POLYPHENOL COMPOSITIONS AND SUGARS INCLUDING VINASSE AND/OR DIGESTATE AND METHODS OF THEIR PREPARATION

Abstract

The present invention provides a polyphenol composition comprising sugarcane bagasse and/or sugarcane vinasse. Methods of lowering the odour of the polyphenol composition using activated carbon. Methods of preparing the polyphenol composition following fermentation and distillation of a sugarcane derived product. Sugars and foods/beverages comprising the polyphenol composition and methods of preparing those.

Claims

1-119. (canceled)

120. A food grade polyphenol composition comprising liquid or powdered sugar cane vinasse and/or sugar cane digestate, wherein the vinasse or digestate has 500 to 15,000 mg GAE/100 g polyphenols.

121. The polyphenol composition of claim 120, wherein the sugar cane vinasse and/or sugar cane digestate is a sugar juice vinasse/digestate or a molasses vinasse/digestate or a combination thereof.

122. The polyphenol composition of claim 120, wherein the sugar cane vinasse and/or sugar cane digestate is a sugar cane digestate.

123. The polyphenol composition of claim 122, wherein the sugar cane digestate is a sugar cane vinasse digestate.

124. The polyphenol composition of claim 120, wherein the vinasse or digestate has: (i) no odour; or (ii) no odour when combined with white refined sucrose to produce a 100 mg GAE polyphenol/100 g carbohydrate sugar; or (iii) no odour or reduced odour because: (a) the odour is less that the odour of the sugar cane vinasse and/or sugar cane digestate used to prepare the polyphenol composition; (b) the odour is less than the usual odour of sugar cane vinasse and/or sugar cane digestate; (c) the polyphenol composition has an odour intensity of 0-3 according to the VDI 3882-1 olfactometry standard; (d) the polyphenol composition has no odour when combined with white refined sucrose or raw mill sugar to produce a 10 mg GAE polyphenol/100 g carbohydrate sugar; (e) the volatile organic content of the sugar cane vinasse and/or sugar cane digestate is reduced; or (f) combinations of (a) to (e).

125. The polyphenol composition of claim 120, wherein the composition comprises sugar cane vinasse and/or sugar cane digestate formed during preparation of bioethanol, rum, amino/organic acid, yeast propagation or a combination thereof.

126. The polyphenol composition of claim 125, wherein the vinasse and/or digestate have a Chemical Oxygen Demand (COD) of 30-150 g O.sub.2/L and/or a biochemical oxygen demand (BOD) of about 30-80% of the COD.

127. The polyphenol composition of claim 122, wherein the digestate is anaerobic digestate.

128. The polyphenol composition of claim 120, wherein (i) the vinasse is sugarcane juice vinasse, massecuite vinasse and combinations thereof; and (ii) the digestate is sugar cane vinasse digestate.

129. The polyphenol composition of claim 120, wherein digestate also had less than 1 mg/kg arsenic, less than 0.30 mg/kg antimony; less than 0.03 mg/kg cadmium, 1.9 mg/kg selenium; less than 0.004 mg/kg mercury; and 0.12 mg/kg lead.

130. The polyphenol composition of claim 120, wherein the digestate and/or vinasse is soluble.

131. The polyphenol composition of claim 120, wherein (i) the digestate is powdered and is <40% w/v plant fibre, <35% w/v ash and/or >7% w/v polyphenols; or (ii) the digestate is concentrated or diluted to 40% solids in liquid.

132. The polyphenol composition of claim 131, wherein the digestate includes one or more of (i) more than 7000 mg GAE/100 g polyphenols; the powdered digestate is less than 5% moisture by weight; and the powdered digestate has a pH 9.0 to 9.6 when 1 g of powder is dissolved in 10 ml water.

133. The polyphenol composition of claim 120, wherein the polyphenol composition is 5,000 to 15,000 mg GAE/100 g polyphenols.

134. The polyphenol composition of claim 120, wherein the polyphenol composition is <5 Pol % w/w.

135. The polyphenol composition of claim 120, wherein the polyphenol composition is not an extract.

136. The polyphenol composition of claim 120, wherein the polyphenol composition is an affinity filtration filtrate and/or an ion-exchange resin filtrate.

137. A method of preparing a powdered polyphenol composition of claim 1 comprising spray drying liquid sugar cane digestate and/or sugar cane vinasse.

138. The method of claim 137, wherein prior to spray drying the high boiling point fraction of the digestate/vinasse is removed under reduced pressure.

139. A method of preparing a liquid polyphenol composition comprising: (a) preparing a powdered polyphenol composition in accordance with claim 137; and (b) reconstituting the powder of (a) as a liquid by combining the powder with water, affination syrup, molasses or combinations thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0215] FIG. 1 depicts the GI of several of the samples prepared in Example 6.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0216] Reference will now be made in detail to certain embodiments of the invention. While the invention will be described in conjunction with the embodiments, it will be understood that the intention is not to limit the invention to those embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents, which may be included within the scope of the present invention as defined by the claims.

[0217] Further aspects of the present invention and further embodiments of the aspects described in the preceding paragraphs will become apparent from the following description, given by way of example.

[0218] All of the patents and publications referred to herein are incorporated by reference in their entirety.

[0219] For purposes of interpreting this specification, terms used in the singular will also include the plural and vice versa.

[0220] One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0221] The inventors of the present invention have developed a high concentration polyphenol composition and methods for its production. The polyphenol composition is high concentration and low odour. The composition is prepared from byproducts of the sugar cane industry and from ethanol from sugar cane or products sourced from sugar cane. There are environmental advantages to the repurpose of these byproducts in the minimisation of difficult to dispose waste and increasing the renewability of resources.

[0222] The odour reduction of the compositions of the invention is the result of affinity removal of an organic fraction via carbon or resin. There are environmental advantages to processes that do not require an extraction step such as minimising waste by avoiding the use of large quantities of additional solvents including ethanol and requiring less infrastructure to perform the process.

[0223] The polyphenol composition, vinasse or digestate are used to add polyphenols to polyphenol containing sugar products. These sugars (particularly sucrose sugars) are either crystalline or amorphous and are healthier than traditional white sucrose sugar. Some of these sugars are described in International patent specification numbers PCT/SG2019/050416 and PCT/SG2019/050057 and Singapore patent specification number 10201902102Q. Processes for preparing these sugars is described in these specifications and also in International patent specification numbers PCT/SG2019/050377.

[0224] It will be understood that various terms employed in the specification, examples and claims have meanings that will be understood by one of ordinary skill in the art. However, certain terms are defined below.

[0225] As used herein, except where the context requires otherwise, the term “comprise” and variations of the term, such as “comprising”, “comprises” and “comprised”, are not intended to exclude further additives, components, integers or steps.

[0226] The term “affination syrup” refers to a syrup used at a sugar refinery in the preparation of affined sugar. Raw sugar and water are combined, slowly agitated and heated (eg up to 45° C.) to soften the raw sugar syrup layer. This is then centrifuged. Syrup passes through the centrifuge screens and the sugar crystals (affined sugar) remain behind. The crystals are washed with water and that water often combined with the syrup to form affination syrup, which is heated (eg to about 76° C.) before combination with a new batch of raw sugar.

[0227] The term “amorphous” refers to a solid that is largely amorphous, that is, largely without crystalline structure. For example, the solid could be 80% or more amorphous, 90% or more amorphous, 95% or more amorphous or about 100% amorphous.

[0228] Amorphous sugars can be prepared as described in Singapore patent specification number 10201902102Q.

[0229] The term “anaerobic” refers to something living, active, occurring, or existing in the absence of free oxygen. For example, an anaerobic digestate is a digestate prepare in anaerobic conditions by anaerobic microorganisms.

[0230] The term “bagasse” refers to sugar fibre either from sugar cane or sugar beet. It is the fibrous pulp left over after sugar juice is extracted. Bagasse products are commercially available, for example, Phytocel is a sugar cane bagasse product sold by KFSU.

[0231] The term “Biochemical Oxygen Demand” (BOD) is a measure of the amount of food (or organic carbons) that bacteria can oxidize. BOD (biochemical oxygen demand) indicates the amount of biodegradable matter in effluent.

[0232] The term “brown sugar” refers to a food grade sugar of brown colour.

[0233] The term “chemical oxygen demand” (COD) is an indicative measure of the amount of oxygen that can be consumed by reactions in a measured solution. It is the total measurement of all chemicals in the water that can be oxidized but provides no information on their biodegradability.

[0234] The terms “efficacious” or “effective amount” refer to an amount that is biologically or chemically effective. In this context, one example is an effective amount of polyphenols in a sugar product to achieve a low glycaemic sugar.

[0235] The term “endogenous” refers to something originating from within an organism. In the context of the present invention, it refers to something originating from within sugar cane, for example, a phytochemical including monophenol or polyphenol and polysaccharide can be endogenous because the compound originated from within the sugar cane.

[0236] The term “extract” refers to a composition extracted or removed from something else. For example, an extract can be prepared by solubilising a soluble fraction from an original product into an extraction solvent. An extract can also be prepared by precipitating an insoluble fraction from an original product out of an extraction solvent. The extract is a product of combining the original product (usually plant based material) and extraction solvent (commonly ethanol). The fraction of the original product is solubilised into the extraction solvent to form a supernatant, the supernatant is separated from the remnants of the original product, and removed with the extraction solvent. The extract is the supernatant including the solubilised portion in the extraction solvent, or a concentrate or dried version thereof. The remnants of the original product, such as plant matter, left behind and/or precipitated following extraction is not an extract.

[0237] The term “filtrate” refers to a composition that has been filtered, for example, by size or affinity filtration.

[0238] The term “food grade” refers to products suitable for human consumption including products suitable for combination with other products to prepare a food.

[0239] The term “high glycaemic” refers to a food with a glucose based GI of 70 or more.

[0240] The term “low glycaemic” refers to a food with a glucose based GI of 55 or less.

[0241] The term “massecuite” refers to a dense suspension of sugar crystals in the mother liquor of sugar syrup. This is the suspension that remains after concentration of the sugar juice into a syrup by evaporation, crystallisation of the sugar and removal of molasses.

[0242] The massecuite is the product that is washed in a centrifuge to prepare bulk sugar crystals.

[0243] The term “medium glycaemic” refers to a food with a glucose based GI of 56 to 69.

[0244] The term “odourless” refers to no discernible odour. In particular, odour is not discerned by a human in close proximity (eg 10 cm away).

[0245] The term “prebiotic” refers to a food ingredient that stimulates the growth and/or activity of one or more beneficial gastrointestinal bacteria. Prebiotics may be non-digestible foods or of low digestibility. A prebiotic can be a fibre but not all fibres are prebiotic. Oligosaccharides with a low degree of polymerisation ie are thought to better stimulate bacteria concentration than oligosaccharides with higher degree of polymerisation.

[0246] The term “phytochemical” refers generally to biologically active compounds that occur naturally in plants.

[0247] The term “polyphenol” refers to chemical compounds that have more than one phenol group. There are many naturally occurring polyphenols and many are phytochemicals. Flavonoids are a class of polyphenols. Polyphenols including flavonoids naturally occur in sugar cane. In the context of the present invention the polyphenols that naturally occur in sugar cane are most relevant. Polyphenols in food are micronutrients that are of interest because of the role they are currently thought to have in prevention of degenerative diseases such as cancer, cardiovascular disease or diabetes.

[0248] The term “raw sugar” refers to a food grade sugar of light brown colour.

[0249] The term “reduced” refers to an amount lower than a reference amount. Reduced odour can be an odour lower than the previous odour of a specific sample of vinasse and/or digestate. Alternatively, the reduced odour can be lower than the usual odour for the vinasse and/or digestate. A polyphenol composition (ie vinasse or digestate of the invention) has a reduced odour when there is no odour following combining the polyphenol composition with white refined sucrose or raw mill sugar to produce a 10, 15, 20, 30, 40 or 50 mg GAE polyphenol/100 g carbohydrate sugar/polyphenol blend.

[0250] The term “reducing sugar” refers to any sugar that is capable of acting as a reducing agent. Generally, reducing sugars have a free aldehyde or free ketone group. Glucose, galactose, fructose, lactose and maltose are reducing sugars. Sucrose and trehalose are not reducing sugars.

[0251] The term “refined white sugar” refers to fully processed food grade white sugar that is essentially sucrose with minimal reducing sugar content and minimal phytochemicals such as polyphenols or flavonoids.

[0252] The term “sugar” refers to a solid, the majority of which is sucrose, glucose or fructose (ie a sucrose sugar could be 80%, 90%, 95%, 99% w/w sucrose). The sugar may contain one or more other low molecular weight sugars. The sugar may be solid or liquid. Solid sugar may be crystalline or amorphous. The sugar may contain other ingredients such as polyphenols and various minerals. Common sucrose sugars are prepared from sugar cane or from sugar beet.

[0253] The term “sugar cane digestate” refers to digestate containing sugar cane polyphenols. The digestate can be prepared from a feed stock ultimately sourced from sugar cane such as sugar cane juice, sugar cane molasses, massecuite, raw or brown sugar cane, or sugar cane vinasse. Sugar cane vinasse is preferred.

[0254] The term “sugar cane vinasse” refers to vinasse containing sugar cane polyphenols. The vinasse can be prepared from a feed stock ultimately sourced from sugar cane such as sugar cane juice, sugar cane molasses, massecuite, or raw or brown sugar cane sugar. Sugar cane juice or molasses are preferred.

[0255] The term “sugar juice” refers to the syrup or liquid produced from sugar-rich plant feedstocks, such as the juice produced following crushing/pressing sugar cane or the liquid exiting a diffuser during the processing of sugar beets.

[0256] The term “cane juice” or “sugar cane juice” refers to the syrup produced from pressed and/or crushed peeled sugar cane. Ideally sugar cane juice is at least 60° Brix.

[0257] The term “beet juice” refers to the liquid exiting a diffuser after the beet roots have been sliced into thin strips called cossetes and passed into a diffuser to extract the sugar content into a water solution.

[0258] The term “very low glycaemic” refers to a food with a glucose-based GI of less than half the upper limit of low GI (ie the GI is in the bottom half of the low GI range).

[0259] Vinasse or Dunder

[0260] Vinasse and dunder are both terms for an effluent byproduct of the sugar and ethanol industries. It is produced when carbohydrate is fermented to produce alcohol (during, for example bioethanol production or the production of rum) or amino/organic acid (eg ascorbic acid) or to propagate yeast. Specifically, it is a byproduct of the distillation step subsequent to fermentation of carbohydrates obtained from different sources of saccharides materials (eg sugarcane, sugar beet, starchy materials and lignocellulose materials). For example, vinasse is the liquid left in the boiler after distilling a batch of rum. Vinasse contains high organic matter concentrations. For the purposes of this invention the carbohydrate source includes polyphenols such as sugar cane juice or sugar cane molasses. As bioethanol production increases, the amounts of vinasse increase and alternate methods to dispose of the vinasse are in need.

[0261] Vinasse is the remaining biomass and yeasts after distillation of bioethanol produced through fermentation. Commercial fermentation is performed on a carbohydrate-rich feedstock where the carbohydrate is easily accessible. Fermentation is a process that typically begins aerobically but becomes anaerobic, leading to production of ethanol. This production of ethanol selects microorganisms (mainly yeasts) that are able to grow in an alcoholic environment. Most yeasts can only tolerate a maximum alcohol percentage of 10-15%. Accordingly, the yeast cells die before the population can move on to fully digest the food sources in the mixture that are less readily accessible than easily accessible carbohydrates.

[0262] Vinasse can be obtained from sugar and bioethanol producers.

[0263] Fermentation of Molasses

[0264] Molasses is optionally transferred from storage tanks to a fermentation tank where it is tested for pH, bacterial, and essential mineral levels. Sulfuric acid is used to adjust and maintain the pH level during fermentation. Water and yeast are added to the molasses mixture to start fermentation. The molasses mixture is allowed to ferment (eg for 12-40 hours) for (i) ethanol propagation for rum and industrial grade alcohol, biofuel; (ii) yeast propagation; and (iii) amino/organic acid production. Ammonium compounds or yeast extracts may also be used to raise the nitrogen level of the solution to required levels necessary for fermentation.

[0265] A similar process can be used to ferment sugar cane juice.

[0266] Alcohol Distillation After fermentation, the yeast, which has settled to the bottom of the tank, is separated from the liquor mixture. The separated liquor mixture is fraction distilled.

[0267] Batch distillation may be performed with various types of well-known equipment.

[0268] The simplest form is a single simple pot still. The fermented product (often referred to as beer) is heated in a pot fitted with a vapor pipe, which leads to a condenser coil immersed in a water tank. As the beer is heated, the alcohol and other volatile congeners are distilled off, condensed, and run into a storage tank. The process is continued until most of the alcohol has been distilled out.

[0269] The residue, or stillage, is emptied out of the pot and the distillate is optionally returned from the storage tank to the pot to be redistilled to increase the proof. The stillage is also known as vinasse or dunder. This vinasse can be included in sugars of the invention. The vinasse can also be further processed to vinasse digestate and/or to reduce its odour to prepare a polyphenol composition of the invention.

[0270] Fermentation and distillation of sugar cane can generate ten times the volume of vinasse to the ethanol produced.

[0271] Yeast Propagation

[0272] Yeast propagation can also follow the fermentation process.

[0273] Yeast cells are grown in a series of fermentation vessels. Yeast fermentation vessels are operated under aerobic conditions. Once the optimum quantity of yeast has been grown, the yeast cells are recovered at the final fermentation stage by centrifugal yeast separators. Vinasse is the remains of the feedstock after the final fermentation stage.

[0274] Vinasse can then be used as feedstock in biofuel production, as an agricultural fertilizer and soil conditioner, to prepare polyphenol containing sugars, or further processed to vinasse digestate and/or to reduce its odour.

[0275] Vinasse Composition

[0276] Vinasse is mainly of plant origin, with some microbial residue (yeast). It components vary based on the starting material used. The components of vinasse are readily metabolized and utilized by microorganisms as energy sources. Vinasse is a dark brown liquid that can have a a boiling point of over 100° C., a relative density at 20° C. (kg/I) of about 1.33, a viscosity of about 100 cps at 20° C. and forms an infinite aqueous solution is water.

[0277] The vinasse from sugar cane juice is acidic (reported as pH 3.5-5.4 but also as 5.4 to 6.8), predominantly soluble (eg about 80% soluble or more) and has an organic matter concentration (Chemical Oxygen Demand (COD) of 50-150 g O.sub.2/L). The biochemical oxygen demand (BOD) can be about 75% of the COD. COD/BOD ratios can be about 1.3 indicating high biodegradability. This makes vinasse suitable for anaerobic digestion. Vinasse can have 1230±630 mg/L of nitrogen, 190±35 mg/L phosphorus, 3500±2500 mg/L sulfate (See Naspolini 2017). Vinasse also contains the macronutrients required for microorganisms in anaerobic digestion. Vinasse can be processed to prepare digestate.

[0278] The vinasse assessed in Ahmed 2013 at 28.33° C. was 11.02° Brix, 22.9 μs/com conductivity, pH 4.31 10500 mg/I dissolved solids, 4633 mg/I suspended solids and contained 1735 mg/I calcium, 86 mg/I copper, 17 mg/I Iron (Fe), 14 mg/I manganese, 0.01 mg/I aluminium, 820 mg/I sulphate (SO.sub.4), 78 mg/I phosphate (PO.sub.4), and 600 mg/I nitrate (NO.sub.2) but was devoid of all microbial groups, which is expected following the distillation processing temperatures. Molasses vinasse had a COD 48 g O.sub.2/L and BOD 25.8 g O.sub.2/L. The COD/BOD ratio was then 1.86 with lower biodegradability than the Naspolini 2017 sugarcane juice vinasse. The vinasse was about 82% moisture, about 10% ash, about 6% protein and under 1% carbohydrates. In Brazil vinasse from sugar cane juice and molasses has been reported as having about 15% and about 20% ash respectively indicating that content can vary based on the region from which the sugarcane juice, molasses or other carbohydrate used to produce ethanol is sourced.

[0279] According to Devia-Orjuela (2019), vinasse is characterized by low pH values, high chemical oxygen demand (COD: 32,000-109,700 mg/L), and biological oxygen demand (BOD: 13,414-87,700 mg/L). Vinasse is composed mainly of water; organic solids; and minerals like potassium, calcium, and magnesium. Powdered vinasse was found to have 23% lingnin, 12.7% cellulose and 8.7% hem icellulose.

[0280] Vinasse includes some sugars (for example, fructose, and/or galactose) and sugar alcohols (for example, mannitol, xylitol, and/or dulcitol). Sugar alcohols are produced as byproducts of the ethanol-producing yeasts used to generate the ethanol and vinasse. Vinasse can be, for example, 8-12 or 10° Brix. This makes vinasse suitable for further anaerobic digestion.

[0281] Commercially Available Vinasse/Dunder

[0282] BioDunder® is a liquid by-product of ethanol that is produced using the Biostil fermentation/distillation process at Wilmar BioEthanol's Sarina Distillery, Queensland, Australia. BioDunder, (which contains approximately 30-40% solids) is the end product of molasses fermentation (following distillative removal of alcohol) and comprises vegetable matter (yeast biomass) containing potassium, sodium, nitrogen, calcium, magnesium, phosphorous and sulphur. Biodunder® is a dark, brown/black viscous liquid generally containing 5-35% protein, 11-65% ash, 5-25% carbohydrates, 1-6% glycerol, and 30-80% water by weight. It generally has a pH of 4.0-4.5 and a specific gravity of about 1.12 at 20° C. The glycerol is added to the raw dunder to prepare the commercial product. For the purposes of this invention, it is preferable to source the dunder prior to addition of the glycerol and/or remove the glycerol prior to use of the dunder in the methods of the invention or to prepare the vinasse/digestate of the invention.

[0283] Digestate

[0284] Digestate is the liquid remnants of the original input (ie feedstock) material following microbial digestion including, in particular, anaerobic digestion. Polyphenols remain in the digestate.

[0285] Any product of anaerobic digestion of biodegradable feed stocks (including vinasse) could be considered a digestate. However, in industry, digestate often, in context, refers to the product of an anaerobic digestion of feed stocks lacking high levels of easily accessible carbohydrate. The digestate remaining following digestion of a low accessible carbohydrate feedstock (such as vinasse) is different to vinasse. Digestion in low carbohydrate/low alcohol conditions leads to different volatile products being produced through anaerobic digestion, mainly biogas. As opposed to the formation of alcohol during fermentation of compositions high in easily digestible carbohydrates (such as sugar cane juice).

[0286] Digestate pH varied between 6.5 to 9.5 whereas vinasse pH may be as low as 3.5 due to higher content of residual organic and volatile acetic acids from fermentation including acetic, fulvic and aconitic acids which form very stable mineral chelates of boron, calcium, copper, iron, manganese, zinc.

[0287] Anaerobic Digestion

[0288] Anaerobic digestion is widely used as a source of renewable energy. The process produces a biogas, consisting of methane, carbon dioxide, and traces of other ‘contaminant’ gases. This biogas can be used directly as fuel, in combined heat and power gas engines or upgraded to natural gas-quality biomethane. Anaerobic digestion also produces a nutrient-rich digestate byproduct that can be used as fertilizer. Anaerobic digestion is used as part of the process for treating biodegradable waste. Anaerobic digesters can also be fed with purpose-grown energy crops, such as sugar cane.

[0289] Anaerobic digestion is a staged process. The stages are as follows: [0290] Hydrolysis: breakdown of complex insoluble organic matter into simple sugars, fatty acids, and amino acid. [0291] Acidogenesis: further breakdown of simple sugars, fatty acids, and amino acids into alcohols & volatile fatty acids (VFAs). [0292] Acetogenesis: conversion of VFAs and alcohols into acetic acid, CO.sub.2, and hydrogen. [0293] Methanogenesis: acetic acid and hydrogen are converted into methane and CO.sub.2 by methanogenic bacteria.

[0294] There are mesophilic, acidophilic and thermophilic anaerobic digestion systems.

[0295] The digestion process begins with bacterial hydrolysis of the input materials (for example vinasse) in order to break down insoluble organic polymers such as carbohydrates (eg lignin, cellulose and hem icellulose) and make them available for other bacteria. Acidogenic bacteria then convert the sugars and amino acids into carbon dioxide, hydrogen, ammonia, and organic acids. Acetogenic bacteria then convert these resulting organic acids into acetic acid, along with additional ammonia, hydrogen, and carbon dioxide. Finally, methanogens convert these products to methane and carbon dioxide.

[0296] Various versions of anaerobic digestion are in commercial use and are considered suitable to prepare digestates of the invention.

[0297] Batch or Continuous

[0298] Anaerobic digestion can be performed as a batch process or a continuous process. In a batch system, biomass is added to the reactor at the start of the process. The reactor is then sealed for the duration of the process. In its simplest form batch processing needs inoculation with already processed material to start the anaerobic digestion.

[0299] In continuous digestion processes, organic matter is constantly added (continuous complete mixed) or added in stages to the reactor (continuous plug flow; first in—first out). Here, the end products are constantly or periodically removed, resulting in constant production of biogas. A single or multiple digesters in sequence may be used. Examples of this form of anaerobic digestion include continuous stirred-tank reactors, upflow anaerobic sludge blankets, expanded granular sludge beds, and internal circulation reactors.

[0300] In preferred embodiments, a continuous digestion process is used to prepare digestates of the invention.

[0301] Vinasse Digestate

[0302] Vinasse digestate is the non-volatile materials that remain after fermentation and distillation to produce vinasse and then further anaerobic digestion. Optionally, the further anaerobic digestion is after the initial volatiles have been removed. This process involves anaerobic digestion in an environment with substantial alcohol and then anaerobic digestion in a substantially-alcohol free or low alcohol environment.

[0303] In preferred embodiments, the polyphenol composition of the invention is sugar cane vinasse digestate. Either sugar cane juice or molasses is fermented and then distilled to produce ethanol. The vinasse by-product of this process is then fed into an anaerobic digestor to produce biogas. Optionally, this is a continuous digestor that only requires inoculation at the beginning of ethanol production season.

[0304] The anaerobic digestor converts vinasse into biogas (gas), digestate (liquid) and sludge (solid). Some sugar cane mills that produce ethanol also have a digestor onsite that is essentially a covered lagoon. Biogas is captured under the covers and continuously vented from the digestor, dried and compressed to use as a fuel source. Digestate overflows into a digestate well and is pumped out to a storage facility. Sludge builds up in the digestor and is removed as needed. This can be as infrequently as once at the end of the season.

[0305] Digestors of 140,000 m.sup.3 can be used at a consistent incoming flowrate of 100 kL/day per digestor. No water is added, however the incoming total solids from vinasse production can vary. Temperature is ambient, which during the 4-5 month long season is 25-35° C. Residence time in the lagoon is approximately 100 days.

[0306] Drying and Reconstituting

[0307] If needed, the vinasse and/or digestate is optionally concentrated to 30-50° Brix (eg about 40° Brix).

[0308] The vinasse and digestate can both be spray dried, for example, for easy transport. The powdered form is relatively stable and easier to transport. Dried vinasse and/or digestate can be combined with a sugar to increase polyphenol content. Alternatively, the vinasse and/or digestate powder concentrate can be reconstituted into liquid form with either water or affination syrup (for example, in a 1:10 ratio or to about 10° Brix) and then combined with the sugar.

[0309] Polyphenol Content Measurement

[0310] Polyphenol content can be measured in terms of its catechin equivalents or in terms of its gallic acid equivalents (GAE). Amounts in mg CE polyphenols/100 g can be converted to mg GAE polyphenols/100 g by multiplying by 0.81 ie 60 mg CE polyphenols/100 g is 49 mg GAE polyphenols/100 g.

[0311] A laboratory method for determining polyphenol content is described in Kim, Dae-Ok (2003).

[0312] Odour Measurement

[0313] There are various methods for measuring odour. These methods are both sensory and instrumental. The testing used in this specification has been informal sensory testing and is explained in the examples, definitions and throughout the specification as appropriate. More formal testing can be conducted by dynamic olfactometry.

[0314] Dynamic olfactometry is a sensorial method standardized by the European Standard EN13725:2003 (Standard EN 13725:2003. Air Quality-Determination of Odour Concentration by Dynamic Olfactometry; CEN:Brussels, Belgium, 2003), which provides the odour concentration of a sample, referring to the sensation that it causes in a panel of opportunely selected people directly exposed to that odour.

[0315] The odour concentration, expressed in European odour units per cubic meter (ouE/m.sup.3), represents the number of dilutions with neutral air that are necessary to bring the concentration of the sample to its odour detection threshold (OT), i.e., the threshold at which the odour is perceived by 50% of the examiners. To put it in the simplest manner, if the sample needs to be diluted 100 times with clean air so that the panel cannot perceive the odor anymore, this means that the sample has a concentration of 100 ouE/m.sup.3.

[0316] The analysis is carried out by presenting the sample to the examiners (i.e., panelists) at increasing concentrations by means of a dilution device, called an olfactometer, which dilutes the samples according to given ratios with reference air, which is made odour- and humidity-free through filtration with active carbon or silica gel.

[0317] In order to ensure reliable and repeatable results, the EN 13725:2003 fixes precise criteria for panel selection based on individuals' threshold for n-butanol in nitrogen (between 20 and 80 ppb) and the standard deviation of the individual's responses, which are verified periodically.

[0318] German guidelines VDI 3882 Part 1 (Blatt 1: 1992, Olfactometry; determination of odour intensity) and VDI 3882 Part 2 (1994, Olfactometry—determination of hedonic odour tone), describe how to apply olfactometric measurements for the determination of odour intensity and odour pleasantness/unpleasantness (hedonic tone). Odour intensity is expressed in a scale from 0 (not perceptible) to 6 (extremely strong). Hedonic tone is measured in a scale from −4 (extremely unpleasant) to +4 (extremely pleasant) (FIG. 1).

[0319] The odour intensity scale is not perceptible (0), very weak (1), weak (2), distinct (3), strong (4), very strong (5), or extremely strong (6). The sugar cane vinasse or sugar cane digestate starting material can be distinct, strong, very strong or extremely strong in odour (ie a 3-6). The reduced odour polyphenol compositions of the invention are not perceptible, very weak, weak, or distinct in odour (ie a 0-3) in the odour intensity scale. The original digestate, vinasse or vinasse digitate is strong, very strong, or extremely strong in odour (ie 4-6).

[0320] Glycaemic Response (GR)

[0321] GR refers to the changes in blood glucose after consuming a carbohydrate-containing food. Both the GI of a food and the glycaemic load (GL) of an amount of a food are indicative of the glycaemic response expected when food is consumed.

[0322] GI

[0323] The glycaemic index is a system for classifying carbohydrate-containing foods according to the relative change in blood glucose level in a person over two hours after consuming that a food with a certain amount of available carbohydrate (usually 50 g). The area under the two hour blood glucose response curve (AUC) is divided by the AUC of a glucose standard, where both the standard and the test food must contain an equal amount of available carbohydrate. An average GI is usually calculated from data collected from 10 subjects. Prior to a test the person would typically have undergone a twelve hour fast. The glycaemic index provides a measure of how fast a food raises blood-glucose levels inside the body. Each carbohydrate containing food has a GI. The amount of food consumed is not relevant to the GI. A higher GI means a food increases blood-glucose levels faster. The GI scale is from 1 to 100. The most commonly used version of the scale is based on glucose. 100 on the glucose GI scale is the increase in blood-glucose levels caused by consuming 50 grams of glucose. High GI products have a GI of 70 or more. Medium GI products have a GI of 55 to 69. Low GI products have a GI of 54 or less. These are foods that cause slow rises in blood-sugar.

[0324] Those skilled in the art understand how to conduct GI testing, for example, using internationally recognised GI methodology (see the Joint FAO/WHO Report), which has been validated by results obtained from small experimental studies and large multi-centre research trials (see Wolever et al 2003).

[0325] In vitro GI testing is now also available, see Example 6.

[0326] GL

[0327] Glycaemic load is an estimate of how much an amount of a food will raise a person's blood glucose level after consumption. Whereas glycaemic index is defined for each type of food, glycaemic load is calculated for an amount of a food. Glycaemic load estimates the impact of carbohydrate consumption by accounting for the glycaemic index (estimate of speed of effect on blood glucose) and the amount of carbohydrate that is consumed. High GI foods can be low GL. For instance, watermelon has a high GI, but a typical serving of watermelon does not contain much carbohydrate, so the glycaemic load of eating it is low.

[0328] One unit of glycaemic load approximates the effect of consuming one gram of glucose. The GL is calculated by multiplying the grams of available carbohydrate in the food by the food's GI and then dividing by 100. For one serving of a food, a GL greater than 20 is high, a GL of 11-19 is medium, and a GL of 10 or less is low.

[0329] Cane Juice

[0330] Cane juice contains all the naturally occurring macronutrients, micronutrients and phytochemicals present in the syrup extracted from pressed and/or crushed peeled sugar cane that are normally removed in white refined sugar, which is 99.9% sucrose.

[0331] Molasses

[0332] Is a viscous by-product of sugar preparation, which is separated from the crystallised sugar. The molasses may be separated from the sugar at several stages of sugar processing. Molasses contains the same compounds as cane juice but is a more highly concentrated source of phytochemicals.

[0333] Spray Drying and Other Drying Methods

[0334] Spray drying operates on the principle of convection to remove the moisture from the liquid feed, by intimately contacting the product to be dried with a stream of hot air. The spray drying process can be broken down into three key stages: atomisation of feedstock, mixing of spray and air (including evaporation process) and the separation of dried product from the air. Other appropriate drying methods include fluidized bed drying, ring drying, freeze drying and low temperature vacuum dehydration.

[0335] Atomisation

[0336] In order to ensure that the particles to be dried have the maximum surface area available to contact the hot air stream, the liquid feed is often atomised, producing very fine droplets ultimately leading to more effective drying. There are several atomiser configurations that exist, the most common being the wheel-type, pneumatic and nozzle atomisers.

[0337] Evaporation and Separation

[0338] The second stage of the spray drying process involves the evaporation of moisture by using hot gases which flow around the surface of the particles/droplets to be dried.

[0339] There are notably three different types of air-droplet contacting configurations that exist: co-current, counter-current and mixed flow, all of which have differing applications depending on the product to be dried.

[0340] Both co-current and counter-current drying chambers are able to be used for heat sensitive materials, however the use of mixed-flow drying chambers is restricted to drying materials that are not susceptible to quality degradation due to high temperatures.

[0341] The final stage of the spray drying process is the separation of the powder from the air stream. The dry powder collects at the base of the drying chamber before it is discharged or manually collected.

[0342] Glass Transition Temperature

[0343] The glass transition temperature (Tg) is the substance-specific temperature range at which a reversible change occurs in amorphous materials from the solid, glassy state to the supercooled liquid state or the reverse. The glass transition temperature becomes very important for the production of dried products, particularly in relation to the processing and storage stages of manufacture. The glass transition temperature of the powders can be determined via differential scanning calorimetry (DSC).

[0344] ICUMSA

[0345] ICUMSA is a sugar colour grading system. Lower ICUMSA values represent less colour. ICUMSA is measured at 420 nm by a spectrophotometric instrument such as a Metrohm NIRS XDS spectrometer with a ProFoss analysis system. Currently, sugars considered suitable for human consumption, including refined granulated sugar, crystal sugar, and consumable raw sugar (ie brown sugar), have ICUMSA scores of 45-800. Sugars with scores above 800 are currently used for cosmetics or other non-edible purposes, but require further processing to be fit for human consumption. Consequently, the food grade sugars of the invention with ICUMSA values of 500 to 5000 ICUMSA are unexpected.

[0346] The sugar particles of the present invention may optionally be prepared using the methods and systems described in Australian Provisional Patent Application No 2016902957 filed on 27 Jul. 2016 with the title “Process for sugar production” or International Patent Publication number WO 2018/018089 with the same title.

[0347] Arsenic Reduction

[0348] If the arsenic levels of the vinasse or digestate are too high, they can be reduced by known methods including use of calcium-alginate beads (see Bezbaruah 2014).

Preparation of a Sugar Using the Polyphenol Composition of the Invention

[0349] Beet sugar does not contain polyphenols and neither does refined white sugar contain more than trace amounts of polyphenols. Polyphenols can be added to either to lower the glycaemic index of the sugar and prepare, for example, a low glycaemic or very low glycaemic sugar. Sugars prepared by controlled washing of sugar cane massecuite, for example, using the controlled washing method as described in patent publication numbers WO 2018/018090 and/or WO 2018/018089 to produce a sugar of 20 to 45 mg CE polyphenols/100 g carbohydrate and suitable reducing sugar content, further polyphenols can be added by combining that sugar with vinasse, digestate or a polyphenol composition of the invention to the sugar.

[0350] The further polyphenols may be added to the sugar in a powdered or liquid form. One option is to spray the liquid or powdered polyphenols onto the sugar. The process for adding the polyphenol additive onto the sugar can be completed as described in international patent application number PCT/SG2019/050377.

[0351] It is preferred that the polyphenols added to the sugar are polyphenols that, even if not sourced from sugar cane, are present in sugar cane. The polyphenols can be sourced from sugar cane, for example, from sugar cane vinasse (for example following distillation of sugar cane juice or molasses) or sugar cane digestate (following anaerobic digestion of sugar cane, sugar cane juice, molasses or sugar cane vinasse) or sugar cane vinasse/digestate diluted with or dissolved in water, affination syrup or molasses. Optionally, two or more polyphenol sources are added to the sugar. Preferably, at least one polyphenol source is a polyphenol composition according to the first or second aspects or the invention, their options, embodiments or alternatives. A further polyphenol source is optionally a sugar cane waste stream such as dunder or molasses.

[0352] The polyphenol content is then determined and, if needed, additional polyphenols added to achieve the desired amounts such as about 46 mg CE polyphenols/100 g carbohydrate to about 100 mg CE polyphenols/100 g carbohydrate or about 10 mg CE polyphenols/100 g carbohydrate to about 100 mg CE polyphenols/100 g carbohydrate.

REFERENCES

[0353] Ahmed, 0 et al, Physiochemical, Chemical and Microbiological Characteristics of Vinasse, a by-product from ethanol industry, American Journal of Biochemistry (2013) 3(3):80-83. [0354] Australian provisional patent application number 2016902957. [0355] Bezbaruah, A. N. et al, Ca-alginate-entrapped nanoscale iron: arsenic treatability and mechanism studies, Journal of Nanoparticle Research (2014) 16:2175. [0356] Devia-Orjuela, J. S. et al, Evaluation of press mud, vinasse powder and extraction sludge with ethanol in a pyrolysis process, Energies (2019) 12:4145. [0357] International patent publication numbers WO 2018/018090 and WO 2018/018089. [0358] International patent application numbers PCT/SG2019/050377, PCT/SG2019/050416 and PCT/SG2019/050057. [0359] Joint FAO/WHO Report. Carbohydrates in Human Nutrition. FAO Food and Nutrition. Paper 66. Rome: FAO, 1998. [0360] Kim, Dae-Ok, et al, Antioxidant capacity of phenolic phytochemicals from various cultivars of plums, Food Chemistry (2003) 81, 321-26. [0361] Naspolini, B. F. et al, Bioconversion of sugarcane vinasse into high-added value products and energy, BioMed Research International (2017) article ID 8986165. [0362] Singapore patent application number 10201902102Q. [0363] Wolever TMS et al. Determination of the glycemic index values of foods: an interlaboratory study, European Journal of Clinical Nutrition (2003) 57:475-482.

[0364] A copy of each of these is incorporated into this specification by reference.

EXAMPLES

Example 1—Properties of Liquid Dunder & Digestate Compositions

[0365] Liquid sugar cane juice vinasse digestate commercially sourced from a bioethanol manufacturer in Thailand was analysed for phenolic content by both CE and GAE methods. The results were 1.000% w/w as catechin equivalents (1000 mg CE/100 g) or 0.810% w/w as gallic acid equivalents (809.8 mg GAE/100 g).

[0366] Several samples of brown to dark brown liquid digestate from different batches were obtained. The digestate used had 9-11% solids as determined by a Brix Meter. The digestate also had 0.94 mg/kg arsenic, less than 0.30 mg/kg antimony; less than 0.03 mg/kg cadmium, 1.9 mg/kg selenium; less than 0.004 mg/kg mercury; and 0.12 mg/kg lead. The digestate has a six month to one year shelf life depending on storage conditions.

Example 2— Spray Drying Liquid Digestate or Dunder

[0367] Sugar cane juice vinasse digestate was commercially sourced from a bioethanol manufacturer in Thailand in liquid form and spray dried to powder form (inlet temperature 160° C. & outlet temperature 80° C.).

[0368] The spray drying can be either before or after further processing with activated carbon as described in Example 3 below.

[0369] The spray dried digestate was pH tested. The results for two samples was pH 9.2 & 9.5 when 1 g of powder was dissolved in 10 ml water.

[0370] The properties of the vinasse digestate powders D1 to D3 are in Table 1. The three digestate samples were assessed for sucrose sugar content by polarisation using HPLC and near infra-red (NIR pol), for conductivity using a conductivity meter, for ash levels using NIR (NIR ash), for colour levels using NIR (NIR colour), for polyphenol levels using a Folin-Ciocalteu laboratory method with UV detection at 760 nm (adapting method from Kim et al for determining mg GAE polyphenols/100 g carbohydrates (Lab GAE). The NIR results were all determined using standard titration methods to correlate NIR reading with ash, colour, polyphenol and other results.

TABLE-US-00001 TABLE 1 Properties of a number of digestate powders Conduct- NIR NIR Lab GAE NIR Pol ivity Ash Color mg GAE/ Sample ID % w/w μs/cm % w/w ICUMSA 100 g Digestate BDL 21.9 11 11106 6818 composition 1 (D1) Digestate <1 23.2 13 7805 10789 composition 2 (D2) Digestate BDL 24.3 11 7981 11273 composition 3 (D3)

[0371] The spray dried digestate of the invention is preferably soluble in water. Where the dunder or digestate used to prepare the spray dried powder has an insoluble fraction, the insoluble fraction is preferably removed before spray drying.

[0372] Spray dried sugar cane juice vinasse digestate powder was provided to the National Measurement Institute in Australia for analysis. The powder had a standard plate count of 12,000 CFU per g, less than 10 yeast CFU per g, less than 40 mould CFU per g and 1,500 aerobic thermophilic bacterial spores CFU per g.

[0373] Based on the results to date, the product specifications for spray dried digestate of the invention are proposed to be as set out in Table 2 below.

TABLE-US-00002 TABLE 2 Polyphenol composition specifications Chemical Name Proportion Plant Fibre <40% w/v Ash <35% w/v Polyphenols  >7% w/v Parameter Specification Appearance Fine Powder Moisture ≤5.0% pH 10% w/v 9-9.5 Polyphenols >7000 mg/100 g GAE Heavy Metals <2 mg/kg Microbiological Properties Total Plate Count ≤15,000 CFU/g Yeast and Mould  ≤1,500 CFU/g Salmonella Absent

[0374] The spray dried powder generally has less than 2% w/w moisture.

[0375] The brown to dark brown fine powder has 2.0 mg/kg arsenic, cadmium and selenium; 1.0 mg/kg mercury; and 0.2 mg/kg lead. The powder is 6.0% and has a bulk density of 0.7-0.85 kg/L. The powder has the following microbiological properties: total plate count of 15,000 CFU/g, yeast of 500 CFU/g and no salmonella.

[0376] Following the above invention, spray drying was also conducted with sugar cane digestate concentrated to 25 brix. The concentrate is passed through an evaporator at 80° C. for 1 minute and then pumped into the spray dryer. This process produces acceptable dry powder.

Example 3— Further Processing Digestate by Activated Carbon to Reduce Odour

[0377] The effect on odour and GAE levels for polyphenols upon contact of digestate with powdered activated carbon (PAC) was studied.

[0378] The loose PAC was pre-washed prior to the tests. 400 g of PAC was pre-washed with 1 L of distilled water to remove carbon dust. The water was decanted and discarded and replaced with fresh distilled water at regular intervals over a 48 hour period until the water was no longer coloured. At this point, the water was discarded and carbon was dried in an oven at 100° C. until the carbon was desiccated to powder form.

[0379] Spray dried digestate powder (20 g) was mixed with distilled water and made up to 100 ml volumetrically. 5×5 ml aliquots of this mixture were taken. 0.4 g of washed PAC was added to four of these aliquots. All five aliquots were made up to 100 ml volumetrically. All five diluted flasks were placed on a mechanical stirrer and agitated for mixing according to an assigned PAC contact time (0, 15, 30, 45, 60 min). The sample with a PAC contact time of 0 min was the sample where PAC was not added.

[0380] Upon mixing and after the nominated PAC contact time had elapsed, the samples were filtered through Whatman 41 (0.22 μm) filter paper. Filtered subsamples were analysed for polyphenol content according to Kim et al. Unfiltered samples with a PAC content time of 0 min were also analysed for polyphenol content according to Kim et al. The odour of the filtered subsamples was also assessed by a panel of 4 people.

[0381] The results of this testing is tabulated below in Table 3. Odour was found to decrease with increased PAC time. GAE levels did not greatly decrease with increased PAC time.

TABLE-US-00003 TABLE 3 PAC contact time vs GAE levels and sensory profile PAC Sensory Sam- contact Dilution GAE mg/ Analysis ple time Filtered for assay 100 g (4 people) 1  0 min No 1/100 11423 earthy smell 2  0 min No 1/200 10073 earthy smell 2  0 min Yes 1/200 10202 earthy smell 3 15 min Yes 1/200 9329 moderate earthy smell 4 30 min Yes 1/200 10566 slight earthy smell 5 45 min Yes 1/200 10022 no odour 6 60 min Yes 1/200 9842 no odour

[0382] Spray dried activated carbon processed digestate stored in clear, sealed, plastic bags has a shelf life of 6 months to 1 year.

Example 3a-Further Processing of Digestate to Reduce Odour

[0383] The effect of activated carbon on odour and turbidity for sugar cane juice vinasse digestate was further studied. The testing was conducted by SLS Global Technical Support of Pall Corporation in Bangkok, Thailand.

[0384] Liquid vinasse digestate of 44° and 10° brix was commercially sourced as starting material. These samples had a dark brown colour and strong odour from organic compounds. The 44° brix sample had high viscosity. The 10° brix sample had low viscosity.

[0385] Reduction in Turbidity

[0386] The 10° brix sample was pre-filtered using Seitz® K900 sheet filter at a flow rate of 15 ml/min (Flux rate 400 LMH) at 25° C. with filtrate volume 700 ml at differential pressure 2.0 bar. Seitz® K900 sheet filters are 4.3 mm thick sheets made of cellulose, diatomaceous earth and perlite. The K900 is 46% ash with water permeability of 1700 L/m.sup.2/m in. The turbidity value decreased from 70.5 NTU to 50.6 NTU with no effect on the odour. Odour was inspected manually by the conductor of the test smelling the sample. See Table 3a below.

TABLE-US-00004 TABLE 3a Result of trial with 10° brix sample pre-filtered using Seitz ® K900 sheet filter at a flow rate of 15 ml/min (Flux rate 400 LMH) at 25° C.: Differential Pressure Turbidity Value Volume (ml) (bar) (NTU) Start 0 70.5 100 0.5 n/a 200 0.9 n/a 300 1.2 n/a 400 1.4 n/a 500 1.5 n/a 600 1.7 n/a 700 2.0 50.6

[0387] Reduction in Odour

[0388] The pre-filtered 10° brix vinasse digestate was then filtered using Seitz® Stax depth filters AKS2 or AKS4 at a flow rate of 10 ml/min (Flux rate 272 LMH) at 25° C. with filtrate volume 300 ml at differential pressure 2.0 bar.

[0389] The AKS2 targets 400-1000 dalton contaminates and is recommended for use in high efficiency decolourisation having 1.4 kg PAC in the 12 inch module. The AKS4 targets 400-1500 dalton contaminants and is a low efficiency general purpose filter having 0.7 kg PAC in the 12 inch module.

[0390] A larger filter is recommended for filtering 700 ml volume or filtering in 600 ml volumes or less at a time.

[0391] Odour was inspected manually by the conductor of the test smelling the sample. The results are presented in Tables 3b and 3c below.

TABLE-US-00005 TABLE 3b Result of trial with 10° brix sample filtered using Seitz ® AKS2 at a flow rate of 15 ml/min (Flux rate 400 LMH) at 25° C.; following pre-filtration as described above: Differential Turbidity Value Volume (ml) Pressure (bar) (NTU) Odour Test Start 0 50.6 Strong odour 100 0.8 n/a Not detected 200 1.5 n/a Not detected 300 2.0 41.6 Not detected

TABLE-US-00006 TABLE 3c Result of trial with 10° brix sample filtered using Seitz ® AKS4 at a flow rate of 15 ml/min (Flux rate 400 LMH) at 25° C.; following pre-filtration as described above: Differential Turbidity Value :Volume (ml) Pressure (bar) (NTU) Odour Test Start 0 50.6 Strong odour 100 0.5 n/a Light odour 200 1.0 n/a Light odour 300 2.0 40.5 Light odour

[0392] Filtration of the 10° brix sample using a Seitz® AKS2 filter following pre-filtration resulted in a sample where odour could not be detected. Filtration of the 10° brix sample using a Seitz® AKS2 filter following pre-filtration resulted in a sample where only light odour could be detected. The odour could potentially be further removed by a second filtration step with this filter.

[0393] The 44° brix sample blocked the Seitz® AKS2 and AKS4 filters at both 25° C. and 60° C. due to high particle counts and the high viscosity of the sample. Therefore, viscosity reduction, for example by dilution or increase in temperature, is recommended before filtering.

[0394] It is expected that increasing the temperature of filtration from 25° C. to a temperature such as to 60° C. will decrease the viscosity of digestate samples, including the 10° brix sample. It is expected that this will increase the lifetime of filters used in connection with the samples.

[0395] Similarly, it is expected that the retention of more particles in the pre-filtration stage would also increase the lifetime of the filters used subsequently. This could be achieved by using a finer grade pre-filter, such as a Supradisc™ II X700 depth filter module. This module could be regenerated through operation in the reverse direction.

Example 3a-Pilot Scale Processing of Digestate to Reduce Odour

[0396] Pilot scale carbon filtration was tested as follows.

[0397] Pre-filtration using a Supradisc™ II X700 depth filter module was performed at a flux rate of 200 LMH (L/hr/m.sup.2) and a flow rate of 110 L/h on:

[0398] 10° Brix digestate obtained via dilution of a more viscous digestate with water

[0399] (Sample A); and 10° Brix digestate obtained by dissolution of an evaporated sample of digestate in water (Sample B).

[0400] The pre-filter was cleaned by backwash, ie reverse direction water flow, between each filtration.

[0401] The pre-filtered samples were then carbon filtered through a Supradisc™ AKS4 module at a flux rate of 200 LMH (flow rate 110 L/h) to reduce odour. The results for Samples A and B are in Tables 3d and 3e, respectively. It is expected that use of a coarser pre-filter such as a Supradisc™ IIT 1000 would be successful and increase the filtration cycle time before backwash.

TABLE-US-00007 TABLE 3d Result of filtration trials of 10° brix sample obtained via dilution of liquid concentrate Initial dP Final dP Time Volume Turbidity (bar) (bar) (min) (L) (NTU) Pre-filtration of Sample A Start 0.1 2.5 10 20 200 1.sup.st fraction 0.1 2.5 10 20 40.6 filtration 2.sup.nd 0.1 2.5 10 20 41.0 fraction filtration 3.sup.rd 0.1 2.5 10 20 42.2 fraction filtration 4.sup.th 0.1 2.5 10 20 40.5 fraction filtration 5.sup.th fraction 0.1 2.5 10 20 41.3 filtration 6.sup.th fraction 0.1 2.5 10 20 41.5 filtration 7.sup.th fraction 0.1 2.5 10 20 40.2 filtration Carbon filtration of Sample A Filtration 0.1 0.1 60 160 32.7

[0402] The odour was strong before and after pre-filtration. Following the carbon filtration, the odour was reduced to slight.

TABLE-US-00008 TABLE 3e Results of pre-filtration and filtration of 10° brix sample dissolved from evaporate Initial dP Final dP Time Volume Turbidity (bar) (bar) (min) (L) (NTU) Pre-filtration of Sample B Start 0.1 2.5 10 20 220 1.sup.st fraction 0.1 2.5 10 20 40.0 filtration 2.sup.nd 0.1 2.5 10 20 40.2 fraction filtration 3.sup.rd 0.1 2.5 10 20 41.1 fraction filtration 4.sup.th 0.1 2.5 10 20 40.4 fraction filtration Carbon filtration of Sample B 1.sup.st 0.1 0.1 60 100 35.5 Filtration

[0403] The sample had a strong odour both before and after pre-filtration. Following filtration only a slight odour remained.

[0404] Following the deodorising of the sugar cane digestate, the digestate was concentrated to about 25 brix and spray dried at 200-260° C. The resulting dry powder had the following composition:

TABLE-US-00009 Properties Unit of Unit of of measure- Properties of measure- NIX Result ment NIX Result ment Crude 18.70 % of dry Calcium 27000 mg/kg protein matter (ppm) Crude fibre 0.10 % Phosphorus 1800 mg/kg (ppm) Crude fat 0.70 % Magnesium 20000 mg/kg (ppm) Ash 30.27 % Potassium 100000 mg/kg (ppm) Dry matter 96.50 % Sodium 3100 mg/kg (ppm) Moisture 3.50 % Sulphur 8900 mg/kg (ppm) Total sugars 0.11 % Iron 360 mg/kg (ppm) PH 9.00 Zinc 45 mg/kg (ppm) Polyphenols 10 000 mg/100 g Selenium 1.9 mg/kg GAE Total plate <10 000 cfu/g Antimony ND mg/kg count Yeast <100 cfu/g Arsenic 0.94 mg/kg Mould <100 cfu/g Cadmium <0.03 mg/kg Salmonella ND In 25 g Lead 0.12 mg/kg E.coli <3 MPN/g Mercury ND mg/kg

[0405] The powder also has about 1,000 GAE mg/100 g. This is 5 times the polyphenols of molasses (about 200 GAE mg/100 g).

Example 3b— Polyphenol Levels During Odour Reduction Process

[0406]

TABLE-US-00010 K900 (pre-filter) + AKS2 carbon filter % reduction in sample fraction GAE mg/100 ml polyphenols Sample 10 brix 507.6 — K900 (pre-filter) 509.2 0 AKS2 0-100 ml 357.1 30 AKS2 100-200 ml 477.8 6 AKS2 200-300 ml 482.1 5 Average 13.67

[0407] The carbon filtration only results in a 5-30% reduction in polyphenols.

TABLE-US-00011 K900 (pre-filter) + AKS4 carbon filter % reduction in Sample fraction GAE mg/100 ml polyphenols Sample 10 brix 507.6 — K900 (pre-filter) 509.2 0 AKS4 0-100 ml 404.8 20 AKS4 100-200 ml 463.8 9 AKS4 200-300 ml 424.5 16 Average 15%

[0408] This carbon filtration only resulted in a 9-20% reduction in polyphenols.

[0409] Overall odour reduction was achieved with an average of 14.3% loss of polyphenols (ie 5-30% polyphenol reduction).

Example 4— Further Processing of Digestate

[0410] The high boiling point fraction of the digestate may be removed under reduced pressure. Reduced pressure removes and/or decreases the volatile components. For example, the carboxylic acid (such as acetic acid) and aldehyde components in the vinasse are reduced when the high boiling point fractions is removed.

Example 5— Reconstitution of Polyphenol Composition for Spraying

[0411] The spray dried digestate of Example 2 (optionally processed in accordance with Examples 3 and/or 4) can be reconstituted into a liquid—for example after transport to the facility for preparation of a polyphenol enhanced sugar. The polyphenol composition spray dried powder (optionally 10,000 GAE mg/100 g but varies batch to batch) was combined with affination rehydration syrup of 40° Brix.

[0412] The rehydration syrup has no polyphenols. The syrup and powder were combined at a 10 g/g ratio and sprayed as a liquid onto a base sugar (optionally GAE mg/100 g but varies batch to batch).

[0413] The skilled person is able to calculate the amount of liquid composition to spray onto the base sugar to achieve a desired polyphenol composition in a final sugar.

[0414] The reconstituted polyphenol composition liquid generally contains:

TABLE-US-00012 Chemical Name Proportion Protein <3% w/v Ash <5% w/v Carbohydrates <3% w/v Water 80-90% w/v

Example 5a— Preparation of Spray Dried Reduced Odour Filtrate

[0415] Protocol 1

[0416] 120-150 L raw sugar cane vinasse digestate of 5-8 brix is pre-filtered and then filtered as described in Example 3a. The digestate is evaporated to about 40 brix and then spray dried to powder form.

[0417] Protocol 2

[0418] Raw sugar cane vinasse digestate of 5-8 brix is evaporated to about 40 brix. This concentrate is diluted back to 10 brix and about 120 L, pre-filtered and then filtered as described in Example 3a. The 100-120 L of filtered digestate is then spray dried to powder form.

Example 6— Effect of Polyphenols on GI of Sugar

[0419] The effect of polyphenol content on the GI of sugar was studied. Traditional white sugar (ie essentially sucrose) was used as a control. Sugars with varied quantities of polyphenols were prepared by adding various amounts of polyphenol content to traditional white sugar.

[0420] Table 4 shows the results of testing of an in vitro Glycemic Index Speed Test (GIST) on the sugars prepared. The method involved in vitro digestion and analysis using Bruker BBFO 400 MHz NMR Spectroscopy. The testing was conducted by the Singapore Polytechnic Food Innovation & Resource Centre, who have demonstrated a strong correlation between the results of their in vitro method and traditional in vivo GI testing.

TABLE-US-00013 TABLE 4 Sugar polyphenol content v GI Sample Polyphenol content GI number GI 1 0 mg CE polyphenols/ About 68 Medium 100 g carbohydrate 2 30 mg CE polyphenols/ <55 (about 53) Low 100 g carbohydrate 3 60 mg CE <20 (about 15) Very Low polyphenols/100 g carbohydrate

[0421] While the GI of fructose is 19, the GI of glucose is 100 out of 100. We therefore expect that the as glucose increases in less refined sugars the glycemic response also concurrently increases.

[0422] A second set of sugars were prepared in which reducing sugars (1:1 glucose to fructose) were added to some of the white refined sugar plus polyphenol sugars. The GI of these sugars was also tested using the GIST method and the results are in Table 5.

TABLE-US-00014 TABLE 5 Effect of polyphenol and reducing sugar content on GI Sample # Name of Material/Sample Sample Code GI Banding 1 Sugar + 30 mg CE PP/100 g GI103 Low carbohydrate + <0.16% RS 2 Sugar + 30 mg CE PP/100 g GI104 Medium carbohydrate + 0.3% RS 3 Sugar + 30 mg CE PP/100 g GI105 Medium/High carbohydrate + 0.6% RS (about 70) 4 Sugar + 60 mg CE PP/100 g GI106 Very low carbohydrate + 0% RS (about 15) 5 Sugar + 60 mg CE PP/100 g GI107 Low (about 29) carbohydrate + 0.6% RS *PP = polyphenols; RS = reducing sugars (1:1 glucose:fructose)

[0423] The GI of several samples from Table 3 are graphed in FIG. 3.

Example 7— Analysis of Polyphenol Content in Sugar

[0424] 40 g of sugar sample was accurately weighed into a 100 ml volumetric flask. Approximately 40 ml of distilled water was added and the flask agitated until the sugar was fully dissolved after which the solution was made up to final volume with distilled water. The polyphenol analysis was based on the Folin-Ciocalteu method (Singleton 1965) and was adapted from the work of Kim et al (2003). In brief, a 50 μL aliquot of appropriately diluted raw sugar solution was added to a test tube followed by 650 μL pf distilled water. A 50 μL aliquot of Folin-Ciocalteu reagent was added to the mixture and shaken. After 5 minutes, 500 μL of 7% Na.sub.2CO.sub.3 solution was added with mixing. The absorbance at 750 nm was recorded after 90 minutes at room temperature. A standard curve was constructed using standard solutions of catechin (0-250 mg/L). Sample results were expressed as milligrams of catechin equivalent (CE) per 100 g raw sugar. The absorbance of each sample sugar was determined and the quantity of polyphenols in that sugar determined from the standard curve.

[0425] Where the sugar is a less refined sugar prepared by a limited wash, an alternative method for analysis of the polyphenol content is to measure the amount of tricin in a sample using near-infra red spectroscopy (NIR). In these circumstances, the amount of tricin is proportional to the total polyphenols. Further information on this method is available in Australian Provisional Patent Application No 2016902957 filed on 27 Jul. 2016 with the title “Process for sugar production” or international patent publication number WO 2018/018089.

Example 8— Analysis of the Reducing Sugar Content in Sugar

[0426] There are several qualitative tests that can be used to determine reducing sugar content in a sugar product. Copper (II) ions in either aqueous sodium citrate or in aqueous sodium tartrate can be reacted with the sugar. The reducing sugars convert the copper(II) to copper(I), which forms a copper(I) oxide precipitate that can be quantified.

[0427] An alternative is to react 3,5-dinitrosalicylic acid with the sugar. The reducing sugars will react with this reagent to form 3-amino-5-nitrosalicylic acid. The quantity of 3-amino-5-nitrosalicylic acid can be measured with spectrophotometry and the results used to quantify the amount of reducing sugar present in the sugar product.

Example 9—Preparation of a Sugar Using a Digestate of the Invention for Polyphenol Content

[0428] As discussed above, polyphenol levels in dunder and digestate can vary based upon the source employed. Accordingly, in order to produce a sugar comprising a defined level of polyphenol it is necessary to control the amount of digestate (or PAC treated digestate) added based upon the source employed.

[0429] The amount of digestate required to produce sugars with polyphenol levels of 60 mg GAE/100 g of material were calculated for digestate concentrates (ie digestate filtered and condensed to a desired concentration) featuring 1000 mg GAE/100 g of material. The washed-sugar to which the PAC treated digestate was applied featured 5 mg GAE polyphenols/100 g carbohydrates. The calculated quantities are tabulated below.

[0430] Methods of preparing polyphenol containing sugars are described in PCT/SG2019/050377. The polyphenol compositions of the invention including sugarcane vinasse, sugarcane vinasse digestate and PAC treated sugarcane vinasse digestate polyphenol compositions are suitable for addition to refined white sugar or various incompletely refined sucrose sugars to prepare a sugar as described in that application.

TABLE-US-00015 TABLE 6 Levels of digestate required to produce sugars of defined polyphenol levels Polyphenol level in 1000 mg GAE/100 g 1000 mg GAE/100 g digestate material (ie 10,000 mg material (ie 10,000 mg GAE/Litre) GAE/Litre) Polyphenol level in washed 5 mg GAE/100 g material 10 mg GAE/100 g material starting sugar Polyphenols content target 48 mg GAE/100 g material 48 mg GAE/100 g material Polyphenols needed to 43 mg GAE/100 g material 43 mg GAE/100 g material reach target mL of digestate required to 4.34 g of digestate per 100 2.17 g of digestate per 100 prepare 100 g of a g starting sugar g starting sugar composition containing digestate and washed sugar wherein the polyphenol concentration is about 48 mg GAE/100 g of material

[0431] The skilled person will be able to determine the appropriate quantity based on the quantity of polyphenols in the digestate (or dunder) and the quantity (if any) in the starting sugar. Further, examples are set out in Table 7 below.

TABLE-US-00016 TABLE 7 Grams of polyphenol composition to add to base sugar in various scenarios Polyphenol Grams of Dried Powder Top Up polyphenol Polyphenol Base Sugar Required composition Composition mg GAE/ mg GAE/ required/100 g mg GAE/100 g 100 g 100 g Grams Scenario 1 7900 20 10 0.0013 Scenario 2 7900 10 20 0.0025 Scenario 3 7900 0 30 0.0038 Scenario 4 7900 20 40 0.0051

Example 10— Industrial Application of Polyphenol Composition to a Sugar

[0432] Two different base sugars (partially refined sugars traditionally sent to a refinery for further refining to white sugar) were used as the base for addition of various quantities of various dilutions of various polyphenol compositions. To determine the effect of the additive on the final sugar. The polyphenol compositions were mixed with sugar (alternatively the polyphenol composition can be added into the affination syrup before or after centrifuging) to evenly distribute the polyphenol composition throughout the sugar. The polarity (Pol), reducing sugar content (RS), moisture content, ash content, colour and polyphenol content (GAE) were determined by NIR and for some samples the polyphenol content confirmed by the laboratory method. The characteristics of the base and prepared sugars are in Table 8 and the polyphenol composition, dilution and volume used to prepare each sugar shown in Table 9 below.

TABLE-US-00017 TABLE 8 Characteristics of the base sugar and prepared sugars S1-S24 Lab GAE NIR mg/ NIR Pol RS Moisture NIR Ash Colour NIR GAE 100 g Base sugar 1 99.52 0.13 0.11 0.10 673 20 21 S1 99.33 0.14 0.11 0.13 976 23 24 S2 99.26 0.15 0.13 0.13 1159 23 24 S3 99.05 0.23 0.14 0.17 1197 32 26 S4 99.14 0.22 0.14 0.17 1218 29 27 S5 99.18 0.24 0.13 0.2 1358 35 S6 99.2 0.21 0.13 0.21 1589 33 S7 98.69 0.28 0.15 0.25 2510 47 S8 98.97 0.33 0.12 0.23 2672 38 S9 99.3 0.28 0.14 0.24 1929 36 S10 99.33 0.25 0.11 0.22 1797 32 30 S11 99.16 0.34 0.12 0.2 2180 37 35 S12 99.07 0.34 0.04 0.15 3128 37 54 Base sugar 2 99.35 0.17 0.11 0.11 864 22 23 S17 99.34 0.18 0.09 0.21 1436 24 26 S18 99.38 0.22 0.13 0.24 1982 30 31 S19 99.18 0.26 0.1 0.2 2672 31 39 S20 98.92 0.27 0.11 0.18 3263 37 46 S21 98.88 0.32 0.11 0.16 3684 40 56 S22 98.73 0.33 0.31 0.19 4402 47 S22 99.03 0.28 0.1 0.13 3937 38 61 S23 98.81 0.29 0.07 0.16 3602 41 56 S24 98.65 0.36 0.23 0.31 4176 45 S24 99 0.32 0.06 0.19 3978 86

TABLE-US-00018 TABLE 9 Polyphenol composition, dilution and volume added to the base sugar to prepare sugars S1-S24 ml dilution added to Increase Polyphenol PC GAE Dilution of 100 g base in GAE Composition mg/100 g PC g/ml sugar mg/100 g Base sugar 1 S1 D1 6818 0.2/100 5 4 S2 D1 6818 0.2/100 10 4 S3 D1 6818 0.4/100 5 5 S4 D1 6818 0.4/100 10 6 S5 D1 6818   1/100 5 15 S6 D1 6818   1/100 10 13 S7 D1 6818   7/60 5 27 S8 D2 10789   7/60 2 18 S9 D2 10789   7/60 1 16 S10 D2 10789   7/60 0.5 10 S11 D2 10789   7/60 1.5 15 S12 D2 10789   7/60 2.5 33 Base sugar 2 S17 D2 10789  10/100 0.5 3 S18 D2 10789  10/100 1 8 S19 D2 10789  10/100 2 16 S20 D2 10789  10/100 3 23 S21 D2 10789  10/100 4 33 S22 D2 10789  10/100 5 S22 D2 10789  10/100 5 38 S23 D2 10789  20/100 2.5 33 S24 D2 10789  20/100 5 S24 D2 10789  20/100 5 63

Example 11— Liquid Digestate Testing of Pesticide, Herbicide, Fungicide and Other Levels

[0433] Samples of liquid sugar cane juice vinasse digestate were commercially sourced and provided to the National Measurement Institute in Australia for analysis. The results provided are in Table 10 below.

TABLE-US-00019 TABLE 10 Impurity levels in vinasse Organochlorine (OC) Pesticides Aldrin mg/kg <0.02 BHC-alpha mg/kg <0.02 BHC-beta mg/kg <0.02 BHC-delta mg/kg <0.02 BHC-Total mg/kg <0.02 Chlordane mg/kg <0.02 DDD-o.p. mg/kg <0.02 DDE-o.p. mg/kg <0.02 DDT-o.p. mg/kg <0.02 DDD-p.p. mg/kg <0.02 DDE-p.p. mg/kg <0.02 DDT-p.p. mg/kg <0.02 DDT-total mg/kg <0.02 Dicofol mg/kg <0.02 Dieldrin mg/kg <0.02 Endosulfan-a. mg/kg <0.02 Endosulfan-b. mg/kg <0.02 Endosulfan- mg/kg <0.02 Sulphate Endosulfan- mg/kg <0.02 Total Endrin mg/kg <0.05 HCB mg/kg <0.01 Heptachlor mg/kg <0.01 Heptachlor- mg/kg <0.01 Epoxide Lindane mg/kg <0.02 Methoxychlor mg/kg <0.05 Nonachlor mg/kg <0.05 Trichlorfon mg/kg <0.02 Organophosphate (OP) Pesticides Acephate mg/kg <0.01 Azinphos ethyl mg/kg <0.02 Temephos mg/kg <0.05 Azinphos methyl mg/kg <0.02 Bromophos ethyl mg/kg <0.05 Carbophenothion mg/kg <0.05 Chlorfenvinphos mg/kg <0.05 Chlorpyrifos mg/kg <0.01 Chlorpyrifos mg/kg <0.02 Chlorthal dimethyl mg/kg <0.02 Coumaphos mg/kg <0.01 Demeton-S-Methyl mg/kg <0.01 Diazinon mg/kg <0.02 Dioxathion mg/kg <0.05 Dichlorvos mg/kg <0.02 Dimethoate mg/kg <0.02 Ethion mg/kg <0.05 Fenamiphos mg/kg <0.01 Fenchlorphos mg/kg <0.05 Fenitrothion mg/kg <0.02 Fenthion mg/kg <0.01 Formothion mg/kg <0.05 Malathion mg/kg <0.02 Methacrifos mg/kg <0.05 Methamidophos mg/kg <0.01 Methidathion mg/kg <0.01 Mevinphos mg/kg <0.02 Monocrotophos mg/kg <0.01 Omethoate mg/kg <0.02 Parathion ethyl mg/kg <0.02 Parathion methyl mg/kg <0.02 Phorate mg/kg <0.02 Phosalone mg/kg <0.05 Phosmet mg/kg <0.02 Phosphamidon mg/kg <0.05 Pirimiphos methyl mg/kg <0.02 Profenofos mg/kg <0.05 Prothiofos mg/kg <0.01 Terbufos mg/kg <0.05 Triazophos mg/kg <0.05 Fungicides Benalaxyl mg/kg <0.01 Bistertanol mg/kg <0.05 Boscalid mg/kg <0.01 Captan mg/kg <0.05 Chlorothalonil mg/kg <0.01 Cyproconazole mg/kg <0.01 Cyprodinil mg/kg <0.01 Diclofluanid mg/kg <0.02 Dicloran mg/kg <0.02 Difenoconazole mg/kg <0.01 Dimethomorph mg/kg <0.01 Diphenylamine mg/kg <0.02 Epoxiconazole mg/kg <0.01 Fenarimol mg/kg <0.05 Fludioxonil mg/kg <0.02 Fenpyrazamine mg/kg <0.01 Flusilazole mg/kg <0.01 Hexaconazole mg/kg <0.05 Imazalil mg/kg <0.05 Iprodione mg/kg <0.02 Kresoxim methyl mg/kg <0.02 Mandipropamid mg/kg <0.01 Metalaxyl mg/kg <0.01 Metrafenone mg/kg <0.01 Myclobutanil mg/kg <0.02 Oxadixy 1 mg/kg <0.05 Oxycarboxin mg/kg <0.05 Pacloburtrazol mg/kg <0.01 Penconazole mg/kg <0.02 Piperonyl butoxide mg/kg <0.01 Prochloraz mg/kg <0.05 Procymidone mg/kg <0.02 Propamocarb mg/kg <0.02 Propiconazole mg/kg <0.01 Pyraclostrobin mg/kg <0.01 Pyrimethanil mg/kg <0.01 Quintozene mg/kg <0.01 Tebuconazole mg/kg <0.01 Tolclophos methyl mg/kg <0.01 Tolylfluanid mg/kg <0.05 Triadimefon mg/kg <0.05 Triaimenol mg/kg <0.01 Vinclozolin mg/kg <0.02 Herbicides Atrazine mg/kg <0.01 Bromacil mg/kg <0.01 Carfentrazone Ethyl mg/kg <0.02 Ethofumesate mg/kg <0.01 Isoxaben mg/kg <0.01 Linuron mg/kg <0.01 Methabenzthiazuron mg/kg <0.01 Metolachlor mg/kg <0.01 Metribuzin mg/kg <0.01 Molinate mg/kg <0.05 Oxyfluorfen mg/kg <0.01 Napropamide mg/kg <0.01 Norflurazon mg/kg <0.01 Pendimethalin mg/kg <0.01 Propachlor mg/kg <0.02 Trifluralin mg/kg <0.01 Carbamates Methiocarb mg/kg <0.05 Acaricides Buprofezin mg/kg <0.01 Propyzamide mg/kg <0.01 Simazine mg/kg <0.01 Clofentezine mg/kg <0.01 Disulphoton mg/kg <0.01 Etoxazole mg/kg <0.01 Hexythiazox mg/kg <0.01 Propargite mg/kg <0.01 Tebufenpyrad mg/kg <0.02 Tetradifon mg/kg <0.02 Others Chlorfenapyr mg/kg <0.02 Phenols O-Phenyphenol mg/kg <0.02 Carbamates Aldicarb (incl sulfoxide mg/kg <0.01 & sulfonem) Carbaryl mg/kg <0.01 Pirimicarb mg/kg <0.02 Synthetic Pyrethroids Bifenthrin mg/kg <0.01 Bioresmethrin mg/kg <0.02 Cyfluthrin-b. mg/kg <0.01 Cyfluthrin mg/kg <0.01 Cyhalothrin-1. mg/kg <0.01 Cyhalothrin mg/kg <0.01 Cypermethrin mg/kg <0.01 Deltamethrin mg/kg <0.02 Esfenvalerate mg/kg <0.01 Fenvalerate mg/kg <0.01 Fluvalinate mg/kg <0.01 tau-Fluvalinate mg/kg <0.02 Permethrin mg/kg <0.02 Phenothrin mg/kg <0.02 Pyrethrins mg/kg <0.02 Insecticide Acetamiprid mg/kg <0.01 Fipronil mg/kg <0.01 Chlorantaniliprole mg/kg <0.01 Clothianidin mg/kg <0.01 Emamectin mg/kg <0.01 Fenoxycarb mg/kg <0.02 Flubendiamide mg/kg <0.02 Indoxacarb mg/kg <0.01 Methoxyfenozide mg/kg <0.02 Novaluron mg/kg <0.05 Py ri proxyfen mg/kg <0.01 Spinetoram mg/kg <0.01 Spirotetramat mg/kg <0.01 Thiamethoxam mg/kg <0.01 C5 Residues Azoxystrobin mg/kg <0.01 Vamidothion mg/kg <0.05 Benomyl mg/kg <0.05 Benzyladenine mg/kg <0.0005 Carbendazim mg/kg <0.01 Diuron mg/kg mg/kg <0.05 Fenhexamid mg/kg <0.02 Fenpyroximate mg/kg <0.05 Imidacloprid mg/kg <0.01 Methomyl mg/kg <0.02 Pymetrozine mg/kg <0.01 Soinosad mg/kg <0.01 Tebufenozide mg/kg <0.02 Thiabendazole mg/kg <0.01 Thiacloprid mg/kg <0.02 Trifloxystrobin mg/kg <0.01 Trace Elements Antimony mg/kg 0.021 Arsenic mg/kg 0.18 Cadmium mg/kg <0.01 Copper mg/kg 0.39 Lead mg/kg 0.055 Mercury mg/kg <0.01 Selenium mg/kg 0.099 Tin mg/kg 0.011 Zinc mg/kg 2.1