Food Research International 51 (2013) 510517

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Food Research International

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A rheological characterisation of an olive oil/fatty alcohols organogel

F.R. Lupi, D. Gabriele , V. Greco, N. Baldino, L. Seta, B. de Cindio
University of Calabria, Department of Engineering Modelling, Via P. Bucci - Cubo 39C, I-87036 Rende (CS), Italy

a r t i c l e

i n f o

a b s t r a c t
Policosanol is a mixture of fatty alcohols well known for its nutraceutical effects on consumers' health. In the present paper, the potential ability of this food-grade additive to organogel vegetable oils has been investigated by preparing samples based on virgin olive oil at different policosanol concentrations. The onset of crystallisation temperature (Tco) and the gelation temperature (Tg) were evaluated by carrying out dynamic temperature ramp tests, whereas the rheological characteristics of samples at temperatures lower than the crystallisation onset were studied with frequency sweep tests at 25 C. Experimental data have shown that crystallisation occurs for policosanol concentration larger than 0.1% w/w whereas gelation (i.e. the formation of a structured 3D network) can be observed only for concentrations larger than a critical value ranging between 0.3% and 0.5% w/w. The onset of crystallisation temperature increases nonlinearly with organogelator amount even if an asymptotic trend seems to be present for concentrations larger than 50% w/w. A fractal model was used to t experimental storage modulus data, G, as a function of policosanol amount, and a fractal dimension equal to 2.75, in agreement with literature values, was found. Finally, a phenomenological equation based on a modied fractal model, was proposed to t, with a good agreement, the experimental Tco values as a function of the policosanol fraction. 2013 Elsevier Ltd. All rights reserved.

Article history: Received 3 September 2012 Accepted 5 January 2013 Available online 17 January 2013 Keywords: Organogel Oleogel Policosanol Olive oil Crystallisation Rheology

1. Introduction Organogels are solid-like systems structured in an organic solvent (also named Oleogels when the solvent is an edible oil (Dassanayake, Kodal, & Ueno, 2011; Marangoni & Garti, 2011)); their production is based on the action of low molecular-weight organic gelators (LMOG) or polymeric gelators (PG), which can self-assemble forming threedimensional networks and stabilising the solvent within (Gronwald, Snip, & Shinkai, 2002; Hughes, Marangoni, Wright, Rogers, & Rush, 2009). In both aqueous (hydrogels) and organic (organogels) solvents the gelation process is mainly based on attractive forces occurring among small molecule aggregates (Co & Marangoni, 2012) or bres (Shapiro, 2011). The resulting weak secondary interactions, hydrogen bonding, interaction, van der Waals forces between long alkyl chains, and electrostatic forces, (Hughes et al., 2009; Kim, Kim, Lee, & Lee, 2010) lead to the formation of a structure of nanometre to micrometre dimension. The interest in organogel properties is progressively increasing for different elds and industrial applications (Marangoni & Garti, 2011); among them, the rheological structuration and stabilisation of vegetable oil-based shortenings, emulsions or suspensions are promising technologies adopted to replace saturated and/or trans fats (Duffy et al., 2009;
Corresponding author at: Department of Engineering Modelling, University of Calabria, Via P. Bucci, Cubo 39C, I 87030 Rende (CS), Italy. Tel.: +39 0984 496687; fax: +39 0984 494009. E-mail address: domenico.gabriele@unical.it (D. Gabriele). 0963-9969/$ see front matter 2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.foodres.2013.01.013

Lupi, Gabriele, de Cindio, Snchez, & Gallegos, 2011; Lupi, Gabriele, Baldino et al., 2012; Pernetti, van Malssen, Flter, & Bot, 2007; Rogers, Wright, & Marangoni, 2008; Rogers, Wright, & Marangoni, 2009). Many different fat-based foods are characterised by a typical viscoelastic structure given by the mechanical properties of hard fats and their interactions (Ghotra, Dyal, & Narine, 2002; Marangoni, 2009). Such a structure is very often improved by the saturation of the aliphatic tails of vegetable liquid oils fatty acids throughout catalytic hydrogenation processes (Singh, Rezac, & Pfromm, 2009). This implies the production of trans fatty acids isomers (TFAs) dangerous for consumers' health (Blanco Muoz, 2004; Marangoni et al., 2007). In order to avoid any chemical rearrangement of fatty acids chains, organic gelators are used as an interesting alternative, promoting the formation of a crystalline network able to give the proper rheological properties to foodstuffs and to stabilise them in the meantime, at a very low concentration (~0.52% w/w) (Hughes et al., 2009). According to the literature, a critical challenge for commercial application of such systems is the discovery of new organogelators, especially the food-grade ones (Co & Marangoni, 2012; Rogers et al., 2009). Different organogelator molecules for food oils are under investigation, such as triacylglycerols (TAGs), diacylglycerols (DAGs), monoacylglycerols (MAGs) (Calligaris, Da Pieve, Arrighetti, & Barba, 2010; Da Pieve, Calligaris, Co, Nicoli, & Marangoni, 2010; Lupi et al., 2011; Lupi, Gabriele, Facciolo et al., 2012; Lupi, Gabriele, & de Cindio, 2012; Ojijo, Neeman, Eger, & Shimoni, 2004), fatty acids and fatty alcohols (Schaink, van Malssen, Morgado-Alves, Kalnin, & van der Linden, 2007), sorbitan tri-stearate and/or lecithin (Pernetti, van Malssen, Kalnin,

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& Flter, 2007; Shchipunov, 2001), sterol and -oryzanol (Bot, den Adel, Roijers, & Regkos, 2009), waxes (Toro-Vazquez, Morales-Rueda, Ajay Mallia, & Weiss, 2010; Toro-Vazquez et al., 2007), ethylcellulose (Laredo, Barbut, & Marangoni, 2011; Zetzl, Marangoni, & Barbut, 2012) and others. As far as fatty alcohols are concerned, some authors investigated their potential application as organogelator molecules. Schaink, van Malssen, Morgado-Alves, Kalnin, and van der Linden (2007) tested a mixture of stearic acid and stearyl alcohol, nding that the mixture of fatty acid and alcohols gives a different crystal morphology with respect to pure components, resulting in a better structured network. Gandolfo, Bot, and Flter (2004) investigated the organogelation of different fatty acids and fatty alcohols with aliphatic chain lengths in the range of 2230 carbons; the authors studied systems made by sunower oil and palmityl alcohol, stearyl alcohol, arachyl alcohol and behenyl alcohol respectively. The authors found that, generally, the fatty alcohols yielded harder products than the fatty acids at the same concentration, and that the minimum concentration at which oil structuration was achieved was about 2% (w/w). Moreover, according to Co and Marangoni (2012), the fatty alcohol gels are formed by large crystals, probably arranged as stacks of platelets. If fatty alcohols and fatty acids are mixed in a particular ratio, small needle-like crystals are observed, and they are, maybe, responsible for the rheology of the nal systems. An indirect application of fatty alcohols for oil structuration is the addition of waxes extracted from vegetable sources, such as Candelilla wax or other (Co & Marangoni, 2012); in particular, for example, Candelilla wax is a mixture of high-melting alkanes, sterols, fatty alcohols, fatty acids and waxy esters extracted from Yerba evergreen shrub (Euphorbia cerifera) which was deeply investigated, as organogelator, by Toro-Vazquez et al. (2007). In the present work, a virgin olive oil was structured with a particular category of fatty alcohols, commercially named policosanol. This food-grade additive can be extracted from different vegetable raw materials, such as sugar cane wax, beeswax, rice bran and wheat germ (Francini-Pesenti, Brocadello, Beltramolli, Nardi, & Caregaro, 2008; Irmak, Dunford, & Milligan, 2006). Policosanol is a mixture of fatty primary alcohols characterised by an aliphatic chain in the range of 2036 carbon, mainly composed of docosanol (C22), tetracosanol (C24), hexacosanol (C26), octacosanol (C28), and triacontanol (C30) (Irmak et al., 2006). According to the literature, policosanol exerts several positive effects on human health, most of all in prevention and treatment of cardiovascular diseases (Xu, Fitz, Riediger, & Moghadasian, 2007). In particular, octacosanol should be able to decrease lipid levels, lowering the LDL (low density lipoprotein) in the blood, and, moreover, in the last few years, it has been used as an enhancer for athletic performance (Xu et al., 2007). In the present work, the investigation of the rheological characteristics of organogels produced with policosanol (mainly composed of octacosanol) as the organogelator and a virgin olive oil as the organic solvent was carried out by using Small Amplitude Oscillation Tests (SAOTs). The main aim of the investigation was the characterisation of gels obtained with different amounts of policosanol, aiming at better understanding the role of organogelator. The onset of crystallisation temperature (Tco) was evaluated for each organogel, whereas a gelation temperature (Tg), evidencing the formation of a three-dimensional structure, was observed for samples having a gelator concentration larger than a critical value. Furthermore, the effect of the process temperature applied during the preparation of the samples on their main rheological characteristics was evaluated. 2. Materials and methods 2.1. Sample ingredients and preparation The investigated organogels were produced with a virgin olive oil supplied by a local supermarket (De Santis, Italy) as the solvent, and

policosanol (with a total amount of 60% w/w of octacosanol) as the organogelator molecule (A.C.E.F., Italy). The fatty alcohol composition of the organogelator, extracted by rice bran wax, was supplied by the producer and it is listed in Table 1. According to the manufacturers, virgin olive oil is mainly composed of mono-unsaturated fatty acids (about 75% w/w), the rest being saturated (about 15% w/w) and poly-unsaturated (about 10% w/w). All gels were produced and investigated following a procedure elsewhere adopted for a different organogelator (Lupi et al., 2011; Lupi, Gabriele, Facciolo et al., 2012; Lupi, Gabriele, & de Cindio, 2012): the policosanol was added to the oil previously warmed up, and the rheological properties of the system obtained were analysed cooling the sample in the rheometer cell, with a controlled thermal ramp, from the temperature of preparation down to lower values. Aiming at investigating the effect of the preparation temperature, samples containing 1%, 2% and 3% (w/w) of policosanol were prepared adding the proper amount of gelator to the oil previously warmed up to the different temperatures of 70 C, 85 C and 97 C in a water bath thermostated by a plate heater (Jolly 2, Falc Instruments, Italy). All other samples were prepared at 85 C. The oil phase was continuously mixed with a laboratory stirrer (RW 20, IKA-Werke, Germany) and maintained at the chosen temperature until the test was performed. Compositions and identiers of tested samples are listed in Table 2. 2.2. Rheological characterisation For all samples a rheological characterisation based on Small Amplitude Oscillation Tests (SAOTs) was carried out. Tests were performed with a controlled stress rheometer DSR-200 (Rheometric Scientic, USA) equipped with a parallel plate geometry ( = 40 mm, gap 1.0 0.1 mm) and a Peltier system (0.1 C) acting under the lower plate. Preliminary stress sweep tests were performed at different temperatures from the preparation value (i.e. 70, 85, 97 C) down to 20 C, approximately every 10 C to investigate the potential changes in linear viscoelastic region as a function of temperature; in the range where a transition was observed, a larger number of stress sweep tests were carried out to be able to describe more nely the change of the linear viscoelastic conditions due to the material gelation. Temperature ramp tests were, then, carried out at the xed frequency of 1 Hz, cooling the sample from the temperature of preparation, according to the considered sample, down to 20 C with a cooling rate of 1 C/min. For each test, the applied stress was modied with the temperature, applying different constant values in different temperature range, following the results obtained in the previously described stress sweep. In this way it was possible to adapt the applied stress to the change in material structure, always guaranteeing the linear viscoelastic conditions. In order to better control the thermal history for all tests, the warm organogel was poured on the lower plate of the rheometer previously heated up to the desired temperature of preparation (70, 85 or 97 C). All samples were prepared three times and the presented results are the obtained average values. Differences of measurement are shown by error bar (standard deviation), and the resulting coefcient of variation was typically lower than 10%. Larger deviations (up

Table 1 Fatty alcohol composition of policosanol used for organogels preparation. Fatty alcohols 1-Tetracosanol (C-24) 1-Hexacosanol (C-26) 1-Octacosanol (C-28) 1-Triacontanol (C-30) 1- Dotriacontanol (C-32) 1-Tertatriacontanol (C-34) Composition (% w/w) 19 335 6069 520 0.110 0.110

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Table 2 Samples identication, composition, gelation temperature (Tg), onset of crystallisation temperature (Tco). The grey background indicates samples without signicant difference (p b 0.05) between Tco and Tg.
Samples ID Olive oil (%w/w) Policosanol (%w/w) S1 S2 S3 S4
b

Tco (C) 21.7 0.9 34.2 0.4 38.3 0.1 42.1 0.4 47.65 0.01 50.3 0.5 52.0 0.2 56.6 0.3 65.7 0.1 72.5 0.5 74.49 0.01 76.7 0.1

Tg (C)a N.G. N.G. 26 1 39.5 0.3 45.6 0.3 49.36 0.01 50.1 0.4 54 1 64.0 0.8 71.6 0.7 73.63 0.05 76.26 0.03

99.9 99.7 99.5 99 98 97.5

0.1 0.3 0.5 1 2 2.5 3 5 15 30 40 55

S5b S6 S7 S8 S9 S10 S11 S12

a
b

97 95 85 70 60 45

N.G. indicates the absence gelation in the investigated temperature range. Samples prepared at three different temperatures i.e. 70 C, 85 C and 97 C.

to 15%) were seldom found either for data at high temperature, owing to instrument limits when the measurement of very low viscous systems is required, or for data in the transition crystallisation region, owing to the fast structuring phenomena. The onset of crystallisation (Tco) was individuated as the temperature value at which a sudden increase of complex modulus G* (and a sharp decrease of the loss tangent curve) in temperature ramp tests is evidenced (Lupi, Gabriele, Facciolo et al., 2012; Lupi, Gabriele, & de Cindio, 2012). This condition corresponds to the formation of the organogelator molecule aggregates (Lupi, Gabriele, Facciolo et al., 2012) and indicates the onset of transition. The determination of temperature Tco was based on the same numerical procedure used by Lupi, Gabriele, Facciolo et al. (2012) for similar organogelled systems. If Tco is the temperature at which the onset of crystallisation occurs, the clusters of crystals start to interconnect among each other at a lower value elsewhere dened as the gelation temperature (Tg) (Lupi, Gabriele, Facciolo et al., 2012). According to a simplied criterion, already used for similar systems (Lupi, Gabriele, Facciolo et al., 2012), the gelation value was identied as the crossover between dynamic moduli or, in the same way, as the temperature at which the loss tangent (i.e. the ratio between G and G) is equal to unity and, therefore, the phase angle () is 45. Rheological characteristics of selected samples (S3S7) were further investigated at 25 C with frequency sweep tests in linear conditions. Aiming at carefully controlling the thermal history of all samples, a temperature ramp test was performed down to 25 C, following the procedure previously described, and after a rest period of 100 s (to guarantee that equilibrium conditions were obtained) the frequency sweep was performed on the same sample. A statistical analysis, when necessary, was carried out using t-student test (Statgraphics Centurion XV, USA) and differences among compared samples were considered signicant at p-value b 0.05 (interval of condence of 95%). All data tting was performed through Table Curve 2D Software (Jandel Scientic, USA). 3. Results and discussion 3.1. Effect of temperature preparation on sample rheological properties The temperature of sample preparation could affect the nal properties of the organogels, in fact, the gelator should be added to the oil

at a temperature where a good solubility can be obtained to obtain the best results. For example, in order to investigate the organic gelation of vegetable oils with ethylcellulose, Zetzl et al. (2012) modied the temperature of gel preparation according to the molecular weight of the polymeric organogelator (estimated with an indirect analysis of the viscosity of a 5% ethylcellulose solution). Starting from these considerations a preliminary analysis on the effects of the preparation temperature was carried out on samples S4, S5 and S7. It is known (de Lucas, Garca, Alvarez, & Gracia, 2007) that melting temperature of pure policosanol ranges between 80 and 82 C depending on the potential differences in composition due to the raw material characteristics, therefore samples were prepared at three different temperatures (70 C, 85 C and 97 C), lower and higher than the expected melting point, trying to nd the preparation temperature more adequate to the characteristics of this organogelator. Temperature ramp tests were adopted to investigated samples manufactured in different conditions and their characteristics were compared in terms of complex modulus, G*, and phase angle, . Fig. 1 shows both rheological properties for samples S4, S5 and S7 at different temperatures of preparation. Obtained results evidence signicant similarities with the rheological characteristics of organogels based on different gelators (see for example monoglycerides oleogels prepared by Lupi, Gabriele, Facciolo et al. (2012)) investigated in similar conditions (temperature ramp tests). It can be seen that, at high temperature, the system is in the molten state and it exhibits a pure viscous behaviour (phase angle value approximately 90 within the instrument limits) with complex modulus characterised by very small values. Cooling the sample, down to a critical temperature, elsewhere dened as Tco (Onset of Crystallisation Temperature) (Lupi, Gabriele, Facciolo et al., 2012), G* shows a sudden increase, contemporarily with phase angle decreases, indicating the beginning of crystallisation. At lower temperature, the system behaves as a solid-like gel material. As far as sample S4 is concerned, no signicant difference in G* can be noticed when the sample is prepared at 85 C or 97 C whereas slight differences arise at low temperatures (mainly during the transition period) when the sample prepared at 70 C is considered. The phase angle seems to be more affected by the preparation temperature even though the greatest differences are observed, also in this case, when the sample prepared at 70 C is compared to the others. When samples S5 and S7 (produced with higher amounts of policosanol) are considered, no difference is observed for both G* and phase angle at any investigated temperature of preparation (even if very scattered data are obtained for sample S7 at low temperature, owing to the relevant consistency of the sample). According to the results obtained it seems that, even for very low gelator concentration, the temperature of preparation does not modify the organogel characteristics in a relevant way if a threshold value of 85 C is exceeded. As a consequence all samples were prepared at 85 C: this temperature seems high enough to ensure the complete melting of the organogelator, avoiding possible rheological differences below the Tco. In addition, it should minimise the oxidative phenomena of the vegetable oil as much as possible, enhanced by the process temperature increase (Pristouri, Badeka, & Kontominas, 2010). This should yield healthier products having better organoleptic properties (Ambrosone, Mosca, & Ceglie, 2006; Mosca, Ceglie, & Ambrosone, 2008). 3.2. Effect of organogelator concentration: Tco and Tg Aiming at investigating the crystallisation phenomena induced by the organogelator, looking for a potential percolation threshold where the gelation of the system occurs, tests at low concentrations of policosanol were carried out (samples S1S4). The same behaviour, already described in the previous section, is observed also for these samples (see Fig. 2ab): at high temperature, all samples behave as viscous systems (as conrmed by the phase angle values close to

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Fig. 1. Temperature ramp test at 1 Hz for samples prepared respectively at 70 C (grey symbols), 85 C (white symbols) and 97 C (black symbols). Data for samples S4, S5 and S7 are shown, respectively, in terms of complex modulus (a, c, e) and phase angle (b, d, f) versus temperature. Bars represent standard deviations.

90), and complex modulus increases with decreasing temperature only because of the decrease in molecular kinetic energy (and therefore in molecular mobility), even though no structural change occurs. A huge increase of G* occurs as soon as the Tco is reached, according to the different amounts of policosanol added to the system and, nally,

Fig. 2. Complex modulus G* (a) and phase angle (b) for samples characterised by the lowest amount of policosanol: S1 (circles), S2 (triangles), S3 (diamonds), S4 (squares). Bars represent standard deviations.

at lower temperatures an almost constant trend is found. Fig. 2b does not evidence any relevant change in phase angle for samples S1 and S2, even though for the latter a more regular trend, with smaller deviations, can be observed at the onset of crystallisation, owing to the increase in elasticity caused by the crystal formation; anyway quite large values are still measured, evidencing a liquid-like behaviour. On the other hand, when the policosanol amount is at least 0.5% (w/w) (sample S3) the material gelation, i.e. the formation of a threedimensional network, is observed at the temperature value (Tg) of 261 C where the phase angle becomes approximately 45 (and the crossover between G and G occurs). As a consequence it seems that between 0.3% and 0.5% (w/w) there exists a critical value dening the transition between a suspension (made of policosanol and fat crystal aggregates) and a gelled system with interconnected unities having a solid-like behaviour dependent on the organogelator amount, as evidenced by the different trends in both G* and observed for samples S3 and S4. Therefore, for samples with a policosanol concentration higher than 0.5% (w/w), both Tco and Tg values were evaluated and they were listed in Table 2. It can be seen that, increasing the organogelator amount, the difference between the critical temperatures becomes smaller. A statistical analysis (t-student test) was carried out and it was determined that Tg is signicantly different (p> 0.05) from Tco up to a policosanol amount equal to 2.5% (w/w), if a fraction larger than 3% (w/w) is added to the organogel no difference is encountered between the two temperatures. This range is very close to that found by Lupi, Gabriele, Facciolo et al. (2012) for monoglycerides organogels (where the range 2.33% w/w was estimated). Moreover, with an increasing amount of policosanol both temperatures seem to approach an asymptotic behaviour as already observed also for olive oil/monoglycerides organogels (Lupi, Gabriele, Facciolo et al., 2012) and for sunower oil/waxes organogels, where the melting temperature of the organogel was evaluated with DSC techniques (Hwang, Kim, Singh, Winkler-Moser, & Liu, 2012). The rheological parameter G was analysed to evaluate the effects of the organogelator amount on the development of the organogel network structure with temperature; in order to obtain results independent of potential kinetic effects, data at a xed distance from the onset of crystallisation were used, therefore G was analysed at 5, 10 and 15 C below the Tco, i.e. at the temperature values of T5,

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T10 and T15 (where T5 = TcoT, T10= Tco2 T, T15= Tco3 T and T = 5 C). It is worth noticing that when high amounts of policosanol are used (samples S8S12) the analysed material becomes so rigid, at temperature lower than Tg, that, owing to instrument limits, it is not possible to carry out the test further. Moreover, as previously discussed, only samples produced with a policosanol amount larger than 0.5% (w/w) gel and they show a network structure, as evidenced by the crossover between G and G moduli in a temperature ramp test. As a consequence it was possible to perform the analysis of storage modulus G, at the different temperatures, only for samples S3S7. Fig. 3 shows G as a function of the policosanol fraction Xp (grammes of policosanol divided by grammes of total oil phase, oil and policosanol); it is worth noticing that storage modulus increases in a non-linear way, with respect to the policosanol fraction, for all the investigated temperatures of analysis, as also found, for monoglycerides of fatty acids organogels, by Lupi, Gabriele, Facciolo et al. (2012) and Ojijo et al. (2004). It can be noticed that, for all samples, G increases with the distance from the onset of crystallisation, conrming that the elastic properties increase, owing to structuring phenomena promoted by both lower temperature and longer measure time. Nevertheless, it is evident that data at T10 and T15 are almost the same, except for sample S3, suggesting that an almost constant value is obtained when the temperature is at least 10 C lower than Tco. A different behaviour is exhibited by S3: probably, owing to the low policosanol amount (0.5% w/w), structuring phenomena are slower and require longer times (or lower temperatures) to yield a plateau value. 3.3. Interpretation of rheological data: the fractal model The non-linear relationship between storage modulus and organogelator content is similar to the behaviour typically observed in fat crystal networks when G is plotted as function of the solid fat content; according to the literature, owing to the fractal nature of these colloidal networks, a fractal model (Tang & Marangoni, 2007) can be adopted to relate the storage modulus to the dispersed particle volume fraction : G =3D
1

Marangoni (2007), but it is worth noticing that parameter can be considered an indirect measurement of the network strength, while D could be considered as a measure of the network microstructure. The original fractal model was further modied to take into account an apparent change in fractal dimension with the range of solid fat content (Tang & Marangoni, 2007), this apparent effect was attributed to the heterogeneous stress distribution due to the non-uniformity of crystal ocs and the presence of clusters not included in the network. As a consequence an effective volume fraction e was used, instead of the parameter , and the following nal equation, working in the whole range of solid fraction content, was obtained (Tang & Marangoni, 2007, 2008):  1 = b 3D k G 1e 2

where k and b are tting constants. In the present work it was not possible to measure the solid fraction during the rheological tests, therefore, according to the literature (Ojijo et al., 2004), a direct correspondence between and the organogelator volume fraction in the mixture was assumed which, in turn, can be assumed to be proportional to the policosanol fraction Xp. As a consequence Eqs. (1) and (2) were modied obtaining, respectively:
1 G X p =3D

3 4

 1 = 3D kX b p G 1e

where and k are different from the parameters proposed by Tang and Marangoni (2007). Owing to the limited range of policosanol fraction, in samples where storage modulus was available, Eq. (3) was considered suitable to t the experimental data and the following linearized form was adopted: lnG ln   1 ln X p : 3D 5

where is a constant dependent on the links between fat crystal ocs (it increases if the links become stronger) and D is the fractal dimension of the network. Fractal objects are geometric elements which exhibit a self-similar or self-afne character, therefore geometric pattern of a fractal object is repeated at different length scales. Further information about the model can be found in the above cited paper of Tang and

Fitting parameters are shown in Table 3. It can be seen (Fig. 4) that the proposed model seems to t the experimental trend quite well and larger deviations observed for few points, can be ascribed to the experimental data scattering. It is worth noticing that the tting lines are almost parallel, as better shown by a more or less constant value of the parameter D (Table 3), evidencing that the microstructure of the network does not change signicantly during the cooling process. An average value of D was, therefore, computed and the obtained parameter (D = 2.75) is in good agreement with data obtained in the literature investigating the crystallisation phenomena of different fats as it can be seen in Table 4 where results of the present work are compared to similar literature data (Ojijo et al., 2004; Tang & Marangoni, 2008). Even though little data is available for G, a greater amount of information is available for the onset of crystallisation temperature; this thermodynamic parameter, roughly marking the transition between a viscous and a viscoelastic behaviour, is relevant in organogel design

Table 3 Parameters of Eq. (5). T (C) Fig. 3. Storage modulus G as function of policosanol fraction (Xp) at T5, T10 and T15. Bars represent standard deviations; for few samples they are smaller than the adopted symbols. 5 10 15 ln() 23.8 0.9 23.9 0.9 24.0 0.9 D ( ) 2.76 0.01 2.75 0.01 2.74 0.01 r2 () 0.98 0.98 0.98

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Fig. 4. Fit of the fractal model for G-Xp data at T5, T10 and T15. Experimental data (symbols) and model ttings (lines).

Fig. 5. Tco values as a function of policosanol fraction and data tting with Eq. (5) empirical model (dashed line). Bars represent standard deviations; for all samples they are smaller than the adopted symbols.

because it affects both the process and the usage conditions; therefore it could be useful to relate it to the organogel microstructure. Looking for a potential relationship, it seems reasonable to consider that also thermodynamic properties, such as the crystallisation point, could be related to the fractal structure of the system. In addition, when Tco data are plotted as a function of the policosanol fraction (Fig. 5), a non-linear behaviour characterised by an asymptotic trend at high policosanol fraction can be observed: this trend is, qualitatively, very similar to G- data reported in the literature over a large range of solid fat fraction (Tang & Marangoni, 2007). Starting from these considerations, an empirical equation, similar to Eq. (2) was proposed to t Tco data for policosanol fractions ranging between 0.005 (the lowest value where gelation occurs) and 0.55:  1 = bX c 3D T co a 1e p

3.4. Rheological behaviour of cold organogels: frequency sweep tests A number of organogelled systems were also investigated through frequency sweep tests at 25 C; in order to guarantee a controlled thermal history all samples were cooled down to the nal temperature on the rheometer plate at a constant cooling rate. Fig. 6 shows the results for samples S3S7 in terms of complex modulus G* (Fig. 6a) and phase angle (Fig. 6b). It can be seen that the behaviour of sample S3 is quite different from the others, evidencing a signicant dependence of complex modulus on frequency and a phase

where a, b, c, are tting constants and D is the average value of the fractal dimensions listed in Table 3 (D = 2.75). All Tco values of samples yielding a gel, therefore having a policosanol fraction equal or higher than 0.5% (w/w) (samples S3 S12), were tted: in fact, even if samples prepared with a policosanol amount ranging from 0.5% w/w to 2.5% (w/w) are characterised by a gelation process delayed with respect to the onset of crystallisation, this delay can be referred only to crystallisation kinetic effects. Therefore, the fractal model, in the present case, was considered valid for any organogel showing a network formation. Fig. 5 shows a very good agreement between experimental data and modied fractal model (Eq. (6)) for the discussed samples. The parameters of Eq. (6), for the considered organogels, are: a = 101 8 C, b = 2.9 0.3 (dimensionless) and c = 0.13 0.01 (dimensionless).

Table 4 Comparison among some literature values for the fractal dimension D in different fat systems. SFC means Solid Fat Content. Fractal model D values 2.79a 2.69 0.02 2.88 0.74 2.69 0.20 2.75 0.58 2.75 Fat system Olive oil/monoglyceride High melting fraction of milk fat and canola oil (low SFC) Anhydrous milk fat (low SFC) Palm oil (low SFC) Cocoa butter (Low SFC) Olive oil/policosanol References Ojijo et al. (2004) Tang and Marangoni (2008) Tang and Marangoni (2008) Tang and Marangoni (2008) Tang and Marangoni (2008) This work Fig. 6. Frequency sweep tests for samples S3, S4, S5 and S7 at 25 C in terms of complex modulus (a) and phase angle (b). Bars represent standard deviations.

a This value was computed by data shown by Ojijo et al. (2004) for a different form of the fractal model.

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angle larger than 45 (therefore a liquid-like behaviour) for all tested frequencies. On the contrary, samples S4S7, within the tested frequency range, are characterised by an almost constant G* (increasing with policosanol concentration) and a very low phase angle: therefore they exhibit typical strong gel behaviour (Rao, 1999). These results further conrm data obtained during the temperature ramp tests, in fact, as already discussed in the previous sections, temperature ramp tests showed that the policosanol amount of 0.5% (w/w) is a threshold value for gelation: for lower concentrations only a suspension of crystalline aggregates is obtained whereas for higher amounts a gelation temperature, decreasing with increasing organogelator concentration, was observed. Moreover at 0.5% (w/w) the gel point occurs approximately at 26 1 C, very close to temperature adopted in frequency sweeps. As a consequence, sample S3, being in a transition condition, behaves like a concentrated suspension which is not yet gelled, whereas all other samples (S4S7), with a concentration much larger than the gelation threshold, exhibit a real strong gel behaviour. It is worth noting that frequency sweep tests evidence that rheological properties of these systems seem suitable for potential applications as saturated fat replacers, in fact when samples S6 and S7 (containing, respectively, 2.5% w/w and 3% w/w of policosanol) are considered, it can be noticed that dynamic moduli are quite close to the values observed for traditional commercial products. According to the literature (Rnholt, Kirkensgaard, Pedersen, Mortensen, & Knudsen, 2012; Vithanage, Grimson, & Smith, 2009) typical values of storage modulus of butter, at 1 Hz and 20 C, range, approximately, between 104 Pa and 3 105 Pa; whereas for different spreadable products and margarine, G or G* values ranging approximately between 2.5 10 4 Pa and 5 106 Pa (Lupi et al., 2011; Vithanage et al., 2009) were measured. Even though commercial fats, such as shortenings or margarine, are usually water in oil emulsions (Vaisey-Genser, 2003) having properties signicantly affected by the emulsication conditions, it is interesting to notice that the potential dispersing oil phase, based on organogels, exhibits a rheological behaviour close to the nal products, even for low organogelator concentrations. On the other hand it can be seen that by increasing the policosanol concentration, the strength of the gel, related to the complex modulus at 1 Hz, is increasing whereas the structuring level, related to the phase angle (i.e. the ratio between the liquid-like and the solid-like behaviour) is almost constant. These results suggest that it is possible to have different materials suitable for fat replacement and having a controlled consistency by changing the organogelator amount. 4. Conclusions Organogels based on olive oil and edible fatty alcohols were prepared and investigated by using rheological methods, aiming at determining both mechanical properties and transition temperatures of the systems considered. The proposed additive, a mixture of fatty alcohols named policosanol, is already well known for its nutraceutical characteristics and in this work its gelling properties were investigated. Temperature ramp tests at a constant cooling rate (1 C/min) evidenced a marked capability of policosanol to structure olive oil even at very low concentrations: it was found that the onset of crystallisation appears at 0.1% (w/w) whereas a gelation threshold for the formation of a network of crystals aggregates was observed for concentrations ranging between 0.3% and 0.5% (w/w). These results were also conrmed by frequency sweep tests, at 25 C, which evidenced a strong gel behaviour of the oil phase, for policosanol amounts greater than 0.5% (w/w). The onset of crystallisation and the gelation temperature do not signicantly differ, at the investigated cooling rate, for a policosanol critical concentration ranging between 2.5% and 3% (w/w); in these

conditions, probably, the organogelator concentration is high enough to promote the formation of a large number of crystals able to immediately aggregate in a three-dimensional network. Storage modulus data, G, at a constant distance from the onset of crystallisation temperature Tco, were well tted by a fractal model which allowed a fractal dimension to be estimated in agreement with the literature values observed for similar systems. Moreover, on the basis of the empirical evidence, a modied fractal model was proposed to t the Tco values as function of organogelator concentration and a very good agreement between the proposed correlation and the experimental data was obtained. Owing to the lack of theoretical basis, this model can be considered, so far, only a phenomenological correlation, nevertheless it could be very useful to relate a macroscopic property (the onset of crystallisation) to the material microstructure (the fractal dimension) helping in designing organogels with controlled properties.

Acknowledgement The authors are grateful for its support to the Food Science & Engineering Interdepartment Centre of the University of Calabria, to L.I.P.A.C., Calabrian Laboratory of Food Process Engineering (Regione Calabria APQ-Ricerca Scientica e Innovazione Tecnologica I atto integrativo, Azione 2 laboratori pubblici di ricerca mission oriented interliera) and to PON R&C (Programma Operativo Nazionale Ricerca e Competitivit 2007 2013) project PON01_00293 Spread Bio Oil. References
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