Method for preparing a colorant composition with improved stability and packaging and / or storage method thereof

Abstract

This disclosure provides a colorant composition comprising N-glutamyl monascorubraminic acid with a reduced generation of the trans-form, by protecting the compound from light. The disclosure also provides products comprising N-glutamyl monascorubraminic acid which are particularly stable to degradation by protecting them from light, as well as methods related thereto.

Description

METHOD FOR PREPARING A COLORANT COMPOSITION WITH IMPROVEDSTABILITY AND PACKAGING AND / OR STORAGE METHOD THEREOFFIELD OF THE INVENTION

[0001] The present invention is generally related to forms of handling and storing colorant compositions to improve their stability. In particular, the invention is related to forms of handling and storing colorant compositions comprising a red pigment to improve the stability of said pigment. More particularly, the invention is related to forms of handling and storing colorant compositions comprising a red pigment obtained from fungi of the Talaromyces genus, even more particularly from Talaromyces atroroseus, to improve the stability of said pigment.BACKGROUND

[0002] The filamentous fungi of the Talaromyces genus are known to produce a variety of pigments, which are azaphilone compounds and include red, orange, and yellow pigments. These pigments, due to being obtained from a natural source, are excellent candidates for preparing natural colorants, which are increasingly being used in a variety of products as a substitute for traditional synthetic colorants.

[0003] In particular, Talaromyces are known to produce a family of red pigments derived from monascorubraminic acid, which incorporate amino acids into their structure, and may generally be referred to as monascus pigments. The different pigments of this family differ between each other according to the incorporated amino acid and they exist as cis- or trans- forms due to a C-C double bond bridging the azaphilone core to the carboxyl group of the base monascorubraminic acid. Of these isomeric forms, the cis- form is the main compound, while the trans- form is considered a byproduct.

[0004] Among these pigments, the one which incorporates glutamyl (N-glutamyl monascorubraminic acid, or TarE) is of particular commercial interest due to its red hue. However, known methods for the production of these pigments, and particularly of TarE, tend to generate significative amounts of the / ra / 7.s-form. Since these compounds have potential as colorants for industries such as the food, pharmaceutical and cosmetic industries, which have strict purity requirements for the compounds used therein, it is of paramount importance to develop methods for their production which reduce the / raw.s-isomerization of the corresponding product.SUMMARY OF THE INVENTION

[0005] It is therefore an aspect of this invention to provide a packaged colorant composition comprising: i) a compound of Formula (I)(I); and ii) a packaging component, wherein the packaging component prevents or reduces the transmission of light.

[0006] In an embodiment of the invention, the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm.

[0007] In an embodiment of the invention, the packaged colorant composition comprises less than 10%, preferably less than 5%, of a compound of Formula (II)(II); wherein the percentage is determined by relative peak area at 520 nm absorbance.

[0008] In an embodiment of the invention, the packaged colorant composition maintains less than 10%, preferably less than 5% of the compound of Formula (II) over a 6-month storage period.

[0009] In an embodiment of the invention, the packaging component reduces the transmission of light or a portion of the wavelength spectrum thereof by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98%.

[0010] In another embodiment of the invention, the packaging component reduces the transmission of light below 500 lux, 400 lux, 200 lux, 100 lux, 75 lux, 50 lux, 25 lux, 10 lux or 5 lux.

[0011] It is another aspect of the present invention to provide a packaged colored end product comprising: i) a colored end product comprising the compound of Formula (I); and ii) a packaging component, wherein the packaging component prevents or reduces the transmission of light.

[0012] In an embodiment of this aspect of the invention, the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm.

[0013] In an embodiment of this aspect of the invention, the colored end product is selected from a foodstuff, a pharmaceutical product, and a cosmetic product.

[0014] It is another aspect of the invention to provide a method for improving the color stability of a colorant composition or a colored end product comprising the compound of Formula (I), themethod comprising handling and storing the colorant composition or the colored end product under protection from light.

[0015] In an embodiment of this aspect of the invention, handling and storing the colorant composition or the colored end product under protection from light comprises packaging the colorant composition or the colored end product with a packaging component which prevents or reduces the transmission of light. Preferably, the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm. In a preferred embodiment, the packaging component reduces the transmission of light or a portion of the wavelength spectrum thereof by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98%. In another preferred embodiment, the packaging component reduces the transmission of light below 500 lux, 400 lux, 200 lux, 100 lux, 75 lux, 50 lux, 25 lux, 10 lux or 5 lux.

[0016] In an embodiment of this aspect of the invention, the hue value of the colorant composition or the colored end product experiences a decrease not greater than 10% over a 6-month storage period.

[0017] A method for producing a colorant composition comprising the compound of Formula (I), the method comprising i) culturing a fungus of the Talaromyces genus in a culture broth in conditions such that the compound of Formula (I) is produced; ii) extracting the compound of Formula (I) produced in step i) from the culture broth, thus obtaining a colorant composition; and iii) packaging the colorant composition with a packaging component which prevents or reduces the transmission of light.

[0018] In an embodiment of this aspect of the invention, the fungus of the Talaromyces genus isT. atroroseus.

[0019] In an embodiment of this aspect of the invention, the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm.

[0020] In an embodiment of this aspect of the invention, the packaging component reduces the transmission of light or a portion of the wavelength spectrum thereof by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98%.

[0021] In another embodiment of this aspect of the invention, the packaging component reduces the transmission of light below 500 lux, 400 lux, 200 lux, 100 lux, 75 lux, 50 lux, 25 lux, 10 lux or 5 lux.BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIG. 1. (A) UV-VIS spectrum of c / .s-TarE. (B) UV-VIS spectrum of / raw.s-TarE.

[0023] FIG. 2. HPLC chromatogram showing the transformation of c / .s-TarE to / raw.s-TarE due to exposure to light at 1000 MW / m2over 3 hours.

[0024] FIG. 3. TLC result for a sample being exposed to light at 1000 MW / m2over 3 hours. The red stain (a) corresponds to c / .s-TarE, while the purple stain (b) corresponds to / raw.s-TarE.

[0025] FIG. 4. Hue variation of a solution of c / .s-TarE over time due to light exposure.

[0026] FIG 5. Effect of different treatments of light exposure over: (A) The El%; (B) the Hue;(C) the % of c / .s-TarE and (D) the % of / ra / 7.s-TarE.DETAILED DESCRIPTION OF THE INVENTION

[0027] The present invention is based on the surprising discovery made by the inventors that the generation of / ra / 7.s-TarE during the production and storage of c / .s-TarE is due to or increased by the exposure of TarE to light, and that moreover, the generation of / ra / 7.s-TarE has a significant impact on the color and thus the utility of the compound.

[0028] Surprisingly and unexpectedly, the production of / raw.s-TarE generates a perceptible color change in the colorant, shifting its hue towards purple while also decreasing its coloring power. Even though some compounds are known to be susceptible to light, there is no teaching in the state of the art that suggests that this is the case for c / .s-TarE, much less that the exposure to light of cv.s-TarE results in the generation of the / ra / v.s-form or any change to the hue or coloring power of the compound. The shift in the properties of the colorant with the generation of the / ra / v.s-form is a non-acceptable outcome generating a problem which the present invention solves in a surprising manner by a cost- effective modification to the methods previously known in the art.

[0029] Correspondingly, it is an aspect of the present invention to provide a packaged colorant composition comprising: i) a compound of Formula (I)(I); and ii) a packaging component, wherein the packaging component prevents or reduces the transmission of light.

[0030] The compound of Formula (I) is the cis- form of N-glutamyl monascorubraminic acid and corresponds to the pigment of the family of monascorubraminic acids which is formed when glutamate is incorporated into the base cv.s-monascorubraminic acid structure. Throughout this description, N- glutamyl monascorubraminic acid may be referred to interchangeably as TarE. Correspondingly, the compound of Formula (I) may be referred to as cv.s-TarE herein.

[0031] The term “colorant composition” is to be understood as referring to a composition which may be readily used as a colorant, such as for coloring a product. More particularly, the term “packaged colorant composition” is to be understood as referring to a colorant composition that is contained within another tangible object, such as a packaged item intended for commercialization, distribution and / or storage.

[0032] The packaged colorant composition of the invention comprises the compound of Formula (I), and may comprise other colored components. The concentrations of colored components in the packaged colorant composition may be ascertained by HPLC with UV / Vis detection at 520 nm, by comparing the peak area observed for each compound, thus expressing the concentrations as percentages determined by relative peak area at 520 nm. By “relative peak area at 520 nm absorbance” it should be understood that the recited percentage for a specific component X corresponds to the value obtained by the following expression:wherein each peak area is obtained from subjecting the packaged colorant composition of the invention to a HPLC analysis in conditions upon which a proper separation of the colored components thereof may be achieved, detecting the compounds with UV / Vis detection at 520 nm. Throughout this description, the concentrations of the colored components which may be present in the packaged colorant composition of the invention will be expressed as relative peak area at 520 nm, unless otherwise is specifically pointed out.

[0033] Correspondingly, in an embodiment, the packaged composition of the invention comprises TarE at a concentration of at least 70%, preferably from 70% to 80%. In some cases, other minor components exhibiting light absorption at 520 nm may be present in the composition, such as the compound of Formula (II) (also referred to throughout this description as / raw.s-TarE),.Formula (II) - / raw.s-TarE

[0034] In an embodiment of the invention, the packaged colorant composition comprises less than10%, preferably less than 5%, of / raw.s-TarE. Due to the protection from light generated by the packaging component, the isomerization of c / .s-TarE to / raw.s-TarE is inhibited, allowing the composition to maintain a low concentration of / raw.s-TarE during its handling and storage. Correspondingly, in an embodiment of the invention, the packaged colorant composition maintains less than 10%, preferably less than 5% of the compound of Formula (II) over a 6-month storage period.

[0035] Other colored components which may be present in the packaged colorant composition of the invention as minor byproducts of the production of c / .s-TarE are, for instance, N-glutamyl monascorubramine, N-glutaminyl monascorubraminic acid, and compounds related thereto. These additional colored components taken together may be present in the packaged colorant composition in a concentration not higher than 10%, preferably not higher than 5%.

[0036] The packaged colorant composition may be in solid form (e.g., as a colored powder) or liquid form (e.g., as a colored solution or suspension). If the composition is in liquid form, it may further comprise a suitable carrier for the compound of Formula (I) to be dissolved or suspended in, and which allows for a proper coloring of the intended product. For instance, such a carrier may be water.

[0037] The packaged colorant composition may further comprise at least one additional colored compound other than the compound of Formula (I), to modify the color hue of the composition asneeded. For example, in some cases a colorant of a blue or yellow hue may be mixed with the packaged colorant composition herein to form additional colorants.

[0038] The packaging component of the packaged colorant composition of the invention may be any packaging means which is suitable for containing the colorant composition, as long as it properly prevents or reduces the transmission of light. For instance, the packaging component may be selected from a container, a vial, a wrapper, a foil, a box, a bin, a box, a protective covering, a canister, a drum or a case. The material of the packaging component may also be selected from any suitable material, including but not limited to plastic, glass, metal, paper, wood, cardboard, or foil.

[0039] The packaging component does not necessarily block the transmission of light completely, as long as it ensures that a properly low amount of / raw.s-TarE is generated during the storage of the packaged colorant composition. For instance, the packaging component may prevent the transmission of light of a particular wavelength range, or may prevent the transmission of a particular percentage of the light the packaged colorant composition is exposed to. In a particular embodiment, the packaging component prevents the transmission of light having a wavelength between 290-800 nm. In another particular embodiment, the packaging component blocks at least 50% of the light, preferably at least 75% of the light, that the packaged colorant composition is exposed to, as measured by, for instance, the comparative light intensity measured by a suitable detector placed outside and inside the corresponding packaging component. In another particular embodiment, the packaging component reduces the transmission of light or a portion of the wavelength spectrum thereof by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98%. In yet another embodiment of the invention, the packaging component reduces the transmission of light below 500 lux, 400 lux, 200 lux, 100 lux, 75 lux, 50 lux, 25 lux, 10 lux or 5 lux.

[0040] The compound of Formula (I) is suitable for its use as a colorant in several industries. It is known that fungus of the Talaromyces genus do not generate mycotoxins such as citrinin, which isproduced by fungus of the Monascus genus. This fact, combined with the fact that the packaged colorant composition according to the present invention is able to maintain low amounts of / raw.s-TarE during handling and storage, makes the packaged colorant composition of the present invention particularly suitable for industries related to products for human use and / or consumption, such as the food, pharmaceutical and cosmetic industries.

[0041] Correspondingly, it is yet another aspect of the present invention to provide a packaged colored end product comprising: i) a colored end product comprising the compound of Formula (I); and ii) a packaging component, wherein the packaging component prevents or reduces the transmission of light.

[0042] The term “colored end product” is to be understood throughout this description as referring to a product colored by the presence of a compound of Formula (I) therein or by the application of the packaged colored composition of the invention thereto, which is ready for human use and / or consumption.

[0043] As mentioned above, the colored end product may be any end product feasible to be colored by the compound of Formula (I). In an embodiment of this aspect of the invention, the colored end product is selected from a foodstuff, a pharmaceutical product, and a cosmetic product. Correspondingly, the colored end product may further comprise additional components depending on the type of product and the use thereof.

[0044] The packaging component of the colored end product according to this aspect of the invention may share the same characteristics with the packaging component of the packaged colorant composition described above.

[0045] By protecting the colored end product described above from light with the packaging component, the amount of / raw.s-TarE generated is kept low during transportation, distribution andstorage thereof. In an embodiment of this aspect of the invention, the packaged colored end product maintains less than 10%, or even less than 5%, of the compound of Formula (II) over a 6-month storage period.

[0046] As mentioned above, the present inventors have surprisingly found that the generation of / raw.s-TarE in a colorant composition due to its exposure to light causes a shift in the color hue of the composition, since / raw.s-TarE has a purple color, different from the red color of cv.s-TarE.

[0047] Correspondingly, it is yet another aspect of the present invention to provide a method for improving the color stability of a colorant composition or a colored end product comprising the compound of Formula (I), the method comprising handling and storing the colorant composition or the colored end product under protection from light.

[0048] The term “color stability” is to be understood as the capability of a colorant composition or a colored end product to maintain their color hue and coloring power unaltered in time. Since there is no suggestion whatsoever in the state of the art about the susceptibility of cv.s-TarE to light, and particularly that the exposure of cv.s-TarE to light could result in the trans- isomerization thereof, with / raw.s-TarE having a purple color, protecting a colorant composition or a colored end product comprising the compound of Formula (I) from light has the surprising effect of minimizing any color change during the handling and storage of said colorant composition or colored end product, thus improving their color stability. The color stability of the colorant composition or colored end product may be measured by techniques known to the person of skill in the art, such as measuring the CIELAB parameters thereof with an appropriate spectrometer, which may then be used to calculate the hue value. The isomerization of cv.s-TarE to / ra / v.s-TarE generates a decrease in the hue value of the colorant. In an embodiment of this aspect of the invention, the hue value of the colorant composition or the colored end product experiences a decrease not greater than 10% over a 6-month storage period.

[0049] In an embodiment of this aspect of the invention, handling and storing the colorant composition or the colored end product under protection from light comprises packaging the colorant composition or the colored end product with a packaging component which prevents or reduces (i.e., significantly reduces) the transmission of light or certain wavelengths thereof. The packaging component used to this end may be a container, a vial, a wrapper, a foil, a box, a bin, a box, a protective covering, a canister, a drum or a case. The material of the packaging component may also be selected from any suitable material, including but not limited to plastic, glass, metal, paper, wood, cardboard, or foil.

[0050] As mentioned above in the present description, c / .s-TarE is generated by culturing a fungus of the Talaromyces genus.

[0051] Correspondingly, it is yet another aspect of this invention to provide a method for producing a colorant composition comprising the compound of Formula (I), the method comprising: i) culturing a fungus of the Talaromyces genus in a culture broth in conditions such that the compound of Formula (I) is produced; ii) extracting the compound of Formula (I) produced in step i) from the culture broth, thus obtaining a colorant composition; and iii) packaging the colorant composition with a packaging component which prevents or reduces the transmission of light.

[0052] Conditions upon which a compound of Formula (I) as described herein may be produced as required in step i) of the method of this aspect of the invention are known in the state of the art. In general, a fungus belonging to the Talaromyces genus must be cultured in an appropriate broth in the presence of L-glutamic acid, or a compound related thereto, such as monosodium glutamate, as a nitrogen source.

[0053] Step i) of the method may involve culturing any fungus species of the Talaromyces genus capable of producing cv.s-TarE. For instance, the fungus of the Talaromyces genus cultured in step i) of the method may be selected from T. albobiverticillius, T. amestolkiae, T. apiculatus, T. assiutensis, T. atroroseus, T. aurantiacus, T. austrocalifomicus, T. bacillisporus, T. barcinensis, T. boninensis, T. brunneus, T. calidicanius, T. cecidicola, T. coalescens, T. convolutus, T. dendriticus, T. derxii, T. duclauxii, T. echinosporus, T. emodensis, T. erythromellis, T. euchlorocarpius, T. flavus, T. juniculosus, T. galapagensis, T. hachijoensis, T. helicus, T. indigoticus, T. intermedins, T. islandicus, T. lagunensis, T. leycettanus, T. loliensis, T. luteus, T. macrosporus, T. malagensis, T. mameffei, T. mimosinus, T. minioluteus, T. muroii, T. palmae, T. panamensis, T. paucisporus, T. phialosporus, T. piceus, T. pinophilus, T. pittii, T. primulinus, T. proteolyticus, T. pseudostromaticus, T. purpureus, T. purpurogenus, T. rademirici, T. radicus, T. ramulosus, T. retardatus, T. rotundus, T. ruber, T. rubicundus, T. rugulosus, T. ryukyuensis, T. sabulosus, T. siamensis, T. stipitatus, T. stollii, T. subinflatus, T. sublevisporus, T. tardifaciens, T. thermocitrinus, T. trachyspermus, T. ucrainicus, T. udagawae, T. unicus, T. variabilis, T. varians, T. verruculosus, T. viridis, T. viridulus, or T. wortmannii. Preferably, the fungus of the Talaromyces genus cultured in step i) of the method is T. atroroseus.

[0054] Step ii) of the method comprises the extraction of the compound of Formula (I) produced in step i) of the method, so as to obtain a colorant composition comprising the compound of Formula (I).

[0055] The extraction process of the produced compound of Formula (I) may comprise: (a) eliminating the biomass of the Talaromyces fungus used for fermentation; (b) eliminating soluble fermentation secondary products; and (c) precipitating the compound of Formula (I). These sub-steps may be carried out by simple precipitation / centrifugation processes, although the present invention also contemplates other means to isolate the compound of Formula (I).

[0056] For instance, the Talaromyces fungus used for fermentation may be eliminated by centrifugation of the fermentation broth after the fermentation process has been completed, separating the supernatant. The supernatant may then be treated to induce the precipitation of soluble fermentation secondary products (such as proteins, peptides and sugars), for instance, by adding ethanol thereto, which may then be removed by centrifugation as well. Once both the fungus and the fermentation secondary products have been removed, the precipitation of the compound of Formula (I) may be achieved by acidification of the medium (for instance, with concentrated sulfuric acid) and an additional centrifugation to assist in the precipitation of the crystals. If necessary, the crystals of the compound of Formula (I) thus obtained may be subjected to a recrystallization to further improve the purity thereof.

[0057] Steps i) and ii) of the method described above may be carried out in metal reactors and piping when production is performed at an industrial scale and thus protected from light during its production and isolation. Steps i) and ii) of the method described above may be carried out in light- protected containers, such as covered flasks and handling steps in reduced light or dark to protect the composition from light.

[0058] Step iii) of the method of this aspect of the invention involves packaging the colorant composition with a packaging component which prevents or reduces the transmission of light, so as to maintain the compound of Formula (I) protected from light to avoid the trans- isomerization thereof. The packaging component used to this end be a container, a vial, a wrapper, a foil, a box, a bin, a box, a protective covering, a canister, a drum or a case. The material of the packaging component may also be selected from any suitable material, including but not limited to plastic, glass, metal, paper, wood, cardboard, or foil. The colorant composition produced by the method may be packaged in a solid form, such as a dried form or powder, or for example, as a slurry or liquid, contained in a package or covered by a protective covering. In addition, the packaged colorant composition or packaged endproduct can be further stored way from a light source, such as stored in a cabinet, secondary container or storage location which prevent or reduces the exposure to light or certain wavelengths thereof.

[0059] The use of an appropriate packaging as indicated in step iii) of the method of this aspect of the invention minimizes the generation of / raw.s-TarE, since said generation is more likely to take place during storage of the colorant composition if it is not properly protected from light. Since there is no suggestion whatsoever in the state of the art about the susceptibility of c / .s-TarE to light, and particularly that the exposure of c / .s-TarE to light could generate the trans- isomerization thereof and thus have a significant unwanted effect on hue and / or coloring power, this feature of the invention has the surprising effect of minimizing the generation of / raw.s-TarE during the storage of the colorant composition comprising the compound of Formula (I) and protecting the desired red hue of the colorant composition.EXAMPLES

[0060] The invention will now be further described based on the following examples. It is to be understood that these examples are intended for illustrative purposes only, and by no means should be construed to be limiting the scope of the invention, which is only defined by the appended claims.Example 1: Production of c / s-TarE

[0061] T. atroroseus conidia were used to inoculate a broth prepared with the following composition per liter: 20 g of glucose, 3 g of sodium nitrate, 2 g of yeast extract, 0.5 g of potassium chloride, 0.5 g of magnesium sulfate heptahydrate, 0.5 g of dipotassium acid phosphate and 10 mg of ferrous sulfate heptahydrate. The pH was adjusted to 5. 50 mb of culture media were inoculated at 2x105conidia per milliliter.

[0062] Flasks were incubated at 30°C and 200 rpm for 48 hours. The seed culture was transferred at 1% to a 2L-bioreactor with color-producing broth with the following composition per liter: 30 g ofstarch, 10 g of glucose, 8.35 g of monosodium glutamate,! g of yeast extract, 5 g of magnesium sulfate,0.5 g of potassium chloride and 40 mg of ferrous sulfate heptahydrate.

[0063] The strain was incubated at 30 °C with aeration of 2 L / min, stirring at 600 rpm and pH was not controlled until it fell below 3.7. After starch depletion, pH was adjusted back to 5 with 4 M sodium hydroxide.

[0064] After the fermentation process (about 70 hours), the culture broth was centrifuged for 10 min at 3000 g to separate the supernatant. 3 volumes of ethanol were added followed by 16 hours incubation at 4°C to induce the precipitation of proteins, peptides, and sugars, which were removed by centrifugation. Concentrated sulfuric acid was added to lower the pH up to 2.5, and the sample was stored at 4°C for 16 hours before centrifugation for 10 min at 4.000 rpm to help precipitate the formed crystals. Potassium hydroxide was used to resuspend the crystals bringing the pH up to 5 or above. The final resuspension was dried by heat or spray drying to obtain cv.s-TarE.Example 2: Effect of light exposure on cvs-TarE

[0065] A 0.1% solution of cv.s-TarE obtained in Example 1 was made by diluting 50 mg of the final powder in 50 ml of water. Aliquots of the solution were exposed to light at 1000 MW / m2inside a Solarbox 1500 (wavelength 290-800) for 30 minutes, 1 hour, 2 hours and 3 hours. Following exposure, the samples were subjected to HPLC and monitored at 520bnm to assess for the presence of the cis- double bond of cv.s-TarE and its transformation to the trans- form.

[0066] HPLC: The following protocol describes the methodology used for identification, assay and purity evaluation of cv.s-TarE. The samples were analyzed using a Thermo Scientific Vanquish Core HPLC system consisting of an autosampler organizer, column manager and heater, quaternary pump and PDA absorbance detector. Enterprise Chromel eon 7.3 software was used for system control and data acquisition. The column used was Column Poroshell 120 Phenyl-Hexyl 150 mm x 2.1 mm, 2.7 pm.

[0067] Instrumental conditions include: Flow: 0.35 mL / min; Detection: 520 nm; Column temperature: 50 °C; Autosampler temperature: 15 °C; Injection volume: 5 pL (loop volume 20 pL); Run time: 33 minutes; Needle wash solvent: MeOH:H2O (20:80). The gradient described in Table 1 was used.Table 1: HPLC solvent gradient

[0068] The mobile phase and solvent solution consist of: Mobile Phase A: 0.1% formic acid, filter;Mobile Phase B: 0.1% formic acid in acetonitrile, filter; Solvent solution (SS): MeOH:H2O (9:1)

[0069] c / .s-TarE and / raw.s-TarE were identified by comparison of the retention times of the peaks in the chromatogram at 520 nm with standard solutions. The two isomers (structures shown below) can be distinguished based on their absorption spectra (FIG. 1).c / .s-TarE / raw.s-TarE

[0070] As shown in FIG. 2, as the time of light exposure increased, the amount of the cis- form decreased and the amount of trans- form increased.

[0071] Surprisingly, the appearance of the trans- form also caused a shift in the colorant hue from red to purple. This can be seen in a color comparison of the samples exposed to light. 3 hours treatment samples were loaded into silica gel 60 plates, and developed with a solvent system comprisingbutanol:water:acetic acid 12:5:3. The appearance of the purple hue also was apparent by TLC (FIG. 3). The hue value of the colorant shifted proportionally over time (FIG. 4), showing a decrease of about 50% over the 3 hours (roughly equivalent to 5 days light exposure at natural conditions, e.g., if exposed to sunlight or ambient light on a benchtop), and causing a shift of the colorant from red to a more purple color.

[0072] For hue measurements of the samples, a solution of the colorant composition was prepared by diluting the colorant composition in a 50 mM dipotassium phosphate buffer having a pH of about 5.0 in order to obtain an absorbance of about 0.5. The CIELAB parameters of the solution were measured using a spectrophotometer Lovibond PFX-Z that automatically calculates L*, a*, and b*. The chroma value (C) was calculated from a* and b* using the equation [(a*)2+(b*)2]1 / 2. The hue value (co) was calculated from a* and b* using the equation tan-l(b* / a*).Example 3: Effect of light exposure on the compound of Formula (I) in powder and liquid

[0073] The effect of light was determined on c / .s-TarE in powder form and in liquid form. Three samples of 150 mg of powder and three samples of a 10% solution in water were exposed to light for 4 hours using the conditions described in Example 2. Control samples of the colorant powder and solution were incubated in the dark an equivalent amount of time. The amount of cis- and trans- forms was assessed by HPLC as described in Example 2 using 2-3 technical replicates for each sample. The amount of cis- and trans- forms was calculated as a percentage of all compounds in total composition, and the change (delta values) was calculated as a percentage increase or decrease of that percentage representation. Results are shown in Table 2. Light exposure generally resulted in about 100% increase in the trans- form as compared to the starting amount in the sample, whereas the dark incubated samples generally had a much smaller change (Table 2). The effect of light exposure was similar for powder and liquid forms.Table 2: Change in trans- form in light and dark conditions

[0074] Coloring power was also examined for the same samples exposed to light for 4 hours or protected in the dark for an equivalent amount of time. Briefly, for each sample, a colorant solution was prepared with 5 mg of cv.s-TarE added to 5 mL distilled water. Color was measured using a spectrophotometer Lovibond PFX-z . A small volume of the colorant solution was added to a 5 mL cuvette with water, so the initial reading was about value of 0.3 AU. Small amounts of the colorant solution described above, or the 10% solution studied, were added in order to increase absorbance by 0.1 AU. These steps were repeated until 0.7 AU was reached, and all the volumes added were recorded. The increase in absorbance was plotted against the % of colorant of the corresponding dilution and the slope of this plot represents the coloring power. Samples were measured in triplicate and the average values are shown in Table 3. Both the liquid solution and powder exhibited about a 15% decrease in coloring power from the 4-hour incubation as compared to the protected samples, which maintained their coloring power.Table 3: Coloring power of light-exposed and protected samplesExample 4: Stability Assay for the Shell life of the colorant powder

[0075] A stability assay was conducted to evaluate the shelf life of the colorant powder by assessing its photolytic degradation under different light exposure conditions. The experiment aimed to determine the extent of degradation over time using spectrophotometric and chromatographic analyses.

[0076] The pigment was subjected to three different treatments, a) Complete light protection; b) exposed to natural light; and c) accelerated degradation using a SolarBox 1500E solarbox exposition at 480 W / m2(where 1 hour exposure is equal to 21 hours of natural sunlight).

[0077] At different times the colorant powder was used to prepare a 10% solution in distilled water. Stability was monitored by measuring El% (coloring capacity) (see Fig. 5A), CIELAB color parameters (in particular the Hue; see Fig. 5B) and High-Performance Liquid Chromatography (HPLC) to measure the amount of c / .s-TarE and / raw.s-TarE.

[0078] The stability assay showed a conversion of c / .s-TarE to / raw.s-TarE over time when exposed to light (see Fig. 5C and Fig. 5D). This conversion explains the loss of coloring capacity and increase in Hue observed. The results indicate that a decrease of 10% of c / .s-TarE corresponds to an increase of about 6% of / raw.s-TarE, which correlates with an increase of 10% of Hue and a decrease of 15% of coloring capacity.

Claims

CLAIMS1. A packaged colorant composition comprising: i) a compound of Formula (I)(I); and ii) a packaging component, wherein the packaging component prevents or reduces the transmission of light.

2. The packaged colorant composition of claim 1, wherein the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm.

3. The packaged colorant composition of claim 1 or 2, wherein the packaged colorant composition comprises less than 10% of a compound of Formula (II)(II); wherein the percentage is determined by relative peak area at 520 nm absorbance.

4. The packaged colorant composition of claim 3, wherein the packaged colorant composition comprises less than 5% of the compound of Formula (II).

5. The packaged colorant composition of claim 3 or 4, wherein the packaged colorant composition maintains less than 10% of the compound of Formula (II) over a 6-month storage period.

6. The packaged colorant composition of claim 3 or 4, wherein the packaged colorant composition maintains less than 5% of the compound of Formula (II) over a 6-month storage period.

7. The packaged colorant composition of any one of claims 1-6, wherein the packaging component reduces the transmission of light or a portion of the wavelength spectrum thereof by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98%.

8. The packaged colorant composition of any one of claims 1-6, wherein the packaging component reduces the transmission of light below 500 lux, 400 lux, 200 lux, 100 lux, 75 lux, 50 lux, 25 lux, 10 lux or 5 lux.

9. A packaged colored end product comprising: i) a colored end product comprising the compound of Formula (I); and ii) a packaging component, wherein the packaging component prevents or reduces the transmission of light.

10. The packaged colored end product of claim 9, wherein the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm.

11. The packaged colored end product of claim 9 or 10, wherein the colored end product is selected from a foodstuff, a pharmaceutical product, and a cosmetic product.

12. A method for improving the color stability of a colorant composition or a colored end product comprising the compound of Formula (I), the method comprising handling and storing the colorant composition or the colored end product under protection from light.

13. The method of claim 12, wherein handling and storing the colorant composition or the colored end product under protection from light comprises packaging the colorant composition or the colored end product with a packaging component which prevents or reduces the transmission of light.

14. The method of claim 13, wherein the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm.

15. The method of claim 13 or 14, wherein the packaging component reduces the transmission of light or a portion of the wavelength spectrum thereof by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98%.

16. The method of claim 13 or 14, wherein the packaging component reduces the transmission of light below 500 lux, 400 lux, 200 lux, 100 lux, 75 lux, 50 lux, 25 lux, 10 lux or 5 lux.

17. The method of any one of claims 12-16, wherein the hue value of the colorant composition or the colored end product experiences a decrease not greater than 10% over a 6-month storage period.

18. A method for producing a colorant composition comprising the compound of Formula (I), the method comprising: i) culturing a fungus of the Talaromyces genus in a culture broth in conditions such that the compound of Formula (I) is produced; ii) extracting the compound of Formula (I) produced in step i) from the culture broth, thus obtaining a colorant composition; and iii) packaging the colorant composition with a packaging component which prevents or reduces the transmission of light.

19. The method of claim 18, wherein the fungus of the Talaromyces genus is T. atroroseus.

20. The method of claim 18 or 19, wherein the packaging component prevents or reduces the transmission of light having a wavelength between 290-800 nm.

21. The method of any one of claims 18-20, wherein the packaging component reduces the transmission of light or a portion of the wavelength spectrum thereof by at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95% or at least 98%.

22. The method of any one of claims 18-20, wherein the packaging component reduces the transmission of light below 500 lux, 400 lux, 200 lux, 100 lux, 75 lux, 50 lux, 25 lux, 10 lux or 5 lux.

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