RO138074A2 - Prebiotic regenerative antioxidant composition and process for preparing the same - Google Patents
Prebiotic regenerative antioxidant composition and process for preparing the same Download PDFInfo
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- RO138074A2 RO138074A2 ROA202200662A RO202200662A RO138074A2 RO 138074 A2 RO138074 A2 RO 138074A2 RO A202200662 A ROA202200662 A RO A202200662A RO 202200662 A RO202200662 A RO 202200662A RO 138074 A2 RO138074 A2 RO 138074A2
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- infusion
- prebiotic
- composition
- nanocellulose
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Abstract
Description
OFICIUL DE STAT PEi-TRU INVLW j ti Șl MĂRCIPEi-TRU STATE OFFICE INVLW j ti Șl MARCI
Cerere de brevet de Invenție Nr OlInvention patent application No. Ol
Data depozit..Ξ1ϊ£'.ζΕΞΊDeposit date..Ξ1ϊ£'.ζΕΞΊ
COMPOZIȚIE PREBIOTICĂ, REGENERATIVĂ SI ANTIOXIDANTĂ SI PROCEDEU » ’ » ’PREBIOTIC, REGENERATIVE AND ANTIOXIDANT COMPOSITION AND PROCEDURE » ’ » ’
DE OBȚINERE A ACESTEIA 9TO OBTAIN IT 9
Prezenta invenție se referă la o compoziție prebiotică și antioxidantă destinată utilizării ca ingredient activ în suplimente nutritive, produse cosmetice și dispozitive medicale, ca și la un procedeu de obținere a acestei compoziții.The present invention relates to a prebiotic and antioxidant composition intended for use as an active ingredient in nutritional supplements, cosmetic products and medical devices, as well as to a process for obtaining this composition.
Sunt cunoscute diferite compoziții cu activitate prebiotică și antioxidantă. Cererea de brevet WO2022130294A1 se referă la o compoziție de carbohidrați, care cuprinde oligozaharide pectice și acid oligo-galacturonic din coaja de fructe, utilizabilă ca aditiv pentru formularea produselor alimentare. Compoziția de carbohidrați este utilă ca înlocuitor pentru zaharurile convenționale, care contribuie la menținerea caracteristicile reologice (de textură) ale produselor alimentare, fără a crește conținutul caloric al acestora. în plus, compoziția de carbohidrați descrisă are potențial bioactiv, având activitatea antioxidanta și proprietăți prebiotice.Various compositions with prebiotic and antioxidant activity are known. Patent application WO2022130294A1 relates to a carbohydrate composition comprising pectic oligosaccharides and oligo-galacturonic acid from fruit peel, usable as an additive for the formulation of food products. The carbohydrate composition is useful as a substitute for conventional sugars, which helps to maintain the rheological (textural) characteristics of food products without increasing their caloric content. in addition, the carbohydrate composition described has bioactive potential, having antioxidant activity and prebiotic properties.
Cererea de brevet US2022233559 A1 revendică o compoziție pe bază de xilooligozaharide care includ și porțiunile terminale ale xilozei. Aceste porțiuni sunt esterificate cu acizi hidroxicinarnici. Compoziția, cu efecte antioxidante și pre- și postbiotice este destinată utilizării ca supliment nutritiv și/sau ingredient alimentar funcțional, pentru îmbunătățirea sănătății digestive.Patent application US2022233559 A1 claims a composition based on xylooligosaccharides that also include the terminal portions of xylose. These moieties are esterified with hydroxycinaric acids. The composition, with antioxidant and pre- and postbiotic effects, is intended for use as a nutritional supplement and/or functional food ingredient to improve digestive health.
Cererea de brevet RO135698 A0 protejează o compoziție nutraceutică cu efect antioxidant și prebiotic utilizată ca aditiv alimentar pentru o gamă de produse finite din industria alimentară. Compoziția, conform invenției, este constituită în procente masice din 0,1...3% lactoferină bovină, 0,1...3% zahăr de cocos, 0,1...8% inulină, 0,5% fosfat de calciu, procentele fiind raportate la 100 g produs alimentar prelucrat gata de consum, pe bază de cereale (în stare solidă pentru micul dejun), băuturi pe bază de lapte (gata de consum), băuturi pe bază de lapte fermentat (inclusiv băuturi de tip iaurt), băuturi nealcoolice (aromatizate), produse pe bază de iaurt, produse pe bază de brânză (nematurată), înghețată, prăjituri și produse de patiserie, bomboane, gumă de mestecat.Patent application RO135698 A0 protects a nutraceutical composition with an antioxidant and prebiotic effect used as a food additive for a range of finished products in the food industry. The composition, according to the invention, consists in mass percentages of 0.1...3% bovine lactoferrin, 0.1...3% coconut sugar, 0.1...8% inulin, 0.5% sodium phosphate calcium, the percentages being reported per 100 g of ready-to-eat processed food product, cereal-based (in a solid state for breakfast), milk-based beverages (ready-to-eat), fermented milk-based beverages (including yogurt), non-alcoholic beverages (flavored), yogurt-based products, cheese-based products (unripened), ice cream, cakes and pastries, candy, chewing gum.
Cererea de brevet RO135559 A0 prezintă un aliment sub formă de baton alimentar hipoglucidic cu efecte antioxidante și prebiotice amplificate utilizat ca adjuvant în dieta hipocalorică și la un procedeu de obținere. Alimentul conform invenției este constituit din 15...35% pulbere din fructe de pădure cu un conținut de 15...20% afine, 1,5...3,5% merișor, 2,0...3,5% cătină, 2,0...3,0% coacăze negre, cu substanța uscată de minim 20%, 50...70% bază de extracție formată din sirop de malț și miere în raport de 3 : 1, 5...15% semințe de chia, 0,5...1,5% sâmburi de caise și alune măcinate, 1,0...2,0% glazură formată din unt de cacao, ulei de cocos și masă de cacao în raport 1:1:1, 2,0...3,0% pudră de cacao care înglobează semințele de chia și 0,001...0,005% sare de Himalaya, alimentul condiționat sub formă de batoane glazurate având următoarele valori nutriționale pentru 100g baton: 380...450 kcal valoare energetică, 20...40% glucide, 10...22% grăsimi, 4...14% fibre, 1...15% proteine și 0...0,3% sare. Procedeul conform invenției are următoarele etape: a) obținerea amestecului format din fructe de pădure cu substanță uscată 15...30% și sare de Himalaya, macerarea acestuia timp de 24..48 ore și uscarea la o temperatură cuprinsă între 4O...45°C timp de 1...3 zile, urmată de mărunțire mecanică până la stadiul de pulbere fină, b) extracția polifenolilor dintr-o altă șarjă de reziduu umed de fructe de pădure cu 10 volume de etanol 50...70% timp de 24...48 ore la temperatura camerei, urmată de extracția polizaharidelor cu 10 volume de apă distilată timp de 3...6 ore la 90...100°C, concentrarea la 60°C și purificarea polizaharidelor prin precipitare cu etanol 70...80%, centrifugare la 10000...15000 rpm și în final concentrare prin evaporare la 37°C, c) înglobarea amestecului de polifenoli și polizaharide, d) prepararea bazei de extracție, e) obținerea compoziției vâscoase, f) porționarea compoziției sub formă de batoane, glazurare, ambalare în pungi și depozitare la temperaturi de sub 10°C.Patent application RO135559 A0 presents a food in the form of a hypoglycemic food bar with enhanced antioxidant and prebiotic effects used as an adjuvant in the hypocaloric diet and in a method of obtaining it. The food according to the invention consists of 15...35% powder from forest fruits with a content of 15...20% blueberry, 1.5...3.5% cranberry, 2.0...3.5 % sea buckthorn, 2.0...3.0% blackcurrants, with a dry substance of at least 20%, 50...70% extraction base consisting of malt syrup and honey in a ratio of 3:1, 5.. .15% chia seeds, 0.5...1.5% apricot kernels and ground hazelnuts, 1.0...2.0% glaze consisting of cocoa butter, coconut oil and cocoa mass in the ratio 1:1:1, 2.0...3.0% cocoa powder that includes chia seeds and 0.001...0.005% Himalayan salt, the conditioned food in the form of glazed bars having the following nutritional values per 100g bar: 380...450 kcal energy value, 20...40% carbohydrates, 10...22% fat, 4...14% fiber, 1...15% protein and 0...0.3% salt . The process according to the invention has the following stages: a) obtaining the mixture consisting of forest fruits with 15...30% dry matter and Himalayan salt, macerating it for 24...48 hours and drying it at a temperature between 4O... 45°C for 1...3 days, followed by mechanical comminution to the fine powder stage, b) extraction of polyphenols from another batch of wet berry residue with 10 volumes of 50...70% ethanol for 24...48 hours at room temperature, followed by extraction of the polysaccharides with 10 volumes of distilled water for 3...6 hours at 90...100°C, concentration at 60°C and purification of the polysaccharides by precipitation with ethanol 70...80%, centrifugation at 10000...15000 rpm and finally concentration by evaporation at 37°C, c) incorporation of the mixture of polyphenols and polysaccharides, d) preparation of the extraction base, e) obtaining the viscous composition, f ) portioning of the composition in the form of bars, glazing, bagging and storage at temperatures below 10°C.
Cererea de brevet CN108611242 A expune o compoziție de băuturi bogată în seleniu, care este un amestec de infuzie de ceai negru cu oțet de mere și pireu de banane, ca și o metodă de preparare a acesteia. Băutura este preparată din următoarele substanțe în procent din greutate: 3-8% în greutate pudră de ceai negru bogat în seleniu, 20-30% în greutate banane, 20-40% în greutate oțet de mere și restul de apă de băut.Patent application CN108611242 A discloses a selenium-rich beverage composition, which is a mixture of black tea infusion with apple cider vinegar and banana puree, as well as a method of preparing the same. The drink is prepared from the following substances by weight: 3-8% by weight selenium-rich black tea powder, 20-30% by weight bananas, 20-40% by weight apple cider vinegar and the rest drinking water.
Cererea de brevet RO134936 A2 descrie o compoziție de hidrogel cu rol cosmeceutic pentru reglarea microbiomului pielii. Compoziția, conform invenției, este constituită din 20...25% o fază internă de particule solide de alginat acoperite cu chitosan cu un conținut minim de 1010 unități formatoare de colonii din Lactobacillus paracasei, eventual în amestec cu L. rhamnosus, și 75...80% o fază externă de hidrogel format din 2,5% hidrolizat proteic de pește, 1% acid ascorbic, 1,5% prebiotic, 0,3% clorură de calciu, 1% glicerină, 1% PEG 6000, 0,7% carbomer, 1% conservant natural din extract concentrat de sâmburi de grapefruit, 0,3% sorbat de potasiu, respectiv, trietanolamină, 0,5% fenoxietanol, respectiv, ulei esențial de imortele, precum și apă distilată.Patent application RO134936 A2 describes a hydrogel composition with a cosmeceutical role for regulating the skin microbiome. The composition, according to the invention, is made up of 20...25% of an internal phase of solid alginate particles covered with chitosan with a minimum content of 1010 colony-forming units of Lactobacillus paracasei, possibly mixed with L. rhamnosus, and 75. ..80% an external hydrogel phase consisting of 2.5% fish protein hydrolyzate, 1% ascorbic acid, 1.5% prebiotic, 0.3% calcium chloride, 1% glycerin, 1% PEG 6000, 0, 7% carbomer, 1% natural preservative from concentrated grapefruit seed extract, 0.3% potassium sorbate respectively triethanolamine, 0.5% phenoxyethanol respectively immortelle essential oil and distilled water.
Cererea de brevet RO 132749 A2 se referă la o compoziție mucoadezivă pentru menținerea și/sau re-echilibrarea microflorei vaginale. Compoziția, conform invenției, este constituită în părți în greutate din 50...80 părți soluție apoasă formată din 20...60% copolimer de tip Poloxamer P407, 1...5% nanoceluloză bacteriană, eventual, 5...10% chitosan, respectiv 0,1...5% pulbere de cactus, 5...20 părți în greutate compuși post biotici de tip acid lactic și acid citric, eventual 2...6 părți în greutate uleiuri esențiale alese dintre ulei esențial de cimbru și arbore de ceai, respectiv 5...10 părți în greutate oligozaharide prebiotice alese dintre inulină și fructo-oligozaharide, și 4...10 părți în greutate extracte hidrogliceroalcoolice de concentrație 50% alese dintre merișor și semințe de struguri.Patent application RO 132749 A2 refers to a mucoadhesive composition for maintaining and/or re-balancing the vaginal microflora. The composition, according to the invention, consists in parts by weight of 50...80 parts aqueous solution consisting of 20...60% Poloxamer P407 type copolymer, 1...5% bacterial nanocellulose, possibly 5...10 % chitosan, i.e. 0.1...5% cactus powder, 5...20 parts by weight post biotic compounds such as lactic acid and citric acid, possibly 2...6 parts by weight essential oils chosen from essential oil of thyme and tea tree, respectively 5...10 parts by weight prebiotic oligosaccharides chosen from inulin and fructo-oligosaccharides, and 4...10 parts by weight hydroglyceroalcoholic extracts of 50% concentration chosen from cranberry and grape seeds.
Brevetul US9808497 B2 dezvăluie o compoziție care cuprinde un concentrat de prune și/sau prune și una sau mai multe oligozaharide prebiotice, nedigerabile, solubile în apă. Opțional, unul sau mai mulți îndulcitori cu conținut scăzut de calorii, un antioxidant, senozide de calciu sau extract din Senna sp., PEG-3350, lemn dulce, cacao, cafea, arome de ceai; se adaugă arome de fructe și condimente, gelatină, agaragar, caragenan, pectină sau pudră de cacao. Alte ingrediente pot fi prezente sub formă de suplimente alimentare, cum ar fi vitaminele solubile în apă, vitaminele solubile în lipide; aminoacizi, maltodextrină, resveratrol, cofeină, suplimente minerale sau produse cunoscute ca având un efect de adjuvant natural pentru inducerea somnului sau pentru utilizare ca laxativ. Sunt posibile o varietate de formulări, cum ar fi premix-uri, sau produse nutraceutice de tipul un laxativ + băutură energizantă, sau batoane energizante, sau ca supliment prebiotic în probiotice de tip iaurturi, sau sub formă de gume, ciocolată, bomboane și deserturi.Patent US9808497 B2 discloses a composition comprising a plum and/or plum concentrate and one or more prebiotic, non-digestible, water-soluble oligosaccharides. Optionally, one or more low-calorie sweeteners, an antioxidant, calcium sennosides or Senna sp. extract, PEG-3350, licorice, cocoa, coffee, tea flavors; fruit flavors and spices, gelatin, agar agar, carrageenan, pectin or cocoa powder are added. Other ingredients may be present in the form of dietary supplements, such as water-soluble vitamins, fat-soluble vitamins; amino acids, maltodextrin, resveratrol, caffeine, mineral supplements or products known to have a natural adjuvant effect for inducing sleep or for use as a laxative. A variety of formulations are possible, such as premixes, or nutraceutical products such as a laxative + energy drink, or energy bars, or as a prebiotic supplement in yogurt-type probiotics, or in the form of gums, chocolates, candies and desserts .
Dezavantajul comun al compozițiilor descrise mai sus este limitarea lor la o singură aplicație, ca supliment nutritiv sau ca ingredient activ al unor produse cosmetice sau dispozitive medicale cu rol în reglarea sănătății pielii și/sau a mucoaselor.The common disadvantage of the compositions described above is their limitation to a single application, as a nutritional supplement or as an active ingredient of some cosmetic products or medical devices with a role in regulating the health of the skin and/or mucous membranes.
Sunt cunoscute și diferite alte compoziții care au utilizări multiple, ca ingrediente în special compoziții pe bază de polifenoli, acizi hidroxicinamici, antociani și/sau flavonoide. Polifenolii sunt larg utilizați în formulări de produse cosmetice (de Lima Cherubim et al., 2020; Zillich et al., 2015), ca și în suplimente nutritive, nutraceutice și alimente funcționale (Cory et al., 2018). Acizii hidroxicinamici au atât aplicații cosmeceutice (Taofiq et al., 2017), cât și aplicații nutraceutice (Silva & Batista, 2017). Antocianii se regăsesc ca ingrediente active în nutraceutice (Salehi et al., 2020; Zhu et al., 2016) și cosmetice (Diaconeasa et al., 2020). Flavonoidele au de asemenea efecte benefice ca ingrediente active în suplimente nutritive și nutraceutice (Egert & Rimbach, 2011; Li et al., 2022) și în produse cosmetice (Obaid et al., 2021). Compozițiile prebiotice și antioxidante cu polifenoli / acizi fenolici / flavonoide și, eventual, oligo/polizaharide prebiotice au de obicei aplicații multiple.Various other compositions are also known which have multiple uses, as ingredients especially compositions based on polyphenols, hydroxycinnamic acids, anthocyanins and/or flavonoids. Polyphenols are widely used in cosmetic formulations (de Lima Cherubim et al., 2020; Zillich et al., 2015), as well as in nutritional supplements, nutraceuticals and functional foods (Cory et al., 2018). Hydroxycinnamic acids have both cosmeceutical (Taofiq et al., 2017) and nutraceutical (Silva & Batista, 2017) applications. Anthocyanins are found as active ingredients in nutraceuticals (Salehi et al., 2020; Zhu et al., 2016) and cosmetics (Diaconeasa et al., 2020). Flavonoids also have beneficial effects as active ingredients in nutritional supplements and nutraceuticals (Egert & Rimbach, 2011; Li et al., 2022) and in cosmetic products (Obaid et al., 2021). Prebiotic and antioxidant compositions with polyphenols / phenolic acids / flavonoids and possibly prebiotic oligo/polysaccharides usually have multiple applications.
Brevetul EP2999687 B1 descrie un procedeu de extracție a acidului ferulic prezent într-o fază apoasă, obținut prin tratarea a cel puțin unui material vegetal și care conține, de asemenea, polizaharide, procesul menționat cuprinzând cel puțin următoarele etape: 1) tratamentul respectivului material vegetal urmat de o separare solid / lichid pentru a recupera o fază solidă și o fază lichidă apoasă cuprinzând acidul ferulic și respectivele polizaharide, 2) tratamentul fazei lichide menționate pentru a separa selectiv, pe de o parte, polizaharidele și, pe de altă parte, acid ferulic prezent într-o fracție apoasă, 3) concentrația fracției apoase menționate care conține acidul ferulic astfel încât să recupereze un flux concentrat de acid ferulic, 4) recuperarea acidului ferulic sub formă solidă.Patent EP2999687 B1 describes a process for extracting ferulic acid present in an aqueous phase, obtained by treating at least one plant material and which also contains polysaccharides, said process comprising at least the following steps: 1) treatment of said plant material followed by a solid/liquid separation to recover a solid phase and an aqueous liquid phase comprising ferulic acid and the respective polysaccharides, 2) treatment of said liquid phase to selectively separate, on the one hand, polysaccharides and, on the other hand, acid ferulic acid present in an aqueous fraction, 3) concentration of said aqueous fraction containing ferulic acid so as to recover a concentrated stream of ferulic acid, 4) recovery of ferulic acid in solid form.
Cererea de brevet RO135149 A0 prezintă un procedeul alcătuit din următoarele etape: măcinarea umedă a materialului vegetal, determinarea activității antioxidante de captare a radicalului 2,2-difenil-1-picrililhidrazil (DPPH), față de care se dozează un amestec de enzime de extracție format din pectinază și feruloil-esterază, ultrasonicare repetată a substratului vegetal cu enzime, urmată de extracția sub-critică în contracurent, separarea materialului vegetal de extractul alcoolic și concentrarea până la 35% substanță uscată cu recuperarea etanolului, din care rezultă un extract având un conținut minim de 2,7 g resveratrol, 12,2 g acizi hidroxicinamici totali și 18,3 g oligozaharide pectice la 100 ml extract etanolic concentrat și o activitate antioxidantă de captare a radicalului DPPH de cel puțin 3,5 mg echivalent Trolox per g extract. Compoziția rezultată este destinată utilizării pentru produse cosmetice și nutraceutice.The patent application RO135149 A0 presents a process consisting of the following steps: wet grinding of the plant material, determination of the antioxidant activity of capturing the radical 2,2-diphenyl-1-picrylylhydrazyl (DPPH), against which a mixture of extraction enzymes is dosed consisting of pectinase and feruloyl-esterase, repeated ultrasonication of the plant substrate with enzymes, followed by sub-critical countercurrent extraction, separation of the plant material from the alcoholic extract and concentration up to 35% dry substance with ethanol recovery, resulting in an extract having a minimum content of 2.7 g resveratrol, 12.2 g total hydroxycinnamic acids and 18.3 g pectic oligosaccharides per 100 ml concentrated ethanolic extract and a DPPH radical scavenging antioxidant activity of at least 3.5 mg Trolox equivalent per g extract . The resulting composition is intended for use in cosmetic and nutraceutical products.
Dezavantajul comun al unor astfel de compoziții este durata lor limitată de acțiune în timp. Pentru a crește durata de acțiune a diferitelor extracte au fost realizate formulări cu eliberare controlată. Cererea RO134737 A2 prezintă un procedeu de obținere a unor compoziții de ingrediente multifuncționale cu eliberare controlată pentru utilizare în industria alimentară. Procedeul, conform invenției, constă în gelifierea termică a compușilor activi din extractul din pieliță de struguri, având conținut ridicat în compuși flavonoidici într-o matrice de microîncapsulare din proteine de zer cu funcționalitate ridicată, modificare prin denaturare termică și reticulare enzimatică, rezultând hidrogeluri cu eficiență ridicată de încapsulare pentru compușii antocianici, având un conținut de antociani monomerici totali de 6,69...9,82 mg echiv. malvidin 3glucozid/100 g s.u. și activitate oxidantă de 38,74...45,19 mMol Trolox/g s.u.The common disadvantage of such compositions is their limited duration of action over time. To increase the duration of action of the various extracts, formulations with controlled release were made. Application RO134737 A2 presents a process for obtaining compositions of multifunctional ingredients with controlled release for use in the food industry. The process, according to the invention, consists in the thermal gelation of the active compounds from the grape skin extract, having a high content of flavonoid compounds in a microencapsulation matrix of whey proteins with high functionality, modification by thermal denaturation and enzymatic cross-linking, resulting in hydrogels with high encapsulation efficiency for anthocyanin compounds, having a total monomeric anthocyanin content of 6.69...9.82 mg equiv. malvidin 3glucoside/100 g s.u. and oxidizing activity of 38.74...45.19 mMol Trolox/g s.u.
Cererea de brevet US20200368168A1 descrie un procedeu de preparare a unor particule compozite prin electropulverizare, pentru a îmbunătăți stabilitatea polifenolilor, care include următoarele etape: pregătirea unui compozit pectină-silice; adăugarea de compuși polifenolici în compozitul pectină-silice pentru a prepara o soluție de bază; și electropulverizarea coaxială a soluției de bază și a soluției de acoperire pentru a prepara particule electropulverizate încărcate cu polifenoli.Patent application US20200368168A1 describes a process for preparing composite particles by electrospraying, to improve the stability of polyphenols, which includes the following steps: preparation of a pectin-silica composite; adding polyphenolic compounds to the pectin-silica composite to prepare a base solution; and coaxially electrospraying the base solution and the coating solution to prepare electrosprayed particles loaded with polyphenols.
Deși eliberează controlat polifenolii, compozițiile descrise până în prezent nu au o acțiune prelungită pentru că sunt îndepărtate de pe mucoase. Problema tehnică pe care o rezolvă această invenție este de a realiza o compoziție prebiotică, regenerantă și antioxidantă cu acțiune prelungită datorită adezivității sale la mucoase.Although they release polyphenols in a controlled manner, the compositions described so far do not have a prolonged action because they are removed from the mucous membranes. The technical problem that this invention solves is to make a prebiotic, regenerating and antioxidant composition with prolonged action due to its adhesiveness to mucous membranes.
Compoziția prebiotică și antioxidantă conține per 100 g produs 12,2 - 12,8 grame de nanoceluloză bacteriană, 0,164 - 0,216 grame de polifenoli totali, exprimați ca echivalent acid galic, 90-92 mg de siliciu solubil exprimat ca H4SiO4, 14,7 - 14,85 mg seleniu, predominant sub formă de nanoparticule cu seleniu zerovalent, și are o activitate antioxidantă de captare a radicalului DPPH, 2,2-difenil-1 -picrililhidrazil, de cel puțin 1,2 mg echivalent Trolox per g compoziție, un index de mucoadezivitate determinat reologic de minim 55 și un scor al activității prebiotice față de Lactobacillus johnsonii ATCC 33200 de cel puțin 1,05.The prebiotic and antioxidant composition contains per 100 g of product 12.2 - 12.8 grams of bacterial nanocellulose, 0.164 - 0.216 grams of total polyphenols, expressed as gallic acid equivalent, 90-92 mg of soluble silicon expressed as H 4 SiO 4 , 14 .7 - 14.85 mg of selenium, predominantly in the form of zero-valent selenium nanoparticles, and has a DPPH radical scavenging activity, 2,2-diphenyl-1-picrylylhydrazyl, of at least 1.2 mg Trolox equivalent per g composition, a rheologically determined mucoadhesive index of at least 55 and a prebiotic activity score against Lactobacillus johnsonii ATCC 33200 of at least 1.05.
Procedeul de obținere a acestei compoziții este alcătuit din următoarele etape:The process of obtaining this composition consists of the following stages:
Prepararea unei suspensii de frunze de ceai negru, în raport de 7 -10 g în 1000 ml apă adusă la 9O....95°C și micronizarea frunzelor de ceai și uniformizarea infuziei prin amestecarea viguroasă într-un omogenizator de înaltă presiune cu piston, prevăzut cu o valvă de tip muchie de cuțit, două cicluri la 250 MPa;Preparation of a suspension of black tea leaves, at the ratio of 7 -10 g in 1000 ml of water brought to 9O...95°C and micronization of the tea leaves and uniformity of the infusion by vigorous mixing in a high-pressure piston homogenizer , fitted with a knife edge valve, two cycles at 250 MPa;
Filtrarea infuziei rezultate și dizolvarea de zahăr cristal, în cantitate de 8 g la 100 ml de infuzie și răcirea infuziei îndulcite la temperatura camerei;Filtering the resulting infusion and dissolving crystal sugar, in the amount of 8 g per 100 ml of infusion and cooling the sweetened infusion to room temperature;
Adăugarea de 15 mg de selenit și 50 g de polen multifloral la 1000 ml de infuzie și inocularea infuziei îndulcite și tratate cu biofilm / peliculă de cultură de Kombucha / SCOBY, 5 părți biomasă umedă la 100 de părți infuzie îndulcită și cultivare statică a consorțiului SCOBY timp de 14 zile la temperatura de 25-28°C;Addition of 15 mg of selenite and 50 g of multifloral pollen per 1000 ml infusion and inoculation of the sweetened and treated infusion with biofilm / Kombucha / SCOBY culture film, 5 parts wet biomass to 100 parts sweetened infusion and static cultivation of the SCOBY consortium for 14 days at a temperature of 25-28°C;
Recoltarea peliculei de Kombucha / SCOBY, îndepărtarea melanoidinelor prin spălare repetată cu două soluții alcaline, respectiv 1M si 4M NaOH, asistată de ultrasonare la o amplitudine de 60 pm și cu un aport specific de energie de 25 J, urmată de clătirea intensivă cu apă distilată a peliculelor până la un pH apropiat de neutru (7,5 8), mărunțirea preliminară a peliculelor utilizând un tocător electric cu cuțite, prelucrarea mecanică avansată utilizând o moară coloidală pentru obținerea de microfibrile; fracționarea microfibrilelor celulozice la presiune de 1500 bari, utilizând un microfluidizator cu cameră diamantată în formă de Z, cu scopul obținerii de nanofibrile celulozice;Kombucha / SCOBY film harvesting, melanoidin removal by repeated washing with two alkaline solutions, respectively 1M and 4M NaOH, assisted by ultrasound at an amplitude of 60 pm and with a specific energy input of 25 J, followed by intensive rinsing with distilled water of the films to a pH close to neutral (7.5 8), preliminary comminution of the films using an electric chopper with knives, advanced mechanical processing using a colloidal mill to obtain microfibrils; fractionation of cellulose microfibrils at a pressure of 1500 bars, using a microfluidizer with a Z-shaped diamond chamber, in order to obtain cellulose nanofibrils;
îndepărtarea grăuncioarelor de polen prin filtrare, neutralizarea filtratului la pH=7 și omogenizarea nanocelulozei cu filtratul de Kombucha neutralizat, în raport de 1 parte suspensie nanoceluloză la 10 părți supernatant, folosind un omogenizator cu piston, la o presiune de 500 bari, 2 treceri și uscarea prin pulverizare a suspensiei rezultate, la o temperatură de intrare de 140 - 145°C și la o temperatură de ieșire de 70-75°C.removing the pollen grains by filtration, neutralizing the filtrate to pH=7 and homogenizing the nanocellulose with the neutralized Kombucha filtrate, in a ratio of 1 part nanocellulose suspension to 10 parts supernatant, using a piston homogenizer, at a pressure of 500 bar, 2 passes and spray drying the resulting slurry at an inlet temperature of 140-145°C and an outlet temperature of 70-75°C.
Avantajele invenției sunt următoarele:The advantages of the invention are as follows:
Pulberea obținută are utilizări diverse, ca supliment nutritiv sau ca ingredient activ al unor produse cosmetice sau dispozitive medicale cu rol în reglarea sănătății pielii și/sau a mucoaselor;The obtained powder has various uses, as a nutritional supplement or as an active ingredient of some cosmetic products or medical devices with a role in regulating the health of the skin and/or mucous membranes;
Efectul prebiotic și antioxidant este amplificat de interacțiunea dintre nanoparticulele de seleniu și polifenoli;The prebiotic and antioxidant effect is amplified by the interaction between selenium nanoparticles and polyphenols;
Seleniu determină inhibarea producerii de melanoidine de către bacteriile acetice, iar acest faptfacilitează procesul de purificare a celulozei bacteriene, ca și procesul ulterior de obținere a nanocelulozei prin nanofibrilare;Selenium inhibits the production of melanoidins by acetic bacteria, and this fact facilitates the purification process of bacterial cellulose, as well as the subsequent process of obtaining nanocellulose through nanofibrillation;
Siliciu eliberat din grăuncioarele de polen împreună cu nanoceluloza bacteriană are efect regenerativ;Silicon released from pollen grains together with bacterial nanocellulose has a regenerative effect;
Produsele pe baza pulberii obținute auo acțiune prelungită datorită mucoadezivitătii recunoscute a nanocelulozei, care favorizează aderenta la diferitele tipuri de mucoase, intestinală, vaginală, nazală, ca și pe piele.The products based on the obtained powder have a prolonged action due to the recognized mucoadhesiveness of nanocellulose, which favors adhesion to different types of mucous membranes, intestinal, vaginal, nasal, as well as on the skin.
Prezenta invenție este ilustrată prin următoarele exemple, care ilustrează invenția fără a o limita. 1The present invention is illustrated by the following examples, which illustrate the invention without limiting it. 1
Exemplu 1. Se prepară într-un vas de sticlă termorezistentă o suspensie de frunze de ceai negru,7 g în 1000 ml apă adusă la 9O....95°C. Suspensia de frunze de ceai se micronizează și se uniformizează infuzia, prin amestecarea viguroasă într-un omogenizator de înaltă presiune cu piston (de ex. GEA Niro Soavi Arriete NS2006, Parma, Italia), prevăzut cu o valvă de tip muchie de cuțit, două cicluri la 250 MPa. Se filtrează infuzia rezultată pe filtru cu hârtie de filtru (Whatman™ nr. 1, Cytiva, Marlborough, MA, SUA). în suspensia rezultată se dizolvă zahăr cristal, în cantitate de g la litru de infuzie filtrată. Infuzia se răcește la temperatura camerei. Se adaugă 15 mg de selenit la 1 litru de infuzie și 50 grame polen multifloral. Se inoculează cu biofilm / peliculă de cultură de Kombucha / SCOBY, 5 părți biomasă umedă la 100 de părți infuzie îndulcită. Se cultivă consorțiul SCOBY timp de 14 zile la temperatura de 25°C. Se recoltează biofilmul / pelicula de Kombucha, care este format(ă) din celuloză bacteriană. Se îndepărtează melanoidinele prin spălare repetată cu două soluții alcaline, respectiv 1M si 4M NaOH, asistată de ultrasonare la o amplitudine de 60 pm și cu un aport specific de energie de 25 J (asigurate de ex. procesor ultrasonic UP50H, Hielscher, Teltow, Germania). Se clătesc intensiv cu apă distilată pelicule până la un pH apropiat de neutru (7,5 - 8). Se mărunțesc peliculele Kombucha utilizând un tocător electric cu cuțite (ca de ex. blende Bosch, Gerlingen, Germania). Se continuă prelucrarea mecanică avansată, utilizând o moară coloidală (ca de ex. MK 2000, IKA, Staufen, Germania), pentru obținerea de microfibrile de celuloză bacteriană. Microfibrilele de celuloză se fractioneazăla nanofibrile celulozice, utilizând un microfluidizator (de ex. LM10 Microfluidizer®, Microfluidics, Westwood, MA, SUA),cu cameră diamantată în formă de Z, operat la o presiune de 1500 bari.Example 1. A suspension of black tea leaves, 7 g in 1000 ml of water brought to 9O...95°C, is prepared in a heat-resistant glass vessel. The tea leaf suspension is micronized and the infusion homogenized by vigorous mixing in a high-pressure piston homogenizer (eg GEA Niro Soavi Arriete NS2006, Parma, Italy) fitted with a knife-edge valve, two cycles at 250 MPa. Filter the resulting infusion on filter paper (Whatman™ No. 1, Cytiva, Marlborough, MA, USA). crystal sugar is dissolved in the resulting suspension, in the amount of g per liter of filtered infusion. The infusion is cooled to room temperature. Add 15 mg of selenite to 1 liter of infusion and 50 grams of multifloral pollen. Inoculate with biofilm / Kombucha culture film / SCOBY, 5 parts wet biomass to 100 parts sweetened infusion. Cultivate the SCOBY consortium for 14 days at 25°C. The Kombucha biofilm is harvested, which consists of bacterial cellulose. The melanoidins are removed by repeated washing with two alkaline solutions, respectively 1M and 4M NaOH, assisted by ultrasound at an amplitude of 60 pm and with a specific energy input of 25 J (provided by e.g. ultrasonic processor UP50H, Hielscher, Teltow, Germany ). Rinse the films intensively with distilled water until the pH is close to neutral (7.5 - 8). Shred the Kombucha films using an electric chopper with knives (eg Blende Bosch, Gerlingen, Germany). Advanced mechanical processing continues, using a colloidal mill (eg MK 2000, IKA, Staufen, Germany), to obtain bacterial cellulose microfibrils. Cellulose microfibrils are fractionated to cellulose nanofibrils using a microfluidizer (eg LM10 Microfluidizer®, Microfluidics, Westwood, MA, USA), with a Z-shaped diamond chamber, operated at a pressure of 1500 bar.
Se îndepărtează grăuncioarele de polen prin filtrare. Filtratul obținut, care este de culoare roșiatică, datorită formării nanoparticulelor de seleniu zerovalent, se neutralizează cu hidroxid de sodiu până la pH-7. Se amestecă filtratul cu nanoceluloza obținută prin fibrilare. Se adaugă 100 grame de nanoceluloză, la 1000 ml supernatant de Kombucha. Se omogenizează folosind un omogenizator cu piston (de ex. GEA Niro Soavi Arriete NS2006, Parma, Italia), operat la o presiune de 500 bari. Se fac două treceri, pentru a fi omogenizate și nanoparticulele de seleniu în amestecul final. Compoziția rezultată se usucă prin pulverizare (de ex. pe un uscător prin pulverizare (ca de ex.Mini Spray Dryer S-300. Buchi, Flawil, Switzerland), la o temperatură de intrare de 140 - 145°C și la o temperatură de ieșire de 70-75°C.The pollen grains are removed by filtration. The filtrate obtained, which is reddish in color due to the formation of zerovalent selenium nanoparticles, is neutralized with sodium hydroxide to pH-7. Mix the filtrate with the nanocellulose obtained by fibrillation. Add 100 grams of nanocellulose to 1000 ml of Kombucha supernatant. Homogenize using a piston homogenizer (eg GEA Niro Soavi Arriete NS2006, Parma, Italy), operated at a pressure of 500 bar. Two passes are made to homogenize the selenium nanoparticles in the final mixture. The resulting composition is spray-dried (e.g. on a spray dryer (such as Mini Spray Dryer S-300. Buchi, Flawil, Switzerland), at an inlet temperature of 140 - 145°C and at a temperature of output of 70-75°C.
Se ia circa un 1 gram din compoziția rezultată, care se cântărește cu precizie și se reconstituie cu 25 ml apă, prin ultrasonare (procesor ultrasonic UP50H, Hielscher, Teltow). în compoziția reconstituită rezultată se determină polifenolii totali cu reactiv Folin Ciocâlteu (Ainsworth & Gillespie, 2007), celuloza totală prin metoda colorimetrică rapidă (Bauer & Ibânez, 2014), siliciul solubil prin metoda colorimetrică (Coradin et al., 2002), seleniul - prin ICP-MS (Constantinescu-Aruxandei et al., 2018). Se exprimă per 100 grame produs. Se determină o cantitate de 12,2 grame de nanoceluloză bacteriană, 0,164 grame de polifenoli totali, exprimați ca echivalent acid galic, 90-92 mg de siliciu solubil exprimat ca hUSiCM, și 14,7 mg seleniu pe 100 grame de produs.About 1 gram of the resulting composition is taken, which is accurately weighed and reconstituted with 25 ml of water by ultrasonication (Ultrasonic processor UP50H, Hielscher, Teltow). in the resulting reconstituted composition, total polyphenols are determined with the Folin Ciocâlteu reagent (Ainsworth & Gillespie, 2007), total cellulose by the rapid colorimetric method (Bauer & Ibânez, 2014), soluble silicon by the colorimetric method (Coradin et al., 2002), selenium - by ICP-MS (Constantinescu-Aruxandei et al., 2018). It is expressed per 100 grams of product. An amount of 12.2 grams of bacterial nanocellulose, 0.164 grams of total polyphenols, expressed as gallic acid equivalent, 90-92 mg of soluble silicon expressed as hUSiCM, and 14.7 mg of selenium per 100 grams of product is determined.
Se determină în compoziția reconstituită activitatea antioxidantă prin metoda capacității antioxidante relative (Cheng et al., 2006). Rezultă o activitate de 1,25 mg echivalent Trolox per g compoziție. Se determină reologic (pe un reometru hibrid HR 30, TA Instruments, New Castle, DE, USA), la o rată de forfecare de 33.9 s-1, indicele de mucoadezivitate, conform metodei descrise recent (Cernohlâvek et al., 2021). Indicele de mucoadezivitate este 55,1. Compoziția rezultată este evaluată pentru activitatea prebiotică, conform metodei de raportare a creșterii pe mediu cu 1% compoziție prebiotică față de creșterea pe mediu cu 1% glucoză (Huebner et al., 2007), și se determină unscor al activității prebiotice față de Lactobacillus johnsonii ATCC 33200, de 1,05.The antioxidant activity is determined in the reconstituted composition by the relative antioxidant capacity method (Cheng et al., 2006). This results in an activity of 1.25 mg Trolox equivalent per g composition. The mucoadhesiveness index is determined rheologically (on a HR 30 hybrid rheometer, TA Instruments, New Castle, DE, USA), at a shear rate of 33.9 s -1 , according to the recently described method (Cernohlâvek et al., 2021). The mucoadhesiveness index is 55.1. The resulting composition is evaluated for prebiotic activity according to the method of reporting growth on medium with 1% prebiotic composition versus growth on medium with 1% glucose (Huebner et al., 2007), and a prebiotic activity score against Lactobacillus johnsonii is determined ATCC 33200, of 1.05.
Se determină efectul regenerativ al compoziției rezultate prin utilizarea metodei leziunilor din culturi de celule - scratch-wound assay (Cory, 2011). Se pune în evidență efectul regenerativ al compoziției realizate.The regenerative effect of the resulting composition is determined by using the method of injuries from cell cultures - scratch-wound assay (Cory, 2011). The regenerative effect of the created composition is highlighted.
Exemplu 2. Se lucrează la fel ca în exemplul 1, cu singura diferență că se folosesc 10 grame de ceai negru. Compoziția rezultată conține per 100 g produs 12,8 grame de nanoceluloză bacteriană, 0,216 grame de polifenoli totali, exprimați ca echivalent acid galic, 92 mg de siliciu solubil exprimat ca HUSiCU, 14,85 mg seleniu, predominant ca nanoparticule de seleniu zerovalent, și are o activitate antioxidantă de captare a radicalului DPPH, de 1,35 mg echivalent Trolox per g compoziție, un index de mucoadezivitate determinat reologic 58,2 și un scor al activității prebiotice față de Lactobacillus johnsonii ATCC 33200 de 1,09. Compoziția are, de asemenea, un efect regenerativ.Example 2. Work the same as in example 1, with the only difference being that 10 grams of black tea are used. The resulting composition contains per 100 g of product 12.8 grams of bacterial nanocellulose, 0.216 grams of total polyphenols, expressed as gallic acid equivalent, 92 mg of soluble silicon expressed as HUSiCU, 14.85 mg of selenium, predominantly as zerovalent selenium nanoparticles, and has a DPPH radical scavenging antioxidant activity of 1.35 mg Trolox equivalent per g composition, a rheologically determined mucoadhesive index of 58.2 and a prebiotic activity score against Lactobacillus johnsonii ATCC 33200 of 1.09. The composition also has a regenerative effect.
Fișă bibliograficăBibliographic sheet
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