Citrullus extraction method and use

Abstract

This invention relates to a zero-waste bioprocess for producing one or more Citrullus extracts from a Citrullus plant. The invention further relates to one or more Citrullus extracts prepared according to the method of the invention and to methods of use of the one or more Citrullus extracts prepared according to the method of the invention including nutritional, medical device, cosmetic and agrobiological applications of use.

Description

[0001] CITRULLUS EXTRACTION METHOD AND USE

[0002] FIELD OF THE INVENTION

[0003] This invention relates to a zero-waste bioprocess for producing one or more Citrullus extracts from a Citrullus plant. The invention further relates to one or more Citrullus extracts prepared according to the method of the invention and to methods of use of the one or more Citrullus extracts prepared according to the method of the invention including nutritional, medical device, cosmetic and agrobiological applications of use.

[0004] BACKGROUND TO THE INVENTION

[0005] Plants from the Citrullus genus have been shown to have various medicinal properties, particularly through the use of processed by-products such as rind and seed of the fruit of the plant. These have been shown to include various essential phytochemicals including saponin, alkaloids, fatty acids, phenolics, citrulline, lycopene, coumarin, and various minerals, among others.

[0006] Citrullus rind and / or seeds have been used in the treatment of conditions including ulcers, diabetes, cardiovascular and respiratory disorders and hypertension as well as various cancers. Citrullus extracts contain high concentrates of L-citrulline, a non- essential amino acid, which is converted by the kidneys into L-arginine and nitric oxide which helps to build protein. Citrullus is therefore also useful for wound treatment as well as treatment of skin diseases and to prevent skin aging through dermatological disorders, environmental abuse (wind, air conditioning, central heating) or through the normal aging process (chronoaging) which may be accelerated by exposure of skin to sun (photoaging).

[0007] However, conventional Citrullus processing methods do not utilise the entire plant and typically result in waste streams comprised of discarded roots, rind, and seed byproducts. Moreover, conventional Citrullus extractions employ aqueous pressing, high-temperature infusions, ion-exchange resins, or supercritical fluids, which can dilute, thermally stress, or require post-processing of actives and are often expensive and complex processes. Furthermore, thermal extraction methods used conventionally to process watermelon causes thermolabile actives (NAD+, L-citrulline) to degrade.

[0008] It would be useful for a simpler, more cost effective and more sustainable processing method that could be used for processing the entire plant including the whole fruit, roots, and also the leaves, stems and flowers, if desired, that would typically be disposed of in waste streams to be developed, including for generation of thermolabile actives such as NAD+, and L-citrulline that would usually be degraded with the use of convention thermal extraction methods. Particularly, it would be useful if such a method integrated full biomass utilization and generated a broad selection of possible value products for use in the fields of nutrition, medical device, cosmetics and agrobiologicals.

[0009] SUMMARY OF THE INVENTION

[0010] According to a first aspect of the invention is provided a method of producing one or more Citrullus plant extract(s) comprising or consisting of the following steps:

[0011] (i) providing harvested Citrullus plant material;

[0012] (ii) washing and optionally surface sanitising the harvested Citrullus plant material;

[0013] (iii) macerating the washed Citrullus plant material to form a slurry;

[0014] (iv) optionally fermenting the slurry;

[0015] (v) optionally subjecting the slurry to freeze-thaw lysis either prior to or after the fermentation step;

[0016] (vi) subjecting the slurry to one or more extraction processes selected from the group comprising: extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration to separate a purified liquid extract from colloidal oleosomes and a fermented solid cake.

[0017] According to a second aspect of the invention is provided a method of producing one or more Citrullus plant extract(s) comprising or consisting of the following steps: (i) providing harvested Citrullus plant material;

[0018] (ii) washing and optionally surface sanitising the harvested Citrullus plant material;

[0019] (iii) macerating the washed Citrullus plant material to form a slurry;

[0020] (iv) fermenting the slurry;

[0021] (v) optionally subjecting the slurry to freeze-thaw lysis either prior to or after the fermentation step;

[0022] (vi) subjecting the slurry to one or more extraction processes selected from the group comprising: extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration to separate a purified liquid extract from colloidal oleosomes and a fermented solid cake.

[0023] According to a third aspect of the invention is provided a method of producing one or more Citrullus plant extract(s) comprising or consisting of the following steps:

[0024] (i) providing harvested Citrullus plant material;

[0025] (ii) washing and optionally surface sanitising the harvested Citrullus plant material;

[0026] (iii) macerating the washed Citrullus plant material to form a slurry;

[0027] (iv) fermenting the slurry;

[0028] (v) subjecting the slurry to freeze-thaw lysis either prior to or after the fermentation step;

[0029] (vi) subjecting the slurry to one or more extraction processes selected from the group comprising: extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration to separate a purified liquid extract from colloidal oleosomes and a fermented solid cake.

[0030] According to a fourth aspect of the invention is provided a method of producing one or more Citrullus plant extract(s) comprising or consisting of the following steps:

[0031] (i) providing harvested Citrullus plant material;

[0032] (ii) washing and optionally surface sanitising the harvested Citrullus plant material;

[0033] (iii) macerating the washed Citrullus plant material to form a slurry; (iv) optionally fermenting the slurry;

[0034] (v) subjecting the slurry to freeze-thaw lysis either prior to or after the fermentation step;

[0035] (vi) subjecting the slurry to one or more extraction processes selected from the group comprising: extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration to separate a purified liquid extract from colloidal oleosomes and a fermented solid cake.

[0036] The Citrullus plant material may be from any one or more plants selected from the Citrullus genus, including Citrullus lanatus, Citrullus colocynthis, and hybrids thereof.

[0037] In one possible embodiment, the method of any one of the first to fourth aspects of the invention is performed with no added water being required to be used, constituting a zero-water process.

[0038] The maceration may be performed by any mechanical maceration technique known to those skilled in the art, more particularly selected from the group comprising chopping or cutting, mincing or milling such as with a hammer mill, or using a high-shear maceration technique including by hydraulic, pneumatic or manual means, or by means of a worm screw. The resultant slurry may be from about 0.1-5 mm in particle size.

[0039] The fermentation may be performed with any one or more sugars or sugar alcohols fermentable by a yeast or bacteria. In a preferred embodiment of the invention, the fermentation may comprise addition of honey (5-30% w / w) and / or food-grade glycerine (5-40% w / w) as well as a fermentation starter culture or endogenous microbiota. The honey may be raw honey. Endogenous microbiota in the context of fermentation refers to the naturally occurring microorganisms — such as bacteria, yeasts, and molds — that are already present in raw materials (i.e. in this case the Citrullus plant material) or the environment. Typically the fermentation is performed for between about 7-14 days at between about 18-30 °C and the pH is controlled at between about 3.0-4.5. The fermentation may further comprise addition of additives or co-substrates to the slurry including aloe, marshmallow, chamomile, licorice, nettle, baobab, citrus zest, seed flours, oats, or enzyme sources such as papain or bromelain, depending on the desired functionality of the final Citrullus plant extract product.

[0040] The step of freeze-thaw lysis may be performed by multiple freezing (for example to between -40 °C to about -80 °C) and thawing cycles to disrupt cellular structures and enhance release of intracellular components. It is to be appreciated that the freezethaw lysis when performed prior to fermentation will enhance NAD+retention and bioactive yield.

[0041] The slurry from step (iii) above, or the purified liquid extract may be subjected to cryopreservation for storage. For example, cryopreservation may be at about -20 °C to -40 °C for about <3 months, thereby to preserve NAD+and L-citrul line.

[0042] The solid cake may be dried and milled to yield a powdered Citrullus plant extract. The drying is preferably performed at < 50 °C. The powdered Citrullus plant extract may be in the form of a flour or protein / fibre concentrate.

[0043] The clarification may comprise addition of bentonite.

[0044] The method may further comprise addition of one or more preservatives to the final Citrullus plant extract product.

[0045] The method may further comprise one or more additional steps selected from the group comprising concentrating the Citrullus plant extract, pasteurising the Citrullus plant extract, and / or homogenising the Citrullus plant extract.

[0046] In one possible embodiment of the first aspect of the invention, the fermentation step is not optional.

[0047] In one possible embodiment of the first aspect of the invention, the freeze-thaw lysis step is not optional.

[0048] In one possible embodiment, the plant material consists of the whole fruit (including the rind, flesh, juice and seeds), <10 % (w / w) of the roots and optionally any one or more of the remaining stems, leaves, and flowers of the Citrullus plant. Inclusion of roots yields elevated osmolyte content including ectoine which may be beneficial for use in barrier-protective formulations comprising the Citrullus plant extract. In an alternative possible embodiment, the plant material consists of the whole Citrullus plant.

[0049] The one or more preservatives may comprise use of any one or more preservatives known to those skilled in the art, but in one possible embodiment comprise the use of a combination of dehydroacetic acid and benzyl alcohol (or Geogard® 221) at about 0.5 - 2 % w / w and / or a combination of phenoxyethanol and ethylhexylglycerin (or PEHG) at about 0.5 - 2 % w / w and / or potassium sorbate at about 0.5-2 % w / w.

[0050] The step of concentrating, if performed, may comprise one or more of freeze drying, spray drying, solvent extraction including with an organic solvent such as alcohol, acetone, chloroform, water or any combination thereof, or centrifugal concentration.

[0051] The step of pasteurising the Citrullus plant extract may be performed using conventional low temperature pasteurisation at between about 63 °C to about 65 °C for about 30 mins.

[0052] The drying and milling step may in one alternative embodiment of the invention comprise the following steps: a. dehydrating the solid cake and fermented solid cake and optionally grinding the dehydrated solid cakes; b. providing a ribbed frusto-conical extraction apparatus comprising: i. a ribbed frusto-conical support having side walls disposed at a 60-degree angle from a base of the support, the side walls further comprising a set of raised grooves spiralling towards the base on an interior surface of the support, the base defining an aperture, and the support being adapted to seat over a vessel, optionally the support further comprising a set of handles; and ii. a frusto-conical filter including a paper filter adapted to line the interior of the frusto-conical support when placed therein; c. placing the frusto-conical filter into the frusto-conical support to form the ribbed frusto-conical extraction apparatus; d. optionally, rinsing the frusto-conical filter in the ribbed frusto-conical extraction apparatus with water having a temperature of from between about 60 to 65°C; e. placing the ribbed frusto-conical extraction apparatus onto a collecting vessel and adding the dehydrated Citrullus plant solid cakes from step a. above to the ribbed frusto-conical extraction apparatus; f. adding water having a temperature of from between about 60 to 65°C to the ribbed frusto-conical extraction apparatus and collecting a purified secondary Citrullus plant liquid extract filtered through the frusto- conical filter in the collecting vessel.

[0053] The ribbed frusto-conical support may comprise internal spiral ribs at a 60° angle base relative to the base of the support for laminar flow extraction.

[0054] The dehydration step a. may comprise drying by means of the sun, a dehydrator, an oven, or a low heat drum coffee roaster such as a PROBAT low heat drum coffee roaster. It is to be appreciated that any low heat means for drying may be similarly used.

[0055] The ribbed frusto-conical support may be comprised of materials selected from the group comprising ceramic, plastic, glass, stainless steel, and copper.

[0056] It is to be appreciated that the size of the aperture in the base of the frusto-conical support may be selected depending on the desired flow rate and brewing time.

[0057] The frusto-conical filter may be Whatman No.1 filter paper.

[0058] The step f. may be performed in one or more steps, wherein if performed in more than one step, an initial volume of water may be added, followed by a waiting period of between about 30 seconds to 1 minute, followed by adding a further volume of water until the desired volume of purified liquid Citrullus plant extract is collected in the collecting vessel. Typically, the ratio for step f. is 1 g of dried Citrullus plant extract to 15 g of water.

[0059] Typically, the total time period for step f. may be between about 3 to about 4 minutes or until no further liquid extract is produced. The method may further comprise a final step g. of concentrating the purified liquid Citrullus plant extract collected after step f. above. The step of concentrating may comprise one or more of freeze drying, spray drying, solvent extraction including with an organic solvent such as alcohol, acetone, chloroform, water or any combination thereof, or centrifugal concentration.

[0060] In one embodiment of the first aspect of the invention, the ribbed frusto-conical extraction apparatus may have up to a five-litre capacity.

[0061] The purified Citrullus plant extract collected after step f. above may further be pasteurized. The step of pasteurising may be performed using conventional low temperature pasteurisation at between about 63 °C to about 65 °C for about 30 mins.

[0062] Additional processing methods may further be applied if required, such as homogenization.

[0063] According to a fifth aspect of the invention, there is provided a Citrullus plant liquid and / or solid extract produced by the method of any one of the first to fourth aspects of the invention. The Citrullus plant liquid and / or solid extract may comprise the following bioactive compounds:

[0064] A. L-citrulline;

[0065] B. quercetin;

[0066] C. NAD+; and

[0067] D. optionally ectoine and / or oleosomes.

[0068] According to a sixth aspect of the invention, there is provided a Citrullus plant liquid and / or solid extract produced by the method of any one of the first to fourth aspects of the invention for use as a bioactive composition, a medical device, a cosmetic composition, a nutritional composition including a nutraceutical, or an agrobiological.

[0069] The term “agrobiological” means a composition that improves plant health, stimulates growth or increases yield or agricultural productivity. The agrobiological composition may further comprise an agriculturally acceptable carrier, for foliar or soil application. The agrobiological composition may be for use to improve plant health, stimulate growth or increase yield or agricultural productivity. In a first possible example of the sixth embodiment of the invention, the bioactive composition may comprise weight / weight (w / w) about 1 - 10 % L-citrulline, about 0.1 - 5 % ectoine, about 0.01 -2 % quercetin, about 1 - 15 % oleosomes, and about 93.93 % ACE inhibitory polysaccharides including pectic polysaccharides, hemicellulosic polysaccharides and neutral polysaccharides.

[0070] In a second possible example of the sixth embodiment of the invention, the nutritional composition may comprise the liquid and / or powdered Citrullus plant extract of the invention formulated into a food or drink-additive powder, a fortified food or drink, a supplement including a sports supplement and / or a capsule. In an optional alternative embodiment of the invention the liquid and / or powdered Citrullus plant extract of the invention may be used directly as a supplement for addition to food or liquid consumables, including as a tea.

[0071] The medical device or cosmetic composition may comprise the liquid Citrullus plant extract.

[0072] In a third embodiment example of the sixth aspect of the invention, the Citrullus slurry fermented for 10 days at 25 °C and pH 3.6) is filtered to 50 pm and extracted in 30% glycerine and 10% fynbos honey to generate a clarified extract. The clarified extract is combined with 0.3% hyaluronan, 2% niacinamide, and natural preservatives to produce a leave-on cosmetic serum for improving hydration and barrier metrics.

[0073] In a fourth embodiment example of the sixth aspect of the invention the medical device is a hydrogel wound dressing or surgical sealant device comprising or consisting of (A) an anionic polymer solution containing pectin (DE 30-60%), alginate and / or gelatin / modified starch, 5-20% Citrullus liquid extract in a citrate buffer (w / w), and (B) a multivalent cation solution (e.g., CaCI2, Ca-gluconate). Components (A) and (B) may be sterilized by filtration and filled into a dual chamber syringe or spray cartridge using aseptic fill conditions. The filled dual chamber syringe or spray cartridge may be sterilized by gamma or e-beam radiation. Gelation of components (A) and (B) occurs upon mixing during application with the dual chamber syringe or spray cartridge, forming the hydrogel wound dressing or surgical sealant. More particularly, in one possible example of a hydrogel wound dressing, component (A) may comprise or consist of 2-4% (w / w) pectin (DE 50-55%), 0.5-1.5% alginate (w / w), 10% glycerine (w / w), and 5-20% Citrullus liquid extract in a citrate buffer (w / w), and compound (B) may comprise or consist of 1-10% calcium salt (w / w). Further, more particularly, in one possible example of a surgical sealant component (A) may comprise or consist of

[0074] 4-6% (w / w) pectin (DE 50-55%), 0.5-1.5% alginate (w / w), 10% glycerine (w / w), and

[0075] 5-20% Citrullus liquid extract in a citrate buffer (w / w), and compound (B) may comprise or consist of 1-10% calcium salt (w / w) at 0.1 - 2.0 M, sufficient to induce gelation upon mixing. The surgical sealant may be comprised in a spray cartridge. The spray cartridge may be single-use, with luer-lock adapters for atomization or bead-line application across bleeding surfaces to assist hemostasis and seal small leaks.

[0076] The hydrogel wound dressing exhibits strong adherence to moist tissue and maintains a humid wound environment for improved healing.

[0077] In a fifth embodiment example of the sixth aspect of the invention, the cosmetic composition may be formulated as a dermatological composition further comprising xanthan gum, niacinamide, vitamin E, and multi-MW hyaluronic acid for use in treating psoriasis and / or eczema. Alternatively, in the fifth embodiment example of the sixth aspect of the invention, the dermatological composition may comprise the liquid Citrullus plant extract, ivermectin, botanical oils such as rosemary, jojoba, and / or castor oil, vitamin B3 and vitamin E for use in treating psoriasis and / or eczema. In a sixth embodiment example of the sixth aspect of the invention, the cosmetic composition may comprise the liquid Citrullus plant extract, vitamin B3, vitamin E and one or more botanical oils such as rosemary, jojoba, and / or castor oil as a moisturiser.

[0078] The dermatological composition or cosmetic composition may be formulated for topical administration. The dermatological composition or cosmetic composition may be for use in improving skin hydration and / or barrier function. For example, the topical formulation may be as a lotion, a cream, a serum, a gel, a foam, a shampoo, or a massage oil. The dermatological composition or cosmetic composition may further comprise a dermatologically acceptable carrier.

[0079] The dermatological composition or cosmetic composition may include additional active ingredients selected from the group comprising any one or more of the following ingredients: i) Mauritia flexuosa fruit oil; ii) Elaeis guineessis (palm) oil;

[0080] Hi) Gossypium herbaceum seed oil; iv) Linum usitatissimum (flax) seed oil; v) tocopherol; vi) natural albumen vii) octyldodecyl olivate; viii) hydrolyzed quinoa, ix) jojoba esters x) Camellia sinensis leaf extract; xi) hydrogenated lecithin; xii) ubiquinone; xiii) bisabolol; xiv) terapeptide-30; xv) palmitoyl dipeptide-5 diaminobutyroyl hydroxythreonine; and xvi) Aloe vera extract.

[0081] In one possible embodiment, the dermatological composition or cosmetic composition further includes natural albumen, octyldodecyl olivate, hydrolyzed quinoa, and tocopherol.

[0082] In another possible embodiment, the dermatological composition or cosmetic composition further includes jojoba esters, Camellia sinensis leaf extract, hydrogenated lecithin, ubiquinone, bisabolol, terapeptide-30, palmitoyl dipeptide-5 diaminobutyroyl hydroxythreonine, and Aloe vera extract.

[0083] According to a seventh aspect of the invention there is provided use of the liquid and / or powdered Citrullus plant extract of any one of the first to fourth aspects of the invention in the manufacture of a bioactive composition, a medical device, a cosmetic composition, a nutritional composition including a nutraceutical, or an agrobiological composition comprising (a) the liquid and / or powdered Citrullus plant extract; and (b) one or more nutraceutically, cosmetically or agrobiologically acceptable carriers or excipients. The medical device or cosmetic composition may be formulated for topical administration.

[0084] In a first embodiment example of the seventh aspect of the invention the medical device comprises a hydrogel wound dressing or a surgical sealant comprising or consisting of (A) an anionic polymer solution containing pectin (DE 30-60%), alginate and / or gelatin / modified starch, 5-20% Citrullus liquid extract in a citrate buffer (w / w), and and (B) a multivalent cation solution (e.g., CaCI2, Ca-gluconate).

[0085] More particularly, the medical device comprising the hydrogel wound dressing may be manufactured by:

[0086] A) providing 2-4% (w / w) pectin (DE 50-55%), 0.5-1.5% alginate (w / w), and 10% glycerine (w / w) in a citrate buffer containing 5-20% Citrullus liquid extract (w / w) to a first chamber of a dual chamber syringe or spray cartridge comprising a static mixer mechanism; and

[0087] B) providing 1-10% calcium salt (w / w) solution to a second chamber of the dual chamber syringe or spray cartridge, wherein upon application by the dual chamber syringe or spray cartridge comprising the static mixer mechanism, the contents of A) and B) above are mixed to form a cohesive gel hydrogel wound dressing.

[0088] Alternatively, the medical device comprising the surgical sealant may be manufactured by:

[0089] A) providing 4-6% (w / w) pectin (DE 50-55%), 0.5-1.5% alginate (w / w), and 10% glycerine (w / w) in a citrate buffer containing 5-20% Citrullus liquid extract (w / w) to a first chamber of a dual chamber syringe or spray cartridge comprising a static mixer mechanism; and

[0090] B) providing 1-10% calcium salt (w / w) at 0.1 - 2.0 M, sufficient to induce gelation upon mixing solution to a second chamber of the dual chamber syringe or spray cartridge, wherein upon application by the dual chamber syringe or spray cartridge comprising the static mixer mechanism, the contents of A) and B) above are mixed to form a cohesive gel surgical sealant.

[0091] According to second embodiment example of the seventh aspect of the invention, the nutraceutical composition may be for consumption as a dietary supplement, including as a drink or a food comprising the liquid and / or powdered Citrullus plant extract of the invention formulated into a food or drink-additive powder, a fortified food or drink such as a tea, a sports supplement and / or a capsule.

[0092] According to an eighth aspect of the invention there is provided a method of treating a subject with the liquid and / or powdered Citrullus plant extract of any one of the first to fourth aspects of the invention. The liquid and / or powdered Citrullus plant extract may be in the form of a bioactive composition, a medical device, a cosmetic composition, a nutritional composition including a nutraceutical, or an agrobiological composition comprising (a) the liquid and / or powdered Citrullus plant extract; and (b) one or more nutraceutically, cosmetically or agrobiologically acceptable carriers or excipients. The treatment may be by topical administration.

[0093] In one possible embodiment example of the eighth aspect of the invention, the treatment may comprise sealing a bleeding tissue surface, including during surgery with the medical device in the form of a dual chamber syringe or spray cartridge comprising a static mixer mechanism, wherein:

[0094] A) an anionic polymer solution containing pectin (DE 30-60%), alginate and / or gelatin / modified starch, 5-20% Citrullus liquid extract in a citrate buffer (w / w) are contained in a first chamber of the dual chamber syringe or spray cartridge; and

[0095] B) a multivalent cation solution (e.g., CaCI2, Ca-gluconate) is contained in a second chamber of the dual chamber syringe or spray cartridge, wherein upon application to the bleeding tissue surface by the dual chamber syringe or spray cartridge the static mixer mechanism combines the contents of A) and B) to form a cohesive gel that seals the bleeding tissue surface.

[0096] More particularly, the medical device may comprise a hydrogel wound dressing wherein A) comprises or consists of: 2-4% (w / w) pectin (DE 50-55%), 0.5-1.5% alginate (w / w), and 10% glycerine (w / w) in a citrate buffer containing 5-20% Citrullus liquid extract (w / w); and B) comprises or consists of 1-10% calcium salt (w / w). Alternatively, the medical device may comprise a surgical sealant wherein A) comprises or consists of: 4-6% (w / w) pectin (DE 50-55%), 0.5-1.5% alginate (w / w), and 10% glycerine (w / w) in a citrate buffer containing 5-20% Citrullus liquid extract (w / w); and B) comprises or consists of 1-10% calcium salt (w / w) at 0.1 - 2.0 M, sufficient to induce gelation upon mixing.

[0097] According to a ninth aspect of the invention, there is provided a kit comprising: AA) the dual chamber syringe or spray cartridge device comprising the Citrullus extract of fifth or sixth aspect of the invention; and BB) instructions for mixing and applying the components to form a hydrogel on a target site.

[0098] DETAILED DESCRIPTION

[0099] The invention relates to methods for producing one or more extracts from plants of the genus Citrullus. The methods convert harvested plant material into a purified liquid extract and, in certain embodiments, a fermented solid cake and / or a dried, milled extract. The processes disclosed herein may be conducted with whole fruit and, optionally, additional plant tissues, and can incorporate maceration, optional or mandatory fermentation, optional or mandatory freeze-thaw lysis, and one or more extraction and clarification operations that separate a purified liquid extract from colloidal oleosomes and solid residues. The methods are compatible with food-grade inputs and processing conditions, and are suitable for producing extracts for use in nutritional, medical device, cosmetic and agrobiological applications of use.

[0100] The following description of the invention is provided as an enabling teaching of the invention, is illustrative of the principles of the invention and is not intended to limit the scope of the invention. It will be understood that changes can be made to the embodiment / s depicted and described, while still attaining beneficial results of the present invention. Furthermore, it will be understood that some benefits of the present invention can be attained by selecting some of the features of the present invention without utilising other features. Accordingly, those skilled in the art will recognise that modifications and adaptations to the present invention are possible and can even be desirable in certain circumstances, and are a part of the present invention.

[0101] As used herein, “endogenous microbiota” in the context of fermentation refers to the naturally occurring microorganisms — such as bacteria, yeasts, and molds — present in the raw Citrullus plant material and / or ambient environment. As used herein, “freeze-thaw lysis” refers to repeated freezing and thawing of a plant slurry to disrupt cellular structures and enhance release of intracellular components.

[0102] In this application, the term “about” when referring to a numerical value means that the value may vary by ±10% unless otherwise specified or dictated by the context.

[0103] EXAMPLES

[0104] Plant Material

[0105] Citrullus plant material is used in the method of the invention, and may include C. lanatus, C. colocynthis as well as any one or more edible subspecies or varieties of this genus, including subsp. vulgaris, mucosospermus, lanatus (including the var. caffer or tsamma tsamma, karkoer, bitterboela, and the var. citroides or citron melon, makataan).

[0106] The plant material may comprise the whole fruit including rind, flesh, juice, and seeds. In one embodiment, the plant material further comprises up to 10% (w / w) of roots and optionally any one or more of the remaining stems, leaves, and flowers of the Citrullus plant. Inclusion of roots can yield elevated osmolyte content, including ectoine, which may be beneficial for barrier-protective formulations comprising the extract. In an alternative embodiment, the plant material consists of the whole Citrullus plant.

[0107] Processes

[0108] General process overview

[0109] In representative embodiments, the method includes: providing harvested Citrullus plant material; washing and optionally surface sanitising the plant material; macerating the washed material to form a slurry; optionally or mandatorily fermenting the slurry; optionally or mandatorily subjecting the slurry to freeze-thaw lysis either prior to or after fermentation; and subjecting the slurry to one or more extraction processes selected from extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration to separate a purified liquid extract from colloidal oleosomes and a fermented solid cake.

[0110] In certain embodiments the fermentation step is optional, in other embodiments it is performed. In certain embodiments the freeze-thaw lysis step is optional, in other embodiments it is performed. The order of the fermentation and freeze-thaw lysis steps may be varied as described herein.

[0111] Washing and optional surface sanitising

[0112] Harvested plant material is washed to remove soil and debris. The washing step may be followed by an optional surface sanitisation step using sanitising agents and conditions known to the skilled person, to reduce surface bioburden prior to maceration.

[0113] Maceration

[0114] The washed plant material is macerated to form a slurry. Maceration may be performed by any mechanical technique known in the art, including chopping or cutting, mincing or milling (e.g., using a hammer mill), or high-shear maceration by hydraulic, pneumatic, or manual means, or by means of a worm screw. The resultant slurry typically has a particle size of about 0.1-5 mm.

[0115] Fermentation

[0116] In certain embodiments the slurry is fermented. Fermentation may be conducted with one or more sugars or sugar alcohols fermentable by yeast or bacteria. In preferred embodiments, honey at about 5-30% w / w and / or food-grade glycerine at about 5- 40% w / w is added, together with a fermentation starter culture or relying on endogenous microbiota. The honey may be raw honey. Fermentation is typically performed for about 7-14 days at about 18-30 °C with pH controlled at about 3.0-4.5. The fermentation may further include additives or co-substrates depending on the desired functionality of the final product, including aloe, marshmallow, chamomile, licorice, nettle, baobab, citrus zest, seed flours, oats, or enzyme sources such as papain or bromelain.

[0117] In certain embodiments fermentation is optional. In other embodiments fermentation is mandatory. Where fermentation is performed, freeze-thaw lysis may be applied before or after fermentation as described below.

[0118] Freeze-thaw lysis The slurry is optionally or mandatorily subjected to freeze-thaw lysis. This step may comprise multiple freezing cycles to a temperature between about -40 °C and about -80 °C followed by thawing, thereby disrupting cell structures and enhancing release of intracellular components. Freeze-thaw lysis may be conducted prior to fermentation, after fermentation, or both.

[0119] Extraction and separation

[0120] Following maceration and any optional or mandatory fermentation and / or freeze-thaw lysis, the slurry is subjected to one or more extraction processes. Suitable processes include extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration. These processes are applied in an order and combination effective to separate a purified liquid extract from colloidal oleosomes and a fermented solid cake. Clarification may comprise the addition of bentonite.

[0121] The purified liquid extract obtained after separation may be further processed as described herein, including concentration, pasteurisation, and / or homogenisation.

[0122] Preservation, pasteurisation, and optional concentration

[0123] The method may include the addition of one or more preservatives to the final Citrullus extract. Preservatives may be selected from those known to the skilled person. In one embodiment, the preservative system comprises a combination of dehydroacetic acid and benzyl alcohol at about 0.5-2% w / w. In another embodiment, the preservative system comprises phenoxyethanol and ethylhexylglycerin at about 0.5-2% w / w. In a further embodiment, potassium sorbate is employed at about 0.5-2% w / w.

[0124] The extract may be pasteurised, for example using conventional low-temperature pasteurisation at about 63-65 °C for about 30 minutes. The extract may be concentrated by one or more of freeze drying, spray drying, solvent extraction including with alcohol, acetone, chloroform, water or any combination thereof, or centrifugal concentration.

[0125] Cryopreservation

[0126] The slurry obtained after maceration and / or the purified liquid extract may be cryopreserved for storage. Cryopreservation may be conducted at about -20 °C to -40 °C for a period of up to about three months to preserve labile constituents including NAD+and L-citrulline.

[0127] Solid cake processing and powdered extracts

[0128] The separated solid cake may be dried and milled to yield a powdered Citrullus extract. Drying is preferably performed at a temperature of <50 °C. The dried and milled extract may be in the form of a flour or a protein / fibre concentrate.

[0129] In an alternative embodiment, production of a purified secondary liquid extract from the dehydrated solid cake is carried out using a conical brewing operation. This embodiment may include the following steps. The solid cake and fermented solid cake are dehydrated and optionally ground. A ribbed frusto-conical extraction apparatus is provided comprising a ribbed frusto-conical support adapted to seat over a collecting vessel and a frusto-conical filter including a paper filter adapted to line the interior of the support. The ribbed frusto-conical support may comprise side walls disposed at a 60-degree angle from the base, the side walls including raised grooves spiralling towards the base, and the base defining an aperture. The support may optionally include a set of handles. The frusto-conical filter may comprise a paper filter such as a Whatman No. 1 filter paper.

[0130] The frusto-conical filter is placed into the support to form the ribbed frusto-conical extraction apparatus. In certain embodiments, the filter is optionally rinsed in situ with water at about 60-65 °C. The extraction apparatus is placed onto a collecting vessel and the dehydrated Citrullus plant solid cakes are added to the extraction apparatus. Water at about 60-65 °C is then added and the purified secondary liquid extract filtered through the frusto-conical filter is collected in the vessel. The addition of water may be performed in one or more steps. Where multiple additions are used, an initial volume is added, followed by a waiting period of about 30 seconds to about one minute, and then a further volume is added until the desired volume is collected. A typical ratio is about 1 g of dried Citrullus plant extract to about 15 g of water. The total brewing time is typically about 3-4 minutes or until no further extract is produced.

[0131] After collection, the purified secondary liquid extract may be pasteurised, for example at about 63-65 °C for about 30 minutes. The secondary liquid extract may be further concentrated by one or more of freeze drying, spray drying, solvent extraction including with an organic solvent such as alcohol, acetone, chloroform, water or any combination thereof, or centrifugal concentration.

[0132] The dehydration step may be performed by any low-heat drying method, including sun drying, a dehydrator, an oven, or a low-heat drum coffee roaster such as a PROBAT low-heat drum coffee roaster. The frusto-conical support may be made from ceramic, plastic, glass, stainless steel, or copper. The size of the aperture in the base of the frusto-conical support may be selected according to the desired flow rate and brewing time. In certain embodiments, the ribbed frusto-conical extraction apparatus may have a capacity of up to five litres.

[0133] Additional processing

[0134] The method may further comprise homogenisation of the liquid extract and / or intermediate streams to improve consistency, stability, and mouthfeel or sensorial properties of the final product.

[0135] Embodiments and variations

[0136] In one embodiment, fermentation is optional and freeze-thaw lysis is optional. In another embodiment, fermentation is performed and freeze-thaw lysis is optional. In a further embodiment, fermentation is performed and freeze-thaw lysis is performed. In another embodiment, fermentation is optional and freeze-thaw lysis is performed. In each embodiment, the extraction processes are selected from extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration, in an order and combination effective to achieve separation of a purified liquid extract from colloidal oleosomes and a fermented solid cake.

[0137] The skilled person will appreciate that the foregoing parameters, sequences, and operating ranges may be adapted based on the characteristics of the starting material, the target composition profile, and the intended application of the extract, while remaining within the scope of the methods disclosed herein.

[0138] Citrullus plant extract bioactive compounds

[0139] The following bioactive compounds are known to be present in the Citrullus plant extract produced according to the method of the invention: A. L-citrulline;

[0140] B. quercetin;

[0141] C. NAD+; and

[0142] D. optionally ectoine and / or oleosomes.

[0143] L-citrulline is converted in the body to L-arginine, which boosts nitric oxide production. Nitric oxide helps relax blood vessels, improving circulation and lowering blood pressure and may therefore be used in the treatment of hypertension, prehypertension or mild hypertension. In addition, L-citrulline is associated with an improvement in anaerobic performance and increase in endurance during sporting activities by increasing blood flow to muscles. More specifically, L-citrulline though the increase in nitric oxide production, has been associated with an increase in VO2max.

[0144] Quercetin is a plant-derived flavonoid with a wide range of medical and therapeutic applications, largely due to its antioxidant, anti-inflammatory, antiviral, anti-cancer, and cardioprotective properties. For example, quercetin acts to reduce oxidative stress in the treatment of eczema and / or psoriasis. Furthermore, quercetin helps plants cope with abiotic stresses such as salinity, drought, and oxidative stress and is a biostimulant enhancing physiological and biochemical processes as well as being a natural antimicrobial I antifungal compound.

[0145] Ectoine has been identified to provide cytoprotective, anti-inflammatory, anti-oxidative, and stabilizing applications and acts to increase hydration in animals and humans. Additionally, foliar application of ectoine (0.5 mM) was shown to significantly improve growth attributes, gas exchange, and nutrient content in plants. It has been indicated to reduce oxidative stress in plants and to work synergistically with arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) to enhance root development; improve soil nutrient availability and act as a buffer against salinity- induced damage.

[0146] Oleosomes have been used in cosmetic applications to improve skin hydration and barrier repair, as well as photoprotection and as an anti-inflammatory in treatment of psoriasis and eczema. Oleosomes have also been associated with wound healing, in reducing inflammation at wound sites. Furthermore, oleosomes are natural emulsifiers and can replace fat and are associated with controlled nutrient release in nutraceuticals. Applications for use of the Citrullus plant extract

[0147] The Citrullus plant extracts have application as bioactives, medical devices, cosmetics, nutritional supplements or nutraceuticals, or agrobiologicals to improve plant health, stimulate growth or increase yield or agricultural productivity.

[0148] Nutraceutical formulations

[0149] The Citrullus plant extract of the invention can be formulated into a food or drinkadditive powder, a fortified food or drink, a sports supplement and / or a capsule or may be used directly as a supplement for addition to food or liquid consumables, including as a tea.

[0150] More specifically, the Citrullus plant extract may be incorporated into the following products as a nutraceutical supplement.

[0151] 1) Smoothies: Add a spoonful of Citrullus extract or powder to a smoothie for a nutritional boost. The powder adds vitamins, minerals, and antioxidants without altering the flavor significantly.

[0152] 2) Hydrating drinks or sports drinks: Incorporate Citrullus extract or powder into a drink for added nutrition. The drink may comprise additional flavourings or additives as desired.

[0153] 3) Energy Bars: Incorporate Citrullus extract or powder into energy bars for added sweetness and nutrition. The energy bars may comprise additional flavourings or additives as desired.

[0154] 4) Baking: Use the Citrullus extract or powder as an ingredient in baking, such as in muffins, pancakes, or bread, to infuse them with extra nutrients.

[0155] 5) Supplements: The Citrullus extract or powder can be packaged and sold as a standalone dietary supplement, marketed for its high vitamin and antioxidant content. The Citrullus extract or powder can be packaged as a pill through pill pressing or encapsulated in a capsule.

[0156] Cosmetic formulations

[0157] Formulation:

[0158] The purified Citrullus extract may be incorporated into the following cosmetic products. 1) Lotions and Creams: Combine the Citrullus extract with other natural ingredients like shea butter, Aloe vera, and essential oils. Emulsifiers and preservatives can be added to create a stable lotion or cream. The Citrullus extract can serve as a hydrating, soothing, and antioxidant-rich ingredient, while the cold-pressed oil provides moisturization.

[0159] 2) Serums: Use the Citrullus extract in a concentrated form for serums, adding to it hyaluronic acid or other skin-benefiting ingredients. This creates a product that can be applied directly to the skin for intense hydration and protection.

[0160] 3) Face Masks: Mix Citrullus extract powder with clay and other beneficial ingredients to create a face mask. The Citrullus, when activated with water, releases its nutrients, providing a refreshing and revitalizing experience for the skin.

[0161] 4) Packaging and Storage: Ensure that the products are packaged in airtight containers, preferably opaque to protect the active ingredients from light exposure, which can degrade them.

[0162] Benefits:

[0163] 1 ) Hydration: The Citrullus natural water content and vitamins make it an excellent hydrating agent, ideal for lotions and creams.

[0164] 2) Anti-Aging: The antioxidants in Citrullus help fight free radicals, making the products suitable for anti-aging skincare lines.

[0165] 3) Soothing: Citrullus’s cooling properties can be utilized in products aimed at soothing irritated or sensitive skin.

[0166] Skincare products would appeal to consumers seeking natural and plant-based beauty solutions.

[0167] Filaggrin-Mediated Hydration and Barrier Support

[0168] Without being bound by theory, it is believed that the bioactive composition derived from Citrullus according to the present invention enhances epidermal barrier integrity and hydration through modulation of the filaggrin-natural-moisturizing-factor (NMF) pathway. The preserved L-citrulline within the glycerine-honey-extracted bioslurry serves as a biochemical precursor for NMF synthesis and contributes to up-regulation of filaggrin expression in keratinocytes. In combination with NAD+and quercetin antioxidants and lipid-rich oleosomes, the extract promotes aggregation of keratin filaments, maintains corneocyte cohesion, and supports enzymatic conversion of filaggrin into hygroscopic amino-acid derivatives that retain water within the stratum corneum. The net cosmetic effect is improved moisture balance, reduced transepidermal water loss (TEWL), and reinforcement of the skin’s protective barrier, producing a visibly smoother, more elastic surface.

[0169] Medical Device Applications

[0170] Medical devices in the form of hydrogels comprising the Citrullus plant extract of the invention, suitable for use as wound dressings or surgical sealants may be manufactured. In general, the devices comprise an anionic polymer component and a multivalent cation component configured to form a cohesive gel upon mixing at the point of use. In certain embodiments, the anionic polymer solution contains pectin having a degree of esterification (DE) within the range of 30-60%, alginate and / or gelatin or modified starch, and a citrate buffer containing 5-20% (w / w) of a Citrullus liquid extract produced according to the method of the invention. The multivalent cation component may comprise a calcium salt, such as calcium chloride or calcium gluconate. The components can be provided in a dual chamber syringe or spray cartridge fitted with a static mixer so that, upon application, the components mix and gel in situ.

[0171] In a first embodiment the medical device comprises a hydrogel wound dressing or a surgical sealant comprising or consisting of: (A) an anionic polymer solution containing pectin (DE 30-60%), alginate and / or gelatin / modified starch, and 5-20% (w / w) Citrullus liquid extract in a citrate buffer; and (B) a multivalent cation solution, for example a solution of CaCl2 or calcium gluconate. Upon mixing components (A) and (B), ionic crosslinking occurs to form a cohesive gel suitable for topical or surgical application.

[0172] In certain embodiments relating to a hydrogel wound dressing, the device may be manufactured by providing to a first chamber of a dual chamber syringe or spray cartridge equipped with a static mixer: 2-4% (w / w) pectin having a DE of 50-55%, 0.5-1 .5% (w / w) alginate, and 10% (w / w) glycerine in a citrate buffer containing 5-20% (w / w) Citrullus liquid extract. A second chamber of the syringe or cartridge may be charged with a 1-10% (w / w) solution of a calcium salt. During application, actuation of the device advances the contents of the chambers into the static mixer, thereby combining the anionic polymer solution with the calcium salt solution to form a cohesive hydrogel wound dressing at the site of delivery.

[0173] In alternative embodiments relating to a surgical sealant, the device may be manufactured by providing to a first chamber of a dual chamber syringe or spray cartridge equipped with a static mixer: 4-6% (w / w) pectin having a DE of 50-55%, 0.5-1 .5% (w / w) alginate, and 10% (w / w) glycerine in a citrate buffer containing 5-20% (w / w) Citrullus liquid extract. A second chamber of the syringe or cartridge may be charged with a 1-10% (w / w) solution of a calcium salt. During application, the contents of the chambers are mixed through the static mixer to form a cohesive gel surgical sealant.

[0174] In some embodiments, the anionic polymer solution further comprises gelatin or a modified starch in place of or in addition to alginate. The calcium salt solution may consist essentially of calcium chloride or calcium gluconate in water, optionally buffered, and may include pharmaceutically acceptable excipients to adjust viscosity, osmolarity, or stability. The citrate buffer and glycerine may be selected to provide desired handling characteristics, humectancy, and pH control for compatibility with the pectin and alginate components and to promote consistent gelation kinetics upon contact with the multivalent cations.

[0175] The dual chamber syringe or spray cartridge can be configured to deliver a predetermined volumetric ratio of the anionic polymer solution to the calcium salt solution. The static mixer may comprise one or more helical mixing elements sized to achieve homogeneous mixing under manual or mechanical actuation forces ordinarily applied in clinical settings. The resulting gel exhibits cohesion and adherence suitable for wound coverage or tissue sealing depending on the relative concentrations of pectin and alginate in the anionic polymer solution and the concentration of the calcium salt in the cation solution.

[0176] The embodiments described herein may be used independently or in combination. Unless otherwise specified, concentration percentages are weight by weight, ranges are inclusive, and individual values within the disclosed ranges are contemplated as if expressly recited. The terms “comprise,” “comprising,” “contain,” and “containing” are intended to be open-ended and non-limiting.

Claims

CLAIMS1 . A method of producing a Citrullus extract, the method comprising or consisting of: a) providing harvested Citrullus plant material; b) washing and optionally surface sanitising the harvested plant material; c) macerating the washed plant material to form a slurry; d) optionally fermenting the slurry with one or more sugars or sugar alcohols and a fermentation starter culture or endogenous microbiota; e) optionally subjecting the slurry to freeze-thaw lysis before and / or after the fermenting; and f) subjecting the slurry to one or more extraction processes selected from extraction through a glycerine-honey matrix, decanting, clarification, centrifugation, concentration, and filtration to separate a purified liquid extract from colloidal oleosomes and a solid cake.

2. The method of claim 1 , wherein the step of fermenting is not optional.

3. The method of claim 1 or 2, wherein the step of freeze-thaw lysis is not optional.

4. The method of any preceding claim, wherein step d) comprises fermenting the slurry for 7-14 days at 18-30 °C and pH 3.0-4.5 in the presence of 5-30% w / w honey and / or 5-40% w / w food-grade glycerine and a starter culture or endogenous microbiota.

5. The method of any preceding claim, wherein the freeze-thaw lysis comprises multiple freezing cycles of between -40 °C to -80 °C followed by thawing.

6. The method of any preceding claim, further comprising cryopreserving the slurry and / or the purified liquid extract at -20 °C to -40 °C for up to 3 months.

7. The method of any preceding claim, wherein the plant material comprises the whole fruit including rind, flesh, juice and seeds, and optionally up to 10% w / w roots and any of stems, leaves and flowers.

8. The method of any preceding claim, further comprising drying and milling the solid cake at a temperature <50 °C to yield a powdered Citrullus extract.

9. The method of any preceding claim, wherein the one or more extraction processes comprise extraction through a matrix comprising 10-40% w / w glycerine and 5-30% w / w honey.

10. The method of any preceding claim, further comprising pasteurising the purified liquid extract at 63-65 °C for 30 minutes.

11. A method of producing a purified secondary liquid extract of Citrullus, comprising:A) dehydrating the solid cake obtained by the method of any one of claims 1-12 and optionally grinding the dehydrated cake;B) placing the dehydrated cake into a ribbed frusto-conical extraction apparatus comprising a ribbed frusto-conical support with an interior frusto-conical paper filter and a base aperture; andC) adding water at 60-65 °C to the ribbed frusto-conical extraction apparatus at a ratio of about 1 g dried cake to 15 g water and collecting filtrate for 3-4 minutes or until no further extract is produced.

12. A Citrullus extract obtainable by the method of any one of claims 1-11.

13. The extract of claim 12, comprising the bioactive compounds L-citrulline, quercetin, NAD+, and optionally ectoine and / or oleosomes.

14. The extract of claim 12 or 13, which is a liquid extract.

15. The extract of claim 12 or 13, which is a powdered extract including in the form of a flour or a protein / fibre concentrate.

16. A bioactive composition comprising the extract of any one of claims 12-15 and a nutraceutically, cosmetically or agrobiologically acceptable carrier or excipient.

17. A cosmetic composition comprising the extract of any one of claims 12-15 and a cosmetically or dermatologically acceptable carrier.

18. The cosmetic composition of claim 17, further comprising one or more of xanthan gum, niacinamide, vitamin E, and hyaluronic acid, or comprising niacinamide and hyaluronic acid.

19. The cosmetic composition of claim 17 or 18, formulated as a lotion, cream, serum, gel, foam, shampoo or massage oil.

20. A nutritional composition comprising the extract of any one of claims 12-15 formulated as a food or drink additive powder, a fortified food or drink, a supplement including a sports supplement, a capsule or a tea.

21. A medical device composition comprising:I) an anionic polymer component comprising pectin having a degree of esterification of 30-60%, alginate and / or gelatin or modified starch, a citratebuffer and 5-20% w / w of a Citrullus liquid extract according to any one of claims 12-15; andII) a multivalent cation component comprising a calcium salt solution.

22. The medical device composition of claim 21 , wherein the anionic polymer component comprises 2-4% w / w pectin having a degree of esterification of 50- 55%, 0.5-1.5% w / w alginate, and 10% w / w glycerine.

23. The medical device composition of claim 21 or 22, wherein the calcium salt is calcium chloride or calcium gluconate at 1-10% w / w.

24. A dual-chamber syringe or spray cartridge device comprising: aa) a first chamber containing the anionic polymer component of any one of claims 21-22; and bb) a second chamber containing the multivalent cation component of claim 21 , wherein the device is configured to mix the components upon actuation to form a cohesive hydrogel.

25. Use of the extract of any one of claims 12-15 in the manufacture of a medical device, a cosmetic composition, a nutritional composition including a nutraceutical, or an agrobiological composition comprising the liquid and / or powdered Citrullus plant extract; and (b) one or more nutraceutically, cosmetically or agrobiologically acceptable carriers or excipients.

26. An agrobiological composition comprising a Citrullus extract according to any one of claims 12-15 for use as a foliar or soil application to improve plant health, stimulate growth or increase yield or agricultural productivity.

27. A cosmetic composition comprising a Citrullus extract according to any one of claims 12-15 for use in improving skin hydration and / or barrier function.

28. The cosmetic composition according to any one of claims 17 to 19 or the cosmetic composition for use of claim 27 which is administered topically.

29. A Citrullus extract according to any one of claims 12-15 for use as an active ingredient in a hydrogel wound dressing or surgical sealant.

30. A kit comprising: AA) the dual-chamber device of claim 24; and BB) instructions for mixing and applying the components to form a hydrogel on a target site.

31. The method of any one of claims 1-11 , wherein the fermentation step comprises addition of one or more additives or co-substrates selected fromaloe, marshmallow, chamomile, licorice, nettle, baobab, citrus zest, seed flours, oats, papain and bromelain.

32. The method of any one of claims 1-11 , further comprising homogenising the purified liquid extract.

33. The method of any one of claims 1-11 , wherein the plant material consists of the whole Citrullus plant.

34. The cosmetic composition of any one of claims 17-19 or the cosmetic composition for use of claim 27, further comprising one or more of: Mauritia flexuosa fruit oil, Elaeis guineensis oil, Gossypium herbaceum seed oil, Linum usitatissimum seed oil, tocopherol, albumen, octyldodecyl olivate, hydrolysed quinoa, jojoba esters, Camellia sinensis leaf extract, hydrogenated lecithin, ubiquinone, bisabolol, tetrapeptide-30, palmitoyl dipeptide-5 diaminobutyroyl hydroxythreonine and Aloe vera extract.

35. The method of any one of claims 1-11 , or 31 to 33 which is a zero-water process with no added water being required to be used.