BRPI1002604A2 - flavored, whey-based, lutein enriched milk drink and its manufacturing and use process - Google Patents

Abstract

ACIDIFIED MILK DRINK, BASED ON MILK AND MILK SERUM, AROMATIZED, ENRICHED WITH LUTEINE AND ITS MANUFACTURING AND USE PROCESS. The present invention patent provides for a liquid food product, ready for consumption, composed of whey derived from the manufacture of enzyme-coagulated cheeses, standard mines, fresh mines, rennet and others at (40 to 70%) acidified with pH below 4.5 with 50% citric, lactic or tartaric acid (1.5% to 3%), plus homogenized milk (20 to 60%), sugar (8 to 20%), sodium citrate (0, 01 to 0.5%), stabilizer (0.1 to 1%), fumaric acid (0.01 to 0.3%), potassium sorbate (0.01 to 0.3%), lutein extract (0.001 to 0.0 1%) and natural aromas of açaí, banana, kiwi, acerola, grape, peach, orange, lemon, apple, pineapple, tangerine and identical to the natural citrus fruit (0.02 to 0.2%), it can be made in light, diet and lactose reduction forms. Additionally, the present application is intended for the manufacturing process of said product and its use in the dairy and food industries.

Description

"ACIDIFIED MILKWOOD-BASED MILKWOOD DRINK ENRICHED WITH LUTEIN AND ITS MANUFACTURING AND USE PROCESS".

Field of the invention

The present invention provides a ready-to-eat liquid food product consisting of whey from the manufacture of enzymatically coagulated cheeses, standard mines, fresh mines, rennet and others (40 to 70%) acidified to pH below 4.5 with 50% citric, lactic or tartaric acid (1.5% to 3%) plus homogenized milk (20 to 60%), sugar (8 to 20%), sodium citrate (0.01 stabilizer (0.1 to 1%), fumaric acid (0.01 to 0.3%), potassium sorbate (0.01 to 0.3%), lutein extract (0.001 to 0.01%) and natural aromas of acai, banana, kiwi, acerola, grape, peach, orange, lemon, apple, pineapple, mandarin and identical to the natural of citrus fruits (0.02 to 0.2%). Additionally, the present application is intended for the manufacturing process of said product and its use in the dairy and food industries.

2. State of the Art - Background of the Invention

Internal and external economic pressures in the dairy industry are growing increasingly worldwide. Internally due to the process of concentration between producers and externally due to the global economic situation. Along with these pressures is the need to develop new products that are more competitive, with lower manufacturing costs, more nutritious, with greater health benefits (PAULA, 2005).

Milk is considered a healthy food and consumption of dairy products is associated with a quality diet. Milk provides an affordable matrix rich in a variety of essential nutrients such as easily digestible minerals, vitamins and proteins with balanced amino acid profiles, and is an important support for all body functions (STEIJNS, 2008). Despite increasing recognition of the benefits associated with their consumption, research data suggest that milk and dairy product intake among adolescents has declined over the last decades, and this consumption is well below official US Department of Health and Human Services recommendations ( BOWMAN, 2002; LARSON5 et al., 2006).

Brazil is the sixth largest milk producer with a volume of 25,327 thousand tons, accounting for 4.52% of world production in 2007. In 2008 the estimated production of cheeses produced under inspection was 640 thousand tons, with an increase of 10, 34% of the 2007 production. However, considering that the production of informal cheese (without inspection) is 40% of the total, it can be concluded that the volume produced reached 896 thousand tons in 2008 (EMBRAPA, 2008). However, considering the volume of cheese produced in 2008, approximately 8 million liters of whey were generated, which due to its organic matter content can be a serious environmental problem. This fact supports the need for constant search for alternative methods for its rational and sustainable use, avoiding that this raw material is considered another component of the effluents of the dairy industry.

Due to the increasing awareness of the importance of healthier living and the growing concern to change their eating habits, companies have been looking for alternatives to win over consumers by developing new, lower-priced quality products that bring claims of functional properties. . In this sense, improving a food that already has excellent nutritional characteristics by nature, providing a simple and inexpensive technology and still realizing the rational and sustainable ecological use of whey, is a strategic advantage of industrial competitiveness that is intended with the present invention.

The market for functional foods grows at a rate of 10% per annum, while the other food industries in the range of 2-3% per annum. Consumers have demonstrated purchase intent and monetary appreciation for functional attributes that relate food and health. Thus, several efforts have been made to consolidate this niche market and launch new products with health claims (BARREIRO-HURLE, COLOMBO et al., 2008).

The present invention provides a low cost technology that aims at harnessing whey in the cheese industries and popularizing access to a nutritious food containing a bioactive function component, proposes the development, characterization and evaluation of the quality and stability of an acidified, flavored, pasteurized and lutein-added milk drink stored at room temperature and refrigerated.

2.1. LITERATURE REVIEW

2.1.1. Importance of whey as a co-product

Whey is the aqueous portion of milk that separates from the mass during conventional cheese or casein manufacturing, and retains approximately 55% of milk's nutrients (Kosikowski, 1979). Approximately 85 to 90% of the volume of milk used to make cheese results in whey, which contains around half of the milk solids represented by proteins, minerals, vitamins and especially lactose (GONZFILEZ SISO, 1996). Whey has approximately 0.8% high biological value proteins, which are exceptionally rich in lysine and tryptophan, well balanced in sulfur amino acids, thanks to the high levels of cysteine that is a precursor of methionine, its quality is superior to casein. . Whey proteins are attributed great nutritional value because of their composition rich in essential amino acids with high bioavailability. Whey is a source of several macroelements, the most abundant of which are calcium, sodium, magnesium, potassium and phosphorus, and contains most of the water-soluble vitamins in milk and is particularly rich in B vitamins (MELLO, 1989). ).

The nutritional and functional characteristics of whey are related to its composition, structure and biological function. In the last decades, there has been a growing interest in the nutritional quality of these proteins for their use in nutritional applications, such as enteral formulas, in the form of whole or pre-digested proteins, aiming at weight gain and recovery of the nutritional status of several types of patients (MELLO, 1989; DE WIN 1998; CAPITANI 2004). Extraction of its main nutrients (lactose and whey proteins) through the manufacture of Ricotta, dairy drinks, use in animal feed and transformation into concentrate or powder are the main means of use of whey.

2.1.2. Acidified milk drink

In Brazil, Normative Instruction No. 16 of August 23, 2005, which proves the Technical Regulation of Identity and Quality of Milky Beverage, defines dairy drink as the milk product resulting from the mixture of milk (in natura, pasteurized, sterilized, UHT). , reconstituted, concentrated, powdered, whole, semi-skimmed or partially skimmed and skimmed) and whey (liquid, concentrated and powdered) whether or not containing added foodstuff (s), vegetable fat, fermented milk (s), selected milk yeasts and other milk products. The dairy base represents at least fifty-one percent (51%) mass / mass (w / w) of the total ingredients of the product (BRAZIL, 2005).

Because acidified dairy drinks are economically viable today, they are an alternative to fermented dairy products such as yoghurt, which often appear in consumers' shopping baskets. Thus, the development of new products in this line is a market opportunity. The acidified dairy beverage manufacturing process allows milk proteins to remain soluble at low pH, which may provide extended shelf life under room temperature or refrigeration. The procedure also allows the use of pasteurization only as a form of microbiological control, eliminating sterilization, avoiding the use of more sophisticated equipment that can inactivate various nutrients in the food. The production of acidic lactic beverages is faster and cheaper compared to fermentation production, a process commonly used in Brazil.

Increased consumption of dairy drinks such as yogurt, fermented milks and milk-like beverages (mix of milk and whey) has been widespread. In Brazil there has been a growing acceptance for milk-like beverages as well as for milk drinks formulated with a mixture of fermented dairy products, representing approximately one third of the yogurt and other dairy market (GALLARDO-ESCAMILLA et al, 2006).

Acidified dairy drinks are produced in various parts of the world and exist in different variations, such as drinks with milk and fruit, drinks with yoghurt, buttermilk, whey drinks, kefir, etc. These products may be described as a liquid protein system acidified with the stability and viscosity of milk.

To such beverages stabilizers are added to prevent sediment formation and phase separation. Caseins in milk pH are present as micelles and these protein particles are stabilized by steric repulsive interactions. When milk pH is lowered, many phenomena happen, modifying the stability of casein micelles. At pH 4.6, calcium is soluble (OULD ELEYA et. Al, 1995; LAURENT and BOULENGUER, 2002) and casein aggregation is irreversible, forming a three-dimensional network consisting of clustered filaments. When the acidified dairy drink is processed, the physical properties of the protein lattice are modified by heat treatment, which contributes to the acceleration of the protein aggregation process and homogenization destroys the gel, producing caseinate fragments. Under these conditions, sedimentation occurs and can only be avoided by adding a stabilizing polymer (TAMIME & ROBINSON, 1999; LAURENT and BOULENGUER, 2002).

In the case of acidified milk drinks, which are normally prepared from stabilized fermented sugared milk or by direct acidification with fruit juices and or addition of weak acids, the pH ranges from 3,4 to 4,6. During acidification at a pH around the isoelectric point (pH 4.6) the micelles aggregate, mainly because of the collapse of the extended layer formed by kappa-casein on the micelle surface (TAMIME & ROBINSON, 1999). At a pH below the isoelectric point, milk proteins are positively charged and stabilizers with negative charges are commonly employed to return the system to a stable state without phase separation. When adequate stabilizer is added before acidification, it adsorbs on the surface of casein micelles by electrostatic interactions and prevents their aggregation (NAKAMURA et al., 2005). The time-temperature heat treatment binomial, which denatures whey proteins in milk and allows them to bind to casein prior to acidification as well as the homogenization process can significantly improve the stabilization system in acidified milk (SEDLMEYER et al., 2004).

2.1.3. Barrier technology and food preservation

In traditionally preserved foods there is a combination of factors to ensure microbiological safety and stability, thus allowing their conservation. The concept of combining various obstacles to controlling microbial growth and thus conserving food was developed in 1995 by Leistner and colleagues. In barrier technology, various factors such as temperature, pH, chemical preservatives, reduced water activity, among others are used to create a series of barriers that ensure the microbiological safety of processed food. The selected barriers to be applied must be high enough to eliminate the microorganism population.

Such barriers must take into account the starting number and type of microorganism to be successful. However, these barriers that normally function properly may be overcome by a larger initial population of microorganisms or may not work well in other nutrient-rich foods. In this case, such barriers then need to be modified in different or increased intensity combinations (FELLOWS, 2006).

The application of several barriers can induce changes in the microbial cell to maintain its viability, which can cause metabolism to deplete, preventing its growth. The metabolic exhaustion of microorganisms, which can cause food "self-sterilization" is accelerated when more barriers are present, probably due to the increased energy demand to keep internal homeostasis under stress (LEISTNER, 2006). 2000). Exposure to different types of stress simultaneously requires energy and shock protein synthesis, which can make microorganisms metabolically depleted (LEISTNER, 2000; BEALES, 2004).

DAVES (1995) reported that maintaining the quality of a food product during storage is mainly due to inhibition of the multiplication of spoilage microorganisms. According to GOLD (1996), the conservation of food is based on the delay of microbial multiplication and, therefore, must act on the extrinsic and intrinsic factors of growth and survival of the microorganism.

Most preservation techniques, currently employed to prevent or delay microbial multiplication in food products, consist of lowering or raising temperature, lowering pH, reducing water activity (Aw) 5 modifying oxirreduction potential (Eh), preservatives and competitiveness among microorganisms (LEISTNER, 2000). According to LEISTNER (1995), conventional techniques are being used together with some alternatives, causing synergism between them.

Whey and milk are excellent culture media for various microorganisms such as filamentous fungi, yeast and bacteria, and need to be thermally processed to ensure their quality. However, most microorganisms multiply better at pH values around 7.0 (6.6 to 7.5), although a few (filamentous fungi and yeast) multiply at pH below 4.0 (JAY, 2005).

In the present invention, heat treatment of pasteurization was used in conjunction with acidification and the use of preservative to produce a microbiologically stable, long-life dairy drink at room temperature and refrigeration for up to 90 days.

2.1.4. Functional foods and lutein

In recent decades consumer demand for food production has changed considerably. More and more consumers believe that food contributes to health. Today, food is not only intended to satisfy hunger and provide the necessary nutrients for humans, but can also prevent malnutrition-related illnesses, improve consumers' physical and mental well-being. The growing demand for these foods can be explained by the high cost of health care, the constant increase in life expectancy and people's desire to improve quality of life (SIRÓ et al, 2008).

Functional food can be defined as similar in appearance to conventional food, consumed as part of the usual diet, capable of producing demonstrated metabolic or physiological effects useful in maintaining good physical and mental health and may help to reduce the risk of chronic degenerative diseases. , in addition to its basic nutritional functions (DIAS, 2009).

Allegation of functional property is that relating to the metabolic or physiological role that the nutrient or non-nutrient has in the growth, development, maintenance and other normal functions of the human organism. Allegation of health property is that which states, suggests or implies the existence. of relationship between food or ingredient with disease or health-related condition ANVISA (1999).

Lutein (Tagetes erecta extract) and its zeaxanthin isomer, dihydroxylated carotenoids are natural pigments belonging to the class of yellow colored xanthophylls. They are the main pigments contained in the human macula and retina. They act as antioxidants by protecting cells from oxidative damage and filter out high-energy blue light, improving visual acuity (ALTERNATIVE MEDICINE REVIEW, 2005).

Mammals are biochemically incapacitated for carotenoid biosynthesis, but may accumulate these pigments in adipose tissues and or convert precursors from the diet (DAVIES & MORLAND, 2004). Epidemiological studies have shown an association between high levels of carotenoids in the diet and or in the blood and a protective effect against the development of chronic diseases such as certain cancers, cardiovascular disease, degenerative macula disease and cataracts (EDGE et al, 1997).

Lutein is found in green leafy vegetables, especially dark leafy vegetables such as spinach, kale, watercress and broccoli. In smaller quantity is found in fruits and vegetables such as kiwi, orange, corn, Brussels sprouts, pepper, among others. Diets rich in fruits and vegetables containing carotenoids have been linked to reduced risk of cancer (HANDELMAN et al, 1999). Epidemiological studies show that populations at higher risk of developing macular degeneration have a low lutein and zeaxanthin diet and low plasma levels of these pigments (DAVIES & MORLAND, 2004).

The macula is one of the most metabolically active tissues in the body, has a high concentration of polyunsaturated fatty acids, is very susceptible to damage, with low regeneration capacity (DAVIES & MORLAND, 2004). Of all the carotenoids present in human plasma, lutein and zeaxanthin are the two found in larger amounts deposited in the eyeball in much higher concentrations than in any other human tissue (Krinsky et al, 2003; Ahmed et al, 2005).

Natural compounds containing conjugate double bonds act for their antioxidant effect on free radical scavenging (FONTANA, 2004). Lutein is considered the predominant antioxidant in the eye, protecting tissues from oxidation by filtering blue light and neutralizing free radicals. Increased lutein intake correlates positively with increased macular pigment density, which is inversely related to the risk of macular degeneration (MOLLDREM et al, 2004).

The macula is an area in the retina responsible for central vision, and macular degeneration is a slowly progressive, painless disease that irreversibly affects the macula tissue and inevitably leads to vision loss. Loss of macular function through degeneration can deteriorate central vision, leaving only lateral or peripheral vision intact (WANG et al., 2006).

Once the disease is established, there is no return to normal vision, even with the increase of these carotenoids in the diet; thus, there is only a decrease or stabilization in progressive vision loss when lutein is supplemented or after onset.

The protective effect can be explained by the selective accumulation of these carotenoids in the retina and their density in the macula (LANDRUM et al, 1997; BERENDSCHOT et al, 2000). The daily dietary intake of lutein required to decrease the risk of macular degeneration is approximately 6.0 to 20.0 mg per day (ALTERNATIVE MEDICINE REVIEW, 2005).

Among the main benefits associated with lutein, in addition to the evidence of reduced risk of developing age-related macular degeneration, the beneficial effects in protecting against atherosclerosis, cataracts, cancer and other diseases (ALVES-RODRIGUES and SHAO, 2004; OAK, 2000; DAGNELIE, 2000; DELI et al, 2004; DOWNHAM & COLLINS, 2000).

Lutein can be employed in the functional food industry to enrich fruit products, dairy drinks, cereals, cookies, sauces and soups. As a natural colorant it can be used to color foods such as edible oils, margarine, mayonnaise, mustard, salad dressings, yogurt, cakes, ice cream and dairy products (SOWBHAGYA et al., 2004). Epidemiological studies have confirmed that lutein is safe and is considered as a GRAS (Generally Recognized as Safe) substance. Food enrichment can be successfully performed without changes in the other ingredients and without impacting the sensory properties of the final product (DAGNELIE, 2000).

2.2 - Patent Documents

The invention PI 0600975 is a totally different product from our product and claims "Milk Product Composition and Process for Obtaining" the composition of which is a homogenized and sterilized Dairy Beverage formulation very similar to the Dairy Beverage normally produced in industries. Brazilian women. Our product is the development of an acidified, flavored, pasteurized and lutein-added dairy drink that can be stored for at least 90 days both refrigerated and stored at room temperature. Our product also fits the designation "Dairy with added" as defined by the Technical Regulation of Identity and Quality of Dairy. Our beverage is a technologically viable alternative and can be easily produced and marketed by the dairy industry without the need for major investments as in the case of the aforementioned invention which requires expensive equipment such as sterilizers.

In our case we can take advantage of the facilities available in a dairy industry and still make the rational and sustainable use of whey generated in cheese making. The acidified, flavored, pasteurized and lutein-added dairy drink object of this patent is nutritionally superior to soft drinks because it conveys not only "empty calories" but proteins, vitamins and minerals that are essential for maintaining health. The addition of a bioactive component (lutein) confers considerable antioxidant activity to the formulation of the developed dairy drink. This gives the product a metabolic or physiological role in the protection of the human body against damage caused by oxidative stress, especially in relation to the maintenance of normal macular tissue functions. The acidified dairy beverage manufacturing process allows milk proteins to remain soluble at low pH, which may provide extended shelf life under room temperature or refrigeration. The procedure also allows the use of pasteurization only as a form of microbiological control, eliminating sterilization, avoiding the use of more sophisticated equipment that can inactivate various nutrients in the food. Production of acidic lactic beverages is faster and cheaper than fermentation production.

PI05201314A relates to the preparation of a "High Calcium Low Calorie Non-Fat Milky Drink and Method". The product is characterized in that the non-fat milk, a whey protein solution, or a combination of the two is ultrafiltered or evaporated and chromatographed to remove carbohydrates or to reduce the amount of carbohydrates. It can be hydrolyzed with lactase, fiber, vitamin D and other additives such as flavorings and other vitamins. The mixture is subjected to pasteurization, ultra pasteurization or UHT treatment and then homogenized, added calcium and aseptically filled. This patent is a totally different formulation from our proposed Lutein Flavored Acidified Milky Drink proposal because it contemplates a special-purpose weight loss product and utilizes a complex and costly manufacturing technology that could never be deployed in any dairy industry. . Moreover, the formulation of this product is totally different due to the technological manufacturing process used, especially with regard to the protein content which is quite high and makes the formulation very expensive. Our proposal is to develop a low cost acidified, flavored, pasteurized and lutein-added dairy product that can be stored for at least 90 days both refrigerated and stored at room temperature. Our product also fits the designation "Dairy with added" as defined by the Technical Regulation of Identity and Quality of Dairy. Our beverage is a technologically viable alternative and can be easily produced and marketed by the dairy industry without the need for major investments as in the case of the aforementioned invention which requires expensive equipment such as sterilizers. In our case we can take advantage of the facilities available in a dairy industry and still make the rational and sustainable use of whey generated in cheese making. The addition of a bioactive component (lutein) confers considerable antioxidant activity to the formulation of the developed dairy drink. This gives the product a metabolic or physiological role in the protection of the human body against damage caused by oxidative stress, especially in relation to the maintenance of normal macular tissue functions. It can thus be called a functional product that in addition to nourishing has a proven beneficial function to the body of people who consume the product regularly and maintain healthy lifestyle habits. The acidified dairy beverage manufacturing process allows milk proteins to remain soluble at low pH, which may provide extended shelf life under room temperature or refrigeration. The procedure also allows the use of pasteurization only as a form of microbiological control, eliminating sterilization, avoiding the use of more sophisticated equipment that can inactivate various nutrients in the food. 3. PRODUCT AND MANUFACTURING PROCESS - MATERIAL AND METHODS

The development of the present invention was conducted at the facilities of the Research Center of the "Candido Tostes" Dairy Institute of the Minas Gerais Agricultural Research Company (CEPQ / ILCT / EPAMIG).

Physicochemical, microbiological and sensory analyzes were performed in the milk and dairy research laboratory and in the CEPQ / ILCT / EPAMIG sensory analysis laboratory. The processing, the type of acid used, the ingredients, the binomial heat treatment time / temperature of the product, the manufacturing process and the ideal formulation for the production of the acidified dairy drink were defined with the aid of preliminary tests performed in the research laboratory. and at the industrial core of CEPQ / ILCT / EPAMIG. Based on the definition of all parameters and formulation, the beverage was produced in five repetitions for proximate evaluation and monitoring of shelf life.

The experiment was conducted using a completely randomized design. It was evaluated whether the treatments (storage under refrigeration in a domestic refrigerator at 8 to 10 ° C, and at room temperature at 25 ° C) and the storage period (times 1, 30, 60 and 90 days after manufacture) had significant effect. on physicochemical, microbiological and sensory variables. Data were subjected to analysis of variance and regression analysis, with the aid of Statistical Analysis System (SAS) version 9.1. Descriptive statistics was also used for evaluation, presentation and discussion of results.

3.1. Preliminary Tests

For the manufacture of the acidified dairy drink, 105 preliminary tests were performed in the CEPQ / ILCT / EPAMIG research laboratory. Different types of acids (citric, fumaric, acetic, lactic and mixture), sugar content, different pH levels were tested, seeking the best results in terms of conservation and sensory acceptance of the acidified milk drink, different types. and levels of use of various stabilizers (pectins, guar gum, xanthan gum, carrageenan, carboxymethylcellulose, gelatines) and their mixtures, as well as various time-temperature heat treatment binomials. The acids with the best sensory acceptance and the stabilizers that avoided phase separation were selected, keeping the soluble proteins at pH below 4.5. The choice of flavor to be used in the acidified dairy drink was made by sorting test with a panel of 14 untrained tasters. The 12 tested flavors that had the highest acceptance rate are contained in the present invention.

3.2. Obtaining serum

Whey was obtained from the industrial core of the Cândido Tostes Dairy Institute from the production of Minas Frescal cheese exclusively produced for this purpose. However, whey obtained from the manufacture of other types of cheese such as mines, rennet and others can be used in product development.

For the production of Minas Frescal cheese, 100 liters of pasteurized milk standardized to 3.0% (w / w) fat were used to which calcium chloride (from 25 mL to 45 mL of 50% solution) was added. The milk was then slowly acidified with lactic acid solution (0.75% to 0.95% or 85 ° Dornic) so that the acidity was increased by 0.01 g to 0.03 g lactic acid per 100 mL of milk. The temperature was adjusted to 40 ° C and then coagulated using 4 mL to 6 mL of coagulant (Chy-max®, Chr-Hansen). After 30 min, the curd was slowly cut into large cubes (2 cm edge) and allowed to stand for one minute. The stirring process continued until about 20 minutes after cutting the curds. The serum was then removed to make the acidified dairy drink.

3.3. Product Manufacturing Process

Fresh whey obtained from Minas Frescal cheese is filtered through a desorption filter to eliminate all remaining cheese fines and heat treated at 62 ° C to 65 ° C for 0.50 minutes to 2 minutes. for disposal of any coagulant residue. To the whey is added standardized milk with 3.0% fat and homogenized in two stages: at 140 kgf / cm2 and 35 kgf / cm2. For the reduced lactose product the enzyme β-galactosidase is added at this stage to the whey and milk mixture until the required hydrolysis time. To this mixture of whey and milk is added, under stirring, premixed solid ingredients (sugar, sodium citrate, stabilizer, fumaric acid, potassium sorbate and lutein extract) and the mixture is stirred until complete dissolution of the mixture. all solid ingredients. For the light and diet products at this stage the sugar is reduced by 25% and 100%, respectively, and for the last sugar reduction the sweetener vim can be added which can be sucralose, aspartame, sodium cyclamate or saccharin. or mix of them. And the addition of vitamins is performed. The temperature of the mixture is adjusted to 65 ° C and homogenized at 140 kgf / cm2 in the first stage and 35 kgf / cm2 in the second stage of the homogenizer. The mixture is acidified with 50% (w / v) lactic, citric or tartaric acid to pH below 4.5, and then natural or natural-like aroma is added. The product is heat treated at 68 ° C for 12 minutes, hot packaged in polyethylene bottles of varying volumes, previously sanitized in 200 mg / l active chlorine solution. After filling, the bottles should be capped, inverted for 2 minutes, and then rapidly cooled to 20 ° C in a cold water bath. Properly labeled bottles should be stored refrigerated in a domestic refrigerator (8 ° C to 10 ° C) or at room temperature. Physicochemical, microbiological and sensory analyzes were performed at times 1, 30, 60 and 90 days of storage. Figure 1 shows the manufacturing process of the lutein-enriched acidified dairy drink.

3.4. Determination of shelf life of acidified milk drink

Processing was performed in five repetitions, the bottles being stored at room temperature (25 ° C) and refrigerated (8 to 10 ° C) to evaluate their shelf life. Periodically, at times 1, 30, 60 and 90 days, microbiological, physicochemical, sensory and appearance analyzes are performed to verify possible product changes during storage. To assess the microbiological stability of the beverage, analyzes of the most likely number of coliforms are performed at 30 ° C and 45 ° C, one day after manufacture, and counts of messsophilic aerobic microorganisms and filamentous fungi and yeast at different times chosen until the end. storage at both temperatures.

For physicochemical evaluation of the drink, analyzes of centesimal composition (one day), besides pH, acidity and apparent viscosity (1, 30, 60 and 90 days), as well as the evaluation of the appearance, odors foreign matter and packaging deformation throughout refrigerated storage at room temperature. Sensory analyzes (acceptance tests) were performed during the entire storage period under refrigeration and at room temperature. Table 1 shows the summary of the physicochemical, microbiological and sensory analyzes performed on milk, whey and product during storage under different temperatures.

3.5 Sensory analysis

The evaluations were performed at the CEPQ / ILCT / EPAMIG Sensory Analysis Laboratory. The tests were conducted in individual booths under white light. The samples were presented in a randomized and balanced manner, at refrigeration temperature (5 ° C), properly coded with random three-digit numbers, in 50 mL disposable cups, containing approximately 30 mL of the acidified milk drink.

During the preliminary tests the choice of the aroma to be used in the acidified milk drink was made by ordination test, according to the methodology described by Minim (2006) and Paula (2009). The ordination was performed by a panel of 14 untrained tasters. Twelve different flavors were tested. The tests were conducted in two sessions with six samples at a time. The same type of evaluation form was used in each session.

To determine acceptability, acceptance tests were performed on each repetition of the acidified milk drink, at 1, 30, 60 and 90 days storage under refrigeration and at room temperature. Acceptance was assessed by a minimum of 30 untrained tasters using the nine-point hedonic scale (MINIM, 2006).

To assess whether storage time significantly altered drink acceptance, data were subjected to analysis of variance and linear regression.

3.6. Lutein addition and evaluation of product antioxidant activity

The 5% lutein extract obtained from the Clove flower (Targetes erecta), provided by DSM Nutritional Products Ltd. (Wurmisweg 576, CH-4303 Kiseraugst, Switzeland) was added in an amount corresponding to 0.032 mg lutein. per milliliter of the acidified beverage so as to obtain an amount of 8 mg of pure lutein in a 250 mL bottle of beverage. This value was defined based on the results obtained by Dias (2009) and based on literature data, which indicate the daily dietary intake of lutein required to decrease the risk of macular degeneration from approximately 6.0 to 20.0 mg per day. (ALTERNATIVE MEDICINE REVIEW, 2005).

Antioxidant activity analyzes of the dairy drink were performed at the Natural Dyes Laboratory of the Food Technology Department of the Federal University of Viçosa. Antioxidant activity was evaluated when the drink completed 30 days of storage. After the extraction process was performed, according to the methodology developed and standardized by Dias (2009), the samples were resuspended in 5 mL of 99.9% purity absolute ethanol (Merck®) and submitted to the antioxidant activity analysis.

The evaluation of lutein antioxidant activity in the product was performed using the ABTS (2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) radical cation spectrophotometric technique), according to the methodology described by Cai et al (2004) and Dias (2009), with some modifications, using a Shimadzu 160-Vis UV-Visible spectrophotometer. ABTS '+ radical cation was generated by reacting 7.0 mM ABTS with 2.45 mM potassium persulfate at a ratio of 1: 1 (v / v) in the absence of light after 15 minutes in a water bath. 50 ° C. After this period, the obtained ABTS '+ radical solution was diluted in absolute ethanol to an absorbance of 0.700 ± 0.050 at a wavelength of 734 nm.

Subsequently, 0.5 mL of the lutein sample was mixed with 3.5 mL of the diluted ABTS '+ radical solution. Absorbance readings were taken at 734 nm immediately after mixing (time 0) and at times 20 min, 40 min and 60 min. During this period, the samples were kept under heating (50 ° C) in a water bath. Control and blank were prepared by replacing 0.5 mL of the lutein sample with 0.5 mL of absolute ethanol and 0.5 mL of the non-lutein-added beverage sample, respectively. Trolox was used as a reference standard. A calibration curve was prepared for Trolox concentrations ranging from 0 to 0.135 mM.

The results were expressed as Trolox equivalent antioxidant capacity (TEAC), correlating the reduction in absorbance in the dose response curve with standard antioxidant (Trolox), and percentage inhibition (% i), according to the following equation:

<formula> formula see original document page 19 </formula>

on what:

Ab = white absorbance at time t minutes rest in the dark; and

Aa - sample absorbance at time t minutes of rest in the dark.

4. Conclusions

Based on the conditions employed in this patent, the results obtained in the product development, it can be concluded that the developed dairy drink can be stored for at least 90 days both refrigerated and stored at room temperature.

The product falls under the designation "Milky Drink with Addition" defined by the Technical Regulation of Identity and Quality of Milky Drink. The new beverage is a technologically viable alternative and can easily be produced and marketed by the dairy industry without the need for major investment, taking advantage of the facilities and available whey.

The Milky Drink is nutritionally superior to soft drinks by serving not only "empty calories" but protein, vitamins and minerals that are essential for maintaining health.

The new beverage developed showed physicochemical and microbiological stability over time, demonstrating that the barriers imposed as acidity, preservatives and pasteurization were efficient in its conservation.

The average acceptance of the refrigerated product was slightly higher than the product stored at room temperature, with a reduction in sensory acceptance during storage at both temperatures.

The added lutein imparted considerable antioxidant activity to the developed dairy beverage formulation. This may give the product a metabolic or physiological role in protecting the human body against damage caused by oxidative stress, especially in relation to the maintenance of normal macular tissue functions.

Claims (13)

1. Table of Claims 2. FLAVORED MILK AND MILKY ACIDIFIED MILKIN DRINKS, A ready-to-eat liquid food product characterized by containing whey from the production of enzymatically coagulated cheeses type Minas Gerais, Minas Frescal, Cheese of rennet and others to (40 to 70%) acidified to pH below 4.5 with 50% citric, lactic or tartaric acid (from 1.5% to 3%), plus homogenized milk (30 to 60%) , sugar (8 to 20%), sodium citrate (0.01 to 0.5%), stabilizer (0.1 to 1%), fumaric acid (0.01 to 0.3%), potassium sorbate (0 0.01%), lutein extract (0.001 to 0.01%); AROMATIZED, LUTEIN-ENHANCED MILK SERUM ACIDIFIED MILK SERUM, according to claim 1, characterized in that the combined product is light by reducing at least 25% of the added sugar content; AROMATIZED, LUTEIN-ENHANCED MILKWOOD ACIDIFIED MILK SERUM, according to claim 1, characterized in that the combined product is diet by substituting sugar and adding a sucralose sweetener, aspartame, sodium cyclamate or saccharide or mixture thereof; ACIDIFIED, FLAVORED, MILKED, MILK-BASED MILK-DRY MILK DRINK according to claim 1, 2, and 3, characterized in that the product contains reduced lactose content through whey and milk delactose. by addition of the enzyme β-galactosidase; FLAVORED MILK-SUGAR-MILKED ACIDIFIED MILK SERUM, according to claim 1, 2, 3 and 4, characterized in that the product contains natural aromas of acai, banana, kiwi, acerola, grape, peach, orange, lemon, apple, pineapple and tangerine (0.02 to 0.2%); Acidified milk whey based on whey and milk, enriched with lutein according to claim 1, 2, 3 and 4, characterized in that the product contains the same natural aroma of citrus fruit (0 0.2 to 0.2%); 8. FLAVORED MILK-SUGAR-ACIDIFIED MILK-BASED MILK SERUM, according to claim 1, 2, 3, 4, 5 and 6, characterized in that the product is added with vitamins; 9. AROMATIZED, LATIN-ENHANCED MILKY ACIDIFIED MILK DRY PROCESS "according to claims 1 to 6, comprising" the steps in obtaining the whey from manufacture of fresh, standard mines, rennet and other cheese; whey filtration; thermosing at 65 ° C for 1 minute; addition of standardized milk with 3.0% fat and homogenized at 140 and 35 kgf / cm2; addition of solid ingredients mixed: sugar, sodium citrate, stabilizer, fumaric acid, potassium sorbate and lutein extract; temperature adjustment to 65 ° C; homogenization at 60 ° C (two stages: initial 140.62 kgf / cm and final 35, 15 kgf / cm2); acidification with 50% (w / v) lactic, citric or tartaric acids up to pH below 4.50; addition of natural or natural-like flavors; heat treatment at 68 ° C for 12 minutes; hot filling in 250 ml, 500 ml, 1000 ml, 2000 ml, 25 ml containers. 00 ml and 3000 ml, rapid cooling to 20 ° C; refrigerated storage at 8 to 10 ° C or at room temperature for up to 90 days "; 10. AROMATIZED, LATIN-ENHANCED MILKY ACIDIFIED MILK SERUM DRINKING PROCESS, according to claim 8, because the product is light by reducing at least - 25% of the product. added sugar content; 11. FLAVORED "FLAVORED MILK-SUGAR-BASED MILK-DRINKED MILK-DRINKING" PROCESS, according to claim 8, because the product is dietary through sugar substitution and the addition of a sweetener which may be sucralose, aspartame, sodium cyclamate or saccharide or a mixture thereof; 12. AROMATIZED, LATIN-ENHANCED MILKY ACIDIFIED MILK SERUM DRINKING PROCESS, according to Claim 8, because the product contains reduced lactose content through whey and whey delactose. milk by addition of the enzyme β-galactosidase; 13. AROMATIZED, LATIN-ENHANCED, MILK-SUGAR-BASED MILK DRY PROCESS, according to claim 8, 9, 10 and 11, by adding vitamins;