfuryl alcohol, glycerol, iso-butanol, isopropyl alcohol, methanol, 2-(2-methoxyethoxyl)ethanol, 2-methyl-l -butanol, 2-methyl-l- pentanol, 3-methyl-2 -butanol, neopentyl alcohol. 2-pentanol, 1,3-propanediol. propan-l-ol, propylene glycol, propylene glycol methyl ether, isopropanol, or, any combination thereof. Preferably, the alcohol is isopropyl alcohol.

[000141] Optionally, the neutralised LM amidated pectin is washed with the alcohol one, two. three, four, five, six, seven, eight, nine, ten, or more than ten times. Preferably, the neutralised LM amidated pectin is washed with the alcohol three times.

[000142] Once the neutralised LM amidated pectin is washed, the neutralised LM amidated pectin can undergo a pressing step. The product of the pressing step is a pressed LM amidated pectin.

[000143] Optionally, the neutralised LM amidated pectin can be pressed until the product of the pressing step has a dry substance of from 30 to 70 weight %, preferably from 40 to 60 weight %. more preferably from 45 to 55 weight %, most preferably 50 weight % of the total weight of the product of the pressing step.

[000144] The pressed LM amidated pectin can undergo a drying step. The product of the drying step is a LM amidated pectin.

[000145] Optionally, the drying step removes water and/or alcohol present in the pressed LM amidated pectin.

[000146] Optionally, the pressed LM amidated pectin can be dried until the product of the drying step has a moisture content of from 1 to 30 weight %, preferably from 2 to 20 weight %, more preferably from 7.5 to 15.0 weight 5, even more preferably from 9 to 11 weight %, most preferably 10 weight % of the product of the drying step. [000147] The LM amidated pectin is then ready for quality analysis and further modification. Further modification can include standardisation with sugar to make a commercial product, and/or grinding the LM amidated pectin.

The I. amidated pectin in a food or beverage product

[000148] Another aspect of the present invention relates to the LM ami dated pectin in a food or beverage product.

[000149] The LM amidated pectin may be incorporated into a dairy' product, which includes, but is not limited to. milk, condensed milk, dried milk, milk-drinks, cream, butter, yoghurt, cheese, custard, cottage cheese, cream cheese, curd, frozen dairy products such as frozen custard, frozen yogurt and ice-cream, gelato, powdered milk, evaporated milk, sour cream, soured milk, whey, whey protein or whipped cream.

Method of producing the LM amidated pectin in a food or beverage product

[000150] Another aspect of the present invention relates to a method of making a food or beverage product comprising the LM amidated pectin. The method of making the food or beverage product comprises the steps:

(a)_providing the LM amidated pectin; and

(b) combining the LM amidated pectin with at least one food or beverage product.

EXAMPLES

[000151 ] The following are non-limiting examples that discuss, with reference to tables and figures, the advantages of the present invention. The examples set forth herein are merely examples among other possible examples.

Example 1: A non-limiting example of forming an LMA amidated pectin by the wet amidation process.

[000152] In this non-limiting example, a LM amidated pectin was produced by the wet amidation method according to the first aspect of this disclosure. In this example, the LM amidated pectin was obtained by (a) extracting HM pectin from a citrus peel to form a wet precipitated HM pectin, and, (b) forming a LM amidated pectin by subjecting the wet precipitated HM pectin to amidation.

[000153] In this non-limiting example, the following steps were taken for (a) extracting HM pectin from a citrus peel to form a wet precipitated HM pectin:

1. Mixing citrus peel with acidic, hot water, wherein the temperature of the acidic hot water is from 70 to 80°C, the pH is from 1.90 to 2. 10 and the residence time is from 3 to 3.5 hours.

2. Clarifying the product of step 1 by using centrifugation and filtration.

3. Using a cationic resin to remove the calcium from the product of step 2.

4. Concentrating the product of step 3 until the HM pectin is present in water at a concentration of from 27 to 30 g/1.

5. Modifying the degree of esterification (DE) of the product from step 4 by adding nitric acid. The step is conducted at a temperature of from 30 to 35°C. the nitric acid is present at from 0.80 to 1 .0 weight % of the total weight of the mixture and the residence time is from 72 to 96 hours. The resultant product has a degree of esterification (DE) of from 54 to 58 %. The resultant degree of esterification is measured by titration.

6. Precipitating the HM pectin from the product of step 5 by adding isopropyl alcohol.

7. Washing the product from step 6 with isopropyl alcohol.

8. Pressing the product of step 7 until the product from step 7 has a dry substance of 50 weight % of the total weight of the product of the drying step. The product is a wet precipitated HM pectin. In this non-limiting example, the following steps were taken for forming a LM amidated pectin by subjecting the wet precipitated HM pectin to amidation.

9. Mixing the wet precipitated HM pectin with isopropyl alcohol and ammonia. The alcohol is present at from 60 to 70 weight % of the total weight of the mixture, the ammonia is present at from 4.0 to 6.0 weight % of the total weight of the mixture, the suspension is maintained at a temperature of from 4 to 6 °C. The product from step 8 is in the suspension for from 3 to 6 hours. The reaction converts HM pectin into a LM amidated pectin.

10. Filtering the LM amidated pectin formed in step 9 from the isopropyl alcohol and ammonia.

11. Neutralising the LM amidated pectin with acidulated alcohol, wherein the acidulated alcohol comprises nitric acid in isopropyl alcohol.

12. Washing the product of step 1 1 with isopropyl alcohol three times.

13. Pressing the product from step 12 until the product from step 12 has a dry substance of 50 weight % of the total weight of the product of the pressing step.

14. Drying the product from step 13 until the product from step 13 has a moisture content of 10 weight % of the total weight of the product of the drying step. The resultant product is LM amidated pectin.

[000154] In step 5, the resultant degree of esterification of the HM pectin is measured by titration. The HM pectin is washed with acidified isopropanol (IP A) to remove any cations, salts and sugar present prior to titration. Then the titration is carried out using bromothymol blue at 0. 1 % in ethanol as the indicator. The purified HM pectin is mixed with five drops of bromothymol blue indicator and then titrated with 0. 1 mol/L sodium hydroxide. The ester groups of the HM pectin are saponified with 0.5 mol/L sodium hydroxide, which is then neutralized with exactly the same amount of 0.5 mol/L hydrochloric acid so that the released acid groups can then be titrated with 0. 1 mol/L sodium hydroxide solution. The method described is an example of a JECFA procedure. [000155] Example 2: A non-limiting example of comparing the intrinsic viscosity) of a LM amidated pectin formed via the wet amidation process according to traditional methods 1, 2 and 3 set out above to the LM amidated pectin formed via the wet amidation method of the present invention

[000156] In this non-limiting example, the intrinsic viscosity of the LM pectin prior to amidation was compared to the LM amidated pectins as produced by the traditional methods 1, 2 and 3 set out above and the LM amidated pectin produced by the method set out in example 1.

[000157] In this non-limiting example, a dry HM pectin and a LM amidated pectin produced via the traditional method 1 were compared. The dry HM pectin is called dry HM pectin (1) and the LM amidated pectin is called LM amidated pectin (1) from here on in. The dry HM pectin (1) and the LM amidated pectin (1) were produced via the method set out under the heading begin “Traditional method T’, wherein the dry HM pectin (1) was the product of step (1) of the method and the LM amidated pectin (1) was the product of step (2) of the method.

[000158] In this non-limiting example, a dry HM pectin and a LM amidated pectin produced via the traditional method 2 were also compared. The dry HM pectin is called dr ' HM pectin (2) and the LM amidated pectin is called LM amidated pectin (2) from here on in. The dry HM pectin (2) and the LM amidated pectin (2) were produced via the method set out under the heading begin “Traditional method 2”, wherein the dry' HM pectin (2) was the product of step (1) of the method and the LM amidated pectin (2) was the product of step (2) of the method.

[000159] In this non-limiting example, a dry HM pectin and a LM amidated pectin produced via the traditional method 3 were also compared. The dry HM pectin is called dry HM pectin (3) and the LM amidated pectin is called LM amidated pectin (3) from here on in. The dry HM pectin (3) is the same as dry' HM pectin (2). The dry HM pectin (3) and the LM amidated pectin (3) were produced via the method set out under the heading begin “Traditional method 3”, wherein the dry HM pectin (3) was the product of step (1) of the method and the LM amidated pectin (3) was the product of step (2) of the method.

[000160] In this non-limiting example, a wet precipitated HM pectin and a LM amidated pectin produced via the method according to the present disclosure were also compared. The wet precipitated HM pectin is called wet precipitated HM pectin (4) and the LM amidated pectin is called LM amidated pectin (4) from here on in. The wet precipitated HM pectin (4) and the LM ami dated pectin (4) were produced via the method set out in example 1, wherein the wet precipitated HM pectin (4) was the product of step (8) of the method and the LM amidated pectin (4) was the product of step (14) of the method.

[000161] The intrinsic viscosity of each of dry HM pectin (1), LM amidated pectin (1), dry HM pectin (2), LM amidated pectin (2), dry HM pectin (3), HM amidated pectin (3), wet precipitated HM pectin (4) and HM amidated pectin (4) was measured. The intrinsic viscosity measurements were performed by using a Gel Permeation Chromatography (GPC) system equipped with a Viscotek TDA apparatus with a number of detectors (the detectors being refractive index, right angle laser light scattering (90°) and low angle laser light scattering (9°) and a viscometer). To determine the intrinsic viscosity, each of the samples were separately dissolved at a concentration of 2 mg/ml and eluted with a buffer solution of 0.1 M LiNCh and 1 g/1 EDTA. After complete dissolution, each sample was separately filtered through membranes of 0.2 micrometer porosity: the injection volume was 100 microliters and the flow rate was 0.75 ml/min. The chosen method of analysis was triple point, calibrated with Pullulan provided by the equipment supplier with the MP (the molecular weight of the highest peak) ranging from between 5000 to 250000 g/mol. The measurements were measured at 35 °C.

[000162] The results of the intrinsic viscosity analysis are shown in Table 1 and FIG. 4.

Table 1: The intrinsic viscosity⁷ values of the HM pectin and LM amidated pectin produced via the traditional methods 1-3 and the method set out in Example 1.

*These values were determined by taking the average intrinsic viscosity' difference for each sample.

[000163] As can be seen from Table 1 and FIG. 4, the difference in the intrinsic viscosity values for the HM pectin (4) and LM amidated pectin (4) is lower than for the HM pectins and LM amidated pectins made according to traditional methods. Advantageously, the pectin undergoes much less degradation when subjected to the method of forming the amidated pectin according to the present invention, and therefore the intrinsic viscosity values are much closer.

Claims

2. The method of claim 1, wherein the low methoxyl amidated pectin has a degree of methylation of below 50 %.

3. The method of claim 1 or claim 2, wherein the wet precipitated high methoxyl pectin has a degree of methylation of above 50 %.

4. The method of any one of claim 1 to 3, wherein the wet precipitated high methoxyl pectin has an intrinsic viscosity of from 500 to 650 ml/g. or, from 510 to 640 ml/g, or. from 525 to 615 ml/g.

5. The method of any one of claims 1 to 4, wherein the low methoxyl amidated pectin has an intrinsic viscosity of from 450 to 650 ml/g, or, from 475 to 625 ml/g, or, from 488 to 600 ml/g.

6. The method of any one of claims 1 to 5, wherein the wet precipitated high methoxyl pectin is in the suspension with the one or more alcohols and ammonia at a temperature of from 1 to 12 °C, or. from 2 to 10 °C, or, from 3 to 8 °C, or, from 4 to 6 °C.

7. The method of any one of claims 1 to 6, wherein the wet precipitated high methoxyl pectin is in the suspension with the one or more alcohols and ammonia for a reaction time of from 1 to 8 hours, or, from 2 to 7 hours, or from 2.5 to 6.5 hours, or, from 3 to 6 hours.

8. The method of any one of claims 1 to 7, wherein the wet precipitated high methoxyl pectin has a dry substance content of from 20 to 80 weight %, or, from 30 to 70 weight %, or, from 40 to 60 weight %, or, from 45 to 55 weight %, or, 50 weight % of the total weight of the wet precipitated high methoxyl pectin.

9. The method of any one of claims 1 to 8, wherein the ammonia is present in the suspension at from 1 to 10 weight %. or, from 2 to 8 weight %, or. from 3 to 7 weight %, or, from 4 to 6 weight % of the total weight of the suspension.

10. The method of any one of claims 1 to 9, wherein the one or more alcohols is present in the suspension at from 50 to 80 weight %, or, from 55 to 75 weight %. or, at 60 to 70 weight % of the total weight of the suspension.

11. The method of any one of claims 1 to 10, wherein the one or more alcohols are any one of tert-amyl alcohol, benzyl alcohol, 1 ,4-butanediol, 1, 2, 4-butanetriol, butanol, 1-butanol, 2- butanol. tert-butyl alcohol, diethylene glycol, ethanol, ethylene glycol. 2-ethylhexanol, furfuryl alcohol, glycerol, iso-butanol, isopropyl alcohol, methanol, 2-(2-methoxyethoxyl)ethanol, 2- methy 1-1 -butanol, 2-methyl-l -pentanol, 3-methyl-2-butanol, neopentyl alcohol, 2-pentanol, 1,3- propanediol, propan- l-ol, propylene glycol, propylene glycol methyl ether, isopropanol, or, any combination thereof.