Physicochemical characteristics of anhydrous milk fat mixed with fully hydrogenated soybean oil

Food Research International 132 (2020) 109038

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Physicochemical characteristics of anhydrous milk fat mixed with fully hydrogenated soybean oil

T

Maria Isabel Landim Neves , Mayara de Souza Queirós, Rodolfo Lázaro Soares Viriato, Ana Paula Badan Ribeiro, Mirna Lúcia Gigante ⁎

Faculty of Food Engineering, University of Campinas UNICAMP, 13083-862 Campinas, São Paulo, Brazil

ARTICLE INFO

ABSTRACT

Keywords: Dairy fats Hard fat Crystallization modifiers Structuring oil Fats for the food industry

There is a growing demand for fats that confer structure, control the crystallization behavior, and maintain the polymorphic stability of lipid matrices in foods. In this context, milk fat has the potential to meet this demand due to its unique physicochemical properties. However, its use is limited at temperatures above 34 °C when thermal and mechanical resistance are desired. The addition of vegetable oil hard fats to milk fat can alter its physicochemical properties and increase its technological potential. This study evaluated the chemical composition and the physical properties of lipid bases made with anhydrous milk fat (AMF) and fully hydrogenated soybean oil (FHSBO) at the proportions of 90:10; 80:20; 70:30; 60:40; and 50:50 (% w/w). The increased in FHSBO concentration resulted in blends with higher melting point, which the addition of 10% of FHSBO increase the melting point in 12 °C of the lipid base. Also, FHSBO contributed for a higher thermal resistance conferred by the coexistence of polymorphs β' and β, which remained stable for 90 days. Co-crystallization was observed for all blends due to the total compatibility of milk fat with the fully hydrogenated soybean oil. The results suggest a potential of all blends for various technological applications, makes milk fat more appropriate to confer structure, and improve the polymorph stability in foods. The blends presenting singular characteristics according to the desired thermal stability, melting point, and polymorphic habit.

1. Introduction Milk fat is considered the most complex of all natural fats, with approximately 400 fatty acids varying according to the chain size and the saturation degree, consisting of hundreds triacylglycerols (TAGs) species, which vary in the number of carbon atoms (C24 to C56) (Vanhoutte, Dewettinck, Vanlerberghe, & Huyghebaert, 2003). This unique composition results in fat with an extended melting range (−40 °C to 40 °C), which is found as a mixture of liquid and crystallized fat at most processing, storage, and consumption temperatures (Sonwai & Rousseau, 2010; Vanhoutte et al., 2003; Vanhoutte, Dewettinck, Vanlerberghe, & Huyghebaert, 2002). Milk fat crystallizes predominantly in the polymorphic form β', which is maintained during the storage period probably by the large distribution of triacylglycerol species and the presence of more than 20% of palmitic acid (Herrera, de Leon Gatti, & Hartel, 1999; Lambert et al., 2018; Timms, 1984). The β' crystals tend to be smaller in size conferring softness and good sensory quality (Rousseau, 2000; Sonwai & Rousseau, 2010). Due to its unique texture and flavor characteristics, milk fat is used

⁎

in the manufacture of ice cream, confectionery, and chocolates (Sonwai & Rousseau, 2010; Vanhoutte et al., 2002, 2003). However, some foods can be affected by the ratio between liquid and crystallized fat, thus impairing its application. For example, most TAG in milk fat is crystallized up to 15 °C, which makes it a hard fat, unattractive to the consumer due to the lack of spreadability, such as butter. Thus, to increase the plasticity of milk fat while maintaining a pure milk-based product, the most used methods include the thermal fractionation of fat and the use of the olein phase to make butter (Queirós, Grimaldi, & Gigante, 2016) or the use of high-frequency ultrasound that modifies the fat crystallization behavior (Lee & Martini, 2019). The increase in milk fat plasticity can also be achieved by adding vegetable oils to milk fat, leading to the formation of spreads (Viriato, Queirós, Neves, Ribeiro, & Gigante, 2019). On the other hand, most of the milk fat is in the liquid form above 35 °C, which limits its use when high thermal and mechanical resistance are desired, for example in the manufacture of milk chocolate. A blend made with milk fat and wax (Kerr, Tombokan, Ghosh, & Martini, 2011) or partially hydrogenated vegetable oils (Shen, Birkett, Augustin, Dungey, & Versteeg, 2001) may be an alternative for

Corresponding author. E-mail addresses: [email protected] (M.I.L. Neves), [email protected] (A.P.B. Ribeiro), [email protected] (M.L. Gigante).

https://doi.org/10.1016/j.foodres.2020.109038 Received 20 August 2019; Received in revised form 22 January 2020; Accepted 25 January 2020 Available online 30 January 2020 0963-9969/ © 2020 Elsevier Ltd. All rights reserved.

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the technological applications that require thermal and mechanical resistance and a melting point higher than the body temperature. Among the hydrogenated vegetable oils, the hard fats obtained from the total hydrogenation of vegetable oils can be a technological alternative (Shen et al., 2001). Hard fats were introduced on the market due to the need to replace partially hydrogenated fats, which were associated with the risk of cardiovascular diseases and other metabolic and functional disorders due to the presence of trans fatty acids (Enig, 1996; FDA, 2015; Hunter, 2005; Mensink & Katan, 1990; Zevenbergen, Houtsmuller, & Gottenbos, 1988). Despite the ability to modulate the crystallization process (Omonov, Bouzidi, & Narine, 2010), the use of hard fats is limited, due to the incompatibility of their blends with vegetable oils, which can exhibit different melting point, molecular size, and polymorphic behavior (Guedes et al., 2014; Masuchi, Grimaldi, & Kieckbusch, 2014; Ribeiro, Grimaldi, Gioielli, & Gonçalves, 2009). The research hypothesis is that milk fat, which is naturally plastic and has unique physicochemical properties, can be modified by the addition of hard fats to form compatible and differentiated lipid bases. Soybean oil was selected among the hard fats, once it is available at low cost and consists predominantly (> 80%) of stearic acid (C18: 0), which has a neutral metabolic effect in relation to serum cholesterol (Lottenberg, 2009; Ribeiro, Basso, & Kieckbusch, 2013). Therefore, the aim of the study was to obtain and characterize blends made with different proportions of anhydrous milk fat (AMF) and fully hydrogenated soybean oil (FHSBO) to provide stable lipid bases with greater thermal and mechanical resistance for different technological applications.

2.1.2. Solid fat content (SFC), compatibility and melting point (MP) of control and blends The control and blends were melted at 80 °C for 30 min to completely erase the crystal memory. The samples were maintained in a dry thermostatic bath with temperature controlled by the Peltier Tcon 2000 system (Duratech, Garden Grove, USA) and subjected to tempering according to the AOCS Method Cd 16b-93 direct method I, with serial readings (AOCS, 2009). The SFC readings were performed in duplicate at 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, and 70 °C, with tempering for unstabilized fats, in a low-resolution pulsed Nuclear Magnetic Resonance spectrometer (NMR), Bruker pc120 Minispec (Silberstreifen, Rheinstetten, Germany). The compatibility of the blends was then evaluated from those data using a compatibility diagram by correlating the FHSBO ratio and the solid fat content at 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, and 65 °C as previously described in Quast, Luccas, Ribeiro, Cardoso, and Kieckbusch (2013). The melting point was calculated for the temperature corresponding to 4% solids content, obtained from the SFC curve given by NMR (Karabulut, Turan, & Ergin, 2004). 2.1.3. Thermal behavior The thermal analyses were performed by differential scanning calorimetry (DSC) (DSC Q2000 - TA Instruments, USA) using a calibration factor determined with indium, according to (AOCS, 2009). For that, 10 mg of sample were weighed on aluminum pans. An empty, hermetically sealed aluminum pan was used as reference. For the crystallization behavior, the analysis conditions were: isotherm at 80 °C for 10 min followed by cooling at a rate of 2 °C/min to −40 °C. For the melting behavior, the conditions were isotherm at −40 °C for 30 min, followed by a heating rate of 5 °C/min to 80 °C. The parameters onset temperature (To), final temperature (Tend), peak temperature (Tp), crystallization enthalpy (ΔH) and melting enthalpy were determined (AOCS, 2009; Biliaderis, 1983).

2. Materials and methods 2.1. Preparation and characterization of the blends The AMF and FHSBO were provided by Cargill Foods, Brazil. AMF was in accordance with the Brazilian legislation (Brasil, 1996, 2005). The binary AMF:FHSBO blends were prepared at the AMF: FHSBO ratios (w/w) of 90:10, 80:20, 70:30, 60:40, and 50:50. The AMF: FHSBO blends, AMF, and FHSBO controls were melted at 80 °C and stirred for 30 min to ensure complete melting and homogeneous blending. Subsequently, both the controls and blends were stored at 5 °C in an incubator chamber (Marconi, MA415/S) until analysis. The lipid bases were melted at 80 °C for 30 min to erase crystal memory prior to each analytical determination. They were then stabilized according to the protocols required for each determination.

2.1.4. Crystallization kinetics The samples were subjected to isothermal crystallization at 25 °C in a nuclear magnetic resonance spectrometer (NMR) equipment (Bruker pc120 Minispec, Germany), using high dry bath (0–70 °C ± 0.1 °C) (TCON 2000 - Duratech, EUA) and a Lauda circulator heater (E200 Ecoline-star edition). The blends were melted at 80 °C for 30 min to erase the crystal memory (Ribeiro, Grimaldi, Gioielli, dos Santos, et al., 2009). The crystallization kinetics was based on the non-linearized Avrami equation, as a function of the induction period, maximum solids content, and the crystallization stabilization time (Campos, 2004).

SFC(t) =1 SFCmax

2.1.1. Fatty acid composition The fatty acid composition was determined by gas chromatography using the Agilent 6850 GC USA chromatograph system (Santa Clara, CA, USA) after esterification as reported by (Hartman & Lago, 1973). The fatty acid methyl esters were methylated and separated according to the AOCS Ce 1f-96 method (AOCS, 2009) using a DB-23 AGILENT capillary column (50% cyanopropyl-methylpolysiloxane, 60 m long, internal diameter of 0.25 mm and 0.25 μm film thickness) using the following analysis conditions: flow rate of 1.0 mL/min; linear speed of 24 cm/s; detector temperature of 280 °C; injector temperature of 250 °C; oven temperature of 110 °C for 5 min, 110–215 °C at a rate of 5 °C/min, and 215 °C for 24 min; carrier gas: helium; volume injected: 1.0 μL, split ratio of 1:50. The qualitative composition was determined by comparing the peak retention times with those of the respective Sigma fatty acid standards. The quantitative composition was calculated by area normalization and expressed as mass percentage. The analysis was performed in triplicate.

e

kt n

(1)

where SFC (t) is the solid fat content (%) as a function of time (t), SFCmax is the limit solid fat content, k is the Avrami constant (min−1), which considers both nucleation and growth rate, and n is the Avrami exponent, which indicates the crystal growth mechanism (Wright, Hartel, Narine, & Marangoni, 2000). 2.1.5. Microstructure Crystal morphology was determined by polarized light microscopy (Olympus BX51, USA) coupled to a digital camera (Media Cybernetic, USA). The raw materials and blends were melted at 80 °C for 30 min in an oven, and a drop was placed on a preheated glass slide at 80 °C for 30 min with the aid of a capillary tube, which was covered with a cover slip. The slides were maintained in an incubation chamber (Marconi, MA415/S) at 25 °C and 40 °C for 24 h. Three images were captured using ImagePro Plus software version 7.0 (Media Cybernetic, USA), with polarized light and 20× magnification. Then, a quantitative

2

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analysis was performed using the following parameters: total number of crystalline elements, average density, and crystal diameter (Gamboa & Gioielli, 2006).

(C16: 0) and stearic acids (C18: 0) (Jensen, 2002; MacGibbon & Taylor, 2006) while FHSBO exhibited homogeneity in fatty acids with very high concentration (> 85%) of stearic acid, as can be seen in Table 1. Stearic acid is naturally present in soybean oil, and is also derived from the total hydrogenation from oleic (C18: 1), linoleic (C18: 2) and linolenic (C18: 3) fatty acids (Ribeiro et al., 2013). The addition of FHSBO to AMF resulted in a significant increase (p < 0.01) in the saturated fatty acid concentration (Table 1), and the proportions of 10 and 50% FHSBO resulted in an increase of 4.6 and 17.5% of total saturated fatty acids in the blends, respectively. This increase is mainly due to the higher concentration of stearic acid (C18: 0).

2.1.6. Polymorphism The polymorphic form of the fat crystals was determined after 7, 50, and 90 days of stabilization at 25 °C by X-ray diffraction (XRD), according to the AOCS Cj 2-95 method (AOCS, 2009). The analyses were carried out on a Philips PW 1710 diffractometer (PANalytical, Almelo, The Netherlands), using Bragg Brentano geometry (θ: 2θ) with Cu kα radiation (λ = 1.54178 Å, 40 KV voltage, and 30 mA current). Measurements were obtained using a step of 0.02° in 2θ and an acquisition time of 2 s, with scans of 5 to 30° (2θ scale). The identification of the polymorphic form was performed using the characteristic interplanar distances or short spacings of each crystal type. The relative proportions of the different crystal types were estimated by the relative intensity of short spacings (AOCS, 2009; Schenk & Peschar, 2004).

3.2. Physical properties As shown in Fig. 1, the addition of FHSBO to AMF was accompanied by a gradual and significant increase (p < 0.01) in the solid fat content, with compatibility for all AMF: FHSBO blends at the temperatures studied. Compatibility is represented by the linear evolution of the solid fat content as a function of the FHSBO concentration (Fig. 1b). The increase in the solids content was accompanied by an increase in the melting point of the blends (Table 2). The addition of 10% FHSBO resulted in a temperature increase of 12 °C in the melting point of the lipid base, which decreased with increasing the FHSBO concentration, with no changes observed for the melting point of the blends AMF: FHSBO 60:40 and 50:50. The AMF presents a plastic fat behavior, characterized by a complex fatty acids composition and presence of the long, medium, and short-chain fatty acids, which results in a slow decrease in solid fat content with increasing the temperature. On the other hand, FHSBO has a homogeneous fatty acid composition (~86% C18: 0) and maintains practically 100% solids at all temperatures, with an abrupt response to the decrease in solids fat at the temperature near to the melting point of stearic acid, since its composition has majority stearic acid (Fig. 1a). The increase in the FHSBO concentration altered the proportion of fatty acids, with no changes in the heterogeneity in fatty acids composition. The proportion between stearic acid (C18: 0)

2.2. Statistical analysis Analysis of Variance (ANOVA) was used to analyze the effects of the addition of fully hydrogenated soybean oil to anhydrous milk fat on the sum of saturated and unsaturated fatty acids composition, melting point, and solid fat content of the lipid bases. In case of difference, the averages were compared by the Tukey’s test at a significance level of 1%. Data were analyzed using STATISTICA 7.0 software (StatSoft Inc., USA). 3. Results and discussion 3.1. Chemical composition As previously reported in the literature, AMF presented heterogeneity in fatty acids composition, with high saturated fatty acid concentration (~73%) and predominance of myristic (C14: 0), palmitic

Table 1 Fatty acid composition (g/100 g of total fatty acids) of anhydrous milk fat (AMF) fully hydrogenated soybean oil (FHSBO) and their blends (90:10; 80:20; 70:30; 60:40; 50:50 AMF:FHSBO % w/w). Mean values ± standard deviation (n = 2). Different letters in the same lines show differences by Tukey’s test at 95% significance (p < 0.05). Fatty Acid

AMF:FHSBO (%w/w) AMF

90:10

80:20

70:30

60:40

50:50

FHSBO

C4:0 C6:0 C8:0 C10:0 C12:0 C14:0 C15:0 C16:0 C16:1 C17:0 C17:1 C18:0 ∑C18:1 t C18:1 cis 9 ∑C18:2 t C18:2 cis 9. 12 ∑C18:3 t C18:3 cis 9. 12. 15 C18:4 C20:0 C20:1 C22:0 C24:0

1.01 1.76 1.28 2.91 4.89 13.00 1.35 35.20 2.08 0.86 0.30 10.24 2.59 17.52 1.05 1.51 0.20 0.67 1.23 0.19 0.10 0.08 0.00

1.06 ± 0.13 1.83 ± 0.16 1.30 ± 0.07 2.78 ± 0.04 4.58 ± 0.03 11.82 ± 0.05 1.22 ± 0.01 31.73 ± 0.21 1.91 ± 0.03 0.79 ± 0.01 0.25 ± 0.01 18.59 ± 0.1 2.24 ± 0.12 15.08 ± 0.18 0.94 ± 0.03 1.41 ± 0.01 0.14 ± 0.02 0.60 ± 0.00 1.14 ± 0.02 0.26 ± 0.00 0.16 ± 0.02 0.12 ± 0.01 0.05 ± 0.02

0.72 ± 0.11 1.36 ± 0.16 0.99 ± 0.05 2.20 ± 0.08 3.77 ± 0.11 10.01 ± 0.39 1.05 ± 0.04 29.49 ± 0.16 1.61 ± 0.07 0.71 ± 0.01 0.24 ± 0.02 27.36 ± 0.32 2.01 ± 0.09 13.95 ± 0.65 0.79 ± 0.04 1.24 ± 0.03 0.17 ± 0.03 0.54 ± 0.04 0.98 ± 0.06 0.32 ± 0.00 0.146 ± 0.02 0.19 ± 0.00 0.08 ± 0.00

0.81 ± 0.03 1.02 ± 0.34 0.82 ± 0.07 2.05 ± 0.93 3.41 ± 0.04 8.95 ± 0.62 0.94 ± 0.01 26.57 ± 0.17 1.42 ± 0.06 0.63 ± 0.00 0.17 ± 0.0 34.78 ± 1.02 2.27 ± 0.018 12.05 ± 0.87 0.71 ± 0.05 1.13 ± 0.16 0.12 ± 0.0 0.45 ± 0.03 0.87 ± 0.08 0.37 ± 0.03 0.06 ± 0.01 0.20 ± 0.07 0.10 ± 0.008

0.59 ± 0.11 0.83 ± 0.05 0.65 ± 0.03 1.62 ± 0.09 2.72 ± 0.16 7.14 ± 0.38 0.76 ± 0.03 24.36 ± 0.28 1.07 ± 0.02 0.56 ± 0.01 0.14 ± 0.01 44.47 ± 1.35 1.75 ± 0.11 9.72 ± 0.56 0.57 ± 0.02 0.94 ± 0.03 0.09 ± 0.01 0.38 ± 0.01 0.68 ± 0.04 0.43 ± 0.01 0.09 ± 0.00 0.25 ± 0.01 0.099 ± 0.03

0.52 ± 0.03 0.66 ± 0.02 0.54 ± 0.02 1.36 ± 0.04 2.35 ± 0.01 6.12 ± 0.07 0.65 ± 0.01 21.98 ± 0.28 0.92 ± 0.05 0.48 ± 0.01 0.11 ± 0.01 49.87 ± 1.10 1.49 ± 0.03 9.15 ± 0.90 0.47 ± 0.01 1.28 ± 0.18 0.08 ± 0.01 0.32 ± 0.01 0.56 ± 0.02 0.48 ± 0.01 0.03 ± 0.00 0.32 ± 0.02 0.14 ± 0.01

– – – – – 0.10 0.14 10.88 – 0.20 – 86.01 – – – – – – – 0.69 – 0.48 0.17

∑Saturated ∑Unsaturated

72.76 27.23

76.14e 23.85a

78.30d 21.69b

80.71c 19.28c

84.53b 15.46d

85.53a 14.46e

100.00 0.00

3

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100

(a)

80

90:10 80:20

60

70:30

40

60:40

20 0

50:50 FHSBO 10

100

Solid fat content (SFC) (%)

energy. The melting event of FHSBO shows successive endothermic, exothermic and endothermic events due to polymorphic changes occurring on heating. The melting event started at 49.83 °C and finished at 57.79 °C, and the recrystallization started in 55.72 °C until 59.96 °C. Finally, a other melting curve started in 58.53 °C and finished in 68.85 °C. The three events releasing 141.65 J/g. Additionally, a peak exceeding the baseline was observed, which represents the crystallization of SSS TAGs during the melting event (Fig. 3g). Such behavior is characteristic of high melting point fats that are not yet fully stabilized (Humphrey & Narine, 2004). The addition of FHSBO to the AMF changed the thermal behavior of the lipid bases. The addition of 10% FHSBO (AMF: FHSBO 90:10) resulted in a displacement of approximately 15 °C in the initial crystallization temperature, from 18.13 to 33.31 °C (Fig. 2b and Table 3), with little changes observed for the other FHSBO concentrations (Table 3). The blends containing 40 and 50% FHSBO (AMF: FHSBO 60:40 and 50:50) exhibited two distinct crystallization peaks (Fig. 2e, f), with the first peak corresponding to the blend of AMF and FHSBO, and a second peak corresponding to the crystallization of low melting TAGs from AMF. Similarly, the addition of 10% FHSBO to the AMF increased the final melting temperature by approximately 15 °C, ensuring a greater thermal resistance to the lipid base (Fig. 3). Similar behavior was observed by Ribeiro et al. (2013) in lipid blends containing hard fat of Crambe, soybean, cotton, palm kernel, pal, and cocoa butter, and by de Oliveira, Ribeiro, dos Santos, Cardoso, and Kieckbusch (2015) in blends containing hard fat of palm, palm kernel, soybean, cotton and palm oil. Both research groups reported that the narrow melting peaks were due to the high concentration of saturated long-chain fatty acids. As can be seen in Fig. 4, the AMF showed a spherulite structure at 25 °C, corresponding to the aggregation of crystalline lamellae growing radially from the same central nucleus (Narine & Humphrey, 2004; Rousset, 2002), while the FHSBO presented crystals known as Maltese cross, at 25 and 40 °C, which is a well-defined spherulite, characteristic of lipid bases with homogeneous TAGs composition (Narine & Humphrey, 2004). Both the addition of FHSBO and the parameter temperature modified the structure of the crystal lattice. At 25 °C, the crystal lattice structure is composed of crystals from both AMF and FHSBO, while only crystals from FHSBO were observed at 40 °C, due to the complete melting of AMF at this temperature. It is clearly seen in the micrographs (Fig. 4) that, at both temperatures, the increase in FHSBO concentration resulted in the formation of a denser crystal lattice and with fewer crystalline elements with a lower number of crystalline suprastructures. Fig. 5 shows the diffractograms at 25 °C after 7, 50, and 90 days of stabilization, respectively, while Table 4 shows the polymorphic forms of the raw materials and blends over time. It is observed that AMF showed a polymorphic habit in β' at all periods evaluated (peaks 4.2 and 3.8 Å) (Fig. 5a). The peak 4.0 Å observed is probably related to the olein TAG crystals present in AMF (Truong, Morgan, Bansal, Palmer, & Bhandari, 2015). The polymorphic habit of milk fat is due to the diversity of the TAG structure (Lambert et al., 2018; O’brien, 2008; Wright et al., 2000). The β-form is commonly found in milk fat (Lambert et al., 2018), although other possible forms can be found, such as γ, α, and β depending on the type of stabilization (Lopez & Ollivon, 2009; Ten Grotenhuis, Van Aken, Van Malssen, & Schenk, 1999). After 7 and 50 days of stabilization, FHSBO exhibited a polymorphic habit in β' (Fig. 5g, Table 4). However, a low-intensity peak, typical of β-form, appeared after 90 days of stabilization (Fig. 5g, Table 4). Hard fat of soybean crystallizes predominantly in the β-form, and may rarely develop the β'-form (Shukla, 1995). When analyzing together, both the melting event and the polymorphic behavior can explain the crystalline behavior of FHSBO. In the melting process, a recrystallization peak (Fig. 2) was clearly observed, suggesting instability of the polymorphic form in FHSBO, leading to the formation of β' crystals, as observed by X-

AMF

20

30

40

50

60

Temperature (C°)

70 100

(b)

90

90

10 °C 15 °C

80

80

20 °C

70

70

25 °C

60

60

30 °C

50

50

35 °C

40

40

30

30

20

20

10

10

0

0

0

10

20

30

% FHSBO in the blend

40

50

40 °C 45 °C 65 °C 50 °C 55 °C 60 °C

Fig. 1. Solid fat content (%) (a) and compatibility diagram (b) of anhydrous milk fat and the blends (90:10; 80:20; 70:30; 60:40; 50:50 AMF:FHSBO % w/w) at different temperatures. Table 2 Melting points of AMF, FHSBO and blends (90:10; 80:20; 70:30; 60:40; and 50:50 AMF:FHSBO % w/ w). Blend

Melting Point (C°)

AMF 90:10 80:20 70:30 60:40 50:50 FHSBO

34.34 46.86 53.83 59.53 64.99 64.74 69.40

± ± ± ± ± ± ±

0.03ª 0.04b 0.02c 0.03d 0.01e 0.06e 0.04f

and the other fatty acids in AMF increased from 0.10 to 0.19, 0.27, and 0.34 in the blends AMF: FHSBO 90:10, 80:20, and 70:30, respectively. Thus, the blend AMF: FHSBO 70:30 presented a melting point close to a typically homogeneous fat (~60 °C), containing 32 and 57% liquid fat at 25 °C and 37 °C, respectively (Fig. 1a). The crystallization behavior and the melting points of raw materials and lipid bases are shown in Figs. 2 and 3, respectively. It is observed that the AMF presented crystallization (Fig. 2a) and melting (Fig. 3a) curves spreading on a wide range of temperatures due to the variety of short, medium, and long-chain fatty acids. The crystallization event started at 18.13 °C and finished at −25.41 °C, releasing 51.76 J/g of energy to crystallize, while the melting event started at −18.17 °C and finished at 48.71 °C, consuming 65.71 J/g of energy. The melting peak is divided into three temperature ranges (Fig. 3a), as follows: melting temperatures from − 40 to 12 °C correspond to crystals formed by low melting point TAGs; from 12 to 25 °C correspond to medium melting point TAGs, and temperatures from 25 to 40 °C represent the high melting point TAGs (Timms, 1984). The AMF melting point was 34 °C. The FHSBO presented narrow crystallization (Fig. 2g) and melting curves (Fig. 3g), reflecting its homogeneous TAGs composition, mainly characterized by SSS (S = stearic acid). The crystallization event of FHSBO started at 51.07 °C, and finished at 39.51 °C, with the release of 116.40 J/g of 4

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Fig. 2. Thermal behavior of crystallization of AMF, FHSBO and the blends (90:10, 80:20, 70:30, 60:40, 50:50 AMF:FHSBO %w/w).

ray diffraction. The occurrence of a low-intensity peak in β after 90 days suggests that, even slowly, the FHSBO stabilized in its most stable polymorph form (β). Except for the blend AMF: FHSBO 90:10, which showed β' crystals after 7 days, the other blends presented crystals in the β' and β forms in different intensities for all periods evaluated (the peaks 4.6 or 4.5, and 3.8, 3.7 or 3.6 Å represent the β form). This result is unexpected for β'-stabilized raw materials, once they should result in blends crystallized in β' form. The crystallization behavior in the β 'and β forms observed for all blends (AMF: FHSBO 90:10, 80:20, 70:30, 60:40, and 50:50) was probably due to the cocrystallization of milk fat and fully hydrogenated soybean oil, which accelerated the polymorphic stabilization process by providing activation energy for the polymorphic transition from β' to β (Narine & Humphrey, 2004).

equilibrium between the liquid and solid fat (Fig. 1a) which characterizes the desirable plasticity from AMF. At body temperature (~37 °C) the blends AMF: FHSBO 90:10 and 50:50 exhibited approximately 40 and 85% liquid fat, respectively. In contrast, although the blends AMF: FHSBO 70:30, 60:40, and 50:50 exhibited good plasticity, their high melting points of approximately 60–70 °C (Table 2) can be used as seeds to modulate crystallization processes. Lipid bases to produce bakery shortenings, such as bread, puff pastry, and cakes, should present 30 and 10% solid fat contents at 25 °C and 40 °C, respectively, suggesting the suitability of the blend AMF: FHSBO 90:10 for this purpose (Ab Latip et al., 2013). The blend AMF: FHSBO 90:10 has the potential for application in cookies and fillers once it exhibited thermal and mechanical resistance at room temperature and β' crystals that promote better creaming (Wilderjans, Luyts, Brijs, & Delcour, 2013). In addition, the presence of liquid fat in the system can provide softness and a pleasant sensation on the palate (Shahidi & Zhong, 2005). The blend AMF: FHSBO 80:20, with melting point near 54 °C and polymorphic stabilization in β′ and β forms suggests its application potential for the manufacture of milk chocolate to inhibit or delay the onset of fat bloom, a defect associated with the increase in molecular mobility of the system and consequent polymorphic transition of the cocoa butter crystals from the βV to βVI form (Lonchampt & Hartel, 2004). The blend AMF: FHSBO 80:20 may increase the physicochemical stability of the system and avoid the molecular mobility necessary for the transition βV → βVI and consequent onset of fat bloom (Hodge & Rousseau, 2002; Sonwai & Rousseau, 2010) Another application potential is the use of the blends AMF: FHSBO 90:10 and 80:20 as wall materials for the microencapsulation of functional ingredients. Both blends showed thermal stability, which favors

3.3. Technological applications in the food industry The addition of FHSBO to AMF modified the composition and the physical properties of milk fat, thus improving its technological potential by increasing both the melting point and the thermal resistance due to the coexistence of polymorphs β′ and β. In general, β′ crystals tend to be smaller and to confer softness, whereas β crystals give greater stability to lipid bases. The polymorphism parameter, associated with the solid fat content, demonstrates the functionality of fat for application in the food industry (Mattice & Marangoni, 2017). The solid fat has a crystalline form, indicates its behavior regarding the appearance, texture, oil exudation, air incorporation, structural integrity, mouth feel, and physical stability. From the technological point of view, taking the body temperature (~37 °C) as a reference, all blends (AMF: FHSBO 90:10, 80:20, 70:30, 60:40, and 50:50) showed 5

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Fig. 3. Thermal behavior of melting of AMF, FHSBO and the blends (90:10, 80:20, 70:30, 60:40, 50:50 AMF:FHSBO %w/w). Table 3 Onset temperature (To), end temperature (Tend), peak temperature (Tp), and enthalpy (ΔH) of crystallization and melting of AMF. FHSBO and blends (90:10; 80:20; 70:30; 60:40; and 50:50 AMF:FHSO % w/w). Event

Blend

Peak

T(o) (°C)

Crystallization

AMF 90:10 80:20 70:30 60:40

– – – – 1 2 Total 1 2 Total –

18.13 33.71 37.98 41.01 43.31 19.12

– – – 1 2 Total 1 2 Total 1 2 Total 1 2 Recrystallization Total

50:50 FHSBO Melting

AMF 90:10 80:20 70:30 60:40 50:50 FHSBO

T(end) (°C)

T(p) (°C)

ΔH (J/g)

−24.51 −19.87 ± 3.46 −17.47 ± 1.47 (−)14.17 ± 0.33 19.12 ± 0.33 (−)10.78 ± 1.10

8.67 29.14 ± 0.08 34.68 ± 0.26 37.14 ± 0.41 40.13 ± 0.17 6.08 ± 0.04

44.62 ± 0.22 −2.67 ± 0.22

21.20 ± 1.13 15.51 ± 1.13

41.65 ± 0.29 6.28 ± 0.05

51.07

39.51

49.12

51.76 61.46 ± 3.17 66.41 ± 3.10 68.44 ± 0.52 68.29 ± 1.01 12.16 ± 0.54 80.45 ± 1.55 79.91 ± 3.55 5.37 ± 0.31 85.28 ± 3.86 116.40

−18.17 −14.84 ± 1.98 −7.61 ± 1.08 −5.72 ± 0.14 37.65 ± 1.62

48.71 64.17 68.52 33.42 65.31

4.14 ± 0.25 40.15 ± 0.63

30.30 ± 1.25 66.41 ± 0.27

16.24 ± 0.05 66.25 ± 0.21

3.89 ± 0.12 41.89 ± 0.50

25.82 0.36 67.29 ± 0.75

16.13 ± 0.05 64.24 ± 0.02

49.83 58.53 55.72

57.79 68.85 59.96

54.37 63.92 57.41

± ± ± ± ±

0.28 0.47 0.33 0.66 0.33

6

± ± ± ±

1.76 1.46 1.37 1.37

19.06 17.66 16.80 60.74 16.27

± ± ± ±

0.15 0.32 0.285 0.01

65.71 71.48 ± 1.71 67.26 ± 2.18 21.58 ± 0.71 29.88 ± 1.7 51.43 ± 2.41 15.89 ± 0.74 35.9 ± 0.03 51.79 ± 0.77 10.33 ± 0.98 52.19 ± 0.6 62.52 ± 1.64 52.57 107.60 18.67 141.65

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Fig. 4. Microscopy images under polarized light of AMF, FHSBO and blends (90:10; 80:20; 70:30; 60:40; and 50:50 AMF:FHSBO %w/w) after stabilization at 25 °C and at 40 °C. 20× magnification.

the entrapment of different functional components in the crystal network, such as vitamins, fatty acids, probiotic cultures, polyphenols, and carotenoids (Darragh & Stone, 1975; Okuro, de Matos Junior, & FavaroTrindade, 2013; Viriato et al., 2020) In addition, it is worth emphasizing that the predominance of milk fat in the lipid bases favors its use for applications in the food industry, once milk fat is appreciated by its unique sensory characteristics.

4. Conclusion The blends made with milk fat and fully hydrogenated soybean oil presented structural, thermal, and mechanical stability. The crystalline form observed in the blends was an unexpected and positive result, which characterizes the synergistic effect of both fats. Although both fats exhibited the polymorphic habit in β', the resulting lipid bases 7

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AMF 4.2 Å

1600

4.6 Å

50 days

3.8 Å

90 days

1200 800 400 0

20

25

2θ (degree)

4.6 Å

3.8 Å

90 days

1000 500

15

20

4.6 Å

1800

25

4.2 Å

30

7 days

3.8 Å

50 days

1800

90 days 1200 600 0

30

15

20

2θ (degree)

25

30

50:50

4.2 Å

7 days

3.8 Å

3000

50 days

Intensity (a.u)

Intensity (u.a)

2θ (degree)

25

2θ (degree)

70:30

60:40

2400

90 days

1200 600 0

20

4.6 Å

50 days

1500

0

15

7 days

Intensity (a.u)

Intensity (a.u)

2000

90 days

500

2400

4.2 Å

50 days

1000

30

80:20

7 days 3.8 Å

1500

0 15

4.2 Å

2000

7 days

Intensity (a.u)

Intensity (a.u)

2000

90:10

4.2 Å

7 days 3.8 Å

2400

4.6 Å

50 days 90 days

1800 1200 600

15

20 2θ (degree) 25

0

30

15

20 2θ (degree) 25

30

FHSBO

3000

4.2 Å

7 days

2400 Intensity (a.u)

50 days 3.8 Å

1800

90 days

4.6 Å

1200 600 0

15

20

2θ (degree)

25

30

Fig 5. X-ray diffraction patterns of AMF, FHSBO, and blends at 25 °C at 7, 50, and 90 days.

showed crystals in β and β' form in different intensities up to 90 days of storage. Co-crystallization, which is a basic requirement for technological application, was observed for all blends due to the total compatibility of milk fat with the fully hydrogenated soybean oil. Additions of

up to 20% fully hydrogenated soybean oil were enough to increase the technological significance of milk fat without losing its natural characteristic of solid and liquid domains in the system. A fat system modulated by fully hydrogenated soybean oil and liquid domains of 8

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Table 4 Short spacing and polymorphic forms of the AMF, FHSBO and blends (90:10, 80:20, 70:30, 60:40, 50:50 AMF:FHSBO % w/w) in 7, 50 and 90 days of storage at 25 °C. Blend

Day

Short spacing (Å) 4.6

GAL

4.4

7 50 90

Polymorphic form 4.2

4.1

3.8

3.7

4.2(vs)

4.0(s) 4.1(vs) 4.1(vs)

3.77(s) 3.95(vs) 3.95(s)

3.7(s) 3.7(s)

β′ β′ β′

3.73(vs)

β′ β′ sub β β′sub β

4.2(vs)

90:10

7 50 90

80:20

7 50 90

4.52(m) 4.48(vs) 4.50(vs)

70:30

7 50 90

4.51(vs) 4.48(vs) 4.47(vs)

60:40

7 50 90

4.53(vs)

50:50

7 50 90

4.54(s) 4.46(s) 4.47(s)

HFS

7 50 90

4.44(m) 4.44(m)

4.19(vs)

4.16(vs) 4.16(vs) 4.16(vs)

4.42(s) 4.44(vs)

4.17(vs)

4.18(vs)

4.16(vs) 4.51(w)

4.15(vs) 4.15(vs) 4.13(vs)

4.12(vs) 4.12(vs) 4.1(vs) 4.1(vs) 4.12(vs) 4.09(vs) 4.11(vs) 4.13(vs)

3.77(s) 3.75(vs) 3.76(vs)

3.6

3.74(vs)

β′sub β β′~ β β′~ β

3.76(vs) 3.8(vs) 3.8(vs)

3.65(m) 3.74(vs) 3.74(vs)

β′~ β β′ ~ β β′ ~ β

3.79(s)

3.66(s) 3.72(vs) 3.74(vs)

3.58(m) 3.6(s)

β′~ β β′sub β β′~ β

3.77 (s)

3.65(m) 3.72(vs) 3.72(vs)

3.62(m) 3.59(m)

β′sub β β′ sub β β′ sub β

3.75(vs)

3.78(m) 3.76(s) 3.8(s)

β′ β′ β′sub β

Intensities: vs. very strong; s. strong; m. médium; w. weak. β′ sub β: β′(vs) and β (s/m/w); β′~ β: β′ and β (vs); β′≪ β: β′(s) and β (vs); β′≪ < ≪ β: β′(m) and β(vs).

milk fat can be used under different technological perspectives, especially as a crystallization inductors and as transport of functional compounds.

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CRediT authorship contribution statement Maria Isabel Landim Neves: Conceptualization, Methodology, Writing - original draft, Formal analysis, Investigation. Mayara de Souza Queirós: Conceptualization, Methodology, Writing - review & editing. Rodolfo Lázaro Soares Viriato: Conceptualization, Methodology, Writing - review & editing. Ana Paula Badan Ribeiro: Writing - review & editing, Supervision. Mirna Lúcia Gigante: Writing - review & editing, Supervision, Project administration. Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Acknowledgments The authors thank Cargill Foods for the donation of anhydrous milk fat and fully hydrogenated soybean oil. This study was financed in part by The Coordination for the Improvement of Higher Education Personnel – Brazil (CAPES) – Finance Code 001. The authors Neves and Viriato thank CAPES for granting the Ph.D. scholarship; Queirós thanks National Council for Scientific and Technological Development - Brazil (CNPq) for the post-doctoral scholarship (157961/2018-4); and Ribeiro thanks CNPq for the productivity grant (423082/2018-3). Appendix A. Supplementary material Supplementary data to this article can be found online at https:// doi.org/10.1016/j.foodres.2020.109038. 9

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