- Email: [email protected]
Advantages of microfiltration processing of goat whey orange juice beverage
Journal Pre-proofs Advantages of microfiltration processing of goat whey orange juice beverage Alexandre H. Vieira, Celso F. Balthazar, Jonas T. Guimaraes, Ramon S. Rocha, Mônica M. Pagani, Erick A. Esmerino, Márcia C. Silva, Renata S.L. Raices, Renata V. Tonon, Lourdes M.C. Cabral, Eduardo H.M. Walter, Mônica Q. Freitas, Adriano G. Cruz PII: DOI: Reference:
S0963-9969(20)30085-5 https://doi.org/10.1016/j.foodres.2020.109060 FRIN 109060
To appear in:
Food Research International
Received Date: Revised Date: Accepted Date:
24 November 2019 1 February 2020 2 February 2020
Please cite this article as: Vieira, A.H., Balthazar, C.F., Guimaraes, J.T., Rocha, R.S., Pagani, M.M., Esmerino, E.A., Silva, M.C., Raices, R.S.L., Tonon, R.V., Cabral, L.M.C., Walter, E.H.M., Freitas, M.Q., Cruz, A.G., Advantages of microfiltration processing of goat whey orange juice beverage, Food Research International (2020), doi: https://doi.org/10.1016/j.foodres.2020.109060
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Advantages of microfiltration processing of goat whey orange juice beverage
Alexandre H. Vieira¹, Celso F. Balthazar1, ,Jonas T. Guimaraes¹, Ramon S. Rocha1,2 , Mônica M. Pagani³, Erick A. Esmerino1, Márcia C. Silva2, Renata S. L. Raices2, Renata V. Tonon4, Lourdes M.C. Cabral4, Eduardo H.M. Walter4 Mônica Q. Freitas¹, Adriano G. Cruz2
1Universidade 2Instituto
Federal Fluminense (UFF), Faculdade de Veterinária, 24230-340 Niterói, Brasil
Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ), Departamento de
Alimentos, 20270-021 Rio de Janeiro, Brasil 3Universidade
Federal Rural do Rio de Janeiro (UFRRJ), Instituto de Tecnologia (IT), 23890-000,
Brasil. 4Embrapa
Agroindústria de Alimentos (CTAA), 23020-470, Guaratiba, Rio de Janeiro. Brasil
*Email: [email protected] (A.G. Cruz) Goat whey Orange Juice Beverage
Abstract The objective of this study was to evaluate the microbiological, physicochemical and functional quality of an innovative goat whey orange juice beverage (GOB) processed by microfiltration. The microfiltration (0.2 µm) of the GOBs had a variation on the feed temperature (20, 30, 40, 50°C) and were compared to the conventional heat treatment LTLT (63°C/30 min). Microbiological (aerobic mesophilic bacteria, mold and yeast and lactic bacteria), physicochemical (pH, color, rheology and volatile compounds) bioactive compounds (acid ascorbic, total phenolics) and functional activity (DPPH, ACE, α-amilase and α-glucosidase) analysis were performed. The GOB processed by microfiltration using at least 30°C presented adequate microbial counts (less than 4, 3 and 4 log CFU/mL, for AMB, molds and yeasts and LAB, respectively). In general, the pH, color parameters, volatile and bioactive compounds were not influenced by microfiltration temperature, but presented a difference from the LTLT processing. The rheological parameters were influenced by MF temperature and the utilization of temperatures of 20° and 30°C maintained the consistency similar to the LTLT sample, preserving the compounds responsible for the texture. Therefore, it is suggested a processing of GOB by microfiltration using mild temperatures (between 30° and 40° C) to preserve consistency and also obtain a desirable microbial quality, beyond the preservation of many functional properties and volatile compounds. Keywords: Membrane technology; Functional activity; Dairy product; Microbiology; Rheology.
1. Introduction Nowadays, the modern food processing technologies aims the food safety in production but also the high nutritional quality of the product. In this context, the non-thermal technologies are known as cleaner processes than conventional thermal treatments, energetically efficient, environmental friendly and focus on quality development, maintaining a good acceptability without worsening the product’s safety (Ahmad et al., 2019; Balthazar et al., 2019a). One of the emerging technologies used for milk and dairy products is microfiltration, which is a membrane technology driven by pressure and has been applied to separate compounds of high added value, offering several technological advantages (Castro-Muñoz & Fíla, 2018). The Microfiltration processing presents low energy consumption; high separation efficiency; easy implementation and operation; high productivity; absence of phase transition and non-use of additional solvents, which favor the solute recovery. Therefore, different types of high added value compounds can be separated with membrane technologies, like carbohydrates, pectin, sugars, antioxidant compounds, anthocyanins, proteins and phenolic compounds (Nazir et al., 2019). The membrane processing offer several advantages over the conventional separation methods, like high hydrostatic pressure and high-pressure homogenization, including the mild operational conditions of temperature and pressure, which preserve the functional properties of food products, besides being a cleaner processing due to the absence of chemicals utilization (Soodam. K. & Guinee, 2018). In addition, microfiltration has been used in the dairy industry for several purposes, including the bacterial reduction to produce Extended Shelf Life (ESL) milk (Alegbeleye, Guimarães, Cruz, & Sant’Ana, 2018). Milk and dairy products are known to be healthy since it contains many essential nutrients, like oleic acid, conjugated linoleic acid, Ω-fatty acids, high biological value proteins, vitamins, minerals and bioactive compounds (Balthazar et al., 2017a). Among the consumption of milk of various species, currently, the goat milk consumption has been increasing, because its usually associated to different functional effects, such as health maintenance, risk of chronic diseases reduction, besides other beneficial physiological effects (Clarck & Garcia, 2017). Furthermore, this
dairy matrix is important in nutrition of children, elder and people allergic to bovine milk protein, since goat milk presents proteins of high digestibility and hypoallergenicity (Balthazar et al., 2017a; Verruck et al., 2019). For these reasons, the goat milk can be considered an excellent bovine milk substitute in human nutrition, being also used for manufacturing of functional dairy products (Ranadheera et al., 2019). Among the current chronic diseases affecting the world population, diabetes mellitus has risen dramatically and it is expected to affect 438 million people until 2030, with 70% of the cases occurring in poor countries. The concomitant inhibition of α-amylase and α-glucosidase activities is seen as an effective strategy for the control of this chronic disease (Carrizzo et al., 2018; Saeedi, Hadjiakhondi, Nabavi, & Manayi, 2017). In addition, milk proteins are considered a good source of bioactive compounds with antihypertensive activity (angiotensin-converting-enzyme inhibition), which may be beneficial for cardiovascular health (Marcone, Belton & Fitzgerald, 2017). In recent years, the utilization of whey in foods, such as dairy, bakery, confectionary, meat and canned products (Jeewanthi et al., 2015), edible films and coatings (Chakravartula et al., 2019) and non-food products such as bioplastics, biofertilizers, biofuels (Ahmad et al., 2019) is increasing all around the world. The whey protein has high biological value; however, too much whey is still discarded during manufacturing of dairy products (Ahmad et al., 2019). In food industries, the whey proteins are used as emulsifying, gelling and bulking agent. The antioxidant activity of the whey proteins is well-stablished and it may efficiently inhibit the lipid oxidation (Wen-Qiong et al., 2019). One of the most important aspects in introducing a new goat dairy food into market is to pay attention the public demand and to make specific market surveys to identify those demands in order to reach the new opportunities (Freire et al., 2017). Therefore, the objective of this study was to elaborate an innovative goat whey orange juice beverage processed by microfiltration membrane technology and study the effect of microfiltration feeding temperatures on the microbiological, physicochemical and bioactive compounds and functional characteristics.
2. Material and methods
2.1 Goat whey orange juice beverage (GOB) processing The whey beverages with orange juice (10 L) were manufactured with the following ingredients: 45 % (w/w) of skim goat whey (0.1% fat), 45% (w/w) of fresh orange juice and 10% (w/w) of sugar. This basic formulation was used in five different treatments: Low temperature long time treatment (LTLT - 63°C/30 min) as a control beverage due to conventional thermal treatment applied to beverages; and microfiltration (MF) varying the system feeding temperature: T20 (at 20°C); T30 (at 30°C); T40 (at 40°C) and T50 (at 50ºC). After the processing, the beverages were packaged in sterilized plastic bottles (300 mL) and kept under cold storage (4ºC ± 1) during 15 days, which is the typical shelf-life period for pasteurized dairy products (Prashanth, Jayaprakash, Soumyashree, & Madhusudhan, 2018). The microfiltration was realized in a new ceramic membrane (α-alumina) with 0.2 µm pore (Membralox T1-70 modules, channel diameter 7 mm, length 250 mm, surface area filtration of 0.005 m² (0.054 ft2), housing material 316L SS) and working pressure of 2 kgf/cm². The LTLT and microfiltration treatments were realized in pilot equipment at EMBRAPACTAA laboratory (Guaratiba/RJ, Brazil). The whole experiment was performed in triplicate. 2.2 Microbiological analysis The goat whey orange juice beverages were submitted to the microbiological analysis of Aerobic Mesophilic Bacteria (AMB), Molds and yeasts and Lactic Acid Bacteria (LAB), according to the methodology recommended by “American Public Health Association” (APHA, 2015). The analysis were carried out right after processing (D1), seven days (D7) and 15 days (D15) after cold storage (4 ± 1°C). All the analysis were realized in triplicate. 2.3 pH values and color parameters The pH analysis and instrumental color were carried out right after processing. All the analysis were realized in triplicate. The pH of the samples were measured using a pHmeter (Digimed, model DM-20, São Paulo, Brasil), at 25ºC, inserting the electrode directly into the sample. The samples color were measured using a colorimeter (Colour Quest XE Hunter Lab, Northants, UK) based on CIELAB system (L*, a*, b*) according to the procedure stablished by Balthazar et al. (2015). The chroma (C*, Eq.1), the yellowness index (YI, Eq.2) and the total color
difference (ΔE*, Eq.3) were calculated according to Pathare et al. (2013), through the following formulas:
𝑪 ∗ = (𝒂 ∗ )𝟐 + (𝒃 ∗ )𝟐 𝒀𝑰 =
(Eq.1)
𝟏𝟒𝟐.𝟔 𝒃 ∗
(Eq.2)
𝑳∗
𝟐
𝟐
∆𝑬 ∗ = (∆𝑳 ∗ ) + (∆𝒂 ∗ ) + (∆𝒃 ∗ )
𝟐
(Eq.3)
2.4 Bioactive compounds and functional activity The bioactive compounds evaluated in this study were: total phenolic compounds according to Capatto et al. (2018) and ascorbic acid according to Balthazar et al. (2019b). The measured functional activities were: antioxidant (2,2-diphenyl-1-picrylhydrazyl – DPPH) and antihypertensive (inhibition of angiotensin-converting enzyme – ACE) activities according to Capatto et al. (2018) and antidiabetic activity (α-amilase e α-glucosidase) as described by Balthazar et al. (2019b). The analysis were carried out in samples right after processing and in triplicate. Specifically, the extracts for total phenolic compounds and DPPH analyses were obtained in triplicate, in which approximately 1 g of GOB sample was weighed into beakers (200 mL), and 30 mL mix of water and ethanol (50:50 v/v) was added and stirred at 200 rpm on an orbital table (SL180/D, Solab, Piracicaba, SP, Brazil) for 1 h. The resulting extracts were filtered applying vacuum. The absorbance of total phenolic compounds was measured at 725 nm in a spectrophotometer (Biospectro, SP-220), and the results were obtained using a calibration curve. The results were expressed as gallic acid equivalents per liter of sample (EAG g/L). It was used 1 mL of Folin reagent diluted in distilled water (1:10), which was added to 1 mL of extract and agitated for 1 min. For antioxidant capacity, 2850 μL of a methanolic solution of the DPPH radical (0.06 mM – 700 nm) was mixed to 150 μL of the extract and held in the dark for 60 min. The results of both analysis were expressed as μg Trolox Equivalent/g of sample.
ACE inhibitory activity were performed by filtration of the extracts, the angiotensin converting enzyme inhibitory (ACEI) was determined in a spectrophotometer and the ACE inhibitory activity was calculated as follows: (𝐵 𝐴)
𝐴𝐶𝐸 𝐼𝑛𝑖𝑏𝑖𝑡𝑜𝑟𝑦 𝑎𝑐𝑡𝑖𝑣𝑖𝑡𝑦 (%) = [(𝐵 𝐶) 100]
(Eq. 4)
where, A is the absorbance of ACEI component in the presence of ACE; B is the absorbance without the ACEI component and C is the absorbance without ACE. Ascorbic acid content was determined using the titration method with 2,6-dichlorophenolindophenol, for the extraction. A 5-mL sample was mixed in 25 mL of metaphosphoric acid solution and centrifuged at 13,715 × g for 10 min at room temperature. Then ascorbic acid was quantified using an aliquot of the supernatant (approximately 5 mL), which was diluted in 15 mL of extraction solution and titrated with the indophenol solution and the samples in the absence of light. The ascorbic acid content was expressed in milligrams per gram. The α-amylase and α-glucosidase enzyme inhibition activities were determined α-amylase by measuring absorbance (Abs) at 540 nm and releasing p-nitrophenol at 400 nm, respectively. Solutions without the water-soluble extract and substrate sample as a control and a blank, respectively were used. The inhibition percentage was calculated as follows:
(
Inhibition (%) = 1 ―
Abs sample ― Abs blank Abs control
) × 100
(Eq. 5)
2.5 Volatile compounds The volatile compounds analysis was performed as described by Balthazar et al. (2018). Briefly, the beverage samples were extracted by solid phase micro extraction (SPME) and analyzed by gas chromatography (GC MS, Varian 3800 GC, Milan Italy) coupled to mass spectrometry (MS ion trap VArian 2000, VArian Spa, Milan Italy). The SPME extractions were carried out using 50/30 mm thick DVB/CAR/PDMS (divinylbenzene/carboxen/polydimethylsiloxane) fibres (Supelco, Bellefonte, PA, USA) and 40 mL head space vials in an manual sampler holder (Supelco, Bellefonte, PA, USA). For that, 3 g sample was added with 3 mL saturated NaCl solution maintaining the vial at 40 Cin a dry block (Multi-Blok 2003 Thermo Fisher Scientific, Germany)with a 20 min equilibration time and 30 min extraction time. After extraction, the SPME fiber was manually introduced into the
GCMS for30 min under the following conditions for the thermal desorption of the analytes: injector temperature at 240 °C; splitless injection mode; column CP-Wax 52 CB 60 m, 0.25 mm id, 0.25 mm film thickness; oven temperature programmed from 45 °C for 5 min, then increased to 80 Cat a rate of 10 °C min-1, then to 240 °C at 5 °C min-1 and kept for 30 min; 1 mL min-1 of Helium gas flow, transfer line temperature at 240 °C, electron impact ionization energy at 70 eV, acquisition mass range of 50550 m/z. The volatile compounds were identified by comparison of their experimental spectra with those provided by the National Institute of Standards & Technology (NIST/EPA/NIH MassSpectra Library, NIST11, USA), using the linear retention indices (LRI).LRI values of C8-C40 alkanes standards (Supelco, 40,127-U) were injected under the same chromatographic and mass spectrometric conditions. The analysis was carried out in triplicate. 2.6. Rheology The rheological analysis was carried out as described by Balthazar et al. (2017b). To access the flow curve behavior, a rheometer (model 1500 AR, TA Instruments, New Castle, DE) equipped with cone & plate 140 geometry was used. The flow behavior curves and the mechanical spectrum of the samples were obtained at 25°C. The flow curves were determined using a shear rate varying from 0 to 300 s−1 and the data obtained were adjusted to the power law (Eq. 6):
𝜎 = 𝑘.𝛾𝑛
(Eq.6)
Where 𝜎 is the shear stress (Pa), 𝑘 is the consistency index (Pa sn), 𝛾 is the shear rate (s−1), and
𝑛 is the flow behavior index (dimensionless). The analysis were carried out in triplicate right
after processing. 2.7 Statistical analysis A completely randomized design of the experiment was carried out and conducted in triplicate. The data were analyzed using XLSTAT 2019.2 (Addinsoft, New York, NY, USA). One-way analysis of variance (ANOVA) was used to determine significant differences between sample means, where the level of significance was set at p-value < 0.05. Multiple comparisons of means were performed by using Tukey’s test.
3. Results and discussion 3.1 Microbiological analysis The whey beverages presented Aerobic Mesophilic bacteria (AMB) counts between 1.82 (LTLT) and 5.57 (T20) Log CFU/mL (p< 0.05) after the processing, with no significant difference between the samples treated by microfiltration under different temperature conditions. After 15 days of storage, there was an AMB growth in treatments T30, T40 and T50; and a decrease in treatments LTLT and T20 counts. However, there was not significant growth in relation to the storage time (Figure 1). In relation to the mold and yeast, only the treatments T40 and T50 presented detectable counts (1.95 and 1.37 CFU/mL, respectively) after processing. During the storage period, there was a growth of these microorganisms in T30 (2.57 CFU/mL, p < 0.05) and a decrease in T50 count (1.00 CFU/mL, p < 0.05). The other treatments did not present difference during the storage period. The Lactic acid bacteria (LAB) counts of the microfiltration-processed samples were significantly higher than that treated by LTLT; however, there was no difference in relation to the storage time. In this context, almost all the GOBs met the aerobic mesophilic bacterial counts established by Brazilian legislation for pasteurized whey beverages (Brazil, 2005), which is the maximum limit of 5.18 Log CFU/mL, except the beverage microfiltered at 20°C. Therefore, the microfiltration using at least 30°C allowed the achievement of a product with appropriate microbial count. The microfiltration is the most used technology for juice clarification and spoilage removal in industrial level (Castro-Muñoz et al., 2018). The present study confirm the findings of other studies (Fagnani et al., 2017; Pereira et al., 2015), in which was demonstrated the success of using microfiltration for removal of microorganisms in orange whey beverages. 3.2 pH and Color Parameters The pH results showed a variation between 3.18 (T40) and 4.24 (LTLT) (p <0.05), being the MF-processed beverages at low temperatures (T20 e T30) similar to LTLT (p>0.05). However, the GOB pH were much closer to the orange juice pH (3.45) than the goat whey pH (6.71) independently of the process. The pH of the treatments T40 and T50 had a significant decrease in comparison to the other treatments (p<0.05). In Jørgensen et al. (2016) study evaluating microfiltration of skim milk
with different pore sizes and temperatures, it was observed an association of higher pH with the increased casein micelles present in the permeate. Therefore, our results indicate that the MF using temperatures higher than 40°C may have hampered the permeation of proteins and other fruit compounds responsible for the pH maintaining the pH closer to neutral. The color parameters of goat whey orange juice beverages (GOB) analysis are presented in Table 1. In general, the color parameters L*, a*, b*, C* and YI of the samples processed by MF using different temperatures showed no difference between them, except the parameter a* (Green to red color) at T40 sample (p<0.05), indicating a greater distance from the red color, however it was very subtle. In relation to the LTLT treatment, the MF treatments produced beverages with very different color parameters (p<0.05). The ΔE indicates the total difference of each treatment in relation to an unprocessed GOB (data not shown). The processed beverages showed a small ΔE for LTLT (3 ± 1) and high color differences for MF beverages (23 to 25 ± 1), as observed by Adekunte et al. (2010), indicating that the LTLT process influenced just a little the color of the beverages, different than the Microfiltration process. Low ΔE were already observed for heat treated whey beverages, indicating a mild color change in these cases (Guimarães et al., 2018a). The high values of b* (29± 4), a* (-2.7 ± 0.4) and L* (79.8 ± 0.4) in LTLT, compared to the other treatments (p < 0.05) indicate a bright and yellowish characteristic of this sample (C* =29 ± 4; YI = 53 ± 6, respectively, p< 0.05). Since the microfiltration is usually applied to clarification purposes in juices (Castro-Muñoz et al., 2018), it was suggested that the microfiltration-processed GOBs decreased the beverages turbidity due to the retention of many great particles, which induced the lower intensity of yellow color but a higher lightness. The goat milk does not have carotenoids responsible for the yellowish color, since they are already in its converted form vitamin A (Sant'Ana et al., 2013), therefore, this pigment came from the orange juice. Despite the higher lightness (L*, p < 0.05) of the MF whey beverages, their color intensity was significantly lost (C* = 4.6 - 6) and became less yellowish (YI = 7.4 – 10), which intensely influenced the color difference from unprocessed beverage in relation to LTLT (Table 1). Overall, the GOBs processed by microfiltration presented more lightness, but less intense yellow color, which suggest the need of sensory tests to verify the consumer’s acceptance of these beverages. Fagnani
et al. (2017) observed that carotenoids are the main components responsible for the characteristic color in beverages containing orange juice and Microfiltration usually retain the compounds responsible for the color, due to their great molecular size and interaction between fat soluble substances and membrane particles (Mai et al., 2014). 3.3 Rheology According to Figure 2, three flow curves were performed at a shear rate ranging from 0 to 300 s-1. Data from the third flow curve were fitted to the power law model through non-linear regression analysis (0.879≤R2≤0.999). Consistency index (𝑘) and flow behavior index (𝑛) was calculated and presented in Table 2. In general, the temperature (20 to 50° C) used in microfiltration processes influenced the studied rheological parameters. In microfiltration, from temperature 30°C to 40°C there was a significant decrease in beverage 𝑘 (p<0.05) and an increase in 𝑛 (p<0.05). Beverages processed by LTLT presented a significant higher consistency index (𝑘) than the microfiltered ones (p<0.05). The flow behavior index (𝑛), in general, exhibited low values (𝑛 < 1) indicating a typical pseudoplastic behavior, characteristic of whey beverages, due to the complex interaction between its components (Guimarães et al., 2018b). The pseudoplastic behavior was more pronounced at beverages processed by LTLT (0,69 ± 0.01) than the microfiltered whey beverages (0.73 to 0.78 ± 0.01, p<0.05), except at the microfiltration using a temperature of 30°C (T30, 0.71, p>0.05). These results may be explained since the microfiltration process retains milk and juice compounds, like larger proteins and polysaccharides (Bhattacharjee, Saxena, & Dutta, 2017). Therefore, the retention of suspended solids larger than the pore of the membrane (0.2 µm) resulted in a less consistent beverage tending to a Newtonian behavior, since the presence of a higher concentration of large particles, interacting in a juice, for example, increases its gel-like structure (Dahdouh et al., 2016). In addition, it is reported in literature that the increased temperature of microfiltration increases the permeation flux; however, the increased flux may increase the fouling rate, caused by the insoluble pectic and cellulosic materials present in the fruit juices (De Oliveira, Docê & Barros, 2012), which may decrease the permeation of the other compounds. This fact may have influenced the rheological parameters as observed for 𝑘 and 𝑛 when temperatures higher than 40° C were used.
The rheological results indicated that microfiltration of goat whey orange juice beverage using up to 30°C seems to maintain the rheological characteristics similar to the heat processed (LTLT) beverages. 3.4 Bioactive compounds and functional activity The bioactive compounds content and the functional activities were evaluated in the GOB with the quantification of ascorbic acid, phenolic compounds, and antioxidant, antihypertensive and antidiabetic activities. It was observed that the LTLT samples presented lower values in general, probably due to the high temperature and time of processing, which may have influenced the bioactive peptides integrity and concentration of phenolic compounds and ascorbic acid (Table 3). The beverages processed by microfiltration using temperatures up to 40°C presented significantly higher values of ascorbic acid (T20 = 2.4 mg/g; T30 = 2.5 mg/g and T40 = 2.7 mg/g) than the other treatments (T50 = 0.3 mg/g and LTLT = 0.4 mg/g, p < 0.05). Then, it was suggested that the heat degraded the ascorbic acid in GOB during processing (LTLT and T50), since the ascorbic acid evaluated in the orange juice and goat milk whey (3.8 mg/g and 0.00 mg/g, respectively, data not shown) used for GOB elaboration were proportional to the content found in T20, T30 and T40. The phenolic compounds were retained in high quantity in microfiltration-processed samples (60.2 - 62.2 µg Gallic acid /g) than in LTLT (40.6 µg Gallic acid /g); however, the temperature of processing did not influence the concentration of these compounds in microfiltration (p >0.05). The samples processed by microfiltration had higher antihypertensive and antidiabetic activities than LTLT (p < 0.05). The antihypertensive activity varied between 42.2 % (LTLT) – 57.3 % (T20), the α-amylase between 48.4 % (LTLT) and 62.4 % (T30), the α-glucosidase between 39.3 % (LTLT) and 57.3 % (T20). Therefore, type of processing (LTLT or MF) influenced the activity of the α-amylase and α-glucosidase enzymes and the bioactive peptides content of the manufactured goat whey orange juice beverages. As expected, the process of microfiltration allowed a higher retention of bioactive compounds as observed in other studies (Cassano et al., 2016; Cassano et al., 2018; Castro-Muñoz & Fíla, 2018; Castro-Muñoz et al., 2018; Nazir et al., 2019).
The orange juice contains a high quantity of bioactive compounds, as ascorbic acid, carotenoids and phenolic compounds, which contribute to the total antioxidant capacity of the foods (Escudero-López et al., 2016), however, in some products and types of processing the phenolic compounds seem to contribute more to the antioxidant activity than the ascorbic acid (Guimarães et al., 2019). In this study, the phenolic compounds seemed to contribute more to the DPPH values, since the sample T50 presented low ascorbic acid content and high antioxidant activity. The ascorbic acid is one of the stronger and less toxic natural antioxidants, and may act as a strong free radical scavenger (Khan et al., 2019). The ascorbic acid degradation may occur due to different mechanisms, as temperature, dissolved oxygen, pH, acidity, metallic ions and water activity (Li et al., 2016). Thus, the heat may cause irreversible losses of nutritional compounds, unwanted changings on physicochemical and antioxidant properties (Arjmandi et al., 2017). The antioxidant activity is an indicator of the capacity to inhibit the molecular oxidation caused by free radicals, being important to the shelf life of dairy products and its consumption protect the human body against oxidative damage. The bioactive compounds in foods play a crucial role against the elevation of the oxygen reactive species, like superoxide, hydroxyl and peroxyl radicals, formed by cells under oxidative stress. These bioactive compounds, especially peptides, may give electrons to neutralize the free radicals. Beyond that, the presence of several amino acid residues at the peptide chains may improve the antioxidant properties (Wada, & Lönnerdal, 2015). The milk proteins are important sources of bioactive peptides, which showed to exert beneficial effects on cardiovascular system, as the antihypertensive effect, mainly due to the inhibition of the angiotensin converting enzyme (ACE) (Balthazar et al., 2019). The inhibition of αamylase activity and α-glucosidase together is seen as an effective strategy for controlling diabetes (Saeedi et al., 2017). 3.5 Volatile compounds It were detected 43 compounds in the goat whey orange juice beverages (Table 4), being: 7 acids; 12 alcohols; 4 aldehydes; 3 esters; 7 hydrocarbons; 3 ketones; 5 terpenes; and 2 others. Some
compounds
like
(S)-(+)-3-methyl-1-pentanol,
decanal,
cyclohexene,
3-methyl-6-(1-
methylethylidene)-, naphthalene, 1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-,(1S-cis)-,
β-myrcene, β-phellandrene, were identified only in beverage LTLT, indicating its possible formation due to the heating applied during the processing. Also, dl-menthol, phenol, 2,4-bis(1,1dimethylethyl)-, cyclopropane, propyl-, 4-heptanone, 2,6-dimethyl were identified only at the MF beverages, representing the volatile compounds degraded due to the long exposure to high temperatures in LTLT treatment. Compounds like 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester e methyl isocyanide probably volatized at temperatures higher than 40°C, since they were not identified in samples T40 and T50. Some volatile organic acids are unstable at high temperatures and could be absent in food after heating (Jang et al., 2018). The short and medium chain fatty acids were identified in every treated samples, and their representatives known for providing the goaty flavor are the capric (decanoic acid), caprylic (octanoic acid) and caproic (hexanoic acid) acids (Wang et al., 2019), which were not affected by the heat or microfiltration treatment. Overall, the application of microfiltration avoided the degradation of many thermosensitive volatile compounds.
4. Conclusion The microfiltration of the goat whey orange juice beverage showed to be a process with significant differences from the conventional heat treatment (LTLT), mainly related to the preservation of bioactive compounds and functional activities. The microfiltration was also effective in maintaining the microbiological quality, similar to the LTLT treatment. Overall, the Microfiltration process produced a less consistent or more fluid product, with increased lightness but less intense colors due to the retention of compounds responsible for the yellowish color, like carotenoids by the ceramic membrane, which may change the sensory perception of the consumers, requiring further investigation. Furthermore, the variation in Microfiltration feeding temperature showed to affect some parameters, like microbial reduction and rheology of the beverages, which seemed to be related to the permeation flux and the fouling rate of the membrane at a certain temperature. In general, the pH, color parameters, volatile and bioactive compounds were not influenced by microfiltration temperature. The rheological parameters were influenced by MF temperature and the utilization of
lower temperatures (20° and 30°C) maintained the consistency similar to the LTLT sample, preserving the compounds responsible for the texture. The microfiltration seem to be a promising technique to be applied in dairy industry to produce whey-fruit juice-based beverages with similar rheological characteristics of conventionally treated samples but using lower temperatures, which may be beneficial in relation to the nutritional compounds. In addition, we suggest the processing of GOB by microfiltration using mild temperatures (between 30° and 40° C) to preserve consistency and obtain a desirable microbial quality, beyond the preservation of many functional properties and volatile compounds.
Acknowledgement The authors are grateful to the Rio de Janeiro State Research Support Foundation (FAPERJ), National Council for Scientific and Technological Development (CNPQ) and Higher Education Personnel Improvement Coordination (CAPES) for the financial support.
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All the author had similar contribution in all the steps of the manuscript.
Declarations of interest: none
A 7,00
log CFU/mL
6,00 5,00 4,00
a
3,00
ab b
2,00
b
1,00
LTLT
a
a
T20
b
b
T30 T40 c
T50
c
0,00 Day 1
Day 7
Day 15
B 6,00
log CFU/mL
5,00 LTLT
4,00 3,00 2,00 1,00
T20
a
a a
T30
a
a a b
b
Day 1
Day 7
T40
b
T50
0,00 Day 15
C 6,00
log CFU/mL
5,00 4,00 3,00
a a
2,00 1,00
a a ab
b
a
b c
LTLT
a ab
T20 b
c
T30 T40 T50
0,00 Day 1
Day 7
Day 15
Figure 1. Microbiological counts of goat whey orange juice beverage during refrigerated storage (4°C ±1). A: aerobic mesophilic bacteria; B: molds and yeasts; C: lactic acid bacteria. Different letters in each day means significant difference (p < 0.05) between treatments. No differences were observed during refrigerated storage of 15 days. LTLT: low temperature - long time; T20: treatment at 20°C; T30: treatment at 30°C; T40: treatment at 40°C; T50:treatment at 50°C.
23
9 8
Shear stress (Pa)
7 6 5
L T T T T
4 3 2 1 0 0
50
100
150
200
250
300
Shear rate (s-1) Figure 2. Flow behavior curves of the goat whey orange juice beverages processed by LTLT and microfiltration.
23
24 Table 1. pH and color parameters of goat whey orange juice beverages processed by LTLT and Microfiltration. Treatment
pH
L*
a*
b*
YI
C*
ΔE
LTLT
4.24 ± 0.01a
79.8 ± 0.4 b
-2.7 ± 0.4 c
29 ± 4 a
53 ± 6 a
29 ± 4 a
3± 1c
T20
4.23 ± 0.01a
85.2 ± 0.4 a
1.4 ± 0.2 a
4.4 ± 0.2 b
7.4 ± 0.3 b
4.6 ± 0.2 b
25 ± 1 a
T30
4.22 ± 0.01a
85.3 ± 0.3 a
1.3 ± 0.2 a
4.7 ± 0.4 b
8± 1b
4.9 ± 0.4 b
25 ± 1 ab
T40
3.18 ± 0.01c
85 ± 1 a
0.9 ± 0.2 b
6± 1b
10 ± 1 b
6± 1b
23 ± 1 b
T50
3.91 ± 0.01b
85 ± 1 a
1 ± 0.2 ab
5.6 ± 0.4 b
9± 1b
5.7 ± 0.4 b
24 ± 1 ab
* Results are expressed as mean ± standard deviation. Different letters in the same column means significant difference between beverages (p < 0.05). L*: lightness; a*: green (-) to red (+); b*: blue (-) to yellow (+); YI: yellowness index; C*; chroma; ΔE: color difference. LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
24
25 Table 2. Rheological parameters of goat whey orange juice beverages processed by LTLT and Microfiltration. Treatment
𝒌 (Pa s)
𝒏 (-)
LTLT
0.15 ± 0.01a
0.69 ± 0.01c
T20
0.12 ± 0.01b
0.73 ± 0.01b
T30
0.13 ± 0.01b
0.71 ± 0.01bc
T40
0.073 ± 0.004c
0.78 ± 0.01a
T50
0.070 ± 0.004c
0.77 ± 0.01a
* Results are expressed as mean ± standard deviation. Different letters in the same column means significant difference between beverages (p < 0.05). LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
25
26 Table 3. Bioactive compounds content and functional activities of goat whey orange juice beverages processed by LTLT and Microfiltration. Treatment
Ascorbic acid
DPPH
ACE
α-amylase
αglucosidase
Phenolics
LTLT
0.4 ± 0.1 b
32.2 ± 0.1 b
42.2 ± 0.1 b
48.4 ± 0.1 b
39.3 ± 0.2 b
40.6 ± 0.5 b
T20
2.4 ± 0.2 a
60.3 ± 0.1 a
57.3 ± 0.1 a
62.3 ± 0.1 a
57.3 ± 0.1 a
60.2 ± 0.1 a
T30
2.5 ± 0.4 a
61.4 ± 0.1 a
55.3 ± 0.1 a
62.4 ± 0.1 a
56.2 ± 0.1 a
61.8 ± 0.1 a
T40
2.7 ± 0.2 a
58.3 ± 0.1 a
56.2 ± 0.1 a
61.4 ± 0.1 a
55.5 ± 0.6 a
62.2 ± 0.1 a
T50
0.3 ± 0.1 b
58.0 ± 0.1 a
55.7 ± 0.4 a
61.0 ± 0.3 a
55.4 ± 0.2 a
62.1 ± 0.3 a
* Results are expressed as mean ± standard deviation. Different letters in the same column means significant difference between beverages (p < 0.05). Acid ascorbic is expressed in mg/g. DPPH, ACE, αamylase and α-glucosidase are expressed in % inhibition, phenolics are expressed in µg Gallic acid /g. LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
26
27
Table 4. Volatile compounds of goat whey orange juice beverages processed by LTLT and Microfiltration. LRI T20 T30 T40 T50 LTLT Acids
X X X X X
X X X X X
X X X X X X
X X X X X X X
X X X X
1794 1840 1960 1960 2295
X X X X X
X X X X X X
X X X X X X X
X X X X X X
X X X X -
1354 1478 1509 1536 1687
X -
X -
X
X X -
X X
1268 1449 1486 1664 1680
X
-
X
X
X
1467 1627 1667 1837 1877 2048
Acetic acid Butanoic acid Butanoic acid, 3-methylHexanoic acid Butanoic acid, anhydride Octanoic acid
2153 2258
Nonanoic acid n-Decanoic acid
X
Alcohols 1260 1353 1404 1497
1-Butanol, 3-methyl(S)-(+)-5-Methyl-1-heptanol 3-Hexen-1-ol 1-Hexanol, 2-ethyl2-Propyl-1-pentanol 1,6-Octadien-3-ol, 3,7-dimethyl(S)-(+)-3-Methyl-1-pentanol 1-Octanol 3-Cyclohexen-1-ol, 4-methyl-1-(1-methylethyl)-, (R)dl-Menthol 2-Furanmethanol 1-Hexanol, 4-methyl1-Nonanol 1-Decanol 2,6-Octadien-1-ol, 3,7-dimethyl-, (Z)Geraniol (S)-3-Ethyl-4-methylpentanol (S)-(+)-6-Methyl-1-octanol Phenol, 2,4-bis(1,1-dimethylethyl)-
1497 1552 1561 1561 1609 1643 1656 1658 1659 1758
Aldehydes 4-Pentenal, 2-ethyl3-Furaldehyde Decanal Benzaldehyde Citral Esther Hexanoic acid, ethyl ester Octanoic acid, ethyl ester Acetic acid, octyl ester 6-Octen-1-ol, 3,7-dimethyl-, acetate Hexanoic acid, 3-hydroxy-, ethyl ester
27
28 Cyanic acid, 2,2-dimethylpropyl ester 1,2-Benzenedicarboxylic acid, bis(2-methylpropyl) ester
1758 2530
X X
X
-
-
1072
-
-
-
-
-
1043 1094 1279 1302 1318 1377 1441 1452 1497 1597
1770
X X -
X X -
X X X X -
X -
X X X X X -
1215 1408 1483 1736
X -
X X
X -
X X -
X
1185 1198 1224 1250 1275 1298 1308 1483 1505 1606 1698 1754 1828
X X
X X -
X X -
-
1089 1525 1815
X X
X X
X
X
X X X
Ether Dimethyl ether Hidrocarbons n-Hexane Bicyclo[3.1.1]hept-2-ene, 3,6,6-trimethylBicyclo[2.2.1]heptane, 2,2-dimethyl-3-methylene-, (1R)Cyclohexene, 3-methyl-6-(1-methylethylidene)Cyclohexasiloxane, dodecamethylCyclopropane, propylBenzene, 1,3-bis(1,1-dimethylethyl)Benzene, 4-ethenyl-1,2-dimethyl1-Pentene, 4,4-dimethyl-
1,5-Cyclodecadiene, 1,5-dimethyl-8-(1-methylethenyl)-, [S-(Z,E)]Naphthalene, 1,2,3,5,6,7,8,8a-octahydro-1,8a-dimethyl-7-(1-methylethenyl)1727 , [1R-(1.alpha.,7.beta.,8a.alpha.)]2-Isopropenyl-4a,8-dimethyl-1,2,3,4,4a,5,6,8a-octahydronaphthalene 1733 Alloaromadendrene Naphthalene, 1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-, (1Scis)(-)-.alpha.-Panasinsen
1743 1761
Ketones 4-Heptanone, 2,6-dimethyl2-Nonanone Cyclohexanone, 5-methyl-2-(1-methylethyl)-, cis3,7-Nonadien-2-one, 8-methyl-, (E)-
Terpene 3-Carene .beta.-Myrcene D-Limonene .beta.-Phellandrene .gamma.-Terpinene o-Cymene (+)-4-Carene l-Menthone .alpha.-Cubebene Caryophyllene .alpha.-Terpineol Geranyl acetate Estragole
Others Methyl isocyanide Ethene, fluoroPhenol, 2-nitro-
* LRI – Linear Retention Index.; X , – : presence and absence of the volatile compound . LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
28
29
29
30
Goat whey-orange juice beverage processed by microfiltration;
Microfiltration feeding temperatures were evaluated
Microfiltration preserved the volatile compounds and increased functional activity;
Lower temperatures maintained product consistency but affected microbial reduction
It was suggested a mild microfiltration processing temperature (between 30° and 40°C)
30
S0963-9969(20)30085-5 https://doi.org/10.1016/j.foodres.2020.109060 FRIN 109060
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Food Research International
Received Date: Revised Date: Accepted Date:
24 November 2019 1 February 2020 2 February 2020
Please cite this article as: Vieira, A.H., Balthazar, C.F., Guimaraes, J.T., Rocha, R.S., Pagani, M.M., Esmerino, E.A., Silva, M.C., Raices, R.S.L., Tonon, R.V., Cabral, L.M.C., Walter, E.H.M., Freitas, M.Q., Cruz, A.G., Advantages of microfiltration processing of goat whey orange juice beverage, Food Research International (2020), doi: https://doi.org/10.1016/j.foodres.2020.109060
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Advantages of microfiltration processing of goat whey orange juice beverage
Alexandre H. Vieira¹, Celso F. Balthazar1, ,Jonas T. Guimaraes¹, Ramon S. Rocha1,2 , Mônica M. Pagani³, Erick A. Esmerino1, Márcia C. Silva2, Renata S. L. Raices2, Renata V. Tonon4, Lourdes M.C. Cabral4, Eduardo H.M. Walter4 Mônica Q. Freitas¹, Adriano G. Cruz2
1Universidade 2Instituto
Federal Fluminense (UFF), Faculdade de Veterinária, 24230-340 Niterói, Brasil
Federal de Educação, Ciência e Tecnologia do Rio de Janeiro (IFRJ), Departamento de
Alimentos, 20270-021 Rio de Janeiro, Brasil 3Universidade
Federal Rural do Rio de Janeiro (UFRRJ), Instituto de Tecnologia (IT), 23890-000,
Brasil. 4Embrapa
Agroindústria de Alimentos (CTAA), 23020-470, Guaratiba, Rio de Janeiro. Brasil
*Email: [email protected] (A.G. Cruz) Goat whey Orange Juice Beverage
Abstract The objective of this study was to evaluate the microbiological, physicochemical and functional quality of an innovative goat whey orange juice beverage (GOB) processed by microfiltration. The microfiltration (0.2 µm) of the GOBs had a variation on the feed temperature (20, 30, 40, 50°C) and were compared to the conventional heat treatment LTLT (63°C/30 min). Microbiological (aerobic mesophilic bacteria, mold and yeast and lactic bacteria), physicochemical (pH, color, rheology and volatile compounds) bioactive compounds (acid ascorbic, total phenolics) and functional activity (DPPH, ACE, α-amilase and α-glucosidase) analysis were performed. The GOB processed by microfiltration using at least 30°C presented adequate microbial counts (less than 4, 3 and 4 log CFU/mL, for AMB, molds and yeasts and LAB, respectively). In general, the pH, color parameters, volatile and bioactive compounds were not influenced by microfiltration temperature, but presented a difference from the LTLT processing. The rheological parameters were influenced by MF temperature and the utilization of temperatures of 20° and 30°C maintained the consistency similar to the LTLT sample, preserving the compounds responsible for the texture. Therefore, it is suggested a processing of GOB by microfiltration using mild temperatures (between 30° and 40° C) to preserve consistency and also obtain a desirable microbial quality, beyond the preservation of many functional properties and volatile compounds. Keywords: Membrane technology; Functional activity; Dairy product; Microbiology; Rheology.
1. Introduction Nowadays, the modern food processing technologies aims the food safety in production but also the high nutritional quality of the product. In this context, the non-thermal technologies are known as cleaner processes than conventional thermal treatments, energetically efficient, environmental friendly and focus on quality development, maintaining a good acceptability without worsening the product’s safety (Ahmad et al., 2019; Balthazar et al., 2019a). One of the emerging technologies used for milk and dairy products is microfiltration, which is a membrane technology driven by pressure and has been applied to separate compounds of high added value, offering several technological advantages (Castro-Muñoz & Fíla, 2018). The Microfiltration processing presents low energy consumption; high separation efficiency; easy implementation and operation; high productivity; absence of phase transition and non-use of additional solvents, which favor the solute recovery. Therefore, different types of high added value compounds can be separated with membrane technologies, like carbohydrates, pectin, sugars, antioxidant compounds, anthocyanins, proteins and phenolic compounds (Nazir et al., 2019). The membrane processing offer several advantages over the conventional separation methods, like high hydrostatic pressure and high-pressure homogenization, including the mild operational conditions of temperature and pressure, which preserve the functional properties of food products, besides being a cleaner processing due to the absence of chemicals utilization (Soodam. K. & Guinee, 2018). In addition, microfiltration has been used in the dairy industry for several purposes, including the bacterial reduction to produce Extended Shelf Life (ESL) milk (Alegbeleye, Guimarães, Cruz, & Sant’Ana, 2018). Milk and dairy products are known to be healthy since it contains many essential nutrients, like oleic acid, conjugated linoleic acid, Ω-fatty acids, high biological value proteins, vitamins, minerals and bioactive compounds (Balthazar et al., 2017a). Among the consumption of milk of various species, currently, the goat milk consumption has been increasing, because its usually associated to different functional effects, such as health maintenance, risk of chronic diseases reduction, besides other beneficial physiological effects (Clarck & Garcia, 2017). Furthermore, this
dairy matrix is important in nutrition of children, elder and people allergic to bovine milk protein, since goat milk presents proteins of high digestibility and hypoallergenicity (Balthazar et al., 2017a; Verruck et al., 2019). For these reasons, the goat milk can be considered an excellent bovine milk substitute in human nutrition, being also used for manufacturing of functional dairy products (Ranadheera et al., 2019). Among the current chronic diseases affecting the world population, diabetes mellitus has risen dramatically and it is expected to affect 438 million people until 2030, with 70% of the cases occurring in poor countries. The concomitant inhibition of α-amylase and α-glucosidase activities is seen as an effective strategy for the control of this chronic disease (Carrizzo et al., 2018; Saeedi, Hadjiakhondi, Nabavi, & Manayi, 2017). In addition, milk proteins are considered a good source of bioactive compounds with antihypertensive activity (angiotensin-converting-enzyme inhibition), which may be beneficial for cardiovascular health (Marcone, Belton & Fitzgerald, 2017). In recent years, the utilization of whey in foods, such as dairy, bakery, confectionary, meat and canned products (Jeewanthi et al., 2015), edible films and coatings (Chakravartula et al., 2019) and non-food products such as bioplastics, biofertilizers, biofuels (Ahmad et al., 2019) is increasing all around the world. The whey protein has high biological value; however, too much whey is still discarded during manufacturing of dairy products (Ahmad et al., 2019). In food industries, the whey proteins are used as emulsifying, gelling and bulking agent. The antioxidant activity of the whey proteins is well-stablished and it may efficiently inhibit the lipid oxidation (Wen-Qiong et al., 2019). One of the most important aspects in introducing a new goat dairy food into market is to pay attention the public demand and to make specific market surveys to identify those demands in order to reach the new opportunities (Freire et al., 2017). Therefore, the objective of this study was to elaborate an innovative goat whey orange juice beverage processed by microfiltration membrane technology and study the effect of microfiltration feeding temperatures on the microbiological, physicochemical and bioactive compounds and functional characteristics.
2. Material and methods
2.1 Goat whey orange juice beverage (GOB) processing The whey beverages with orange juice (10 L) were manufactured with the following ingredients: 45 % (w/w) of skim goat whey (0.1% fat), 45% (w/w) of fresh orange juice and 10% (w/w) of sugar. This basic formulation was used in five different treatments: Low temperature long time treatment (LTLT - 63°C/30 min) as a control beverage due to conventional thermal treatment applied to beverages; and microfiltration (MF) varying the system feeding temperature: T20 (at 20°C); T30 (at 30°C); T40 (at 40°C) and T50 (at 50ºC). After the processing, the beverages were packaged in sterilized plastic bottles (300 mL) and kept under cold storage (4ºC ± 1) during 15 days, which is the typical shelf-life period for pasteurized dairy products (Prashanth, Jayaprakash, Soumyashree, & Madhusudhan, 2018). The microfiltration was realized in a new ceramic membrane (α-alumina) with 0.2 µm pore (Membralox T1-70 modules, channel diameter 7 mm, length 250 mm, surface area filtration of 0.005 m² (0.054 ft2), housing material 316L SS) and working pressure of 2 kgf/cm². The LTLT and microfiltration treatments were realized in pilot equipment at EMBRAPACTAA laboratory (Guaratiba/RJ, Brazil). The whole experiment was performed in triplicate. 2.2 Microbiological analysis The goat whey orange juice beverages were submitted to the microbiological analysis of Aerobic Mesophilic Bacteria (AMB), Molds and yeasts and Lactic Acid Bacteria (LAB), according to the methodology recommended by “American Public Health Association” (APHA, 2015). The analysis were carried out right after processing (D1), seven days (D7) and 15 days (D15) after cold storage (4 ± 1°C). All the analysis were realized in triplicate. 2.3 pH values and color parameters The pH analysis and instrumental color were carried out right after processing. All the analysis were realized in triplicate. The pH of the samples were measured using a pHmeter (Digimed, model DM-20, São Paulo, Brasil), at 25ºC, inserting the electrode directly into the sample. The samples color were measured using a colorimeter (Colour Quest XE Hunter Lab, Northants, UK) based on CIELAB system (L*, a*, b*) according to the procedure stablished by Balthazar et al. (2015). The chroma (C*, Eq.1), the yellowness index (YI, Eq.2) and the total color
difference (ΔE*, Eq.3) were calculated according to Pathare et al. (2013), through the following formulas:
𝑪 ∗ = (𝒂 ∗ )𝟐 + (𝒃 ∗ )𝟐 𝒀𝑰 =
(Eq.1)
𝟏𝟒𝟐.𝟔 𝒃 ∗
(Eq.2)
𝑳∗
𝟐
𝟐
∆𝑬 ∗ = (∆𝑳 ∗ ) + (∆𝒂 ∗ ) + (∆𝒃 ∗ )
𝟐
(Eq.3)
2.4 Bioactive compounds and functional activity The bioactive compounds evaluated in this study were: total phenolic compounds according to Capatto et al. (2018) and ascorbic acid according to Balthazar et al. (2019b). The measured functional activities were: antioxidant (2,2-diphenyl-1-picrylhydrazyl – DPPH) and antihypertensive (inhibition of angiotensin-converting enzyme – ACE) activities according to Capatto et al. (2018) and antidiabetic activity (α-amilase e α-glucosidase) as described by Balthazar et al. (2019b). The analysis were carried out in samples right after processing and in triplicate. Specifically, the extracts for total phenolic compounds and DPPH analyses were obtained in triplicate, in which approximately 1 g of GOB sample was weighed into beakers (200 mL), and 30 mL mix of water and ethanol (50:50 v/v) was added and stirred at 200 rpm on an orbital table (SL180/D, Solab, Piracicaba, SP, Brazil) for 1 h. The resulting extracts were filtered applying vacuum. The absorbance of total phenolic compounds was measured at 725 nm in a spectrophotometer (Biospectro, SP-220), and the results were obtained using a calibration curve. The results were expressed as gallic acid equivalents per liter of sample (EAG g/L). It was used 1 mL of Folin reagent diluted in distilled water (1:10), which was added to 1 mL of extract and agitated for 1 min. For antioxidant capacity, 2850 μL of a methanolic solution of the DPPH radical (0.06 mM – 700 nm) was mixed to 150 μL of the extract and held in the dark for 60 min. The results of both analysis were expressed as μg Trolox Equivalent/g of sample.
ACE inhibitory activity were performed by filtration of the extracts, the angiotensin converting enzyme inhibitory (ACEI) was determined in a spectrophotometer and the ACE inhibitory activity was calculated as follows: (𝐵 𝐴)
𝐴𝐶𝐸 𝐼𝑛𝑖𝑏𝑖𝑡𝑜𝑟𝑦 𝑎𝑐𝑡𝑖𝑣𝑖𝑡𝑦 (%) = [(𝐵 𝐶) 100]
(Eq. 4)
where, A is the absorbance of ACEI component in the presence of ACE; B is the absorbance without the ACEI component and C is the absorbance without ACE. Ascorbic acid content was determined using the titration method with 2,6-dichlorophenolindophenol, for the extraction. A 5-mL sample was mixed in 25 mL of metaphosphoric acid solution and centrifuged at 13,715 × g for 10 min at room temperature. Then ascorbic acid was quantified using an aliquot of the supernatant (approximately 5 mL), which was diluted in 15 mL of extraction solution and titrated with the indophenol solution and the samples in the absence of light. The ascorbic acid content was expressed in milligrams per gram. The α-amylase and α-glucosidase enzyme inhibition activities were determined α-amylase by measuring absorbance (Abs) at 540 nm and releasing p-nitrophenol at 400 nm, respectively. Solutions without the water-soluble extract and substrate sample as a control and a blank, respectively were used. The inhibition percentage was calculated as follows:
(
Inhibition (%) = 1 ―
Abs sample ― Abs blank Abs control
) × 100
(Eq. 5)
2.5 Volatile compounds The volatile compounds analysis was performed as described by Balthazar et al. (2018). Briefly, the beverage samples were extracted by solid phase micro extraction (SPME) and analyzed by gas chromatography (GC MS, Varian 3800 GC, Milan Italy) coupled to mass spectrometry (MS ion trap VArian 2000, VArian Spa, Milan Italy). The SPME extractions were carried out using 50/30 mm thick DVB/CAR/PDMS (divinylbenzene/carboxen/polydimethylsiloxane) fibres (Supelco, Bellefonte, PA, USA) and 40 mL head space vials in an manual sampler holder (Supelco, Bellefonte, PA, USA). For that, 3 g sample was added with 3 mL saturated NaCl solution maintaining the vial at 40 Cin a dry block (Multi-Blok 2003 Thermo Fisher Scientific, Germany)with a 20 min equilibration time and 30 min extraction time. After extraction, the SPME fiber was manually introduced into the
GCMS for30 min under the following conditions for the thermal desorption of the analytes: injector temperature at 240 °C; splitless injection mode; column CP-Wax 52 CB 60 m, 0.25 mm id, 0.25 mm film thickness; oven temperature programmed from 45 °C for 5 min, then increased to 80 Cat a rate of 10 °C min-1, then to 240 °C at 5 °C min-1 and kept for 30 min; 1 mL min-1 of Helium gas flow, transfer line temperature at 240 °C, electron impact ionization energy at 70 eV, acquisition mass range of 50550 m/z. The volatile compounds were identified by comparison of their experimental spectra with those provided by the National Institute of Standards & Technology (NIST/EPA/NIH MassSpectra Library, NIST11, USA), using the linear retention indices (LRI).LRI values of C8-C40 alkanes standards (Supelco, 40,127-U) were injected under the same chromatographic and mass spectrometric conditions. The analysis was carried out in triplicate. 2.6. Rheology The rheological analysis was carried out as described by Balthazar et al. (2017b). To access the flow curve behavior, a rheometer (model 1500 AR, TA Instruments, New Castle, DE) equipped with cone & plate 140 geometry was used. The flow behavior curves and the mechanical spectrum of the samples were obtained at 25°C. The flow curves were determined using a shear rate varying from 0 to 300 s−1 and the data obtained were adjusted to the power law (Eq. 6):
𝜎 = 𝑘.𝛾𝑛
(Eq.6)
Where 𝜎 is the shear stress (Pa), 𝑘 is the consistency index (Pa sn), 𝛾 is the shear rate (s−1), and
𝑛 is the flow behavior index (dimensionless). The analysis were carried out in triplicate right
after processing. 2.7 Statistical analysis A completely randomized design of the experiment was carried out and conducted in triplicate. The data were analyzed using XLSTAT 2019.2 (Addinsoft, New York, NY, USA). One-way analysis of variance (ANOVA) was used to determine significant differences between sample means, where the level of significance was set at p-value < 0.05. Multiple comparisons of means were performed by using Tukey’s test.
3. Results and discussion 3.1 Microbiological analysis The whey beverages presented Aerobic Mesophilic bacteria (AMB) counts between 1.82 (LTLT) and 5.57 (T20) Log CFU/mL (p< 0.05) after the processing, with no significant difference between the samples treated by microfiltration under different temperature conditions. After 15 days of storage, there was an AMB growth in treatments T30, T40 and T50; and a decrease in treatments LTLT and T20 counts. However, there was not significant growth in relation to the storage time (Figure 1). In relation to the mold and yeast, only the treatments T40 and T50 presented detectable counts (1.95 and 1.37 CFU/mL, respectively) after processing. During the storage period, there was a growth of these microorganisms in T30 (2.57 CFU/mL, p < 0.05) and a decrease in T50 count (1.00 CFU/mL, p < 0.05). The other treatments did not present difference during the storage period. The Lactic acid bacteria (LAB) counts of the microfiltration-processed samples were significantly higher than that treated by LTLT; however, there was no difference in relation to the storage time. In this context, almost all the GOBs met the aerobic mesophilic bacterial counts established by Brazilian legislation for pasteurized whey beverages (Brazil, 2005), which is the maximum limit of 5.18 Log CFU/mL, except the beverage microfiltered at 20°C. Therefore, the microfiltration using at least 30°C allowed the achievement of a product with appropriate microbial count. The microfiltration is the most used technology for juice clarification and spoilage removal in industrial level (Castro-Muñoz et al., 2018). The present study confirm the findings of other studies (Fagnani et al., 2017; Pereira et al., 2015), in which was demonstrated the success of using microfiltration for removal of microorganisms in orange whey beverages. 3.2 pH and Color Parameters The pH results showed a variation between 3.18 (T40) and 4.24 (LTLT) (p <0.05), being the MF-processed beverages at low temperatures (T20 e T30) similar to LTLT (p>0.05). However, the GOB pH were much closer to the orange juice pH (3.45) than the goat whey pH (6.71) independently of the process. The pH of the treatments T40 and T50 had a significant decrease in comparison to the other treatments (p<0.05). In Jørgensen et al. (2016) study evaluating microfiltration of skim milk
with different pore sizes and temperatures, it was observed an association of higher pH with the increased casein micelles present in the permeate. Therefore, our results indicate that the MF using temperatures higher than 40°C may have hampered the permeation of proteins and other fruit compounds responsible for the pH maintaining the pH closer to neutral. The color parameters of goat whey orange juice beverages (GOB) analysis are presented in Table 1. In general, the color parameters L*, a*, b*, C* and YI of the samples processed by MF using different temperatures showed no difference between them, except the parameter a* (Green to red color) at T40 sample (p<0.05), indicating a greater distance from the red color, however it was very subtle. In relation to the LTLT treatment, the MF treatments produced beverages with very different color parameters (p<0.05). The ΔE indicates the total difference of each treatment in relation to an unprocessed GOB (data not shown). The processed beverages showed a small ΔE for LTLT (3 ± 1) and high color differences for MF beverages (23 to 25 ± 1), as observed by Adekunte et al. (2010), indicating that the LTLT process influenced just a little the color of the beverages, different than the Microfiltration process. Low ΔE were already observed for heat treated whey beverages, indicating a mild color change in these cases (Guimarães et al., 2018a). The high values of b* (29± 4), a* (-2.7 ± 0.4) and L* (79.8 ± 0.4) in LTLT, compared to the other treatments (p < 0.05) indicate a bright and yellowish characteristic of this sample (C* =29 ± 4; YI = 53 ± 6, respectively, p< 0.05). Since the microfiltration is usually applied to clarification purposes in juices (Castro-Muñoz et al., 2018), it was suggested that the microfiltration-processed GOBs decreased the beverages turbidity due to the retention of many great particles, which induced the lower intensity of yellow color but a higher lightness. The goat milk does not have carotenoids responsible for the yellowish color, since they are already in its converted form vitamin A (Sant'Ana et al., 2013), therefore, this pigment came from the orange juice. Despite the higher lightness (L*, p < 0.05) of the MF whey beverages, their color intensity was significantly lost (C* = 4.6 - 6) and became less yellowish (YI = 7.4 – 10), which intensely influenced the color difference from unprocessed beverage in relation to LTLT (Table 1). Overall, the GOBs processed by microfiltration presented more lightness, but less intense yellow color, which suggest the need of sensory tests to verify the consumer’s acceptance of these beverages. Fagnani
et al. (2017) observed that carotenoids are the main components responsible for the characteristic color in beverages containing orange juice and Microfiltration usually retain the compounds responsible for the color, due to their great molecular size and interaction between fat soluble substances and membrane particles (Mai et al., 2014). 3.3 Rheology According to Figure 2, three flow curves were performed at a shear rate ranging from 0 to 300 s-1. Data from the third flow curve were fitted to the power law model through non-linear regression analysis (0.879≤R2≤0.999). Consistency index (𝑘) and flow behavior index (𝑛) was calculated and presented in Table 2. In general, the temperature (20 to 50° C) used in microfiltration processes influenced the studied rheological parameters. In microfiltration, from temperature 30°C to 40°C there was a significant decrease in beverage 𝑘 (p<0.05) and an increase in 𝑛 (p<0.05). Beverages processed by LTLT presented a significant higher consistency index (𝑘) than the microfiltered ones (p<0.05). The flow behavior index (𝑛), in general, exhibited low values (𝑛 < 1) indicating a typical pseudoplastic behavior, characteristic of whey beverages, due to the complex interaction between its components (Guimarães et al., 2018b). The pseudoplastic behavior was more pronounced at beverages processed by LTLT (0,69 ± 0.01) than the microfiltered whey beverages (0.73 to 0.78 ± 0.01, p<0.05), except at the microfiltration using a temperature of 30°C (T30, 0.71, p>0.05). These results may be explained since the microfiltration process retains milk and juice compounds, like larger proteins and polysaccharides (Bhattacharjee, Saxena, & Dutta, 2017). Therefore, the retention of suspended solids larger than the pore of the membrane (0.2 µm) resulted in a less consistent beverage tending to a Newtonian behavior, since the presence of a higher concentration of large particles, interacting in a juice, for example, increases its gel-like structure (Dahdouh et al., 2016). In addition, it is reported in literature that the increased temperature of microfiltration increases the permeation flux; however, the increased flux may increase the fouling rate, caused by the insoluble pectic and cellulosic materials present in the fruit juices (De Oliveira, Docê & Barros, 2012), which may decrease the permeation of the other compounds. This fact may have influenced the rheological parameters as observed for 𝑘 and 𝑛 when temperatures higher than 40° C were used.
The rheological results indicated that microfiltration of goat whey orange juice beverage using up to 30°C seems to maintain the rheological characteristics similar to the heat processed (LTLT) beverages. 3.4 Bioactive compounds and functional activity The bioactive compounds content and the functional activities were evaluated in the GOB with the quantification of ascorbic acid, phenolic compounds, and antioxidant, antihypertensive and antidiabetic activities. It was observed that the LTLT samples presented lower values in general, probably due to the high temperature and time of processing, which may have influenced the bioactive peptides integrity and concentration of phenolic compounds and ascorbic acid (Table 3). The beverages processed by microfiltration using temperatures up to 40°C presented significantly higher values of ascorbic acid (T20 = 2.4 mg/g; T30 = 2.5 mg/g and T40 = 2.7 mg/g) than the other treatments (T50 = 0.3 mg/g and LTLT = 0.4 mg/g, p < 0.05). Then, it was suggested that the heat degraded the ascorbic acid in GOB during processing (LTLT and T50), since the ascorbic acid evaluated in the orange juice and goat milk whey (3.8 mg/g and 0.00 mg/g, respectively, data not shown) used for GOB elaboration were proportional to the content found in T20, T30 and T40. The phenolic compounds were retained in high quantity in microfiltration-processed samples (60.2 - 62.2 µg Gallic acid /g) than in LTLT (40.6 µg Gallic acid /g); however, the temperature of processing did not influence the concentration of these compounds in microfiltration (p >0.05). The samples processed by microfiltration had higher antihypertensive and antidiabetic activities than LTLT (p < 0.05). The antihypertensive activity varied between 42.2 % (LTLT) – 57.3 % (T20), the α-amylase between 48.4 % (LTLT) and 62.4 % (T30), the α-glucosidase between 39.3 % (LTLT) and 57.3 % (T20). Therefore, type of processing (LTLT or MF) influenced the activity of the α-amylase and α-glucosidase enzymes and the bioactive peptides content of the manufactured goat whey orange juice beverages. As expected, the process of microfiltration allowed a higher retention of bioactive compounds as observed in other studies (Cassano et al., 2016; Cassano et al., 2018; Castro-Muñoz & Fíla, 2018; Castro-Muñoz et al., 2018; Nazir et al., 2019).
The orange juice contains a high quantity of bioactive compounds, as ascorbic acid, carotenoids and phenolic compounds, which contribute to the total antioxidant capacity of the foods (Escudero-López et al., 2016), however, in some products and types of processing the phenolic compounds seem to contribute more to the antioxidant activity than the ascorbic acid (Guimarães et al., 2019). In this study, the phenolic compounds seemed to contribute more to the DPPH values, since the sample T50 presented low ascorbic acid content and high antioxidant activity. The ascorbic acid is one of the stronger and less toxic natural antioxidants, and may act as a strong free radical scavenger (Khan et al., 2019). The ascorbic acid degradation may occur due to different mechanisms, as temperature, dissolved oxygen, pH, acidity, metallic ions and water activity (Li et al., 2016). Thus, the heat may cause irreversible losses of nutritional compounds, unwanted changings on physicochemical and antioxidant properties (Arjmandi et al., 2017). The antioxidant activity is an indicator of the capacity to inhibit the molecular oxidation caused by free radicals, being important to the shelf life of dairy products and its consumption protect the human body against oxidative damage. The bioactive compounds in foods play a crucial role against the elevation of the oxygen reactive species, like superoxide, hydroxyl and peroxyl radicals, formed by cells under oxidative stress. These bioactive compounds, especially peptides, may give electrons to neutralize the free radicals. Beyond that, the presence of several amino acid residues at the peptide chains may improve the antioxidant properties (Wada, & Lönnerdal, 2015). The milk proteins are important sources of bioactive peptides, which showed to exert beneficial effects on cardiovascular system, as the antihypertensive effect, mainly due to the inhibition of the angiotensin converting enzyme (ACE) (Balthazar et al., 2019). The inhibition of αamylase activity and α-glucosidase together is seen as an effective strategy for controlling diabetes (Saeedi et al., 2017). 3.5 Volatile compounds It were detected 43 compounds in the goat whey orange juice beverages (Table 4), being: 7 acids; 12 alcohols; 4 aldehydes; 3 esters; 7 hydrocarbons; 3 ketones; 5 terpenes; and 2 others. Some
compounds
like
(S)-(+)-3-methyl-1-pentanol,
decanal,
cyclohexene,
3-methyl-6-(1-
methylethylidene)-, naphthalene, 1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-,(1S-cis)-,
β-myrcene, β-phellandrene, were identified only in beverage LTLT, indicating its possible formation due to the heating applied during the processing. Also, dl-menthol, phenol, 2,4-bis(1,1dimethylethyl)-, cyclopropane, propyl-, 4-heptanone, 2,6-dimethyl were identified only at the MF beverages, representing the volatile compounds degraded due to the long exposure to high temperatures in LTLT treatment. Compounds like 1,2-benzenedicarboxylic acid, bis(2-methylpropyl) ester e methyl isocyanide probably volatized at temperatures higher than 40°C, since they were not identified in samples T40 and T50. Some volatile organic acids are unstable at high temperatures and could be absent in food after heating (Jang et al., 2018). The short and medium chain fatty acids were identified in every treated samples, and their representatives known for providing the goaty flavor are the capric (decanoic acid), caprylic (octanoic acid) and caproic (hexanoic acid) acids (Wang et al., 2019), which were not affected by the heat or microfiltration treatment. Overall, the application of microfiltration avoided the degradation of many thermosensitive volatile compounds.
4. Conclusion The microfiltration of the goat whey orange juice beverage showed to be a process with significant differences from the conventional heat treatment (LTLT), mainly related to the preservation of bioactive compounds and functional activities. The microfiltration was also effective in maintaining the microbiological quality, similar to the LTLT treatment. Overall, the Microfiltration process produced a less consistent or more fluid product, with increased lightness but less intense colors due to the retention of compounds responsible for the yellowish color, like carotenoids by the ceramic membrane, which may change the sensory perception of the consumers, requiring further investigation. Furthermore, the variation in Microfiltration feeding temperature showed to affect some parameters, like microbial reduction and rheology of the beverages, which seemed to be related to the permeation flux and the fouling rate of the membrane at a certain temperature. In general, the pH, color parameters, volatile and bioactive compounds were not influenced by microfiltration temperature. The rheological parameters were influenced by MF temperature and the utilization of
lower temperatures (20° and 30°C) maintained the consistency similar to the LTLT sample, preserving the compounds responsible for the texture. The microfiltration seem to be a promising technique to be applied in dairy industry to produce whey-fruit juice-based beverages with similar rheological characteristics of conventionally treated samples but using lower temperatures, which may be beneficial in relation to the nutritional compounds. In addition, we suggest the processing of GOB by microfiltration using mild temperatures (between 30° and 40° C) to preserve consistency and obtain a desirable microbial quality, beyond the preservation of many functional properties and volatile compounds.
Acknowledgement The authors are grateful to the Rio de Janeiro State Research Support Foundation (FAPERJ), National Council for Scientific and Technological Development (CNPQ) and Higher Education Personnel Improvement Coordination (CAPES) for the financial support.
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All the author had similar contribution in all the steps of the manuscript.
Declarations of interest: none
A 7,00
log CFU/mL
6,00 5,00 4,00
a
3,00
ab b
2,00
b
1,00
LTLT
a
a
T20
b
b
T30 T40 c
T50
c
0,00 Day 1
Day 7
Day 15
B 6,00
log CFU/mL
5,00 LTLT
4,00 3,00 2,00 1,00
T20
a
a a
T30
a
a a b
b
Day 1
Day 7
T40
b
T50
0,00 Day 15
C 6,00
log CFU/mL
5,00 4,00 3,00
a a
2,00 1,00
a a ab
b
a
b c
LTLT
a ab
T20 b
c
T30 T40 T50
0,00 Day 1
Day 7
Day 15
Figure 1. Microbiological counts of goat whey orange juice beverage during refrigerated storage (4°C ±1). A: aerobic mesophilic bacteria; B: molds and yeasts; C: lactic acid bacteria. Different letters in each day means significant difference (p < 0.05) between treatments. No differences were observed during refrigerated storage of 15 days. LTLT: low temperature - long time; T20: treatment at 20°C; T30: treatment at 30°C; T40: treatment at 40°C; T50:treatment at 50°C.
23
9 8
Shear stress (Pa)
7 6 5
L T T T T
4 3 2 1 0 0
50
100
150
200
250
300
Shear rate (s-1) Figure 2. Flow behavior curves of the goat whey orange juice beverages processed by LTLT and microfiltration.
23
24 Table 1. pH and color parameters of goat whey orange juice beverages processed by LTLT and Microfiltration. Treatment
pH
L*
a*
b*
YI
C*
ΔE
LTLT
4.24 ± 0.01a
79.8 ± 0.4 b
-2.7 ± 0.4 c
29 ± 4 a
53 ± 6 a
29 ± 4 a
3± 1c
T20
4.23 ± 0.01a
85.2 ± 0.4 a
1.4 ± 0.2 a
4.4 ± 0.2 b
7.4 ± 0.3 b
4.6 ± 0.2 b
25 ± 1 a
T30
4.22 ± 0.01a
85.3 ± 0.3 a
1.3 ± 0.2 a
4.7 ± 0.4 b
8± 1b
4.9 ± 0.4 b
25 ± 1 ab
T40
3.18 ± 0.01c
85 ± 1 a
0.9 ± 0.2 b
6± 1b
10 ± 1 b
6± 1b
23 ± 1 b
T50
3.91 ± 0.01b
85 ± 1 a
1 ± 0.2 ab
5.6 ± 0.4 b
9± 1b
5.7 ± 0.4 b
24 ± 1 ab
* Results are expressed as mean ± standard deviation. Different letters in the same column means significant difference between beverages (p < 0.05). L*: lightness; a*: green (-) to red (+); b*: blue (-) to yellow (+); YI: yellowness index; C*; chroma; ΔE: color difference. LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
24
25 Table 2. Rheological parameters of goat whey orange juice beverages processed by LTLT and Microfiltration. Treatment
𝒌 (Pa s)
𝒏 (-)
LTLT
0.15 ± 0.01a
0.69 ± 0.01c
T20
0.12 ± 0.01b
0.73 ± 0.01b
T30
0.13 ± 0.01b
0.71 ± 0.01bc
T40
0.073 ± 0.004c
0.78 ± 0.01a
T50
0.070 ± 0.004c
0.77 ± 0.01a
* Results are expressed as mean ± standard deviation. Different letters in the same column means significant difference between beverages (p < 0.05). LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
25
26 Table 3. Bioactive compounds content and functional activities of goat whey orange juice beverages processed by LTLT and Microfiltration. Treatment
Ascorbic acid
DPPH
ACE
α-amylase
αglucosidase
Phenolics
LTLT
0.4 ± 0.1 b
32.2 ± 0.1 b
42.2 ± 0.1 b
48.4 ± 0.1 b
39.3 ± 0.2 b
40.6 ± 0.5 b
T20
2.4 ± 0.2 a
60.3 ± 0.1 a
57.3 ± 0.1 a
62.3 ± 0.1 a
57.3 ± 0.1 a
60.2 ± 0.1 a
T30
2.5 ± 0.4 a
61.4 ± 0.1 a
55.3 ± 0.1 a
62.4 ± 0.1 a
56.2 ± 0.1 a
61.8 ± 0.1 a
T40
2.7 ± 0.2 a
58.3 ± 0.1 a
56.2 ± 0.1 a
61.4 ± 0.1 a
55.5 ± 0.6 a
62.2 ± 0.1 a
T50
0.3 ± 0.1 b
58.0 ± 0.1 a
55.7 ± 0.4 a
61.0 ± 0.3 a
55.4 ± 0.2 a
62.1 ± 0.3 a
* Results are expressed as mean ± standard deviation. Different letters in the same column means significant difference between beverages (p < 0.05). Acid ascorbic is expressed in mg/g. DPPH, ACE, αamylase and α-glucosidase are expressed in % inhibition, phenolics are expressed in µg Gallic acid /g. LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
26
27
Table 4. Volatile compounds of goat whey orange juice beverages processed by LTLT and Microfiltration. LRI T20 T30 T40 T50 LTLT Acids
X X X X X
X X X X X
X X X X X X
X X X X X X X
X X X X
1794 1840 1960 1960 2295
X X X X X
X X X X X X
X X X X X X X
X X X X X X
X X X X -
1354 1478 1509 1536 1687
X -
X -
X
X X -
X X
1268 1449 1486 1664 1680
X
-
X
X
X
1467 1627 1667 1837 1877 2048
Acetic acid Butanoic acid Butanoic acid, 3-methylHexanoic acid Butanoic acid, anhydride Octanoic acid
2153 2258
Nonanoic acid n-Decanoic acid
X
Alcohols 1260 1353 1404 1497
1-Butanol, 3-methyl(S)-(+)-5-Methyl-1-heptanol 3-Hexen-1-ol 1-Hexanol, 2-ethyl2-Propyl-1-pentanol 1,6-Octadien-3-ol, 3,7-dimethyl(S)-(+)-3-Methyl-1-pentanol 1-Octanol 3-Cyclohexen-1-ol, 4-methyl-1-(1-methylethyl)-, (R)dl-Menthol 2-Furanmethanol 1-Hexanol, 4-methyl1-Nonanol 1-Decanol 2,6-Octadien-1-ol, 3,7-dimethyl-, (Z)Geraniol (S)-3-Ethyl-4-methylpentanol (S)-(+)-6-Methyl-1-octanol Phenol, 2,4-bis(1,1-dimethylethyl)-
1497 1552 1561 1561 1609 1643 1656 1658 1659 1758
Aldehydes 4-Pentenal, 2-ethyl3-Furaldehyde Decanal Benzaldehyde Citral Esther Hexanoic acid, ethyl ester Octanoic acid, ethyl ester Acetic acid, octyl ester 6-Octen-1-ol, 3,7-dimethyl-, acetate Hexanoic acid, 3-hydroxy-, ethyl ester
27
28 Cyanic acid, 2,2-dimethylpropyl ester 1,2-Benzenedicarboxylic acid, bis(2-methylpropyl) ester
1758 2530
X X
X
-
-
1072
-
-
-
-
-
1043 1094 1279 1302 1318 1377 1441 1452 1497 1597
1770
X X -
X X -
X X X X -
X -
X X X X X -
1215 1408 1483 1736
X -
X X
X -
X X -
X
1185 1198 1224 1250 1275 1298 1308 1483 1505 1606 1698 1754 1828
X X
X X -
X X -
-
1089 1525 1815
X X
X X
X
X
X X X
Ether Dimethyl ether Hidrocarbons n-Hexane Bicyclo[3.1.1]hept-2-ene, 3,6,6-trimethylBicyclo[2.2.1]heptane, 2,2-dimethyl-3-methylene-, (1R)Cyclohexene, 3-methyl-6-(1-methylethylidene)Cyclohexasiloxane, dodecamethylCyclopropane, propylBenzene, 1,3-bis(1,1-dimethylethyl)Benzene, 4-ethenyl-1,2-dimethyl1-Pentene, 4,4-dimethyl-
1,5-Cyclodecadiene, 1,5-dimethyl-8-(1-methylethenyl)-, [S-(Z,E)]Naphthalene, 1,2,3,5,6,7,8,8a-octahydro-1,8a-dimethyl-7-(1-methylethenyl)1727 , [1R-(1.alpha.,7.beta.,8a.alpha.)]2-Isopropenyl-4a,8-dimethyl-1,2,3,4,4a,5,6,8a-octahydronaphthalene 1733 Alloaromadendrene Naphthalene, 1,2,3,5,6,8a-hexahydro-4,7-dimethyl-1-(1-methylethyl)-, (1Scis)(-)-.alpha.-Panasinsen
1743 1761
Ketones 4-Heptanone, 2,6-dimethyl2-Nonanone Cyclohexanone, 5-methyl-2-(1-methylethyl)-, cis3,7-Nonadien-2-one, 8-methyl-, (E)-
Terpene 3-Carene .beta.-Myrcene D-Limonene .beta.-Phellandrene .gamma.-Terpinene o-Cymene (+)-4-Carene l-Menthone .alpha.-Cubebene Caryophyllene .alpha.-Terpineol Geranyl acetate Estragole
Others Methyl isocyanide Ethene, fluoroPhenol, 2-nitro-
* LRI – Linear Retention Index.; X , – : presence and absence of the volatile compound . LTLT, T20, T30,T40,T50= conventional pasteurization, microfiltration at 20,30,40,50°C respectively.
28
29
29
30
Goat whey-orange juice beverage processed by microfiltration;
Microfiltration feeding temperatures were evaluated
Microfiltration preserved the volatile compounds and increased functional activity;
Lower temperatures maintained product consistency but affected microbial reduction
It was suggested a mild microfiltration processing temperature (between 30° and 40°C)
30