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Innovative Food Science and Emerging Technologies 41 (2017) 411–420

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Innovative Food Science and Emerging Technologies


journal homepage: www.elsevier.com/locate/ifset

Effect of high pressure homogenization (HPH) on microstructure and T


rheological properties of hazelnut milk
Osman Gula, Furkan Turker Saricaoglub,c,⁎, Mustafa Mortasb, Ilyas Atalarb, Fehmi Yazicib
a
Program of Food Technology, Yeşilyurt Demir–Celik Vocational School, Ondokuz Mayis University, Samsun, Turkey
b
Food Engineering Department, Engineering Faculty, Ondokuz Mayis University, Samsun, Turkey
c
Food Engineering Department, Engineering Faculty, Bayburt University, Bayburt, Turkey

A R T I C L E I N F O A B S T R A C T

Keywords: The effect of high pressure homogenization (HPH) on microstructure and rheological properties of hazelnut
High pressure homogenization milks was investigated. Hazelnut milk samples were produced from cold pressed hazelnut cake and homogenized
Hazelnut milk up to 150 MPa pressure. Microstructural and rheological properties of products, except temperature sweep, were
Microstructure greatly affected by HPH treatments. Homogenized samples showed significant reduction in particle size, which
Rheology
turned from bimodal and poly-disperse to monodisperse distributions. HPH decreased the consistency of
Particle size distributions
products from 91.82 to 0.51 Pa.sn and increased flow behavior index from 0.15 to 0.36. All samples showed
higher values G′ than G″, which indicates that samples could be classified as soft-gel network, and hazelnut milk
samples did not obey the Cox-Merz rule without multiplying angular frequency with shift factor. In conclusion,
HPH can be used to reduce the consistency of samples and the friction loss, thus minimizing the amount of
energy required to flow during processing and distribution.
Industrial relevance.
Vegetable based beverages are available at any supermarket as an alternative to dairy products with an
increasing consumer acceptance. Between these beverages hazelnut milk samples are the most noteworthy
products due to important role in human nutrition and health, and moreover due to well accepted and widely
consumed product. Due to its composition, hazelnut milks have very high consistency, and therefore, the energy
consumption for processing and handling is too high. In this research, the high pressure homogenization (HPH)
technology was successfully proposed to be used for improving microstructural properties, and hence reducing
its consistency. The energy consumption during processing and distribution can be minimized by reducing the
consistency and friction losses of product.

1. Introduction have been growing rapidly due to intolerance problems of cow's milk
and changes of food preferences (Bernat, Cháfer, Rodríguez-García,
The rheological properties of foods are important for both con- Chiralt, & González-Martínez, 2015). Vegetable based beverages such as
sumers and manufacturers in terms of giving information about design almond, coconut, peanut and soy milks are the most noteworthy
of processing equipment and the energy consumption of unit operations products available on markets, while there are few products based on
to manufacturers and also about the sensory properties, final product hazelnut. Hazelnut has an important role for human nutrition and
quality and stability to consumers. The steady flow curve is the most health due to including protein, carbohydrates, lipids, minerals,
commonly way to determine the rheological behavior of fluids, but vitamins, dietary fibers, tocopherols, phytosterols and phenolic com-
many phenomena cannot be described by only steady flow curve pounds (Alasalvar, Shahidi, Liyanapathirana, & Ohshima, 2003). Be-
analysis, and thus, elastic behavior must also be taken into considera- sides the health benefits, hazelnut has active taste compounds, such as
tion (Augusto, Ibarz, & Cristianini, 2013). It is important property for amino acids and organic acids, and these compounds make it widely
liquid foods for better understanding of their behavior during storage, consumed and well accepted food (Tey et al., 2011). Hazelnut oil is
processing and consumption. taking consideration due to similar fatty acid composition with olive oil
Vegetable based beverages are diluted emulsions obtained from and extraction is commonly done with chemicals by oil industry.
bean or nuts. In the last few years, the demands for these beverages However, the waste product of extraction cannot be used for human


Corresponding author at: Food Engineering Department, Engineering Faculty, Ondokuz Mayis University, Samsun, Turkey.
E-mail address: furkan.saricaoglu@omu.edu.tr (F.T. Saricaoglu).

http://dx.doi.org/10.1016/j.ifset.2017.05.002
Received 5 May 2016; Received in revised form 30 March 2017; Accepted 12 May 2017
Available online 13 May 2017
1466-8564/ © 2017 Elsevier Ltd. All rights reserved.
O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

nutrition due to chemical residues and it is commonly used for animal Biuret reagent added on 1 mL of hazelnut milk. Then, the absorbance of
feeding. Cold pressing, one of the oldest extraction techniques, is the sample was measured at 550 nm against Biuret reagent as blank using a
unique oil production method without heat treatment or solvent. As spectrophotometer (Cary 60 UV–Vis, Agilent Technologies, USA), and
known, the extraction yield is lower than solvent extraction technique protein solubility was calculated with a standard curve of Bovine Serum
and high-temperature processing, after pressing high quantity of oil Albumin (BSA).
cannot be recovered from the oil source; however, it has a major
advantage of minimizing degradation of nutritive oil constituents 2.2.2. Particle size distribution (PSD)
(Karaman et al., 2015). So, in this research we produced hazelnut milk The PSD of hazelnut milks was measured by light scattering
from hazelnut cake, which is a by-product of cold pressed hazelnut oil (Malvern Mastersizer 2000 with Hydro 2000 G(A), Malvern
industry. Instruments Ltd., UK). Droplets of sample were slowly added into a
High pressure homogenization (HPH) is a non-thermal technology, sample compartment, already filled with distilled water at room
which was initially used as an alternative to thermal processing. The temperature, until obscuration was 2.93%. The mean diameter was
principle of this technology is based on the continuous flowing of a fluid also evaluated based on volume weighted mean diameter (d4,3; Eq. (1))
through a narrow gap valve, which increases the velocity and also and the particle surface area (d3,2; Eq. (2)). This is useful since particles
results in a great pressure drop to atmospheric pressure, resulting in are not ideal spheres, and the (d3,2) is more influenced by the smaller
high shear stress, high turbulence and cavitation. Thus, high mechan- particles, whereas the (d4,3) is more influenced by the larger ones
ical stress is applied to macromolecules and suspended particles in (Bengtsson & Tornberg, 2011; Leite et al., 2014b). Cumulative percen-
fluid, and they became twisted, deformed and even disrupted (Floury, tiles d0.1, d0.5 and d0.9 were also determined, indicating that 10%, 50%
Bellettre, Legrand, & Desrumaux, 2004). In recent years, this technol- and 90% of the particles fell below the specified diameter, respectively.
ogy has been used for improving rheological characteristics of fruit Two replicates were performed for each sample.
juices (Augusto et al., 2013; Leite, Augusto, & Cristianini, 2014b);
∑i ni d i4
enhancing physical properties and stability of almond milk (Bernat d 4,3 =
et al., 2015); and improving the properties of polysaccharides (Wang, ∑i ni d 3i (1)
Li, Wang, Liu, & Adhikari, 2012; Wang, Sun, Zhou, & Chen, 2012) and
proteins (Dong et al., 2011; Liu & Kuo, 2016). ∑i ni d 3i
d3,2 =
The objectives of the present study were (1) to evaluate the effect of ∑i ni d 2i (2)
HPH on steady-state and dynamic oscillatory rheological properties,
particle size distributions and microstructures of hazelnut milk ob-
tained from hazelnut cake; (2) to model experimental data in order to 2.2.3. Solid particle sedimentation (SPS)
explain the effect of HPH on the apparent viscosity of hazelnut milk; Solid particle sedimentation (SPS) was performed according to
and (3) the applicability of the Cox-Merz rule for prediction of steady Valencia-Flores, Hernández-Herrero, Guamis, and Ferragut (2013) with
shear properties from dynamic shear data. slight modification. Homogenized hazelnut milk samples (approxi-
mately 10 g) were transferred to centrifuge tubes and centrifuged
2. Materials and methods (Nüve-Bench Top Centrifuge, NF 1200R, Turkey) at 2500g for 10 min.
The supernatant was weighted and solid deposition at the bottom of the
2.1. Materials and milk preparation tubes was expressed as % (w/w).

Hazelnuts (Corylus colurna) were purchased from Gursoy Hazelnut 2.2.4. Microstructure
Production Factory (Ordu, Turkey) as removed shells-brown skin and The microstructure of the hazelnut milk samples treated with high
the hazelnut oil was removed from hazelnut by using a headed cold pressure homogenization was observed using an optical microscope
press machine (Ekotok 1, Izmir, Turkey). The resulting cold press (Olympus, CX31, Japan) under 20× magnification. The samples were
hazelnut cake including 8.68% moisture, 43.77% protein, 25.20% carefully placed onto a glass slide and covered with a cover slip. The
carbohydrate, 17.38% lipid and 4.97% ash was used for manufacture images were captured at least in triplicate for each sample. The
of hazelnut milk samples. obtained photographic images were commentated with imaging soft-
For preparation of hazelnut milk samples, cold press hazelnut cake ware (Olympus BP20, Japan).
was grounded using a blender (Waring laboratory blender, Conair
Corporation, Stamfold, CT, USA) for 10 min and grounded hazelnut 2.2.5. Rheological measurements
(100 g) was treated with 1 L of water for 10 min using an Ultra-Turrax The rheological properties of hazelnut milk were analyzed with a
homogenizer (IKA-Werke GmbH & Co. KG, Staufen, Germany) at HAAKE Mars III Rheometer (HAAKE Co., Germany) equipped with cone
10,000 rpm. High pressure homogenization (HPH) treatment was and plate geometry (diameter: 35 mm, gap: 0.104 mm and cone
carried out using a high pressure homogenizer (Panda PLUS 2000, angle = 2°). Before measurements, samples were held for 300 s at
GEA Niro Soavi, Parma, Italy) with the pressures of 0 (control), 25, 50, 20 ± 0.1 °C to equilibrate. Rheological results were monitored using a
75, 100 and 150 MPa. Hazelnut milk samples were introduced into the Rheo Win software package. All the rheological parameters were the
homogenizer at 15 °C and after the homogenization samples were mean of two measurements per duplicates of hazelnut milk samples.
quickly cooled using an ice bath.
2.2.5.1. Steady-state shear measurement. The flow behavior of hazelnut
2.2. Methods milk samples was measured by recording shear stress values against
shear rates from 1 to 100 s− 1 through 100 s. The relationship between
2.2.1. pH, °Brix, density and Protein solubility. shear stress and shear rate was described by Ostwald-de-Waele model,
The pH, °Brix and density of hazelnut milk samples were measured Eq. (3), which is the most frequently used model for engineering
using a pH meter (Eutech Cyberscan pH 2700, Ayer Rajah Crescent, applications:
Singapore), refractometer and densitometer, respectively. All measure- ηapp = K × γ n−1
̇ (3)
ments were carried out at 25 °C.
The effect of HPH on protein solubility of hazelnut milks was where ηapp is the apparent viscosity (Pa·s); γ ,̇ the shear rate (s− 1); K, the
determined according to Biuret method (Robinson & Hogden, 1940). consistency index (Pa · sn) and n, the flow behavior index
For this purpose, hazelnut milks were mixed thoroughly, and 1 mL of (dimensionless). These calculations were made using the Rheowin 4

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O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

Fig. 1. The effect of HPH on protein solubility (a) and solid particle sedimentation (b) of hazelnut milk samples in water.

Data Manager software (version 4.20, Haake). 2.2.6. Data & statistical analyses
Analysis of samples were carried out at least in duplicate, and the
2.2.5.2. Dynamic shear measurement. Viscoelastic properties of results were subjected to analysis of variance (ANOVA) using SPSS
hazelnut milk samples were measured using small amplitude 20.0.0 (IBM SPSS Statistics, Chicago, IL, USA). The means were
oscillatory shear tests at 25 °C. The linear viscoelastic region (LVR) of compared with Duncan's multiple range test (significance p < 0.05).
samples was previously determined by running stress sweeps between The modelling of apparent viscosity against PH was performed using
0.01 and 100 Pa, at a frequency of 1 Hz. After that, frequency sweeps at Number Cruncher Statistical System (NCSS, 2007, NCSS, LLC, Utah,
0.1 Pa (obtained from stress sweep in the linear region) were performed USA).
from 0.1 to 100 Hz (0.628–628.3 rad·s− 1). The oscillatory rheological
parameters used to compare the viscoelastic properties of the samples 3. Results and discussions
were storage moduli (G′) and loss moduli (G″). The storage (G′) and loss
(G″) modules were modeled as a power function of oscillatory 3.1. 3.1. pH, °Brix, density and protein solubility.
frequency (ω) (Eqs. (4) and (5)), as commonly used for describing the
viscoelastic behavior of food and dispersions: The pH and °Brix of hazelnut milk samples were significantly
(p < 0.05) changed with homogenization process. pH and °Brix value

G′ = K′(ω)n (4) of control (0 MPa) sample were found as 6.35 and 6.45 and the values
were increased after the homogenization to 6.5 and 7.15, respectively.
″ There was a slight increase in pH values of samples with the increase in
G″ = K″(ω)n (5)
pressure (p > 0.05). These results are in agreement with the report of
′ ″ Bernat et al. (2015). °Brix values of hazelnut milk samples were
where K′ (Pa.sn ) and K″ (Pa.sn ) are constants, n′ and n″ may be referred
significantly (p < 0.05) increased with increasing pressure. The in-
to as the frequency exponents, and ω is the angular frequency (rad·s− 1)
crease in °Brix may be due to increase in soluble protein ratio or release
(Rao & Cooley, 1992).
of starch granules upon mechanical stress during HPH process (Yu,
Temperature sweep was performed at a frequency of 0.1 Hz and
Jiang, Cao, Jiang, & Pan, 2016). Density of the control sample was
0.1 Pa strain (within the linear viscoelastic region) with cone/plate
determined as 1.028 g mL− 1 and a slight decrease (down to
geometry. Samples were loaded onto the surface of the lower plate, and
1.021 g mL− 1) was observed after the homogenization treatment
the upper plate was lowered until it reaches the gap distance. After
(p > 0.05).
loading, the sample was held for 5 min at the initial test temperature
Solubility is an important functional property of proteins, since it
before testing to allow stress relaxation and temperature equilibration.
can affect other functional properties such as rheological, hydrody-
The exposed surfaces of samples were covered with a thin layer of
namic and surface-active properties. Good solubility is crucial for many
silicon oil to avoid dehydration from the sample during heating.
protein-based products. The effect of homogenization pressure (PH) on
Samples were heated from 20 to 85 °C with a heating rate of 10 °C/
protein solubility of hazelnut milk samples is given in Fig. 1a. As can be
min. Storage modulus (G′) and loss modulus (G″) were recorded as a
seen, solubility of hazelnut proteins in water increased as PH increased,
function of temperature.
and after 75 MPa pressure solubility was not significantly affected from
PH. Protein solubility at 100 MPa was determined as 3.17 g/100 mL and
2.2.5.3. Applicability of the cox-Merz rule. The Cox-Merz rule is the most it was insignificantly decreased (3.12 g/100 mL) when the pressure
reliable method to associate with oscillatory and steady shear increased to 150 MPa (p > 0.05). Our results were in accordance with
rheological data. The rule states that the apparent viscosity (ηapp) at a Chen, Xu, and Zhou (2016) for chicken breast myofibrillar proteins and
specific shear rate (γ )̇ is equal to the complex viscosity (η*) at a specific Dong et al. (2011) for peanut proteins, whereas Liu and Kuo (2016)
oscillatory frequency (ω), when γ =̇ ω (Eq. (6)) (Augusto, found that protein solubility of soy flour decreased as PH increased, and
Cristianini, & Ibarz, 2012; Gunasekaran & Ak, 2000). When this rule is this could be related with the differences of protein structures between
valid, rheological properties of hazelnut milk samples can be hazelnut and soy flour.
determined by either oscillatory or steady-state shear experiments.
Thus, the results obtained from steady-state shear and oscillatory shear 3.2. Particle size distribution (PSD)
analyses were used for the specifying of the Cox-Merz rule applicability:
ηapp (γ )̇ = η∗ (ω) γ ̇ = ω (6) The PSD of the samples processed with HPH are shown in Fig. 2, and
derived measurements of average particle diameter (d4,3), particle

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O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

(0 MPa) and homogenized (up to 150 MPa) hazelnut milk samples.


Homogenization process was significantly decreased the SPS value of
hazelnut milk samples (p < 0.05). The higher the SPS, the lower the
stability of hazelnut milk and it was 51.85% for the control sample,
moreover SPS value decreased with increasing homogenization pres-
sure except for 150 MPa. The decreasing SPS value between control and
homogenized samples could be probably associated with the disruption
of large particle aggregates. These large aggregates as seen in Fig. 3,
caused to higher sedimentation and sediment accumulation increased.
Therefore, increasing homogenization pressure led to reduction in
particle size and consequently enhanced the stability of hazelnut milk
Kubo, Augusto, and Cristianini (2013). These results similar to Bernat
et al. (2015) with hazelnut milk, with tomato juice and Cruz et al.
(2007) with soymilk. On the other hand, hazelnut milk homogenized at
150 MPa showed higher sedimentation value than hazelnut milk
treated with 100 MPa, this could be attributed to protein solubility
Fig. 2. Effect of HPH on the PSD of hazelnut milk samples.
which decreased insignificantly between 100 and 150 MPa. The
macromolecular structure and interaction of proteins during HPH was
surface area (d3,2) and cumulative percentiles (d0.1, d0.5, d0.9) are given
changed by using high pressures up to 100 and 150 MPa, and therefore,
in Table 1. As can be seen in Fig. 2, control sample processed at 0 MPa
these pressures caused to higher sedimentation because of enhancing
presented bimodal and poly-disperse distributions, and as the PH
protein-protein interactions (Yu et al., 2016).
increased from 25 to 150 MPa the PSD of hazelnut milk became
monomodal. Similarly, Briviba, Gräf, Walz, Guamis, and Butz (2016)
and Bernat et al. (2015) reported that untreated hazelnut and almond 3.4. Microstructure
milk showed a bimodal particle size distribution due to the complex
composition and preparation process. The finest particle fraction is The microstructures of hazelnut milk samples processed with
probably consisted of proteins, whereas the biggest particles consist of different PH were examined using a typical microscope to explain the
fat droplets, remains of cellular tissues and particle aggregates. How- experimental results. The micrographs of hazelnut milk samples are
ever, some finest particles as seen in the first peak of control sample given in Fig. 3. The micrographs showed that there was a clear
(0 MPa) seem to aggregate since they are not appeared in the peak of difference between control (0 MPa) and homogenized samples (25,
HPH samples. Some of the protein bodies could be unfolded or 50, 75, 100 and 150 MPa) since the homogenization of hazelnut milks
aggregated by high pressure effect (Bernat et al., 2015). The increase considerably reduced the size of the suspended particles. As can be
in PH progressively decreased the mean particle diameter (d4,3), particle seen, hazelnut milk homogenized at 0 MPa consisted mainly of
surface area (d3,2), and other cumulative percentiles (Table 1), and deformable particles with irregular shapes and a number of small
resulted a narrower particle distributions due to the reduction in size of particles, which are thought to be flocculated proteins due to hydro-
oil droplets and plant cell remains. The mean particle diameter and phobic character (Bernat et al., 2015). The great reduction in particle
particle surface area of processed samples were significantly affected by sizes, detected by the light scattering diffraction, can be observed from
all the range of PH (p < 0.05), but cumulative percentiles were not Fig. 3. However, most of the small particles are flocculated through
affected from PH between 100 and 150 MPa since particles could not be protein bridges, which caused to low stability of suspension (as seen in
broken smaller than this range regardless of initial size. Similar results SPS results) despite small particle sizes. The poor stabilizing properties
were also observed by Bernat et al. (2015), Leite et al. (2014b), of the protein, due to its high hydrophobicity and low water affinity, is
Augusto, Ibarz, and Cristianini (2012b), Silva et al. (2010) and the cause of flocculation process and subsequent phase separation
Sentandreu, Gurrea, Betoret, and Navarro (2011), for almond milk, (Bernat et al., 2015).
concentrated orange juice, tomato juice, pineapple pulp and orange Moreover, small particles formed by higher PH values tend to
juice, respectively. aggregate at 50 MPa. It seems that samples treated at 50 MPa exhibited
It can be concluded that the higher the PH value, the less the larger particles than the sample treated at 25 MPa. Similar observations
increase of particle size distribution. This could be explained by a can be made also between samples treated at 75 and 100 MPa. This
physical limitation of the equipment in terms of the gap dimensions and could be explained by increased surface area of the suspended particles
the resultant shear stress, since the smaller particles requires greater with cell disruption and subsequent fragmentation. Cell fragmentation
stress to disrupt than delivered by equipment. released wall materials such as fibers and proteins, and could improve
the particle–particle interactions and result aggregates occurring
(Augusto, Ibarz, and Cristianini (2012b)). Therefore, as the pressure
3.3. Solid particle sedimentation (SPS) of homogenization was increased, a progressive disintegration of
suspension and protein aggregates was observed, allowing for a
Fig. 1b shows the solid particle sedimentation (SPS) for the control completed cell disruption when the suspension was homogenized above

Table 1
Mean and cumulative diameter of hazelnut milks as a function of homogenization pressure (PH).

PH (MPa) d4,3 (μm) d2,3 (μm) d0.1 (μm) d0.5 (μm) d0.9 (μm)

0 183.82 ± 1.80a 27.58 ± 0.86a 9.81 ± 0.06a 142.96 ± 3.42a 444.52 ± 19.97a
25 90.82 ± 0.61b 26.09 ± 0.44b 8.10 ± 0.19b 78.71 ± 2.48b 166.44 ± 17.06b
50 62.59 ± 0.82c 21.79 ± 0.40c 7.19 ± 0.21c 58.39 ± 4.05c 118.84 ± 4.05c
75 50.98 ± 0.72d 18.76 ± 0.42d 6.65 ± 0.32c 44.86 ± 3.18d 104.79 ± 5.47c
100 41.97 ± 0.86e 18.01 ± 0.08d 5.91 ± 0.10d 35.19 ± 3.39e 76.12 ± 5.03d
150 31.40 ± 1.92f 15.23 ± 0.36e 5.53 ± 0.42d 27.25 ± 3.26e 59.34 ± 4.87d

a–f
Values are means ± Standard Deviation. Means within the same column with different letters are significantly different at p < 0.05.

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O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

Fig. 3. Microstructure of hazelnut milk samples processed with HPH.

50 MPa. Significant differences were observed between samples treated Krzemińska, & Mojka, 2013). As can be seen in Fig. 4, the apparent
at 0, 25 and 150 MPa, while samples treated at 50 MPa exhibited the viscosity of all samples at low shear rates decreased insignificantly as
most “undesirable” microstructure. Moreover, larger oil droplets were homogenization pressure increased. This may be related to conforma-
occurred in samples treated at 75–150 MPa than in all the other tional changes of homogenized hazelnut milk samples with increasing
samples. Similar results were also found by Yu et al. (2016) for taro homogenization pressure (Porto, Augusto, Terekhov,
pulp, by Bernat et al. (2015) for almond and hazelnut milk, and by Hamaker, & Cristianini, 2015). These conformational changes occurred
Bengtsson and Tornberg (2011) for potato pulp. The microstructure until no further structural changes, and then the flow behavior
results confirmed the changes in PSD, occurring in the hazelnut milks as remained constant at low shear rates. Apparent viscosities of samples
a result of high pressure homogenization. pressurized at 100 and 150 MPa (Fig. 4) were similar due to changing
water and hazelnut protein interactions, and hence, the apparent
3.5. Rheological behavior viscosities of hazelnut milk samples were also changed.
The data from flow curves of samples could not be fitted to Herschel
3.5.1. Steady-state shear properties Bulkley model due to decreased viscoelasticity of high pressure treated
The flow behavior of the processed hazelnut milks after homo- samples (100 and 150 MPa, data not shown), and therefore, data were
genization pressures of 0, 25, 50, 75, 100 and 150 MPa is shown in fitted the Ostwald-de-Waele model (Eq. (1)), and apparent viscosity at
Fig. 4. It can be seen that the control sample (0 MPa) showed high 50 s− 1 (shear rate in mouth), consistency index and flow behavior
apparent viscosity independent of shear rate. The values of apparent index known as η50, K and n and R2 of Eq. (1) are summarized in
viscosity demonstrated a decreasing trend with increasing shear rates, Table 2. As can be seen, Ostwald-de-Waele model was found to be the
which indicates that all samples exhibited a shear-thinning behavior. At adequate model (R2 = 0.919–0.999) to describe the flow behavior of
high shear rates, the differences in apparent viscosity between samples hazelnut milks. PH led to a decrease in K values as compared with
prepared with different pressures were smaller than those at low shear control, while the flow behavior index (n) showed the opposite
rates. This could be explained with the structural breakdown and behavior. The highest K value (91.82 Pa.sn) was obtained from control
rearrangement induced by the shear rates (Bortnowska, sample and it was decreased as PH increased from 0 MPa to 150 MPa.
On the other hand, the flow behavior index (n) increased from 0.15 to
0.36 in the same range of pressure variation (Table 2). These results are
also similar to those reported for pineapple pulp (Silva et al., 2010),
concentrated orange juice (Leite et al., 2014b) and tomato puree
(Tan & Kerr, 2015), whereas n values of HPH treated hazelnut and

Table 2
Effect of homogenization pressure (PH) on the flow properties and apparent viscosity (η50)
of hazelnut milks: parameters of Ostwald-de-Waele model.

PH (MPa) η50 (Pa.s) K (Pa·sn) n (-) R2

0 0.618 ± 0.080a 91.82 ± 9.74a 0.15 ± 0.030e 0.999


25 0.101 ± 0.024b 1.97 ± 0.46b 0.23 ± 0.010d 0.919
50 0.085 ± 0.007bc 1.66 ± 0.43b 0.25 ± 0.007c 0.971
75 0.069 ± 0.003bc 1.32 ± 0.30b 0.32 ± 0.004b 0.989
100 0.043 ± 0.002c 0.59 ± 0.13b 0.34 ± 0.004b 0.973
150 0.037 ± 0.003c 0.51 ± 0.06b 0.36 ± 0.011a 0.976

Fig. 4. Effect of HPH on apparent viscosity of hazelnut milk samples prepared with Values are means ± Standard Deviation. a–e Means within the same column with
different pressure. different letters are significantly different at p < 0.05.

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O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

almond milk (Bernat et al., 2015) and tiger nuts' milk beverages
(Codina-Torrella, Guamis, Ferragut, & Trujillo, 2017) were close to 1.
The rheological differences between our results and the results of
Bernat et al. (2015) and Codina-Torrella et al. (2017) could be
attributed to utilized raw materials for milk samples' production. In
our research, we used hazelnut cake which is a by-product of cold
pressed hazelnut, and it is also contains less oil than raw hazelnut
which affect the interactions between protein and oil, and hence
rheological properties displayed significant differences.
The relationship between apparent viscosity (at a constant shear
rate of 1/s) and PH was described using an exponential decay (Eq. (7))
and power-sigmoidal function (Eq. (8)) which was studied before by
Harte and Venegas (2010) for polymer suspensions and Augusto, Ibarz,
and Cristianini (2012a) for fruit juice serum model, respectively.
⎛ k + e−k 2 PH ⎞
η = η0 ⎜ 1 ⎟
⎝ k1 + 1 ⎠ (7) Fig. 5. Harte-Venegas and power-sigmoidal function models of hazelnut milk samples
viscosity.
η0
η=
1 + (ax )b (8) Waals interactions or hydrogen bonding, ensure the stability of the
structure. When G′ is greater than the G″, food system shows elastic
where η is the predicted viscosity (Pa.s) after homogenization applied
properties rather than viscous; which corresponds to a gel state. At low
from 0 to 150 MPa, η0 is the viscosity (Pa·s) before homogenization, PH
frequencies, the storage and loss modulus of control sample were higher
is the homogenization pressure, k1 is a constant that determines the
than those of processed with high pressure homogenization. It has been
plateau viscosity after subjecting the hazelnut milks to sufficient
suggested that the mentioned situation based on the relative dense
homogenization pressure(p → ∞), k2 is a constant that determines the
packing particles, inter-particle forces, and entanglements of cells,
initial exponential decay viscosity due to homogenization, where a
fibers and large colloidal molecules, such as proteins (Mert, 2012).
larger k2 indicates a rapid initial reduction in viscosity due to pressure,
The higher storage and loss modulus at low frequencies can also be
a and b are constants of power-sigmoidal function.
explained by the inertial resistance to flow and interaction between the
These models were selected due to exponential decay and a
particles and soluble molecules (Leite, Augusto, & Cristianini, 2014a).
sigmoidal shape of apparent viscosity as a function of pressure.
Similar results were previously reported for many suspensions prepared
Table 3 shows the parameters and determination coefficients of both
at different PH with network-like structures, fruit products (Leite et al.,
models, and Fig. 5 depicts the observed and predicted apparent
2014b; Silva et al., 2010), and tomato products (Augusto et al., 2013;
viscosity values of high pressure homogenized hazelnut milk. As can
Tan & Kerr, 2015). As can be seen in Fig. 6, the slope of G″ for all
be seen, both models could adequately (R2 ≥ 0.965) explain the
samples changed between 0.24 and 0.81, while the slope of G′ were
reduction of viscosity due to PH for hazelnut milk samples, which is
0.12–0.59. The higher the slope, the higher the dependence of
of interest for evaluation and design of HPH processing. k1 and k2
oscillatory frequency is (Karaman, Yilmaz, & Kayacier, 2013). The
values were determined as 0.3655 and 0.0067 for high pressure
slight dependence of G′ on the oscillation frequency is known as
homogenized alginate and 0.5198 and 0.0044 for high pressure
“plateau region”. This region is an intermediate zone of mechanical
homogenized K-carrageenan suspensions, respectively
spectra between the “terminal” and “transition” zones, and is classified
(Harte & Venegas, 2010). Augusto, Ibarz, and Cristianini (2012a)
by a decrease in the slope of both moduli (Dogan, Kayacier, Toker,
analyzed the effect of high pressure homogenization on fruit juice
Yilmaz, & Karaman, 2013). The behavior is typical for highly structured
serum model and reported k1, k2, a and b values as 2.55, 0.0059, 0.0099
materials, classifying the products between true gels (characterized by
and 0.856, respectively. These values are higher than our results, and
covalent cross-linked materials) and concentrated suspensions (char-
this is because of the differences in samples used, applied pressure and
acterized by entanglement networks) (Bayod, Willers, & Tornberg,
unit of apparent viscosity, which is Pa·s at present study while it is
2008).
MPa·s at other studies.
G′ and G″ values of hazelnut milk samples showed a rising tendency
with increasing oscillatory frequency, and thus, it was possible to model
3.5.2. Dynamic shear properties these modulus as a power function of oscillatory frequency (Eqs. (4)
The effect of high pressure homogenization (0–150 MPa) on the and (5)). As can be seen in Table 4, the R2 values of equations were
hazelnut milk samples mechanical spectra is given in Fig. 6. For all the always higher than 0.95, and the lowest R2 values (0.961–0.973) were
products, the storage modulus (G′) was always higher than the loss determined from storage and loss moduli of control sample (0 MPa).
modulus (G″) at all oscillatory frequencies (ω). This indicates that The slopes of storage moduli of hazelnut milk (n′ = 0.12–0.59) were
elastic properties of hazelnut milk are dominant, rather than the viscous relatively independent of frequency and lower than that of loss moduli
ones, and the products can be classified as soft gel network (n″ = 0.24–0.81) which were more dependent on frequency (Table 4).
(Rao & Cooley, 1992). Many food systems can be characterized by a The highest slopes of storage and loss moduli were determined for the
three-dimensional network where weak interactions, such as Wan der

Table 3
Constants for proposed models for describing the change in apparent viscosity of hazelnut milks processed with different homogenization pressure.

⎛ k + e−k 2 x ⎞ η=
η0
η = η0 ⎜ 1 ⎟ 1 + (ax )b
⎝ k1 + 1 ⎠
k1 k2 R2 a b R2

0.0102 ± 0.003 0.1192 ± 0.009 0.997 0.2090 ± 0.007 1.6727 ± 0.241 0.965

Values are means ± Standard Deviation.

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O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

Fig. 6. Effect of HPH on storage modulus, G′ and loss modulus, G″ of hazelnut milk samples, letters a-f refer to applied pressures from 0 to 150 MPa.

Table 4
Modified Cox-Merz rule shift factor and dynamic shear parameters of power-law functions describing storage and loss moduli of hazelnut milks prepared with different homogenization
pressures.

′ ″
PH (MPa) G′ = K′(ω)n G″ = K″(ω)n

K′ n′ R2 K″ n″ R2 αSF

0 4773 ± 325.27a 0.12 ± 0.03d 0.961 849.8 ± 75.16a 0.24 ± 0.02c 0.973 0.018
25 327.4 ± 69.58b 0.13 ± 0.02d 0.991 36.04 ± 0.72b 0.24 ± 0.03c 0.974 0.010
50 50.4 ± 2.80b 0.24 ± 0.01c 0.988 6.09 ± 2.12b 0.47 ± 0.15bc 0.989 0.012
75 14.5 ± 0.23b 0.44 ± 0.08b 0.984 3.03 ± 1.15b 0.52 ± 0.20b 0.982 0.011
100 11.9 ± 0.87b 0.41 ± 0.02b 0.986 0.82 ± 0.05b 0.61 ± 0.05ab 0.981 0.007
150 2.3 ± 0.17b 0.59 ± 0.04a 0.982 0.33 ± 0.06b 0.81 ± 0.01a 0.991 0.025

a–c
Values are means ± Standard Deviation. Means within the same column with different letters are significantly different at p < 0.05.

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O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

Fig. 7. Temperature sweep test of homogenized hazelnut milk samples.

samples processed with 150 MPa homogenization pressures (0.59–0.81, Cankurt, Kayacier, & Sagdic, 2011). The Cox-Merz rule can be used to
respectively) and it was significantly affected from homogenization predict the steady state shear properties of a material from the dynamic
pressure. These results also showed that the higher the slope is, the rheological properties obtained without extensive deformation of
more effective the frequency is. Therefore, it could be stated that the structures (Da Silva, Oliveira, & Rao, 1998). The best of our knowledge,
hazelnut milk processed with different homogenization pressure de- hazelnut milk is a new product and there is no research about
monstrated weak gel like behavior due to the positive slopes of applicability of this rule to hazelnut milk, and for this reason, we
modulus. The highest K′ and K″ values were determined from unpro- investigated whether hazelnut milk samples obey this rule. Hazelnut
cessed hazelnut milk sample in accordance with the steady-state shear cake contains high amount of starch, protein, oil and dietary fibers
results, and decreased as PH increased. which can significantly affect the rheological behaviors of hazelnut
Temperature dependency of the samples was also determined by milk samples. As commonly observed in food products, the magnitude
temperature sweep test applied at constant frequency (0.1 Hz) and of complex viscosity (η∗) is always higher than the magnitude of
stress (0.1 Pa). The effect of temperature on G′ and G″ of hazelnut milk apparent viscosity (ηapp) (Fig. 8).
processed by high pressure homogenization was determined as a The apparent viscosity (ηapp) and complex viscosity (η∗) of hazelnut
function of temperature, and results are given in Fig. 7. The hazelnut milk samples were plotted against shear rate (γ )̇ and angular frequency
milk showed a higher G′ than G″, in the whole range of tested (ω), respectively. As can be seen in Fig. 8, hazelnut milk processed by
temperatures. G′ and G″ values of hazelnut milks were slightly high pressure homogenization did not obey the Cox-Merz rule due to
increased at initial temperatures, and when the temperature reached higher η∗ values than those of ηapp. As a result the Cox-Merz rule was not
to 48 °C, these values increased sharply which indicates the gelation of applicable to the hazelnut milk samples and, as mentioned before,
hazelnut proteins, and the gel point or the temperature (Tgel) that this samples exhibited weak-gel properties because of deviations from Cox-
occurs (Hansen, Hoseney, & Faubion, 1991). The slightly increase of Merz superposition of η∗(ω) versus ηapp(γ )̇ (Karaman et al., 2011). It was
both moduli at initial temperatures can be related to the increase in also reported that biopolymer dispersions and complex food systems
fluidity with increasing temperature and homogenization pressure. This with either high density entanglements or aggregates could not obey
situation may also be attributed to the energy dissipation movement of this rule without applying the shift factor (αSF) (Yoo, Yoo, Kim, & Lim,
the molecules and decrease in intermolecular interactions, which in 2003).
turn decrease the energy needed for the flow, thus decreasing the The hazelnut milk samples had a considerable deviation between ln
interference of the hydrodynamic domains (Lapasin & Pricl, 1995). η∗ versus ln ω and ln ηapp versus ln γ ̇ data. As shown in Fig. 8, the data
Between 40 and 60 °C, an increase was observed for both moduli and set of η∗ versus ηapp was almost parallel for samples and, therefore, the
this situation could be attributed to starch gelatinization or protein deviation could be adjusted by multiplying ω with αSF shift factor
coagulation (Vanin, Michon, & Lucas, 2013), since hazelnut contains (Kim & Yoo, 2006).
both proteins and starch. Moreover, increase in both moduli might be ηapp (γ )̇ = η∗ (αSF ω) (9)
related to swelling of starch granules and protein aggregation or
strengthening of protein network due to thermal coagulation of proteins The shift factor (αSF) values of hazelnut milk samples prepared with
(Turan, Altay, & Çapanoğlu Güven, 2015). It can also be observed that different homogenization pressure is given in Table 4. As can be seen,
both moduli decreased as the temperature increased from 60 to 85 °C, the hazelnut milk samples processed with 0 MPa and 150 MPa pressure
meaning that the strength of the network was decreased due to protein had the highest αSFvalues which were 0.018 and 0.025, respectively. It
denaturation (Gabriele, de Cindio, & D'Antona, 2001). is clear from these results that αSF value generally decreased as the PH
increased, except for 150 MPa pressure. This means that the deviations
3.5.3. Applicability of Cox-Merz rule of hazelnut milk samples from Cox-Merz rule increased as αSF value
An empirical correlation between the apparent viscosity (ηapp) of decreased (Chun & Yoo, 2004; Kim & Yoo, 2006). Therefore, it can be
polymers as a function of shear rate (γ )̇ and the complex dynamic concluded that the deviations of hazelnut milks from the Cox-Merz rule
viscosity (η∗) as a function of angular frequency (ω) was described by was partially pressure-dependent, and the deviation of steady and
Cox and Merz (1958). The Cox-Merz rule is an empirical relationship dynamic viscosity of samples increased as the PH increased.
that equates complex viscosity (η∗) measured with oscillatory shear to
apparent viscosity (ηapp) determined from steady shear flow, where ω is 4. Conclusions
taken as γ ̇ in the oscillatory tests. Applicability of this rule has been
studied for many polymers, solutions and complex food systems HPH processing of hazelnut milk samples increased the solubility of
(Augusto et al., 2013; Basu, Shivhare, Singh, & Beniwal, 2011; Dogan proteins in water as PH increased up to 75 MPa. The mean particle
et al., 2013; Karaman, Yilmaz, Dogan, Yetim, & Kayacier, 2011; diameter of samples decreased as PH increased, but cumulative
Kim & Yoo, 2006; Yaşar, Kahyaoglu, & Şahan, 2009; Yilmaz, Karaman, percentiles did not change between 100 and 150 MPa.

418
O. Gul et al. Innovative Food Science and Emerging Technologies 41 (2017) 411–420

Fig. 8. Comparison of oscillatory and continuous shear viscosities (Cox-Merz rule) of homogenized hazelnut milk samples, letters a-f refer to applied pressures from 0 to 150 MPa.

Microstructural observations revealed that a progressive disintegration rheological properties. The reduction in apparent viscosity reduces the
of suspension and protein aggregates was observed as PH increased up energy consumption during unit operations such as pumping.
to 50 MPa, and after this pressure a completed cell disruption was Therefore, this process may lead to less energy consumption and more
determined. The change in particle size and shape with increasing PH efficient hazelnut milk processing.
altered the steady-state shear properties due to the reducing resistance
to flow of homogenized samples. These changes were discussed and Acknowledgements
modeled well using exponential and sigmoidal functions. Dynamic
shear properties showed that hazelnut milk samples could be classified This research did not receive any specific grant from funding
as soft-gel network, since higher G′ than G″ values was observed in the agencies in the public, commercial, or not-for-profit sectors.
whole range of investigated frequencies and temperatures, and no
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