WO2018039758A1 - Antioxidizing microparticle and uses thereof - Google Patents

Abstract

The present invention relates to a microparticle comprising wall material, a non-ionic surfactant, essential oil from Ocimum gratissimum (basil) and essential oil from Thymus vulgaris (thyme). The invention can be used in the food, pharmaceutical and cosmetic industries, as a means for increasing the shelf life of the product and for antioxidant protection, both in storing the product to avoid oxidation thereof or as an additive in animal feed for the purpose of producing meat with lower propensity for oxidation.

Description

 ANTIOXIDANT MICROPARTRIC AND ITS USES FIELD OF THE INVENTION

The present invention relates to a microparticle with functional and antioxidant properties comprising wall material, nonionic surfactant, Ocimum gratis-sdmum essential oil (alfavae) and Thymus vulgaris essential oil (thyme).

Additionally ¹ , the invention relates to the use of microparticles for the food, pharmaceutical and cosmetic industries as a means to increase the shelf life of meat products by means of antioxidant protection, either in stocking the product avoiding oxidation or as an additive. in animal feed.

 BACKGROUND OF THE INVENTION AND TECHNICAL STATE

 Synthetic antioxidants have been widely used in the food industry and food preservatives due to their effectiveness and low cost, being BHT (butylhydroxytoluene), BHA (butylhydroxyanisol), BHQ (tertiary butylhydroxyquinone) and PG (propyl gallate) the most used. However, such compounds present restriction in some countries, as there is a suspicion that they present potential risks to human health, such as carcinogenic effects. Therefore, there is a growing interest in products that are natural and safer for food applications, with a progressive trend in consumer preference for natural antioxidants.

[004] The search for alternatives to these products has driven research, such as volatile oils present in herbs, which have proven antioxidant activity. These oils, presented in microencapsulated form, will prove to be a promising strategy in the incorporation of poultry diets and also in the production of meat products in order to control iipidic oxidation, be to offer harm to the consumer.

 The state of the art shows that there is a linear correlation between the high percentage of plant phenolic compounds and the high antioxidant capacity, proven by various analytical methodologies (ZHENG; WANG, 2001; KATSUBE et al., 2004; WOJDYLO et al., 2007; DUDONNÉ et al., 2009). Wei; Shibalttot (2010) evaluated the antioxidant effect of 25 types of volatile oils by four different methodologies and classified clove thyme oils with high levels of thyrool. (231) and eugenol (77 $), respectively, as the main antioxidant activity. These results corroborate the work conducted by Kahkonen et al. (1999), Lee et al. (2005) and Woidylo et al. (2007) who identified that thyme essential oil was a potent antioxidant comparable to BHT and alpha-tocopherol.

The eugenol compound is traditionally known to be the largest proportion constituent in clove essential oil [Eugenia caryophyllata]. However, the species Oci is a çratíssimum. , popularly known as alfavacão, also has eugenol as the main compound identified in its composition (SILVA et al., 1999, · VIEIRA et al., 2001). Review by Prabhu et. al, (2009) highlights several recognized and studied properties of 0. gratissímum, highlighting eugenol as the most beneficial component in different parts of the plant, emphasizing its antimicrobial activity and high antioxidant capacity when evaluated in vitro. Dor an et al. (2000) evaluated the in vitro antioxidant capacity of different essential oil compounds and concluded with the highest value for eugenol, being superior to the synthetic antioxidant. BHA

 [007] The tirol and eugenol compounds are recognized as ORAS (generally recognized as safe additives) by the Food and Drug Administration (FDA) and can therefore be used in the preparation of meat products.

 [0 8] However, essential oils are volatile ingredients and their incorporation requires greater care, so the spxay ~ drying process of microencapsulation preserves these compounds efficiently from adverse environmental conditions such as oxygen, light and moisture. for it will be wrapped in a wall material; besides the ease for inclusion and dispersion in meat mixtures,

 That said, the development of a natural product based on microencapsulated essential oils to be added to meat, meat products, and animal diets to control lipid oxidation without harm to the consumer would be to especially for the meat industry.

[010] Abd El-Alim, SSL. , et al. , on 02/99 "Culinary herbs inhibit lipid oxidation in rats and cooked minced meat patties during storage" describe the use of several plants to inhibit lipid peroxidation in chicken and pig meat. The plants were employed: marjoram, wild marjoram, cumin, clove, peppermint, nutmeg, curry, cinnamon basil, saivia, thyme and ginger, the proposed invention differs from the working Abd El-Alim, SSL, et al, mainly because of the. The present invention utilizes a pre-established blend of essential oils in microencapsulated form and which, in addition to application in the formulation of meat products may also be used in poultry feed to obtain benefits in controlling oxidative processes in meat. Furthermore, the present invention is a product with microenepsulation technology, making it more homogeneous and easy to handle and incorporate, as well as the determination of. an optimum level of addition for meat products and poultry feed.

 [OR] Document KR1Q133 & 65S of 08/08/11 refers to a method of producing meat products whose stability is provided by suppressing fat oxidation due to an naturally occurring ethanolic extract that acts as an antioxidant. The proposed invention differs from KR101335658 mainly in that it deals with the use of an ethanolic extract of "chi", an extract obtained from the leaf, trunk and root of the plants. , Saussurea seoulensis and Saussurea grandifolia, for spray application to meat, while the present invention is a combination of two microencapsulated essential oils which is used both as an additive in poultry diets and in meat product formulations and both for the purpose of containing the oxidative processes with well defined application values.

[012] Dawidowicz, A-, et al on 5/21/15 "Does antioxidant properties of te raain component of essential oil reflect its antioxidant properties? The comparison of antioxidant properties of essential oils and their main components" describe a study that discusses the similarities and differences between the antioxidant activities of essential oils and their main components, among which Thymus vulgaris and its component: trmol, aromatic Caryophyllus and its eugenol component. The proposed invention differs from the work of Dawidowicz, A., et al mainly in that the present invention is an association comprising the essential oils of microencapsulated Thymus vulgaris and Ocimum gratissimum, with at least proven antioxidant both in vitro and in vivo. In addition, in vitro antioxidant analyzes were confirmed in vivo, as well as using the meat product matrix by employing appropriate analysis methodologies. In addition, the present invention goes beyond the in vitro evaluation of antioxidant activity, as it validates two ways of applying the product: both as an additive in broiler diets and the subsequent effect on meat conservation, as well as in the preparation mixture. of products here.

[013] The W02 ^document:: Q 143.666.7 of 07/09/12 refers to a composition containing essential oils from aromatic plants of the family Lamiaceae, fungicide action. Essential oils are produced by continuous extraction from Tkymus vulgaris (thyme) and Origanu vulgate (oregano). The oils produced contain a high amount of carvacrol and thymol. The reported product is a mixture of the two plants in their oily form, without stipulating the proportion and just ensuring the boundaries of each major compound. The proposed invention differs from WO201436667 mainly by the form of production and application, as well as by the type of plants used and the proportion of the association. In addition to the fact that the application is for a different purpose, since the present invention validates two forms of antioxidant action (feed additive ¹ and meat product preparation), it also refers to distinct plants, with the technological differentiation of microencapsulation, which transforms the liquid product into microparticles in powder form, facilitating the manipulation and homogenization in the product matrices,

US201107276376 of 07/04/08 relates to a natural antioxidant composition for meat products, characterized in that it is prepared a. from phenolic extracts honey. The application form is by spraying the marinade liquid by spraying or bathing, the purpose of which is to take into account the meat forming a protective film. Therefore, there is no safety limit or level recommendation for product use. The proposed invention differs from document 05201100.76376 mainly in that the proposed invention is an established association of the essential oils of Thymus vulgaris and Ocimum grati & simum in microencapsulated form to be used both as an additive in poultry diets and in meat product formulations, both for the purpose. contain oxidative processes with well defined application values.

[015] Kim, Il-Suk, et al., On 6/21/11 reported in "Ant.ioxida.nt Activities of Hot Water Extracts from Various Spices ^x the use of extracts with antioxidant activity. Among the species evaluated, and Commonly used in meats are: thyme and oregano Kim, Il-Suk, et al. refer only to in vitro antioxidant analyzes, and to affirm the effectiveness and applicability of the substance (s). ) in meat products it is necessary to test in vivo or using the product matrix by appropriate analysis methodologies.The proposed invention differs from the work of Kim, Il-Suk, et al., mainly in that it not only analyzes the activity antioxidant in vitro, but validates two ways of applying the product: both as an additive in broiler diets and the after-effect on meat preservation, as well as in the mixture for the preparation of meat products, having as reference one of the synthetic antioxidants. most commonly used for this purpose.

[016] The document CN1317971C 12.26.15 re fe re the application of essential oils of Thymus vugaris and basil as additives in feed ^■ chicken. 0 percent oil from Thymus vugaris was used in the range of 1.5-2.5% and the percentage of basil in the range of 1, 5-2.5%. The proposed invention differs from CN1317971C mainly in that it is a mixture of various ingredients, including essential oils for use in the diets of poultry and livestock, but without specification or purpose of USE, only the form of manufacture of the product, while that demonstrates from the form of manufacture of the product, microencapsulation technology of the essential oils of Thymus v lgarís and Ocímvm gxatissimxxm, as well as the purpose that this invention will serve,

[017] Dashti, N. D., et al., 12/15 in "Antioxidant Effect of Thyme Essential Hi.1 on Oxidative Stability of Chicken Nuggets" refers to the use of essential oils from thyme species, but greater activity was observed for Eataria uififora. referred to in this paper "as thyme", with. antioxidant function in chicken nuggets. Dashti's work,. D., et al. , is a liquid product so there is evidence of greater difficulty of dispersion and homogeneity in the meat matrix. The proposed invention differs from the work of Dashti, N, D-, et al., Mainly in that only one of the essential oils (same genus of plant but not the same species of Thymus vulgaris) is evaluated in relation to the present invention that composes it. the combination of two essential oils. The product developed in the present invention is an exact 1 to 3 ratio mixture of the oils (Thymus vulgaris and Ocimum gratissimum essentials in microencapsulated form and proven to be effective at a lower level of use than proposed). , since the levels (1000 and 2000 ppm) that showed positive results in the control of the lipid oxidation of meat are much higher than those used in the present invention (f'0 and 140 ppm of the essential oils association) ^: In view of the foregoing, it would be useful if the technique had microparticles comprising essential oils to be added to meat, meat products, and animal diets to control lipid oxidation, alternatively as natural antioxidants in place of synthetics.

 BRIEF DESCRIPTION OF THE INVENTION

 [0153] The present invention relates to an antioxidant microparticle comprising:

 - Wall material: Maltodextrin

 - Emulsifying agent: Polysorbate 80, was a variable proportion of 8 to 12%, preferably 10% by mass referring to total solids;

 - Thyme essential oil and alfavacão essential oil, in the ratio of 1: 3;

 - 4 to 61, preferably 5% Thyme Essential Oil was mass relative to total solids;

 - 14 to 16%, preferably 15% of alfavaça essential oil, by weight relative to total solids.

The obtained microparticle has a size from 1 to 100 µm, the average size being 50 pm in spherical form, and is a release system of thyme and alfavaça oils which are in the concentration between 15 to 25 g of oil / 100 µm. g total solids, preferably 20 g oil / 100 g total solids. Said oils have a percentage of eugenol ranging from 40% to 60%, preferably a minimum of 45%; a percentage of variable carbon between 2% and 10%, preferably a minimum of 5%, a percentage of variable carbon between 3% and 10%, preferably a minimum of 8.0%, and a percentage of variable tirol between 5% and 10%. 15%, preferably at least 7.0%. The microparticle comprises at least 15% thymus plus charcoal and 45% eugenol (or 1: 3 ratio). of these constituents). The maximum imprisonment is 20% of. oil.

 In addition to microparticles, it has application in the food, pharmaceutical and cosmetic industries, with antioxidant action to extend the shelf life of products, particularly in. meat and meat products, as an additive in the feeding of farmed animals, as in animal feed, with action on the quality of meat. Meat products in the present invention are mature fresh meat products, cured meat products, restructured fresh meat products, salted meat products, cured and / or smoked meat products, cured sausage products and. canned and / or potted meat products. More specifically, they are meatballs, hamburgers, nuggets, steak, sausages.

 BKBVB DESCRIPTION OF DRAWINGS

 Figure 1. Root aspect of essential oils of. vulgaris and O, gratissimum visualized under optical microscope. 400x magnification.

 Figure 2. Malonaldehyde (MDA) formation in cooked chicken breast meatballs fed with increasing levels of OEM and stored at 4 ° C for 7 days.

 Figure 3. Malonaldehyde (MDA) formation in cooked drumstick meatballs of broilers fed with increasing OEM levels and stored at 4 ° C for 7 days.

 Figure 4. Malonaldehyde (MDA) levels in cooked breast meatballs of broilers fed with increasing levels of OEM. Average values obtained during the 7 day period. Different letters (A, B) mean statistical difference by Tukey's test (P0,05).

Figure 5. Malonaldehyde (MDA) mve s in cooked drumstick meatballs of broilers fed with increasing levels. OEM Average values - obtained during the period of 7 days. Different letters (A _f B) mean statistical difference by Tukey test (P <0, Q5).

 Figure β. Malonaldehyde (MDA) formation before and after thermal processing of chicken breast meatballs incorporated with ΒΉΤ or two different OEM levels. * Statistical difference compared to raw meat (Durrnet test E <0.05). Different letters (Ά, Β) mean statistical difference between treatments after thermal processing (Tukey test P <0, G5).

 Figure 7. Malonaldehyde (MDA) formation before and after thermal processing of 3ΉΤ or OEM-incorporated chicken drumstick meatballs. * Statistical difference compared to raw meat (Dunnet's test P <0.05). Different letters (A, Bi mean statistical difference between treatments after thermal processing (Tukey test P <0.05).

 DESCRIPTION DET & LE & BA INVENTION

 £ 022] Antioxidant microparticle comprising Thymus vulgaris essential oil; Ocimum essential oil ■ gratiss-magnet? Wall material; and emulsifying agent, wherein. Thymus vulgaris and Qcimum gratissimum essential oils are in a 1: 3 ratio.

[023] The antioxidant microparticle wall material can be selected from calcium alginate; maltodextrin arabic gum; cashew gum; inui; starch; starch derivatives; cellulose; cellulose derivatives; tragacanth gum; Karaia gum; gum mosque; gellan gum; pectin; soy polysaccharides; milk polysaccharides; milk polysaccharide derivatives; casein; gluten gelatin; chitosan waxes; fatty acids, preferably maltodextin. The antioxidant microparticle emulsifying agent may be selected from non-ionic surfactants such as 2- [.4- (2,4-trimethylpentan-2-yl) phenoxy] ethanol and Polysorbate 80, preferably Polysorbate 80.

 [025] In a preferred embodiment the antioxidant microparticle comprises:

 - 4 to 6%, preferably 5% Thymus vulgaris essential oil by mass;

 - 14 to 16, preferably 15% Ocimum gratissimum essential oil by mass .:

 - Maltodextrin as a wall material variable ratio 75% to 85%, preferably 80% by mass for total solids;

 - 8 to 12%, preferably 10% Polysorbate 80 by mass, relative to total solids,

[026] The microparticle comprises 10 to 30% _r preferably 20% essential oils. The combination of the oils of Thymus vnJ.gar.is and Ocimum gratissimum comprises at least thymol; carvacrol; eugenoi, where the ratio of the first two to the last is 1: 3

 [027] Thymus vulgaris oil has a percentage of thymol ranging from 5 to 15%, preferably 7%. Ocimum gratissimum oil, on the other hand, has a percentage of eugenoi ranging from 40 to 60%, preferably 45%.

[028] The association of the essential oils of Thymus v. Garlic and Ocimgrissi presents eugenic percentage ranging from 40 to 60%, preferably 45%, percentage of carvacrol ranging from 2 to 10%., Preferably 5%, _percentage: p-cymene variable between 3 and 10%, preferably 8.0% thymol variable percentage between 5 and 15%, preferably 7.0%. [029] The combination of the essential oils of hymus vulgaris & Ocimum gratissi one presents at least 15% thymol plus carvacrol and at least 45% eugenol.

 The icroparticles, have size between 1 to 100 p.m and average size of 50 μιυ in spherical form.

 The oily composition comprises alpha-thujene; alpha-pinene camphene; beta-pinene; beta-irdrcene alpha-felandrene; delta-3-carene; alpha terpinene; p-cymene; d-liraonene; eucalyptus! beta-ocimene; gamma-terpinene; useful sabinene hydrate; terpinolene; linalool; alloylene; camphor; gammapinpinene; borneoi; terpin-4-ol; alpha terpineol; isobornyl acetate; thymol; carvacrol; alpha-cubenene; eugenol; alpha copaene; beta-bourbonenc; longifolene; z-caryophyllene; beta-gurjune.no; alpha-humulene; gammaururo1ene; germacrene-D; aifururoiene; gamma cadinene; delta-cadinene.

 The oily composition will comprise at least thymol; carvacrol and eugenol, the ratio of the first two to the last being 1: 3.

 [033] In addition, it is an object of the present invention to use the microparticle for the food, pharmaceutical, cosmetic industry and as a protection system for thyme and alfavaça essential oils, as they are highly volatile compounds, allowing the formulation of a product with better antioxidant protection, either during stocking of this product or as feed additive for farm animals, to obtain antioxidant action in the meat of the animal fed with a minimum concentration of 350pprn.

[034] The application of microparticles in meat and meat products to prevent meat oxidation, concentration 350ppra to OOppm, preferably 700ppm. And the addition in animal feed, as in feed, to prevent oxidation of animal flesh is fed at a concentration of 350 to 3500ppm _r 35G0ppm pre circumferentially.

 Embodiment Example

 The Chemical Composition of the Essential Oil Association was determined by GC-MS analyzes prior to the microencapsulation process (Table 1), and the identified major compound was found to be eugenol (46.45% ), followed by carvacrol (8.40%), p-cymene (8.18%) e. thymol (7.05%). The literature reports that the Lamiaceae family plants and especially the comuraente essential oils found in the Tmusus and Ocimum genera are rich in these compounds and have proven antioxidant activity because they have one or more hydroxyl (OH) groups attached to the aromatic ring, - unsaturation and electrons available to be donated.

 Table 1. Chemical composition (% of the essential oil association of T. vnlgaris and O, grâtíssi nm at a 1: 3 ratio.

 Compounds

 Relative IR%

(min) identified

 5.79926 alpha-tu and not 0.5

5,998,933 alpha-pinene 1, 32 o, 38 947 cannon o 0.83

7, 1 976 beta-pyrie 0.17

7.54 990 beta mircene 0.65

 alpha-

8,00 1005 elandreno 0, 48

 delta-3-

8.18 1010 grace 0.13

 alpha-

8, 0 1016 terpine.no 0, 25

 8.72 1025 p-cymene 8.18

 8, 82 1028 d-iimone the X r 01

8, 7 ^: 1032 eucalyptus! 0.58

 eta-z ~

 9.1G 1036 ocimene 1.59

 ga a-

9.86 1057 t and pin 0.33

 hydrate of

 10, 26 1068 lo sahineno 0, 14

10 96 96 terpinolene 0.34

11.49 1102 linalol 1.6 12 55 1127 allocimene 0.73

13, 29 1145 camphor 0, 19

 gamma-

13.76 1157 terpine the 0, 8

1,115,166 bor eol 0.93

14, 63 1178 terpin-4-ol 1.20

 alpha-

15.20 1191 terpineol 0.30

 iso o.rnil

 19.24 1287 acetate 0.57

 19.64 1297 thymol 7.05

20.07 1307 carvacrol 8.40

21.78 134S a.1fa-cubenene 0.22 te, S 53 1366 eugenol 46.45

 22, 7 1376 aifa-copaene 1.66 beta-

23,33; 1385 bourboné the 0, 67

23.63 1392 m = 204 0.43

24, 12 1 04 long.ifolene 0.23

24.76 1419 z-cthiophyllene 4.29 beta-

25, 09 1 27 gurjunene 0, 31

26, 06 1452 a1 fa-humu1ene 0.37 gamma

27, 02 1475 uurolene 0.89

27, 22 1480 germacrene-D 2, 42 alpha-

27, 94 1498 muurolene 0.31

28, 48 1512 gamma-cadynene 0.50 delta-

28.87 1522 cadinene 1.83

TOTAL 37 compounds

Tr - time ^: retention; IR - index, retention

 M croencaps lation of the essential oils association

[-036} The oils of 7. vulgaris and 0. gxâtissimum were combined in 1: 3 ratio and subjected to microencapsulation for use in in vitro and in vivo application assays. As wall material was used altodextrin (brand MorRex 1910® with DE 10) from Corn Products (ogi Guaçu, SP, Brazil). As eraulsificante agent was used Poiysorbate 80. The operating conditions were optimized for raicroencapsulação li'zando ICU is a mini spray çíryer by varying the inlet temperature of 180 ± 5 ° C. The carrier gas used was N2 (RODRIGUES, 2004). In this step, 24 g of maltodextrin was dissolved in 70 g of distilled water with the aid of an Ultra Tur ax (Ά Ά TlQ ^' ). A. A combination of essential oils, comprising a total of 6 g, of which 1, 5 g corresponds to T oil, commonly used, and 4.5 g of O. gratissimum oil plus 3 g of emulsifying agent (Polysorbate 80) was added to the preparation and homogenized, totaling 30% total solids. homogenization, the formulation was passed in the mini spray dryer and the final result was white particles containing, on average, 20% of the association of essential oils, being designated as OEM (microencapsulated essential oils);

 Characterization of the lco-encapsulation process

 [038] Microparticles are white, spherical in shape, similar to white flour and of varying size, averaging 50 μιη as shown in Figure 1. The appearance with the naked eye is uniform and homogeneous, with differences in size perceived only with the use of an electron or optical microscope.

 [039] Quantification of the total phenolic compounds present in the association of óleos essential oils. vulgaris and O. gratissimum in pure and microencapsulated form (OEM.) was determined by the colorimetric method (SINGLETON et al., 1999) using gallic acid as standard. The association of oils in pure (liquid) form was diluted in acetone (0.05 g of OE in 99% acetone Imf}; while, the OEM sample was directly diluted: in Milli-Q water. The color blue produced by phenol reduction in the Folin-Ciocaiteu reagent was measured by spectrophotometer at 740 nm (Shimadzu brand, UV mini 1240 model}. Results were expressed as gallic acid equivalent content (mg GAE / 100 g). .

[040} Antioxidant activity by absorption capacity oxygen radical (ORAC) was determined by quantifying the protective effect of an antioxidant and / or essential oil by assessing the area under the sample fluorescence decay curve when compared to white (maltodextrin), ie , a control without the association of essential oils or another antioxidant agent - Trolox, ura _. Water soluble analog of vitamin E was used as an antioxidant control standard. Fluorescence was monitored and recorded every 56 seconds for approximately 75 min., Optimal Plousta Fluorimeter (BMG LABTECf, Germany) at 37 ° C and with a 4:85 nm excitation filter and 520 nm emission filter. Analyzes were done in triplicate. Results obtained for the combination of essential oils in pure and microencapsulated form (OEM) were expressed as trolox equivalent concentration / mg of the sample.

[041] These analyzes to determine the amount of total phenolics and the antioxidant capacity of the essential oils association were performed before (liquid EO) and after microencapsulation (OE microencapsulated) (Table 2) to verify losses during the microencapsulation process, since the liquid sample composed of volatile oils is subjected to a high temperature process but for a short period. Considering the 100% basis for the net determined values which were respectively 107.77 and 986.76 for total phenolics and ORAC ,. It is observed that the proportion of the results in microencapsulated form was 28 and 21% in natural matter. This value is in accordance with the amount of oil that was incorporated into the microparticle (20%) during the spray-drying process, concluding that the microencapsulation technique did not affect the amount of phenolics and the antioxidant activity of the association. Table 2. Determination of antioxidant activity oxygen uptake capacity (OR&C)

 Nature of the phenolic QR&C total sample (trolox / mg of

(mg AG / mg sample) sample)

 SO 30 ± 1.36

 210 + 29 (21%) microencapstitiated (2SI) *

* ^■ Relative percentage considering the sample in 100% liquid bed form.

 [042] With the association in the microencapsulated form and proven antioxidant activity, the trial began with broilers evaluating increasing levels of inclusion of OEM in diets. Performance was evaluated during the period from 1 to 21 days of age (Table 3) and, at 35 days, the weight (% -> of liver in relation to the live weight of birds (Table 4).

 Experimental management

 [125] One hundred and twenty one day old CobbSOQ broiler chicks were used, all male. On the first day of the experiment, birds with homogeneous weights were housed in cages in a previously cleaned and disinfected room., The cages were housed in a masonry shed, and the ideal room temperature for the birds was obtained by handling curtains, fans and hoods according to the recommendations described in the CobbSOO strain manual. Heating in the first days of life was performed by lamps arranged in each cage.

[044] The experimental period was 35 days, with free water and feed for the birds. ^• The diets of tratamentos-, all in mash form, were composed of corn, soybean meal, soybean oil and mineral vitamin premix for broilers (F30110NB, Cargill). All macro and micro ingredients were present in the same amount between treatments, the only difference being the inclusion of OEM and / or maltodextrin, used as an "inert" ingredient. The diet formulation sought to meet or exceed the requirements of the National Research Council - RC (1999; Experimental diets were formulated without the inclusion of antibiotics, coccidiostats, enzymes and antioxidants, in addition to that contained in preraix (F30110NB, Cargill) to preserve. the vitamins in the diet.

 [045] Treatments with increasing levels of inclusion of the microencapsulated essential oil association (OEM) were designated as: Tl- Negative control diet (no OEM inclusion; T2- Diet with 350 ppm OEM inclusion i corresponding to 70 ppm) of pure association); T3- Diet with inclusion of 700 ppm OEM ((corresponding to 140 ppm of the association); T4- Diet with inclusion of 3500 ppm OEM (corresponding to 700 ppm of pure association), as shown in Tables 3 and 4. .

 Table 3. Bird body weight (PM) at 21 days, feed intake (CR), weight gain (GP) and feed conversion (CA) from 1 to 21 days of age.

 PM21 C5¾ Treatments. GP CA

 1-21 1-21 1-21

Tl- Control 9 9 1301 948 1: 3: 7

T2-350 OEM 1024 1293 974 1,33

T3- 700 OEM 1004 1300 953 1.36

T4- 3500 OEM 1010 1310 959 1.37

Probability 0, 862 0, 933 0, 861 0, 287 hp,% 4.4 4, 6 4.7 2.5 i.0 6 j In the period between 1 and 21 days of age, the treatments were distributed in a randomized block design ⁽ each battery of cages will be a block), with the cages being considered the experimental units. Each treatment consisted of four repetitions of six birds each, totaling 24 birds / treatment. From 21 to 35 days, each bird was considered an experimental unit (repetition), being eight birds / treatment.

 [047] At the end of the experiment (35 days old) all birds from each treatment were slaughtered by cervical dislocation for liver size determination. After weighing, each bird was boned and the skin and cartilage removed. Before packing, they were placed in an ice bucket (chi.I read;), identified and stored in a cold chamber at -20 ° C for 24 h. After this period, a pH analysis was performed with the primary objective of checking if DFD (dark, firm and dried) or? SE (palid, soft, exude ive) changes in meats Parts that showed any of these changes were not used in the shelf life test. determined by direct reading in a previously calibrated digital pH meter with pH 4.0 and 7.0 buffer solutions, by introducing the electrode into the muscle (IAL, 2008.) Six different reading points were taken per sample and the average was taken. The breast and drumstick parts were then frozen at -20 ° C and stored under vacuum in low oxygen permeability plastic bags and duly identified until further analysis,

Table 4 _* Weight relative to body weight (%) of poultry liver at 35 days of age.

 % Liver Treatments

T2- 350 ppm OEM 1.98 ^b

 T3- 700 ppm OEM 1.88 *

T - 3500 ppm OEM 2.40 ^a Probability 0.002

 CV,% π, ι

 a.b different- letters mean statistical differences by cie test Tukey (P¾), G5

 [048] Performance was not significantly affected (P> 0.05) by different levels of inclusion of OEM in broiler diets (Table 3). In this experiment, the main objective was to evaluate the shelf life of the meat of these birds, the performance analysis aimed to verify if there would be no losses, especially in relation to feed intake. One of the reasons for encapsulating the association of essential oils was to soften some organoleptic effects that are exacerbated in these substances and may in some way interfere with the consumption of the products to which they are mixed, in this case the ration. Feed intake was very similar between the different treatments, indicating that there was no interference in the addition of OEMs in the bird diet and being a positive point in the evaluation of this additive category.

[049] The liver percentage of birds was significantly affected (P <8, 05) by the inclusion of OEM levels and their diets (Table 4). The treatment birds that received the highest level of OEM inclusion (3500 pprr were the ones with the largest liver in relation to body weight (P < ^Q , ^Q 5). This liver analysis aimed to verify some production-related stimulation). In spite of the higher liver I of birds from treatment T4, when compared to those from treatment Tl (Control) the difference was not significant, inferring that the difference was not due to the presence of association of essential oils in the body of birds.

Effect of OEM inclusion in chicken feed on lipid oxidation of chicken meat (breast meatballs) thigh) after thermal processing and refrigerated storage

[050] The boneless and boneless meat (breast and drumstick) samples were thawed during the night under refrigeration at 4 ^{<3 °} C. All apparent fat was then removed and the parts ground together in a meat grinder ( Myralux, 12.00 W), being formed by treatment a pool of chest and another of thigh. On average, a total of 5.0 kg of breast and 3.5 kg of thigh were obtained for each treatment.

 For all treatments 0.5% sodium chloride was added to the meat mass which was homogenized and weighed into 301: 1 g portions of meat, and then shaped into meatballs. The meatballs were packed in polyethylene bags with low oxygen permeability and boiled in boiling water at 100 ° C for 8 min, according to Racanicci et al. (2004). They were then cooled in ice water, re-packed in polyethylene bags with high O 2 permeability, and sketched in the cold in a cold chamber at 411 ° C. Each bag was identified and composed of meatballs referring to the experimental treatments.

 [052] For the inclusion dietary QEM assay of poultry, meatballs were evaluated for lipid oxidation by the TBARS test after Q, 2, 4 and 7 days. Six meatballs / day / treatment were analyzed.

 [053] From the Tl poultry pool (control - without addition of any antioxidant additives), a total of 2 kg of breast mass and 1 kg of drumstick were removed for the OEM incorporation test directly into the meat. chicken as described below:

Breast test: For breast meat the following treatments were applied: Tl- control treatment (without the addition of any antioxidant additives); T2- treatment with addition of 150 ppm BHT (99.9); T3- treatment with the addition of 70 ppm OEM (corresponding to 10 ppm pure combination); T4- treatment with the addition of 350 ppm OEM (corresponding to 70 ppm pure association). The analyzes concerning lipid oxidation were performed with the raw meat and soon after cooking, being analyzed 4 meatballs / dressing.

Drumstick test: The following treatments were applied to the drumstick meat: Tl- control treatment (without the addition of any antioxidant additives); T2- treatment with addition of 150 ppm BHT (99.9%) ^' ; T3- treatment with the addition of 700 ppm OEM (corresponding to 140 ppm pure association) ■ The analyzes concerning lipid oxidation were performed with raw meat and immediately after cooking, being analyzed meatballs / treatment.

 Lipid Oxidation - TBARS Analysis

[054] The samples of brisket and drumstick were subjected to oxidative stability assessment by reaction with 2-thiobarbituric acid (TB RS). The methodology was performed according to. Vyncke (1970), with some modifications (SORRENSEN; JORGENSE, 1996). The 5 g meat samples were weighed, plus 15 µl solution containing 7.5% trichloroacetic acid ("TCA), 0.1% pro After the homogenization in Ultra-Turrax for 45 ^: seconds at 13,500 rpm, the aldehydes were extracted with TCA, diluted with EDTA (chelating agent) and GP ( antioxidant) to prevent fat oxidation during exation After this procedure 5 µl of TBA reagent (0.02 M) were added and the TBARS value determined by spectrophotometry at wavelengths 532 and 600 nm, and values are expressed as malonaldehyde (umol / kg meat sample).

 Storage under refrigeration lasted 7 days and an oxygen permeable plastic film was used to pack meatballs and simulate more extreme conditions. Lipid oxidation was developed during the day in all treatments as can be observed in the high TBARS levels in both brisket (Figure 2) and drumstick meat (Figure 3). Therefore, this increase applies mainly to dark meat.

(thigh and drumstick), as they have higher concentrations of polyunsaturated fatty acids than white meat (breast)

(JENSEN, et al., 1997).

 [056] Breast meatballs (Figure 2} from broilers fed OEM levels did not control lipid oxidation compared to control group (0), in contrast, the level of inclusion of 350 ppm in diets significantly worsened. malonaldehyde production (IMDA) (P <0.05), This result does not match those observer in the thigh meatballs

(Figure 3). Thigh meat is three times more fat than brisket meat (Bragagnolo, 2001) and this is an indication of a more evident action of ingestion of essential oils in animal muscle. In meatballs drumstick (Figure 3) from .2 days cooling treatment in which the birds consumed ^• the highest level of OEM (3500 ppm) differed from the control group, which had the worst result among the groups (P 0.05).

[057] The mean values of malonaldehyde (MOA) obtained during the whole refrigeration period (7 days) are: shown in Figure 4 (breast meatballs) and Figure 5 (drumstick meatballs). These results clearly demonstrated the ineffectiveness of essential oils in meat with a lower content of There were significant differences in the accumulated period only between the control group (0) and those supplemented with 350 ppm of OEM n ration, which had the worst result (P <0.05). On the other hand, it was evident when evaluating the thigh meatballs (Figure 5), that the meat of poultry fed with the highest OEM level in their diets (3500 ppm) showed a better control in the formation of oxidation products when compared to the group. control (P <0.05). Treatments with intermediate levels of OEM showed intermediate values of TBARS and may also be considered as a preventive measure of lipid oxidation control.

 Direct incorporation into meat of OEM levels on oxidative oxidation before and after thermal processing

 [058 Meat from. Control birds (without antioxidant additive) were separated and homogenized in equal parts for the incorporation of experimental treatments. Before being subjected to heat treatment a sample was taken representing the treatment of raw meat.

[0.59] Horn observed for brisket meatballs (Figure 6) and drumstick meatballs (Figure 7) Generally thermal processing increased the levels of malonaldehyde in the meat, comparing each post-heat treatment compared to the meat treatment. thermally unprocessed meat - this difference is signaled by between treatments. This indicates that meat processing steps such as grinding and cooking favor the conditions for the development of oxidation products (MITSUMOTO et al., 2005). Sheehy et al. (1993) reported that cooking increased lipid oxidation in liver muscle tissues by 4-10 fold compared to Raw tissues. However _. , in breast meat (Figure 6) the groups with the inclusion of BHT and 350 ppm OEM were efficient in containing the oxidative processes remaining with TBARS levels without significant difference when compared to the raw meat group, ie before undergo thermal processing (P> 0.05). For thigh meat (Figure 7) which has a higher fat content, only the OEM treatment (700 ppm) and not the BHT treatment was able to inhibit the oxidation process caused by heating, not differing from the values of raw meat (PM], 05). Thus, as a conclusion to the impact of thermal heating on TBARS levels, the addition of. 350 ppm. OEM meat was efficient in containing the oxidative process accentuated by this phenomenon in brisket and for thigh meat it was. 700 ppm OEM level.

 Comparing all treatments immediately after thermal processing, both in brisket (Figure 6) and drumstick meat (Figure 7), those with the inclusion of any additives (BHT or OEM) were effective in containing Oxidative processes when compared to the Control group (without additive inclusion) (P <0.05), comparison between treatments after thermal processing, signaled by the different letters "A, B, C" - mean differences between treatments. A highlight is the inclusion of 700 pp OEM in the manufacture of thigh meatballs, as the levels of mideIdehyde were below those produced by BHT treatment. It is important to remember that the total of essential oils present in the microparticles is around 20%, that is, in the amount of 140 ppm in this 700 ppra level of inclusion.

[061] The combination of T. vulgaris and O. grãtissi essential oils in the microencapsulated form proved effective in containing the oxidative processes of chicken meat when mixed with poultry feed. In this case, such effects were observed for meat of poultry fed with the highest inclusion level (3500 ppm), and heat treated drumstick meatballs.

 [062] The association of microencapsulated essential oils when incorporated at the 350 ppm and 700 ppm levels directly into chicken meat were effective in containing the oxidative processes in relation to the control, in the breast and drumstick meatballs, beyond the 700 ppm level. be more efficient than synthetic antioxidant (BHT) in thigh meatballs.

Claims

 1. Antioxidant microparticle. Characterized in that it comprises at least:  - essential oil of Thymus vulgaris;  - Ocimum gratissimu essential oil.  2, Antioxidant microparticle comprising at least:  - essential oil of Thymus vulgaris;  - Ocimum gratissimum essential oil;  - wall material; and  - emulsifying agent,  The essential oils of Thymus vulgaris and Ocimum gratissimum are in the ratio of 1: 3.  3, Antioxidant microparticle comprising at least:  - 4 to 6%, preferably 5% Thymus vulgaris essential oil by mass relative to total solids;  ~ 14 to 16% preferably 15% Ocimum gratissiinum essential oil by mass relative to total solids  - wall material; and  - Age and emuIsifying. Antioxidant microparticle according to claim 2 or 3, characterized in that the wall material may be selected from calcium alginate; arabic gum; maltodextrin; cashew gum; inui; starch-; derivatives of. starch; cellulose; cellulose derivatives; tragacanth gum; Raráía gum; gum mosque; geian gum; pectin; soy polysaccharides; milk polysaccharides; milk polysaccharide derivatives; casein; gelatine; gluten; chitosan; waxes; fatty acids, preferably maltodextrin. Antioxidant microparticle according to any one of claims or characterized in that the emulsifying agent may be selected from non-ionic surfactants; 2- [4- (2, _4-f 4-2-trimethylpentan yDphenoxy] ethanol, polysorbate 80, preferably polysorbate 80.  6. Antioxidant microparticle characterized by at least:  - Maltodextrin as wall material varying proportion from 75% to 85%, preferably from 80% by mass referring to total solids;  - 4% to 6%, preferably 5% of Thymus vulgaris essential oil in iaassa for top solids;  - 14% to 16%, preferably 15% Qcimvm gratissimum essential oil by mass referring to total solids;  S at 12%, preferably 10% Polysorbat 80 by weight relative to total solids.  Microparticle according to any of indications 2, 3 or 6, comprising 10 to 30%, preferably 20% of essential oils. Microparticle according to any one of claims 1, 2, 3 or 6, characterized in that the combination of the oils of Thymus vulgaris and OciMu gratissinrum comprises at least thymol; carvacrol; eugervol, being that the ratio of the first two to the last is 1: 3  Microparticle according to any one of claims 1, 2, 3 or 6, characterized in that the oil of Thym syrupis has a percentage of thymol ranging from 5 to 15%, preferably 7%.  Microparticle according to any one of claims 1, 2, 3 or 6, characterized in that the oil of Occis has a percentage of eugenol ranging from 40 to 60%, preferably 45%.  Microparticle according to any of claims 1,2,3 or 6, characterized in that the combination of the essential oils of Thymus vulga is and Ocimum gramissimum has a percentage of eugenol ranging from 40 to 60%, preferably 45%, percentage of carvacrol. variable between 2 and: 10%, preferably 5%, percentage of p-cyme variable between 3 and 10%, preferably 8.0%, of thymol variable between 5 and 15%, preferably 7.0%.  12. Microparticle, according to. any one of claims 1, 2, 3 or 6, characterized by the combination of the essential oils of Thymus vulga is. and Oçimvm-gratissimu have at least 15% thymol plus carvacrol and at least 45% eugenol.  Microparticle according to any one of claims 1, 2, 3 or 6, characterized in that it has a size between 1 to 100 µm and an average size of 50 µm in spherical form. Antioxidant microparticle comprising: - Maltodextrin as a wall material in varying proportions of 75 to 85%, preferably 80% of total solids;  Poiysorbate S0 as emulsifying agent in varying proportion from 8% to 12%, preferably 10% relative to total solids; and  Oily composition comprising alpha-thujene; alpha pinene; camph. at the; beta-pinene; beta-mircene; alpha-felandrene delta-3-carene; alpha-terpinene p-cymene; d-1 imonerium; euoalyptol; beta-z-ocimene; gamma-terpinene; useful sabinene hydrate; terpinolene; iinalol; allocimene; camphor; gamma-terpinene; borneol; terpin-4-ol; alpha terpineol; isobornyl acetate; tiruol; carvacrol; alpha-cubenene; eugenol; alpha-copasno; beta-bourbonene; iongifolene; z-caryophylline beta-gur juneno; alpha-humuene; gamma muarolene; germacrene-D; alpha-muurolene; gamma cadinene; 11a-cadinene.  Microparticle according to Claim 1A characterized in that the oily composition comprises at least thymol; Carvacrol. eugenol, the ratio of the first two to the last being d 1: 3.  Use of the microparticle as defined in claims 1 to 15, characterized in that it has application in the food, pharmaceutical and cosmetic industry with antioxidant action for stocking products;  Use of the microparticle as defined in claims 1 to 15, characterized in that it has application as a feed additive to obtain antioxidant action in the meat of the fed animal. 18. Use of the microparticle as defined in Claims 1 to 17, characterized in that it has application in meat and meat products to prevent oxidation of meat.  Food composition comprising meat products and microparticles as defined in claims 1 to 18.  Food composition according to claims 7 and 19, characterized in that a. The microparticle is comprised in the range of 350 pp to? 00ppm, preferably 700ppm.  Food composition according to Claims 7 and 17, characterized in that the microparticle is in the range of. 350 ppm at 3500ppm, preferably 3500ppm.