(PDF) Extraction and encapuslation of bioactive compounds

APTEFF, 48, 1-323 (2017) UDC: 633.43:547.562+547.979.8 https://doi.org/10.2298/APT1748261S BIBLID: 1450-7188 (2017) 48, 261-273

Original scientific paper

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EXTRACTION AND ENCAPUSLATION OF BIOACTIVE COMPOUNDS FROM CARROTS

Vanja N. Šeregelj*, Gordana S. Ćetković, Jasna M. Čanadanović-Brunet,

Vesna T. Tumbas Šaponjac, Jelena J. Vulić, Slađana S. Stajčić

University of Novi Sad, Faculty of Technology Novi Sad, Bulevar Cara Lazara 1, 21000 Novi Sad

Carrot is an important root vegetable rich in bioactive compounds like carotenoids and polyphenols with appreciable levels of several other functional components having significant health-promoting properties. Utilization of carotenoids is limited due to their instability. Encapsulation is one of the alternatives used to improve carotenoid stability. The objectives of this study were to optimize the extraction and encapsulation of carote-noids from carrot. Freeze-dried carrots were extracted by conventional solvent extrac-tion (CSE) using four solvents, i.e. ethanol, acetone, ethyl acetate, and hexane. Although CSE using hexane and ethyl acetate resulted in highest carotenoid contents (18.27 and 15.73 mg β-carotene/100 g), acetone and ethanol at a solid to solvent ratio 1:10 w/v with slightly lower carotenoid contents (14.52 and 11.45 mg β-carotene/100 g) were chosen for further studies due to their higher food compatibility and higher polyphenol content (88.86 and 66.21 mg GAE/100 g) than with lower solid to solvent ratio (1:5 w/v). Ethanol and acetone carrot extracts were encapsulated using different carriers by a freeze-drying method in order to obtain the optimum encapsulate with the highest carotenoid encapsu-lation efficiency (EE). Encapsulation using the maltodextrin, whey and soy protein as a wall material yielded an EE of carotenoids ranging from 41.95% to 100%. The en-capsulated β-carotene content was evaluated during two months of storage at ambient temperature under light and dark conditions. Generally, the retention of carotenoids was significantly higher in dark conditions, where maximum retention (65.94-87.32%) occur-red in the samples encapsulated in maltodextrin and soy protein. KEY WORDS: bioactive compounds, carotenoids, extraction, encapsulation, storage

INTRODUCTION In the recent years, the worldwide demand for natural pigments has been increased due to the increased awareness of their health benefits and nutritional properties, and also because of the recognized toxicity of synthetic pigments (1). In view of their provitamin A activity and antioxidant function, carotenoids are one of the most important natural * Corresponding author: Vanja N. Šeregelj, University of Novi Sad, Faculty of Technology Novi Sad, Bulevar

Cara Lazara 1, 21000 Novi Sad, Serbia, e-mail: [email protected]



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food colorants. Epidemiological studies demonstrated that the consumption of diets rich in carotenoids is associated with lower incidence of cancer, cardiovascular diseases, age related macular degeneration and cataract formation (2,3). Carotenoids are red, yellow and orange pigments, which are widely distributed in nature and are especially abundant in yellow-orange fruits and vegetables and dark green leafy vegetables (4). Among these, carrots and carrot-based products are the main source of carotenoids in the human diet. According to Chen et al. (5) β-carotene constitutes a lar-ge portion (60-80%) of the carotenoids in carrots followed by α-carotene (10-40%) and lutein (1-5%). In addition to carotenoids, carrot also contains the appreciable amount of phenolics which have been linked to reducing the risk of major chronic diseases (6). The-refore, studies on the most convenient and effective extraction methods of these bioactive compounds have been attracting the interest of many researchers. There are numerous methods used for extraction of bioactive compounds and some examples include conventional solvent extraction (CSE). However, the extraction effici-ency varies widely due to different polarities of targeted bioactive compounds and sol-vents employed in the extracting process. For example, phenolic compounds are extrac-ted more effectively with highly-polar solvents, while non-polar or low-polar solvents showed higher extraction yields of lipophilic compounds like carotenoids (7,8). Pinelo et al. (9) suggest that the extraction conditions such as solvent concentration, contact time, extraction temperature, solvent to solid ratio and particle size are also important parame-ters affecting the extraction yield of bioactive compounds content and antioxidant ca-pacity of the extracts. Nowadays, the nutritional content of food after preparation and during storage has become an important concern for consumers and manufacturers who have contributed to better food quality and, consequently, to consumers' health. Incorporation of β-carotene in various food systems is limited by its poor water solubility and instability in the pre-sence of light, heat, and oxygen (10). Encapsulation is one of the alternative techniques used for improving carotenoid stability, which entraps a sensitive ingredient inside a coa-ting material (11). The initial step in encapsulating is the selection of a suitable encap-sulation technique and coating material, especially considering their incorporation in food products. Among several methods of encapsulation, freeze drying, also known as lyo-philization, is one of the most useful processes for drying thermosensitive substances such as natural pigments and aromas (12). Different wall materials for coating can be used and a maltodextrin, soy and whey protein are often used. The aim of the present study was to elucidate the effect of different extraction parameters to obtain carrot extracts with high carotenoid and phenolic content, as well as high antioxidant capacity. To protect extracted sensitive bioactive compounds, the obtai-ned and selected extracts were encapsulated by freeze drying method. For this purpose, the effect of different encapsulation agents to obtain the powder with high carotenoid retention was evaluated. The influence of storage and light exposure on carotenoid con-tents in the encapsulates was also examined.



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EXPERIMENTAL

Chemicals and instruments The solvents used for extraction (ethanol, acetone, hexane and ethyl acetate) were purchased from “Zorka” Chemical Co. (Šabac, Serbia). Folin-Ciocalteu reagent, 2,2-di-phenyl-1-picrylhydrazine (DPPH), gallic acid, trichloroacetic acid and Trolox were from Sigma Chemical Co. (St Louis, MO, USA). Maltodextrin was provided by Battery Nu-trition Limited (London, United Kingdom), while soy and whey protein isolate were pur-chased from Olimp Laboratories (Debica, Poland). All other chemicals and solvents used were of the highest commercial grade. Absorbances in spectrophotometrical assays were measured on a MultiskanGO microplate reader (Thermo Fisher Scientific Inc., Waltham, MA, USA) and UV-1800 spectrophotometer (Shimadzu, Kyoto, Japan).

Plant material Fresh carrots were produced by Beoflora d.o.o., Beograd, Serbia, and purchased from the local store. After washing, fresh carrots were chopped in a kitchen blender (B 800 E, Gorenje, Slovenia), dried at -40 °C for 48 h in a freeze-drier (Martin Crist Alpha 2-4, Osterode, Germany), finely ground, and stored at -20 °C until use.

Optimization of the extraction of bioactive compounds from carrots

The extraction of bioactive compounds from carrots implied optimization of the pro-cess considering different polarity of extracting solvents and solid to solvent ratio. The first set of experiments included conventional solvent extraction (CSE) with polar (etha-nol, acetone and ethyl acetate) and non-polar (hexane) solvents at solid to solvent ratio of 1:10 (w/v). The second set of experiments included solvent extraction with ethanol and acetone at a ratio of 1:5 (w/v).

Conventional solvent extraction Freeze-dried carrots were extracted with extracting solvent in two different solid to solvent ratios (1:10 and 1:5 w/v) for 10 minutes, three times with the same volume of solvents. The extraction was performed using a laboratory shaker (Unimax 1010, Hei-dolph Instruments GmbH, Kelheim, Germany) at 300 rpm, under light protection, at room temperature. The obtained three extracts were filtered (Whatman paper No.1), combined, and stored in dark bottles at -20 °C to further analysis.

Determination of bioactive compounds content in carrot extracts

Total carotenoid content (TCar). The content of carotenoids in the carrot extracts was analyzed spectrophotometrically by the method of Nagata and Yamashita (13), using extracting solvent as the blank. The content of TCar was calculated using the equation:



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TCar (mg β-carotene/100ml) = 0.216A663 - 1.22A645 - 0.304A505 + 0.452A453 where A663, A645, A505 and A452 represent the absorbances measured at 663 nm, 645 nm, 505 nm and 453 nm, respectively. The TCar was expressed as mg of β-carotene equiva-lents per 100 g DW. Total phenolics content (TPh). The amount of total phenolics in carrot extracts was determined spectrophotometrically according to the Folin-Ciocalteau method adapted for 96 well microplate (14). The results were expressed as gallic acid equivalents (GAE) per 100 g DW.

Determination of carrot extracts bioactivity

Antiradical activity by DPPH assay (AADPPH). The free radical scavenging effect of carrot extracts on 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical was measured spectro-photometrically based on the modified method by Jiménez-Escrig et al. (15). The ability to scavenge DPPH radicals, i.e. AADPPH, was calculated following the equation:

AADPPH (%) = [(AC − AS) / AC] × 100

where AC is the absorbance of the control, and AS is the absorbance in the presence of the extract. The results were also expressed as µmol Trolox equivalents (TE) per 100 g DW. Reducing power assay (RP). The RP was determined by the method adopted from Oyaizu (16), measuring the reduction of the Fe3+/ferricyanide complexes to the ferrous (Fe2+) form. The absorbances were read at 700 nm against the control. The calibration curve was constructed using Trolox, and the results were expressed as µmol Trolox equi-valents (TE) per 100 g DW. Encapsulation optimization. Three carrier agents, including maltodextrin (MD), soy protein (SP) and whey protein (WP) isolate were used as wall materials for carotenoid encapsulation. Two criteria were considered for optimization of encapsulate formulation: the amount of core material contained in the powders and the retention of core material inside powders affected by different light exposure conditions during storage. Emulsion preparation and freeze-drying. All the powders were prepared according to the method developed by Indrawati et al. (17), with some modifications. MD and WP (7g) were dissolved in 10.5 ml water at 60 °C and kept under stirring until the tempera-ture reached 30 °C, while SP was dissolved in the same way in 40 ml of water. Tween 80 (0.1 g) was added as an emulsion stabilizer. Separately, 10 ml of each extract was com-bined with sunflower oil (1.5 ml), concentrated under reduced pressure on a rotary eva-porator set at 40 °C to remove the organic solvent, and immediately mixed with pre-viously prepared carrier solution. The solutions were vigorously homogenized at 11,000 rpm for 3 min at room temperature, frozen overnight, and subjected to freeze-drying at -40 °C for 48 h. The encapsulated powders were packed in airtight plastic bags and stored at 4 °C until further use. Contents of Surface Carptenoid (SC) and Total Carotenoid (TC) in the encapsulate and Encapsulation Efficiency (EE). SC, TC, and EE were determined by following a modified Barbosa et al. method (11). SCs were determined by direct extrac-



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tion of 0.25 g encapsulate sample with 5 ml acetone on a vortex for 20 s, followed by centrifugation at 5000 rpm (10 min) and supernatant separation. For TC determination, 0.25 g of sample was vortexed with 0.2M PBS (pH 7) for 1 min to break the capsules, extracted with 2 ml of acetone and 3 ml of diethyl ether, and left over night at 4 °C. After separating the pigment layer, extraction was repeated with the same solvent volumes to collect total pigment. The carotenoid quantification was carried out according to the pre-viously described protocols. The encapsulation efficiency was calculated by using the equation:

% EE = [(TC-SC)/TC] x 100 Simultaneously, the control samples, i.e. the carriers without extracts, were prepared in the same way, for the correction of interfering substances originating from the carrier material.

Storage stability tests The control and pigment microcapsules were stored at room temperature in glass and amber bottles for 2 months to determine the effect of time and light exposure on the stability of carotenoids. For that purpose, total carotenoids were determined by described method every month.

RESULTS AND DISCUSSION Previous studies have indicated that carrot extracts represent a rich source of bioactive compounds (18, 19). According to the knowledge that their yield significantly depends on the different extraction conditions, solvent polarity and sample to solvent ratio were evaluated in order to increase the extraction efficiency and choose the most convenient method for further use in food formulations. The effect of different extraction conditions on the total carotenoid and phenolic yields in carrot extracts are shown in Table 1. When using the solvent to solid ratio 1:10, the carotenoid extraction yields obtained under studied conditions were in the range from 0.19 to 18.27 mg β-carotene/100g DW. As expected, the highest content of carotenoids was obtained using hexane as an extrac-ting solvent, according to the empirical rule "like dissolves like". The carotenoid yield in ethyl acetate extract was close to the one obtained with acetone (15.73 and 14.52 mg β-carotene/100 g DW, respectively). Yen et al. (20) reported 24 mg β-carotene/100g DW in freeze-dried carrots successively extracted with acetone and petroleum ether. Using con-ventional solvent extraction employing diacetone alcohol and petroleum ether, and furt-her purification with diacetone alcohol, methanolic KOH and distilled water, Dutta et al. (21) determined lower content of carotenoids (8.4 mg β-carotene/100g DW) compared to the results obtained for conventional solvent extraction in this study. However, the yields of phenolics were significantly different, depending on the sol-vent used for extraction. These varied between 34.85 and 165.52 mg GAE/100 g DW. The results showed that ethyl acetate was superior in isolating phenolic compounds com-



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pared to the other three solvents in the CSE method. This is probably due to the higher polarity and better solubility of phenolic components present in plant material (22). Bys-tricka et al. (23) reported 6-fold lower phenolics content for carrot ethanolic extract, while Cefola et al. (24) reported five times less phenolics in carrot acetone extract (11.37 mg GAE/100g DW and 17.06 mg GAE/100g DW, respectively) compared to the contents in carrot extracts obtained with the same solvents in this study. The selection of a solvent is based on several physicochemical properties along with the cost cost and toxicity. Some solvents, such as ethanol and acetone are designated as "generally recognized and safe" (GRAS) and are the preferred solvents currently used for food products. Since our research focuses on giving priority to the food grade solvents, in order to determine the optimal ratio of plant material/solvent, the extraction of carrots with ethanol and acetone were repeated. From the results presented in Table 1 it follows that using the both ratios (1:10 w/v and 1:5 w/v, respectively) gives approximately equal carotenoid content, with no statistically significant difference (p ≤0.05) in case of etha-nol. The use of ethanol in a ratio of 1:10 w/v, resulted in 2.5% more carotenoids com-pared to the other ratio studied (1:5 w/v), while for acetone extraction the content was by 14.3% lower. In the case of phenolic compounds, a significant increase of the isolation efficiency with higher volumes of extracting solvents was observed. The total phenolic content was by 123.5% higher for ethanolic extract and 154.8% for acetone extract com-pared to the extracts obtained with lower volumes of the same extracting solvents. How-ever, for the encapsulation and storage studies, ethanol and acetone extracts obtained at a 5:1 ratio extraction were chosen because of higher contents of carotenids and the poten-tial of these encapsulates for further use as colorants.

Table 1. Total carotenoid and phenolic contents in carrot extracts

Values with the same letter in the superscript in the same column are not significantly different (p<0.05). TCar - total carotenoid content; TPh - total polyphenol content.

Since carotenoids, along with polyphenols, are the most important phytochemicals in carrots, they are considered as responsible for the antioxidant activity. For the antioxidant activity assessment of carrot extracts, the DPPH free radical scavenging activity as well as reducing power, were monitored spectrophotometrically. The relationship between the carrot extracts concentration and discoloration of DPPH radical solution, reflecting the radical scavenging activity of the analyzed extracts, is shown in Figure 1. The magnitude of the radical scavenging effect of the carrot extracts was ranked as ethanol > acetone > ethyl acetate > hexane. The polarity of the solvents used for the extraction of carrot was different, yielding different composition of the ex-

Extraction method Extraction

solvent TCar

(mgβ-carotene/100g DW) TPh

(mgGAE/100 g DW)

Conventional solvent extraction (10:1 v/w)

Ethanol 11.45±0.52a 66.21±0.15a Acetone 14.52±0.67b 88.86±0.22b

Ethyl acetate 15.73±0.65b,d 165.52±0.12c Hexane 18.27±0.73c 34.85±0.09d

Conventional solvent extraction (5:1 v/w)

Ethanol 11.17±0.51a 29.62±0.02e Acetone 16.60±0.72d 34.85±0.15d



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tracts. The ethanol, acetone and ethyl acetate extracts had the highest total bioactive com-pounds content (i.e. carotenoids and polyphenols), which resulted in the superior DPPH radical scavenging activity in the range from 120.07 to 195.01 µmol TE/100 g DW. On the other hand, the hexane extract did not show any activity towards DPPH radical. Be-sides various antioxidant mechanisms involved in radical scavenging activity, a synergis-tic activity among present bioactive molecules may occur. Cefola et al. (22) investigated the antioxidant activity of various carrot varieties (yellow, orange and purple) on DPPH radicals. The obtained results indicated the highest antioxidant activity of purple carrots, due to the highest content of bioactive phytochemicals, where the total carotenoid and phenolic contents were four times higher than with the other two varieties. The antioxi-dant activity of yellow and orange varieties was similar. The reducing power assay may serve as a significant indicator of potential antioxidant activity, which is evidenced by measuring the ability of the extracts to reduce the Fe3+/ferricyanide complex to the ferrous form. Figure 1b shows the reducing power of the five carrot extracts, where the results of the ethanol and acetone extracts indicate the highest reducing ability (7368.07 and 3167.91 µmol TE/100 g DW, respectively), and ethyl acetate extract the lowest (71.19 µmol TE/100 g DW).

Figure 1. Antiradical activity (a) and reducing power (b) of different carrot extracts

The low bioavailability of carotenoids from natural sources and limited incorporation of β-carotene in food products due to its poor water solubility and instability in the presence of heat, light, and oxygen, has led to the development of encapsulation methods, to improve stability and achieve maximum health benefits. The EE value is considered as an important parameter for evaluating the properties of microcapsules, and it presents the ratio of the pigment bound inside the microcapsules to the total carotenoid content in the microcapsule, comprising the bound and SC. The SC, TE, and EE of carotenoids for all microcapsules are shown in Table 2. The EE value varied considerably with the wall materials and composition of encap-sulating materials. In all acetone extracts. microcapsules, surface and total carotenoid contents were higher than in the ethanol extract microcapsules, indicating better pigment isolation with acetone.



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Table 2. Surface, total carotenoids, and encapsulation efficiency of carotenoids for ethanol and acetone extracts encapsulated in maltodextrin, whey, and soy protein

Sample SC

(mg β-carotene/100g DW) TC

(mg β-carotene/100g DW) EE (%)

ME 0.23 ± 0.02a 0.49 ± 0.04a 53.80 ± 0.29a MA 0.67 ± 0.00b 1.14 ± 0.06b 41.95 ± 2.56a WPE 0.13 ± 0.04a,c 0.64 ± 0.05a 84.87 ± 3.92b WPA 0.16 ± 0.01a,c 1.21 ± 0.06b 85.89 ± 0.88b SPE 0.00 ± 0.00c 0.46 ± 0.06a 100 ± 0.00b SPA 0.51 ± 0.02b 1.14 ± 0.06b 55.36 ± 0.43a

Values with the same letter in superscript in the same column are not significantly different (p<0.05). SC, TC - surface and total carotenoids in microcapsules; EE - encapsulation efficiency; ME, MA - etha-

nol and acetone extract encapsulated in maltodextrin, respectively, WPE, WPA - ethanol and acetone ex-tract encapsulated in whey protein, respectively; SPE, SPA - ethanol and acetone extract encapsulated in soy protein.

The encapsulation efficiency varied from 41.95% up to 100%. The difference bet-ween EE of acetone and ethanol extracts encapsulated in the same carrier was not significantly different, except for the case of soy protein. The maximum efficiency of 100% was obtained for the ethanol carrot extract with soy protein, whereas in the case of acetone extract this value was 55.36%. Since the initial concentration of carotenoids in the ethanol extract was lower, the total amount was successfully encapsulated with soy protein. Generally, both proteins (soy and whey), were found to be better carriers for carotenoids. This is consistent with the reports of Grimme and Brown (25), showing that carotenoids in plants are bound by protein. According to Jafari et al. (26), functional properties of proteins including solubility, film formation, emulsification and stabilization of emulsion droplets, exhibited many de-sirable characteristics for the wall material. These proteins changed their structure during emulsification through unfolding and adsorption at the oil-water interface, and sub-sequently formed a resistant multilayer around oil droplets. That would stabilize the oil-in-water emulsion and improve lipid microencapsulation. In addition, the maltodextrin showed the lowest encapsulation efficiency. This was expected given the fact that powder had the highest amount of unencapsulated β-carotene, which negatively affects its EE. It is well known that carotenoids are unstable against light, oxygen, moisture, and temperature. The degradation rate of carotenoids in microcapsules, under light and dark conditions at room temperature, were monitored by the change in carotenoid retention upon storage for 60 days, as shown in Table 3. As expected, an increase in the storage period in the presence of light resulted in progressive loss of β-carotene in all microcap-sules, confirming instability of carotenoids. For instance, after two months exposed to light, the carotenoids completely vanished from the acetone extract encapsulated in mal-todextrin and from the ethanol extract encapsulated in whey protein. In most of the mic-rocapsules, the decline in the carotenoid content between first and second month was significant (p<0.05). On the other hand, the degradation of encapsulated carotenoids un-der dark conditions was much slower compared to that in the opposite conditions. More-



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over, there was no significant difference (p<0.05) in the carotenoid content after the first and second moth of storage in amber bottles, except in the case of the acetone extract encapsulated in soy protein.

Table 3. Changes in carotenoid content during storage of the microencapsulates

Values with the same letter in superscript in the same column are not significantly different (p<0.05). ME, MA - ethanol and acetone extract encapsulated in maltodextrin, respectively, WPE, WPA - ethanol

and acetone extract encapsulated in whey protein, respectively; SPE, SPA - ethanol and acetone extract encapsulated in soy protein.

n.d. - not detected

Figure 2. Carotenoid retention in the encapsulate samples after one and two months of storage exposed to light (a), and in the dark (b)

Time (months) / Light exposure

Encapsulate sample ME MA WPE WPA SPE SPA

0 0.47±0.04a 1.09±0.10a 0.59±0.09a 1.15±0.11a 0.43±0.06a 1.10±0.09a

1 Light 0.21±0.04b 0.46±0.07b 0.07±0.00b 0.48±0.07b 0.09±0.01b,d 0.53±0.02b Dark 0.41±0.04a 0.86±0.09a,c 0.18±0.01c 0.49±0.07b 0.23±0.03c 0.77±0.03c

2 Light 0.03±0.00c n.d. n.d. 0.03±0.00c 0.04±0.00b 0.29±0.01d Dark 0.37±0.04a 0.82±0.05c 0.17±0.00c,d 0.24±0.02b 0.18±0.00c,d 0.49±0.05b



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The retention of carotenoids, presented in Figure 2, decreased more rapidly during the first 30 days due to exposure to the light for the MD, WP and SP microcapsules with ethanolic extract (to 44.42%, 11.58% and 17.96%, respectively), as well as for the micro-capsules obtained with acetone extract (to 39.94%, 36.38% and 46.57%, respectively).

CONCLUSION In the present study, the elaboration of the effect of different extraction parameters revealed that they have a significant effect on the extraction yield. Pure ethanol and ace-tone are considered as safe solvents, and the high concentration of bioactive compounds from carrots could be isolated with these solvents. Further, the antioxidant potential eva-luation showed that both ethanol and acetone extracts were promising. The major prob-lem of these ingredients is their instability in the presence of heat, light, and oxygen. Hence, the carrot extracts were mixed with three different wall materials (MD, WP and SP) for the evaluation of the encapsulation efficiency and storage stability. All wall ma-terials largely increased the half-life of the encapsulated pigments during storage in dark conditions compared with the samples exposed to light. Generally, the proteins showed the highest encapsulation efficiency, lower degradation rate in both studied conditions, and were defined as the most effective wall material in stabilizing the pigments. Hence, the significance of this research is in demonstrating successful encapsulation and exten-sion of the shelf life of carotenoid content, which can be very important for obtaining attractive and acceptable final products.

Acknowledgements

This research is part of the Project TR 31044 which is financially supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia and by COST Action Eurocaroten CA15136.

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ЕКСТРАКЦИЈА И ИНКАПСУЛАЦИЈА БИОАКТИВНИХ КОМПОНЕНТИ ИЗ ШАРГАРЕПЕ

Вања Н. Шерегељ*, Гордана С. Ћетковић, Јасна М. Чанадановић-Брунет,

Весна Т. Тумбас Шапоњац, Јелена Ј. Вулић, Слађана С. Стајчић

Универзитет у Новом Саду, Технолошки факултет Нови Сад, Булевар Цара Лазара 1, 21000 Нови Сад, Србија

Шаргарепа је коренасто поврће изузетно богато биоактивним једињењима, као што су каротеноиди, полифеноли и друге функционалне компоненте које допри-носе здрављу. Употреба каротеноида је ограничена због њихове нестабилности. Инкапсулација је једна од алтернативних метода која се користи за побољшање њи-хове стабилности. Циљ овог истраживања била је оптимизација екстракције и ин-капсулације каротеноида из шаргарепе. Лиофилизирана шаргарепа екстрахована је конвенционалном методом екстракције (CSE) користећи раствараче различите по-ларности (етанол, ацетон, етил ацетат и хексан). Иако је CSE са хексаном и етил ацетатом резултирала највећим садржајем каротеноида (18,27 и 15,73 mg β-каро-тена/100 g), ацетонски и етанолни екстракти (1:10 m/V) са нешто нижим садржајем каротеноида (14,52 и 11,45 mg β-каротена/100 g), изабрани су за даља истраживања због здравствене прихватљивости и већег садржаја полифенола (88,86 и 66,21 mg β-каротена/100 g). Етанолни и ацетонски екстракти шаргарепе инкапсулирани су ме-тодом лиофилизације на различите носаче, у циљу добијања оптималног инкапсу-лата са највећом ефикасношћу инкапсулације каротеноида. Ефикасност инкапсула-



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ције етанолног и ацетонског екстракта, примењујући малтодекстрин, протеине су-рутке и соје као носаче, била је у распону од 41,95% до 100%. Садржај β-каротена у инкапсулатима тестиран је током 2 месеца складиштења у светлим и тамним усло-вима. Генерално, ретенција каротеноида је значајно већа у мрачним условима скла-диштења, при чему је максимална ретенција (65,94-87,32%) утврђена у узорцима са малтодекстрином и протеинима соје. Кључне речи: биоактивна једињења, каротеноиди, екстракција, инкапсулација,

складиштење

Received: 15 September 2017. Accepted: 01 November 2017.

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(PDF) Extraction and encapuslation of bioactive compounds