BR102022017979A2 - METHOD OF MICROENCAPSULATION OF LACTOBACILLUS HELVETICUS BY SPRAY DRIER AND APPLICATION IN WHITE CHOCOLATE - Google Patents
METHOD OF MICROENCAPSULATION OF LACTOBACILLUS HELVETICUS BY SPRAY DRIER AND APPLICATION IN WHITE CHOCOLATE Download PDFInfo
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- BR102022017979A2 BR102022017979A2 BR102022017979-4A BR102022017979A BR102022017979A2 BR 102022017979 A2 BR102022017979 A2 BR 102022017979A2 BR 102022017979 A BR102022017979 A BR 102022017979A BR 102022017979 A2 BR102022017979 A2 BR 102022017979A2
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- probiotic
- chocolate
- white chocolate
- microencapsulation
- lactobacillus helveticus
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Abstract
A presente invenção descreve um método de microencapsulação de Lactobacillus helveticus por spray drier em matriz polimérica (leite em pó/maltodextrina/trealose/frutoligossacarídeo/amido), investigando a resistência do probiótico encapsulado sob diferentes condições de pH (2 a 7), NaCl (3 a 9%), sacarose (10 a 45%) e temperatura (50ºC) e em matriz de chocolate branco durante o armazenamento a 22ºC. As amostras de chocolate armazenadas foram analisadas quanto à cor (L*, a*, b* e ΔE), pH, acidez, atividade de água, umidade, viabilidade probiótica e modelagem matemática da estimativa da viabilidade do probiótico ao armazenamento. A maior taxa de sobrevivência do probiótico encapsulado foi de 96 % secagem por spray-drier a 110°C. As microcápsulas apresentaram um formato arredondado e com tamanhos similares (2,20 µm ±0,30), sem poros ou rachaduras e com a presença de algumas concavidades ou achatamentos na superfície. O modelo de Weibull permitiu estimar a sobrevivência das células probióticas livres e encapsuladas mostrando serem viáveis em quantidade suficiente para proporcionar potenciais benefícios para a saúde do consumidor (>6 log UFC\g), em matriz de chocolate branco por aproximadamente 90 e 130 dias de armazenamento a 22°C. O chocolate branco pode potencialmente ser usado como carreador de L. helveticus com propriedade probiótica e características físico-químicas semelhantes ao chocolate branco tradicional.The present invention describes a method of microencapsulation of Lactobacillus helveticus by spray drier in a polymeric matrix (milk powder/maltodextrin/trehalose/fructoligosaccharide/starch), investigating the resistance of the encapsulated probiotic under different conditions of pH (2 to 7), NaCl ( 3 to 9%), sucrose (10 to 45%) and temperature (50ºC) and in white chocolate matrix during storage at 22ºC. The stored chocolate samples were analyzed for color (L*, a*, b* and ΔE), pH, acidity, water activity, humidity, probiotic viability and mathematical modeling of the estimation of probiotic viability during storage. The highest survival rate of the encapsulated probiotic was 96% spray-drying at 110°C. The microcapsules had a rounded shape and similar sizes (2.20 µm ±0.30), without pores or cracks and with the presence of some concavities or flattening on the surface. The Weibull model allowed estimating the survival of free and encapsulated probiotic cells, showing that they are viable in sufficient quantity to provide potential benefits to the health of the consumer (>6 log CFU\g), in white chocolate matrix for approximately 90 and 130 days of storage at 22°C. White chocolate can potentially be used as a carrier for L. helveticus with probiotic properties and physicochemical characteristics similar to traditional white chocolate.
Description
[001] A presente invenção descreve um método de microencapsulação de Lactobacillus helveticus em diferentes matrizes (Quitosana, gelatinas, maltodextrina, goma arábica, leite em pó, trealose, frutooligosacarídeo, amido) e proporções a serem utilizadas no encapsulamento do probiótico para posterior aplicação em chocolate branco utilizando spray-drier.[001] The present invention describes a method of microencapsulation of Lactobacillus helveticus in different matrices (Chitosan, gelatins, maltodextrin, gum arabic, milk powder, trehalose, fructooligosaccharide, starch) and proportions to be used in encapsulating the probiotic for subsequent application in white chocolate using spray-drier.
[002] Inúmeras pessoas associam o consumo do chocolate com a sensação de prazer, consumindo o chocolate em diferentes momentos. Este fator se justifica devido à serotonina e feniletilamina, substâncias envolvidas com a excitação dos órgãos e pelo sentimento de euforia e êxtase, que são liberadas quando se consome o chocolate (ANDRADE et al., 2003).[002] Many people associate the consumption of chocolate with the feeling of pleasure, consuming chocolate at different times. This factor is justified due to serotonin and phenylethylamine, substances involved in the excitement of the organs and the feeling of euphoria and ecstasy, which are released when chocolate is consumed (ANDRADE et al., 2003).
[003] O chocolate pode ser definido como uma suspensão de partículas sólidas composta principalmente de açúcar e cacau, envolvidas pela matriz lipídica da manteiga de cacau. Portanto, a manteiga de cacau representa a fase contínua do chocolate, sendo a matriz dispersante para as partículas sólidas do cacau, açúcar e outros ingredientes (LOPES, 2015). Deve fundir rápido e completamente em temperatura próxima à do corpo humano, caso contrário, poderá promover um pobre desprendimento de aroma/sabor e, provavelmente, um residual ceroso (VISSOTTO et al., 1999). Os principais tipos, chocolate escuro, ao leite e branco diferem principalmente no conteúdo de sólidos do cacau, gordura do leite e manteiga de cacau (HUSSAIN et al., 2020).[003] Chocolate can be defined as a suspension of solid particles composed mainly of sugar and cocoa, surrounded by the lipid matrix of cocoa butter. Therefore, cocoa butter represents the continuous phase of chocolate, being the dispersing matrix for solid cocoa particles, sugar and other ingredients (LOPES, 2015). It must melt quickly and completely at a temperature close to that of the human body, otherwise it could promote poor aroma/flavor release and, probably, a waxy residue (VISSOTTO et al., 1999). The main types, dark, milk and white chocolate, differ mainly in the content of cocoa solids, milk fat and cocoa butter (HUSSAIN et al., 2020).
[004] A legislação brasileira define chocolate como produto obtido a partir da mistura de derivados de cacau (Theobroma cacao L.), massa (pasta ou líquor) de cacau, cacau em pó e ou manteiga de cacau, com outros ingredientes, contendo no mínimo 25% (m/m) de sólidos totais de cacau. O produto pode apresentar recheio, cobertura, formato e consistência variados. Chocolate branco é o produto obtido a partir da mistura de manteiga de cacau com outros ingredientes, contendo, no mínimo 20% (m/m) de sólidos totais de manteiga de cacau. O produto pode apresentar recheio, cobertura, formato e consistência variados (BRASIL, 2005).[004] Brazilian legislation defines chocolate as a product obtained from the mixture of cocoa derivatives (Theobroma cacao L.), cocoa mass (paste or liquor), cocoa powder and/or cocoa butter, with other ingredients, containing in the minimum 25% (m/m) total cocoa solids. The product may have different filling, coverage, shape and consistency. White chocolate is the product obtained from mixing cocoa butter with other ingredients, containing at least 20% (w/w) of total cocoa butter solids. The product may have varying filling, coverage, shape and consistency (BRASIL, 2005).
[005] O chocolate branco não contém líquor de cacau, sendo elaborado apenas com açúcar, manteiga de cacau e leite. Além desses ingredientes básicos, também são utilizados nas formulações, emulsificantes e aromatizantes (VISSOTTO et al., 1999).[005] White chocolate does not contain cocoa liquor, being made only with sugar, cocoa butter and milk. In addition to these basic ingredients, emulsifiers and flavorings are also used in formulations (VISSOTTO et al., 1999).
[006] Os tipos de chocolate variam de acordo com os ingredientes adicionados, sua forma e a mistura de cacau utilizada. Se não for adicionado à massa de cacau tem-se o chocolate branco, o qual é constituído apenas de manteiga de cacau com açúcar, emulsificantes e aromatizantes. No chocolate meio amargo não é adicionado leite. Se for substituído a manteiga de cacau por gordura hidrogenada, menos onerosa, tem-se o compound, que pode ser usado como cobertura de bolos e sorvetes, mas se for vendido como chocolate é considerado fraude. Entre as várias formas apresentadas ao consumidor, tem-se o chocolate ao leite ou amargo em tabletes, adicionados de amêndoas, castanha, amendoim, uva-passa e similares, bombons recheados com vários tipos de componentes (OETTERER et al., 2006).[006] The types of chocolate vary according to the ingredients added, their shape and the cocoa mixture used. If it is not added to the cocoa mass, there is white chocolate, which consists only of cocoa butter with sugar, emulsifiers and flavorings. No milk is added to semi-sweet chocolate. If cocoa butter is replaced with hydrogenated fat, which is less expensive, you have the compound, which can be used as a topping for cakes and ice creams, but if it is sold as chocolate it is considered fraud. Among the various forms presented to the consumer, there is milk or dark chocolate in tablets, with the addition of almonds, chestnuts, peanuts, raisins and the like, chocolates filled with various types of components (OETTERER et al., 2006).
[007] Na definição de Vercet (2003), o chocolate branco é composto de açúcar, sólidos de leite, manteiga de cacau, lecitina de soja e vanilina.[007] In the definition of Vercet (2003), white chocolate is composed of sugar, milk solids, cocoa butter, soy lecithin and vanillin.
[008] O termo “alimentos funcionais” foi inicialmente introduzido pelo governo do Japão em meados dos anos 1980, como o resultado de esforços para desenvolver alimentos que possibilitassem a redução dos gastos com saúde pública, considerando a elevada expectativa de vida naquele país (STRINGHETA et al., 2007).[008] The term “functional foods” was initially introduced by the government of Japan in the mid-1980s, as the result of efforts to develop foods that would make it possible to reduce public health spending, considering the high life expectancy in that country (STRINGHETA et al., 2007).
[009] O Brasil (BRASIL, 2008), reconhece até o momento as seguintes alegações de propriedades funcionais aprovadas: ácidos graxos, carotenoides, fibras alimentares, fitoesteróis, proteína de soja, polióis e probióticos. Existem diversos tipos de alimentos funcionais, porém os que recebem maior destaque são os probióticos, os prebióticos e os simbióticos (SAAD et al., 2011).[009] Brazil (BRASIL, 2008) currently recognizes the following claims of approved functional properties: fatty acids, carotenoids, dietary fiber, phytosterols, soy protein, polyols and probiotics. There are several types of functional foods, but the ones that receive the most attention are probiotics, prebiotics and synbiotics (SAAD et al., 2011).
[0010] Segundo BADARÓ et al. (2009) os probióticos são micro organismos vivos que, quando integrados na alimentação, são classificados de alimentos funcionais por beneficiarem a saúde do hospedeiro com risco reduzido de causar doença no consumidor. Probióticos são micro-organismos vivos que oferecem a seu hospedeiro vários benefícios à saúde, mantendo a microbiota intestinal normal, inibindo a adesão de bactérias patogênicas à mucosa intestinal, aumentando a imunidade e reduzindo os níveis de colesterol (LIANG et al., 2020; EVIVIE et al., 2019).[0010] According to BADARÓ et al. (2009) probiotics are live microorganisms that, when integrated into food, are classified as functional foods because they benefit the health of the host with a reduced risk of causing illness in the consumer. Probiotics are live microorganisms that offer their host several health benefits, maintaining normal intestinal microbiota, inhibiting the adhesion of pathogenic bacteria to the intestinal mucosa, increasing immunity and reducing cholesterol levels (LIANG et al., 2020; EVIVIE et al., 2019).
[0011] A definição de probióticos mais aceita internacionalmente é a adotada pela Organização da Agricultura e Alimentação das Nações Unidas (FAO) e Organização Mundial de Saúde (WHO). Segundo a FAO/WHO (2002), probióticos são definidos como “microorganismos vivos que, quando administrados em quantidades adequadas, conferem benefícios à saúde do hospedeiro”.[0011] The most internationally accepted definition of probiotics is that adopted by the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO). According to FAO/WHO (2002), probiotics are defined as “live microorganisms that, when administered in adequate quantities, confer health benefits on the host”.
[0012] Os micro-organismos utilizados como probióticos são usualmente componentes não patogênicos da microbiota humana, tais como bactérias produtoras de ácido láctico: Lactobacillus, Pediococcus, Leuconostoc, Bifidobacterium e alguns Streptococcus; e uma levedura: Saccharomyces boulardii (BARBOSA et al., 2006).[0012] The microorganisms used as probiotics are usually non-pathogenic components of the human microbiota, such as lactic acid-producing bacteria: Lactobacillus, Pediococcus, Leuconostoc, Bifidobacterium and some Streptococcus; and a yeast: Saccharomyces boulardii (BARBOSA et al., 2006).
[0013] Os dois gêneros mais comumente usados como microrganismos probióticos são Bifidobacterium sp. e Lactobacillus sp. (HOJJATI et al., 2020).[0013] The two genera most commonly used as probiotic microorganisms are Bifidobacterium sp. and Lactobacillus sp. (HOJJATI et al., 2020).
[0014] Dentre os micro-organismos que resistem a temperaturas superiores a 35 °C, encontrasse o Lactobacillus helveticus, pois apresenta bom crescimento entre 40 a 45°C, entretanto, podem crescer até 50 - 52°C, não se desenvolvendo em temperaturas inferiores a 15 °C. Geralmente resistem a temperaturas de 60 °C por 90 min (FURTADO, 1990). Sendo que está condição de temperatura possibilita a aplicação em diferentes produtos onde tem-se o calor empregado no processo de fabricação, como é o caso do chocolate.[0014] Among the microorganisms that resist temperatures above 35 °C, Lactobacillus helveticus is found, as it shows good growth between 40 and 45 °C, however, they can grow up to 50 - 52 °C, not developing at temperatures below 15°C. They generally resist temperatures of 60 °C for 90 min (FURTADO, 1990). Since this temperature condition makes it possible to apply it to different products where heat is used in the manufacturing process, as is the case with chocolate.
[0015] Um dos grandes desafios das indústrias de alimentos é manter a estabilidade de compostos como vitaminas, antioxidantes, corantes naturais, conservantes, probióticos entre outros. Pesquisas têm demonstrado que as culturas probióticas podem ser significativamente protegidas através da técnica de microencapsulação. Na área de alimentos tem sido uma alternativa empregada para resolver os problemas de instabilidade e inviabilidade de probióticos (KAILASAPATHY, 2009).[0015] One of the great challenges of the food industry is maintaining the stability of compounds such as vitamins, antioxidants, natural dyes, preservatives, probiotics, among others. Research has demonstrated that probiotic cultures can be significantly protected through the microencapsulation technique. In the food sector, it has been an alternative used to solve the problems of instability and unviability of probiotics (KAILASAPATHY, 2009).
[0016] Dentre as pesquisas realizadas nos últimos anos, com encapsulamento de probióticos tem-se o desenvolvimento de barra de cereal salgada com L. acidophilus LA3, Bifidobarcterium animalis subsp. lactis BL- C1 empregando o método de spray chilling com matriz de óleo de algodão (Bampi et al., 2016). As condições empregadas neste processo foram: homogeneização de 7000 rpm por 60 s, temperatura de aquecimento da matriz lipídica 53 °C e atomização a 5 bar; temperatura da câmara 15 °C ± 2 °C.[0016] Among the research carried out in recent years, with encapsulation of probiotics, there is the development of a salted cereal bar with L. acidophilus LA3, Bifidobarcterium animalis subsp. lactis BL-C1 using the spray chilling method with cottonseed oil matrix (Bampi et al., 2016). The conditions used in this process were: homogenization at 7000 rpm for 60 s, lipid matrix heating temperature 53 °C and atomization at 5 bar; chamber temperature 15 °C ± 2 °C.
[0017] Na pesquisa realizada por Alves et al. (2016), com suco de laranja fermentado em pó contendo Lactobacillus casei NRRL B-442 encapsulado em matriz de maltodextrina (50%) e goma arábica (50%), seguida da secagem em spray dryer 140 e o leito de jus seco a 60, respectivamente. Sendo que as condições de temperatura de entrada do equipamento foi de 140°C e a temperatura de saída de 60 °C, com uma vazão de 0,2 L/h.[0017] In the research carried out by Alves et al. (2016), with fermented orange juice powder containing Lactobacillus casei NRRL B-442 encapsulated in a matrix of maltodextrin (50%) and gum arabic (50%), followed by drying in a spray dryer at 140 and the jus bed dried at 60 , respectively. The equipment's inlet temperature conditions were 140°C and the outlet temperature was 60°C, with a flow rate of 0.2 L/h.
[0018] No estudo realizado por Pereira, Maciel e Rodrigues (2011), com suco de caju e maçã fermentado em pó, também encapsulados pela técnica de spray dryer, usaram como agentes encapsulantes maltodextrina (60%) e goma arábica (40%). A temperatura de entrada do ar de secagem foi 120 °C, com vazão de alimentação de 0,3 L/h.[0018] In the study carried out by Pereira, Maciel and Rodrigues (2011), with powdered fermented cashew and apple juice, also encapsulated by the spray dryer technique, they used maltodextrin (60%) and gum arabic (40%) as encapsulating agents. . The drying air inlet temperature was 120 °C, with a supply flow of 0.3 L/h.
[0019] Na patente EP3593639A1 intitulada Probiotic Fortified Food Products And Methods Of Manufacture é descrito um método de fabricação de um produto alimentício fortificado com probiótico caracterizado pela etapa de aplicação de uma composição inoculada com pelo menos um organismo probiótico em pelo menos uma porção da superfície do produto alimentício, em que a composição inclui uma base que é totalmente láctea. derivados e/ou inclui componentes inerentemente encontrados no leite.[0019] In the patent EP3593639A1 entitled Probiotic Fortified Food Products And Methods Of Manufacture, a method of manufacturing a probiotic-fortified food product characterized by the step of applying a composition inoculated with at least one probiotic organism on at least a portion of the surface is described. of the food product, in which the composition includes a base that is entirely dairy. derivatives and/or includes components inherently found in milk.
[0020] A patente US2018/0271134A1 intitulada Thickened Oil/Water Food Emulsion Packaged Products With Probiotics descreve diferentes composições alimentares emulsionadas embaladas (por exemplo, molhos para salada incluindo uma emulsão de óleo/água) incluindo um probiótico, que pode ser incluído nelas, sem exigir aquecimento durante a embalagem e sem exigir refrigeração para estabilidade de prateleira a longo prazo. O probiótico pode ser formador de esporos. Em uma modalidade, o probiótico compreende esporos que não germinam até serem ingeridos por um consumidor (por exemplo, de modo que os esporos germinem no trato intestinal do consumidor). O probiótico pode compreender esporos de Bacillus coagulans, que podem ser adicionados à emulsão após a adição de um ácido. O molho para salada ou outra composição alimentar pode ser vantajosamente embalado a frio, não necessitando de aquecimento durante a fabricação e sem refrigeração após a embalagem, proporcionando estabilidade de 1 ano ou mais.[0020] Patent US2018/0271134A1 entitled Thickened Oil/Water Food Emulsion Packaged Products With Probiotics describes different packaged emulsified food compositions (for example, salad dressings including an oil/water emulsion) including a probiotic, which can be included therein, without requiring heating during packaging and without requiring refrigeration for long-term shelf stability. The probiotic can be spore forming. In one embodiment, the probiotic comprises spores that do not germinate until ingested by a consumer (e.g., such that the spores germinate in the intestinal tract of the consumer). The probiotic may comprise spores of Bacillus coagulans, which may be added to the emulsion after adding an acid. Salad dressing or other food composition can advantageously be cold packaged, requiring no heating during manufacturing and no refrigeration after packaging, providing stability of 1 year or more.
[0021] A invenção US10897922B2 intitulada Composition and method for germinative compounds in probiotic food and beverage products for human consumption apresenta uma composição e método para ativar esporos probióticos em produtos alimentícios e bebidas, como chás embebidos, café, sopas e molhos. Uma composição nutriente-germinante compreende um ou mais L-aminoácidos, opcionalmente um ou mais tampões para manter o pH da composição quando adicionado à água em uma faixa de cerca de 6-8, opcionalmente D-glicose, D-frutose ou ambos D-glicose e D-frutose e, opcionalmente, um osmoprotetor. A composição nutriente-germinante, uma ou mais espécies de esporos de Bacillus e um produto alimentício ou de bebida podem ser pré-misturados em qualquer combinação. A água é adicionada à mistura e aquecida a uma faixa de temperatura de 42 a 100°C para germinar os esporos probióticos antes de serem consumidos.[0021] The invention US10897922B2 entitled Composition and method for germinative compounds in probiotic food and beverage products for human consumption presents a composition and method for activating probiotic spores in food and beverage products, such as steeped teas, coffee, soups and sauces. A seed-nutrient composition comprises one or more L-amino acids, optionally one or more buffers to maintain the pH of the composition when added to water in a range of about 6-8, optionally D-glucose, D-fructose or both D- glucose and D-fructose and, optionally, an osmoprotectant. The nutrient-germinant composition, one or more species of Bacillus spores and a food or beverage product can be premixed in any combination. Water is added to the mixture and heated to a temperature range of 42 to 100°C to germinate the probiotic spores before they are consumed.
[0022] A patente WO2009/116864A1 intitulada Long-Life Probiotic Food Product apresenta um produto alimentício, como um laticínio ou um produto de frutas de natureza longa vida (ou seja, pode ser armazenado não refrigerado por um longo período de tempo) e que fornece atividade probiótica ao consumidor. Este último é feito pela inclusão de esporos de bactérias probióticas. O produto é particularmente selecionado do grupo que consiste em produtos lácteos (como, por exemplo, bebidas lácteas, produtos alimentícios derivados de lácteos e ingredientes de alimentos lácteos), bebidas de frutas ou seus concentrados.[0022] The patent WO2009/116864A1 entitled Long-Life Probiotic Food Product presents a food product, such as a dairy product or a fruit product, that is long-lived in nature (that is, it can be stored unrefrigerated for a long period of time) and that provides probiotic activity to the consumer. The latter is made by including spores of probiotic bacteria. The product is particularly selected from the group consisting of dairy products (such as, for example, dairy drinks, dairy-derived food products and dairy food ingredients), fruit drinks or concentrates thereof.
[0023] A patente US2021/0369794A1 intitulada Probiotic Formulations for Improving Athletic Performance apresenta uma formulação probiótica onde é fornecida incluindo uma ou mais bactérias, cepas bacterianas ou espécies bacterianas do gênero Veillonella, gênero Faecalibacterium, gênero Phascolarctobacteria, gênero Oscillospira, gênero Ruminococcus, gênero Bacteroides, gênero Blautia, família Christensenellaceae, gênero Dialister ou filo cianobactérias.[0023] Patent US2021/0369794A1 entitled Probiotic Formulations for Improving Athletic Performance presents a probiotic formulation where it is provided including one or more bacteria, bacterial strains or bacterial species of the genus Veillonella, genus Faecalibacterium, genus Phascolarctobacteria, genus Oscillospira, genus Ruminococcus, genus Bacteroides, genus Blautia, family Christensenellaceae, genus Dialister or phylum cyanobacteria.
[0024] A patente intitulada Encapsulated Functional Food Compositions, US2016/0192689A1 apresenta uma composição alimentar funcional compreendendo uma substância comestível ou potável, uma matriz comestível reticulada encapsulando a substância comestível ou potável, um probiótico e um prebiótico, com a condição de que a substância comestível ou potável e a matriz não contenham o probiótico ou ambos contêm o prebiótico. A composição encapsulada oferece certas vantagens em seu transporte, manuseio e consumo. A matriz de membrana comestível pode ser construída para incluir partículas comestíveis que conferem vantagens ao desempenho da matriz de membrana, fornecem nutrição funcional, benefícios de saúde e bem- estar, tratamento terapêutico ao consumidor e/ou aprimoram a experiência gustativa do consumidor. Em aspectos particulares, a composição alimentar funcional fornece benefícios nutritivos e de saúde para o tratamento e prevenção de doenças e manutenção da saúde através da incorporação de certos compostos biologicamente ativos na substância comestível e/ou como partículas na matriz da membrana.[0024] The patent entitled Encapsulated Functional Food Compositions, US2016/0192689A1 presents a functional food composition comprising an edible or drinkable substance, a cross-linked edible matrix encapsulating the edible or drinkable substance, a probiotic and a prebiotic, with the proviso that the substance edible or drinkable material and the matrix do not contain the probiotic or both contain the prebiotic. The encapsulated composition offers certain advantages in transportation, handling and consumption. The edible membrane matrix may be constructed to include edible particles that confer performance advantages to the membrane matrix, provide functional nutrition, health and wellness benefits, therapeutic treatment to the consumer and/or enhance the consumer's taste experience. In particular aspects, the functional food composition provides nutritional and health benefits for the treatment and prevention of disease and maintenance of health through the incorporation of certain biologically active compounds into the edible substance and/or as particles in the membrane matrix.
[0025] A patente US2010/0074994A1 intitulada Dry Food Product Containing Live Probiotic se refere a um sistema de entrega de probiótico que pode ser consumido como lanche ou adicionado a um produto alimentício. Em particular, a divulgação descreve uma guloseima crocante e saborosa que compreende microrganismos probióticos viáveis preservados em uma matriz seca a vácuo de açúcares, proteínas e polissacarídeos. O probiótico permanece viável dentro do tratamento por mais tempo sem a necessidade de revestimento adicional de barreira à umidade. O probiótico também permanece viável no trato gastrointestinal do animal.[0025] Patent US2010/0074994A1 entitled Dry Food Product Containing Live Probiotic refers to a probiotic delivery system that can be consumed as a snack or added to a food product. In particular, the disclosure describes a crunchy and tasty treat comprising viable probiotic microorganisms preserved in a vacuum-dried matrix of sugars, proteins and polysaccharides. The probiotic remains viable within the treatment for longer without the need for additional moisture barrier coating. The probiotic also remains viable in the animal's gastrointestinal tract.
[0026] A patente WO2008/112296 intitulada Compositions And Methods Of Dehydrated Food Fortification apresenta um item alimentar desidratado e não processado (por exemplo, grão, fruta, vegetal ou porção do mesmo) é pelo menos parcialmente revestido com um solvente não aquoso e nutricionalmente aceitável que compreende um organismo probiótico de modo que o item alimentar fortificado tenha um teor total de probiótico de pelo menos 104 a 105 ufc/grama. O prazo de validade de tais composições é notavelmente longo, mantendo uma contagem de UFC relativamente alta, e o sabor e o sabor são notavelmente preservados, mesmo durante longos períodos.[0026] The patent WO2008/112296 entitled Compositions And Methods Of Dehydrated Food Fortification discloses a dehydrated and unprocessed food item (e.g., grain, fruit, vegetable or portion thereof) is at least partially coated with a non-aqueous solvent and nutritionally acceptable that comprises a probiotic organism such that the fortified food item has a total probiotic content of at least 104 to 105 cfu/gram. The shelf life of such compositions is remarkably long, maintaining a relatively high CFU count, and the flavor and flavor are remarkably preserved, even over long periods.
[0027] A patente CN104068187A intitulada Chocolate containing probiotic filling and preparation method for same descreve a invenção de um chocolate contendo um recheio probiótico e um método de preparação para o mesmo. O chocolate é um chocolate contendo um recheio probiótico, em que o recheio probiótico é completamente enterrado em uma cobertura de chocolate; e a cobertura de chocolate é de chocolate branco, chocolate preto e chocolate ao leite. O método de preparação compreende as seguintes etapas de: quando a temperatura do recheio de chocolate em pasta é reduzida para 25-30°C, adicionar pó probiótico e agitar novamente uniformemente, usando um molde de chocolate, envolvendo o recheio de chocolate probiótico produzido no chocolate através do etapas de produção de uma casca, enchimento de um núcleo e vedação de um fundo, em seguida, transporte imediatamente para uma geladeira e condensação e modelagem para obter o produto acabado, formando assim o chocolate recheado contendo probióticos.[0027] Patent CN104068187A entitled Chocolate containing probiotic filling and preparation method for same describes the invention of a chocolate containing a probiotic filling and a preparation method for the same. Chocolate is chocolate containing a probiotic filling, in which the probiotic filling is completely buried in a chocolate coating; and the chocolate coating is white chocolate, dark chocolate and milk chocolate. The preparation method comprises the following steps: when the temperature of the paste chocolate filling is reduced to 25-30°C, add probiotic powder and stir again evenly, using a chocolate mold, enveloping the probiotic chocolate filling produced in the chocolate through the steps of producing a shell, filling a core and sealing a bottom, then immediately transporting to a refrigerator and condensing and shaping to obtain the finished product, thus forming the filled chocolate containing probiotics.
[0028] É descrito na patente US11389499B2 métodos e composições para suplementar a ingestão de fibra dietética através da administração de novos produtos alimentícios de chocolate contendo psyllium. As composições podem ser usadas para suplementar a ingestão de fibra dietética e promover a regulação dietética e o gerenciamento do bem-estar. A composição de chocolate compreendendo proporções particulares de chocolate, psyllium e lipídios, e os métodos de formulação de formação de emulsão aqui divulgados reduzem surpreendentemente a gelificação desagradável na boca/garganta durante a mastigação/deglutição que é frequentemente associada a produtos contendo psyllium. As capacidades protetoras dos ingredientes das presentes composições proporcionam e, consequentemente, permitem uma fabricação adequada e propriedades organolépticas melhoradas que promovem a adoção a longo prazo e o uso consistente pelos sujeitos.[0028] Patent US11389499B2 describes methods and compositions for supplementing dietary fiber intake through the administration of new chocolate food products containing psyllium. The compositions can be used to supplement dietary fiber intake and promote dietary regulation and wellness management. The chocolate composition comprising particular proportions of chocolate, psyllium and lipids, and the emulsion forming formulation methods disclosed herein surprisingly reduce the unpleasant gelling in the mouth/throat during chewing/swallowing that is often associated with psyllium-containing products. The protective capabilities of the ingredients of the present compositions provide and, consequently, allow for proper manufacturing and improved organoleptic properties that promote long-term adoption and consistent use by subjects.
[0029] As patentes e publicações apresentadas não invalidam a presente invenção, pois não há utilização do spray drier na microencapsulação de Lactobacillus helveticus e aplicação em chocolate branco. Portanto, a obtenção do chocolate branco, tendo o probiótico em sua composição torna esse processo tecnicamente e comercialmente atrativo, que apresenta vantagens, como facilidade e rapidez do processo, diferentemente de outros processos de obtenção de alimentos com características similares, quando se tratando de utilização de probióticos.[0029] The patents and publications presented do not invalidate the present invention, as there is no use of the spray drier in the microencapsulation of Lactobacillus helveticus and application in white chocolate. Therefore, obtaining white chocolate, with probiotics in its composition, makes this process technically and commercially attractive, which presents advantages, such as ease and speed of the process, unlike other processes for obtaining foods with similar characteristics, when it comes to use. of probiotics.
[0030] O método descrito na presente patente de invenção envolve a utilização do spray drier na microencapsulação de Lactobacillus helveticus e aplicação em chocolate branco. A obtenção do chocolate branco, tendo o probiótico em sua composição torna esse processo tecnicamente e comercialmente atrativo. O método é vantajoso em relação ao Estado da Técnica por apresentar-se da adição de probiótico em chocolate branco, e da utilização de matrizes composta por 50% maltodextrina + 50% goma arábica e 30% leite em pó + 19% maltodextrina + 19% trealose + 20% frutoligossacarídeo + 10% amido + 2% tween 0,1%, o que nos traz mais uma oportunidade de desenvolvermos um novo produto.[0030] The method described in the present patent involves the use of the spray drier in the microencapsulation of Lactobacillus helveticus and application to white chocolate. Obtaining white chocolate, with probiotics in its composition, makes this process technically and commercially attractive. The method is advantageous in relation to the State of the Art as it involves the addition of probiotics to white chocolate, and the use of matrices composed of 50% maltodextrin + 50% gum arabic and 30% powdered milk + 19% maltodextrin + 19% trehalose + 20% fructooligosaccharide + 10% starch + 2% tween 0.1%, which brings us another opportunity to develop a new product.
[0031] O método proposto na presente patente de invenção tem como objetivo apresentar uma alternativa para a produção de um novo alimento funcional, chocolate branco acrescido com probiótico. Mais especificamente um chocolate branco ascrecido com o probiótico Lactobacillus helveticus microencapsulado pela técnica de spray drier.[0031] The method proposed in the present invention patent aims to present an alternative for the production of a new functional food, white chocolate added with probiotics. More specifically, white chocolate added with the probiotic Lactobacillus helveticus microencapsulated using the spray drier technique.
[0032] A fim de que o presente invento seja plenamente compreendido e levado à prática por qualquer técnico deste setor tecnológico, o mesmo será descrito de forma clara, concisa e suficiente, tendo como base os desenhos, que ilustram análises de diferentes composições do produto e subsidiam abaixo listados: Figura 1. Apresenta as características físico-químicas (atividade água, umidade, pH e acidez) durante a estocagem a 22°C. Figura 2. Apresenta os parâmetros de cor (L*, croma C*, ΔE*ab) do chocolate tradicional (sem o probiótico - Fp) e contendo o probiótico livre (F1) e encapsulado (F2) durante a estocagem a 22°C. Tabela 1. Apresenta os valores dos parâmetros α e β do modelo de Weibull de sobrevivência do L. helveticus livre (F1) e encapsulado (F2) no chocolate branco armazenado a 22°C e análise estatística do ajuste (R2).[0032] In order for the present invention to be fully understood and put into practice by any technician in this technological sector, it will be described in a clear, concise and sufficient manner, based on the drawings, which illustrate analyzes of different compositions of the product and support listed below: Figure 1. Shows the physical-chemical characteristics (water activity, humidity, pH and acidity) during storage at 22°C. Figure 2. Shows the color parameters (L*, chroma C*, ΔE*ab) of traditional chocolate (without the probiotic - Fp) and containing the free (F1) and encapsulated (F2) probiotic during storage at 22°C . Table 1. Presents the values of parameters α and β of the Weibull survival model of free (F1) and encapsulated (F2) L. helveticus in white chocolate stored at 22°C and statistical analysis of the fit (R2).
[0033] A presente invenção descreve um método de microencapsulação de Lactobacillus helveticus em diferentes matrizes (Quitosana, gelatinas, maltodextrina, goma arábica, leite em pó, trealose, frutooligosacarídeo, amido) e proporções a serem utilizadas no encapsulamento do probiótico para posterior aplicação em chocolate branco utilizando spray-drier. As diferentes condições reacionais utilizadas para a preparação do material serão descritas a seguir.[0033] The present invention describes a method of microencapsulation of Lactobacillus helveticus in different matrices (Chitosan, gelatins, maltodextrin, gum arabic, milk powder, trehalose, fructooligosaccharide, starch) and proportions to be used in encapsulating the probiotic for subsequent application in white chocolate using spray-drier. The different reaction conditions used to prepare the material will be described below.
[0034] No preparo do probiótico 1 g da cepa liofilizada foi reativada em 10 mL de caldo MRS (de Man, Rogosa e Sharpe) incubadas em estufa a 37 °C ± 1 °C por 48 h em jarra de anaerobiose. Em seguida, o material foi centrifugado a 4000 rpm por 15 min, 5 °C, lavado com água peptonada e ressuspendido em 10 mL de caldo MRS para obtenção de massa celular (108 - 109 UFC) (Barros et al., 2021).[0034] When preparing the probiotic, 1 g of the lyophilized strain was reactivated in 10 mL of MRS broth (from Man, Rogosa and Sharpe) incubated in an oven at 37 °C ± 1 °C for 48 h in an anaerobic jar. Then, the material was centrifuged at 4000 rpm for 15 min, 5 °C, washed with peptone water and resuspended in 10 mL of MRS broth to obtain cell mass (108 - 109 CFU) (Barros et al., 2021).
[0035] Para o encapsulamento do probiótico utilizou-se 24 g de matriz encapsulante, o qual foi hidratada com 100 mL de solução tampão fosfato de sódio (pH 7,0) por 2 h com agitação magnética. Após a etapa de hidratação foi adicionado 1 g de massa celular do probiótico (108 - 109 UFC/g), mantendo - os sob agitação por 10 min.[0035] To encapsulate the probiotic, 24 g of encapsulating matrix was used, which was hydrated with 100 mL of sodium phosphate buffer solution (pH 7.0) for 2 h with magnetic stirring. After the hydration step, 1 g of probiotic cell mass (108 - 109 CFU/g) was added, keeping them under agitation for 10 min.
[0036] Antes do processo de secagem o spray dryer foi higienizado com água ultrapura e álcool 70%. Posteriormente, realizou-se a secagem em spray dryer, onde a mistura probiótico/matriz encapsulante/solução tampão foi bombeada a uma vazão de 0,08 mL/min, na pressão de 0,08 a 0,12 bar, diâmetro do orifício do bico injetor pneumático de 0,5 mm e temperatura de entrada (câmara de secagem), pré-definidas para cada ensaio, de 110, 120 e 130°C. A amostra em solução (material encapsulante/solução tampão fosfato pH 7,0) foi bombeada até o topo do atomizador, onde entrou em contato com o ar aquecido na câmara de secagem, ocorrendo à evaporação imediata da umidade contida no material. A corrente de ar quente-sólido foi direcionada em um ciclone onde se efetuou a separação do extrato encapsulado em recipiente de vidro (coletor de pó).[0036] Before the drying process, the spray dryer was sanitized with ultrapure water and 70% alcohol. Subsequently, drying was carried out in a spray dryer, where the probiotic/encapsulating matrix/buffer solution mixture was pumped at a flow rate of 0.08 mL/min, at a pressure of 0.08 to 0.12 bar, diameter of the orifice of the 0.5 mm pneumatic injection nozzle and inlet temperature (drying chamber), pre-defined for each test, of 110, 120 and 130°C. The sample in solution (encapsulating material/phosphate buffer solution pH 7.0) was pumped to the top of the atomizer, where it came into contact with the heated air in the drying chamber, resulting in immediate evaporation of the moisture contained in the material. The hot-solid air stream was directed into a cyclone where the extract encapsulated in a glass container (dust collector) was separated.
[0037] Nos ensaios, após a etapa de secagem avaliou-se a contagem de probiótico, a atividade de água, umidade e morfologia das microcápsulas.[0037] In the tests, after the drying stage, the probiotic count, water activity, humidity and morphology of the microcapsules were evaluated.
[0038] As matérias primas utilizadas para fabricação de chocolate branco foram as seguintes: leite em pó integral e leite em pó desnatado, manteiga de cacau, açúcar cristal, vanilina e emulsificantes: lecitina de soja e Poliglicerol poliricinoleato.[0038] The raw materials used to manufacture white chocolate were the following: whole milk powder and skimmed milk powder, cocoa butter, crystal sugar, vanillin and emulsifiers: soy lecithin and polyglycerol polyricinoleate.
[0039] Três formulações foram elaboradas: Fp - Formulação padrão, sem adição do probiótico; F1 - Formulação com adição do probiótico livre; F2- Formulação com adição do probiótico encapsulado.[0039] Three formulations were developed: Fp - Standard formulation, without addition of probiotic; F1 - Formulation with the addition of free probiotic; F2- Formulation with the addition of encapsulated probiotic.
[0040] A seguir serão apresentados exemplos de aplicação do método proposto. Não se têm a intenção de limitar o escopo da invenção, mas apenas de exemplificá-lo. Quaisquer modificações nos procedimentos, desde que sejam mantidas as suas características particulares, e que são essenciais ao conceito inventivo ora revelado e que não fujam do espectro de proteção delineado na solução aqui descritos estão ainda dentro do escopo da invenção.[0040] Examples of application of the proposed method will be presented below. They are not intended to limit the scope of the invention, but only to exemplify it. Any modifications to the procedures, as long as their particular characteristics are maintained, and which are essential to the inventive concept disclosed herein and which do not deviate from the spectrum of protection outlined in the solution described here, are still within the scope of the invention.
[0041] A primeira etapa realizada foi a moagem do açúcar até granulometria de 25 μm, em seguida a pesagem dos demais ingredientes da formulação.[0041] The first step carried out was grinding the sugar to a particle size of 25 μm, followed by weighing the other ingredients of the formulation.
[0042] Inicialmente, realizou-se o derretimento de 1/3 da manteiga de cacau a uma temperatura de 55°C, após a manteiga estar completamente derretida adicionou-se os ingredientes em pó (açúcar, leite em pó integral e desnatado). Neste momento, iniciou-se a etapa de conchagem por 2h a uma temperatura de 55°C. Após 2h adicionou-se o restante da manteiga de cacau, a lecitina de soja e o poliglicerol poliricinoleato. Posteriormente, o produto passou por um moinho de esferas, onde permaneceu entre 30 e 40 min, até que atingir 22 μm, com temperatura controlada de 40 °C a uma rotação de 54 rpm. Após a etapa de moagem realizou-se a etapa de temperagem, onde adicionou-se o probiótico.[0042] Initially, 1/3 of the cocoa butter was melted at a temperature of 55°C, after the butter was completely melted, the powdered ingredients were added (sugar, whole and skimmed milk powder). At this point, the conching stage began for 2h at a temperature of 55°C. After 2h, the remaining cocoa butter, soy lecithin and polyglycerol polyricinoleate were added. Subsequently, the product passed through a ball mill, where it remained for 30 to 40 min, until it reached 22 μm, with a controlled temperature of 40 °C at a speed of 54 rpm. After the grinding stage, the tempering stage was carried out, where the probiotic was added.
[0043] A moldagem do chocolate foi realizada em formas de acrílico e, em seguida, o resfriamento em geladeira convencional, em temperaturas de 5 - 7°C. Para retirada das bolhas de ar das formas, será utilizado um processo manual, onde vibraremos as formas sobre a bancada de mármore até completa eliminação das bolhas de ar. O tempo de resfriamento foi em média 20 min. As amostras foram retiradas da geladeira, desmoldadas e acondicionadas individualmente em embalagens de papel alumínio em sala climatizada a 18 - 20°C.[0043] The chocolate was molded in acrylic molds and then cooled in a conventional refrigerator, at temperatures of 5 - 7°C. To remove air bubbles from the molds, a manual process will be used, where we will vibrate the molds on the marble countertop until the air bubbles are completely eliminated. Cooling time averaged 20 min. The samples were removed from the refrigerator, demolded and individually packaged in aluminum foil packaging in an air-conditioned room at 18 - 20°C.
[0044] As barras de chocolate foram identificadas e armazenadas a 20°C por 7 dias permitindo a formação dos cristais β estáveis. Após este período as amostras foram caracterizadas mediante determinações físico-químicas e contagem do probiótico. Avaliou-se também, a estabilidade do chocolate e do probiótico em amostras de chocolate, em diferentes períodos de armazenamento a 22°C.[0044] The chocolate bars were identified and stored at 20°C for 7 days allowing the formation of stable β crystals. After this period, the samples were characterized by physical-chemical determinations and probiotic counts. The stability of chocolate and probiotics in chocolate samples was also evaluated in different storage periods at 22°C.
[0045] A contagem da bactéria probiótica foi realizada após a etapa de encapsulamento e no chocolate, logo após a elaboração e durante o armazenamento (30, 60, 75, 90 e 100 dias) a 22°C. A análise microbiológica ocorreu conforme metodologia descrita por A análise foi conduzida com 25 g de chocolate adicionada de 225 mL de água peptonada. As amostras foram acondicionadas em sacos estéreis e homogeneizadas em Stomacher (SP Labor - SP190), após foram preparadas diluições (10-1 a 10-11) em citrato de sódio 2 %. Foi inoculado 1 mL de cada diluição em placas de petri estéreis vazias, adicionando, em seguida, o meio de cultura ágar de Man Rogosa & Sharpe (MRS). As placas foram incubadas, invertidas e acondicionadas em jarra com atmosfera anaeróbia, a 35±1°C, por 48 h (SILVA; JUNQUEIRA; SILVEIRA, 2001; GROSSO; FÁVARO-TRINDADE, 2004).[0045] The probiotic bacteria were counted after the encapsulation stage and in the chocolate, immediately after preparation and during storage (30, 60, 75, 90 and 100 days) at 22°C. The microbiological analysis took place according to the methodology described by The analysis was conducted with 25 g of chocolate added to 225 mL of peptone water. The samples were placed in sterile bags and homogenized in a Stomacher (SP Labor - SP190), after which dilutions were prepared (10-1 to 10-11) in 2% sodium citrate. 1 mL of each dilution was inoculated into empty sterile petri dishes, then adding the Man Rogosa & Sharpe (MRS) agar culture medium. The plates were incubated, inverted and placed in a jar with an anaerobic atmosphere, at 35±1°C, for 48 h (SILVA; JUNQUEIRA; SILVEIRA, 2001; GROSSO; FÁVARO-TRINDADE, 2004).
[0046] As amostras de chocolate branco tradicional e com probiótico resultantes foram caracterizadas por atividade água, umidade, pH e acidez) e os parâmetros de cor (L*, croma C*, ΔE*ab).[0046] The resulting traditional and probiotic white chocolate samples were characterized by water activity, humidity, pH and acidity) and color parameters (L*, chroma C*, ΔE*ab).
[0047] A Figura 1 apresenta as características físico-químicas (atividade água, umidade, pH e acidez) e A Figura 2 apresenta os parâmetros de cor (L*, croma C*, ΔE*ab) do chocolate tradicional (sem o probiótico - Fp) e contendo o probiótico livre (F1) e encapsulado (F2) durante a estocagem a 22°C.[0047] Figure 1 presents the physicochemical characteristics (water activity, humidity, pH and acidity) and Figure 2 presents the color parameters (L*, chroma C*, ΔE*ab) of traditional chocolate (without the probiotic - Fp) and containing the free (F1) and encapsulated (F2) probiotic during storage at 22°C.
[0048] A Tabela 1 apresenta os valores dos parâmetros α e β do modelo de Weibull de sobrevivência do L. helveticus livre (F1) e encapsulado (F2) no chocolate branco armazenado a 22°C e análise estatística do ajuste (R2). Os parâmetros fator de forma (α) e fator de escala (β) do CPL e CPM1 não apresentaram variações relevantes, demonstrando um comportamento similar. Os coeficientes estatísticos R2 demonstram um bom ajuste dos dados experimentais e a capacidade preditiva do modelo primário de Weibull.[0048] Table 1 presents the values of parameters α and β of the Weibull model of survival of free (F1) and encapsulated (F2) L. helveticus in white chocolate stored at 22°C and statistical analysis of the adjustment (R2). The shape factor (α) and scale factor (β) parameters of CPL and CPM1 did not show relevant variations, demonstrating similar behavior. The R2 statistical coefficients demonstrate a good fit of the experimental data and the predictive capacity of the primary Weibull model.
[0049] Ao comparar o resultado obtido do L. helveticus livre em chocolate branco (F1) o mesmo manteve-se estável 108 CFU/g por 58 dias de armazenamento a 22°C, sendo considerado superior ao relatado por Gadhiya et al. (2018) que adicionaram L. helveticus livre em chocolate ao leite, but the probiotics viability was maintained at 108 CFU/g only for 15 days when of storage at 10 ± 2 °C.[0049] When comparing the result obtained from free L. helveticus in white chocolate (F1), it remained stable at 108 CFU/g for 58 days of storage at 22°C, being considered superior to that reported by Gadhiya et al. (2018) who added free L. helveticus to milk chocolate, but the probiotics viability was maintained at 108 CFU/g only for 15 days when of storage at 10 ± 2 °C.
[0050] É importante salientar que as descrições e o exemplo não possuem o condão de limitar as formas de execução, as modificações e variações que podem ser realizadas seguindo as formulações, metodologias e produtos do invento ora proposto, mas sim de ilustrar e tornar compreensíveis as inovações conceituais reveladas nesta solução. Desse modo, as descrições devem ser interpretadas de forma ilustrativa e não restritivas, podendo apresentar variações nos equipamentos utilizados nas etapas do processo, proporções, dimensões e demais variáveis, seja do conjunto ou de seus elementos isoladamente, desde que sejam mantidas as suas características particulares, e que são essenciais ao conceito inventivo ora revelado e que não fujam do espectro de proteção delineado na solução proposta.[0050] It is important to emphasize that the descriptions and example do not have the power to limit the forms of execution, modifications and variations that can be carried out following the formulations, methodologies and products of the invention proposed here, but rather to illustrate and make it understandable the conceptual innovations revealed in this solution. Therefore, the descriptions must be interpreted in an illustrative and non-restrictive way, and may present variations in the equipment used in the process stages, proportions, dimensions and other variables, whether of the set or its elements separately, as long as their particular characteristics are maintained. , and which are essential to the inventive concept now revealed and which do not deviate from the spectrum of protection outlined in the proposed solution.
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| BR102022017979A2 true BR102022017979A2 (en) | 2024-03-19 |
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