Food Hydrocolloids 45 (2015) 236e246

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Effects of carboxymethyl cellulose (CMC) and microcrystalline

cellulose (MCC) as fat replacers on the microstructure and sensory
characteristics of fried beef patties
Monika Gibis*, Valerie Schuh, Jochen Weiss
Department of Food Physics and Meat Science, Institute of Food Science and Biotechnology, University of Hohenheim, Garbenstrasse 21/25, 70599 Stuttgart,
Germany

a r t i c l e i n f o a b s t r a c t

Article history: Microcrystalline cellulose (MCC) or carboxymethyl cellulose (CMC) can be used as potential fat replacers;
Received 19 September 2014 both are non-digestible fibers. The objective of this study was, therefore, to assess the impact of added
Accepted 20 November 2014 CMC/MCC on the microstructural/functional characteristics of beef patties. Therefore, low-fat beef patties
Available online 3 December 2014
were formulated with CMC/MCC at concentrations of 0.5e3.0 wt%. For this formulation, 10 wt% of the
ground beef was replaced using a dispersion of CMC or MCC in water. The influence of the addition of
Keywords:
MCC and CMC on the sensory characteristics, such as the texture, color, taste, flavor, and juiciness, of fried
Carboxymethyl cellulose
beef patties was evaluated by using a hedonic scale from 0 to 10 (5 ¼ optimal). The confocal laser
Microcrystalline cellulose
Beef patties
scanning microscopy (CLSM) measurements and sensory evaluation all indicated that addition of CMC
Fibers (>1 wt%) led to destabilization of the microstructure, sensory quality and texture of fried beef patties,
Fat replacer which, upon heating, could no longer be converted into a protein network with coarse meat and fat
particles, a fact that was also revealed in the CLSM images. By contrast, the CLSM images prove that MCC
was highly compatible with the matrix and improved the texture in the sensory evaluation with
increasing concentration compared to the control. The best sensory evaluation achieved was with the
beef patties containing 2 wt% MCC. Generally, hot samples containing MCC had more juiciness than the
control and had a fat-like mouthfeel. Contrarily, CMC is not suitable as a fat replacer in concentrations
more than 0.5 wt%.
© 2014 Elsevier Ltd. All rights reserved.

1. Introduction (Jime
nez-Colmenero et al., 2010), modified starches (Hoffman &

The development of healthier food products with added values
relating to diet and low caloric content has become a key target for
the food industry. The main approaches to reduce the content of fat
in meat products are: (i) the use of lean meat as a raw material to
decrease the amount of fat in the formulation, and/or (ii) replacing
the fat with lower caloric ingredients and/or water (Jime
nez-
Colmenero, 1996; Jime
nez-Colmenero, 2000; Jime
nez-Colmenero,
Carballo, & Cofrades, 2001; Schuh et al., 2013). Alternative com-
ponents that can be used as functional ingredients to replace fat
and meat are hydrocolloids, such as carrageen (Candogan &
Kolsarici, 2003; Hsu & Sun, 2006), xanthan (Ramirez, Barrera,
Morales, & Vazquez, 2001), guar gum (Ulu, 2005), konjac
* Corresponding author. Tel.: þ49 711 459 22293; fax: þ49 711 459 24446.
E-mail address: gibis@uni-hohenheim.de (M. Gibis).
Mellett, 2003), dietary fibers such as rye bran, oat bran and
barley (Desmond, Troy, & Buckley, 1998; Petersson, Godard,
Eliasson, & Tornberg, 2014; Pin ~ ero et al., 2008) or short-chained
polyfructoses (Vandendriessche, 2008), which do not have an
allergen impact compared to non-meat proteins. Microcrystalline
cellulose (MCC) or carboxymethyl cellulose (CMC) can be used as
potential fat replacers; both are non-digestible fibers (Schuh et al.,
2013).
In order to improve its water solubility, CMC is formed from
cellulose after heating with alkali, and this mixture reacts with
monochloroacetic acid leading to an alteration of the glycopyranose
by etherification of the hydroxyl groups with methylcarboxyl
groups. The properties of CMC are dependent on molecular weight,
degree of substitution ( DS) and substitution groups (Shand,
Schmidt, Mandigo, & Claus, 1990). By contrast, the non-water sol-
uble MCC is prepared by partial hydrolysis from wood pulp heated
in hydrochloric acid, and by separation of the non-crystalline part

http://dx.doi.org/10.1016/j.foodhyd.2014.11.021
0268-005X/© 2014 Elsevier Ltd. All rights reserved.
 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246
of cellulose (Lucca & Tepper, 1994). Before its use as an additive in
meat products, it is subjected to severe mechanical shearing,
physically breaking it down into colloidal crystallite aggregates,
which are then co-dried with CMC and/or other functional in-
gredients (Lucca & Tepper, 1994). MCC can be dispersed as insoluble
microcrystals in water under mechanical attrition; 60e70% of the
microcrystals are less than 0.2 mm long. CMC holds the insoluble
cellulose microcrystals together in a network (Lucca & Tepper,
1994). Microcrystalline cellulose is a hydrocolloid belonging to a
wide range of cellulose derivates. It is used for different applica-
tions in cosmetic, pharmaceutical and food products as a filler and
stabilizer due to its viscoelastic properties (Zhao, Kapur, Carlin,
Selinger, & Guthrie, 2011). The addition of MCC to meat products
is not common. Few studies have been made investigating the
impact of MCC on low-fat meat products (Barbut & Mittal, 1996;
Mittal & Barbut, 1993). When replacing fat with alternative in-
gredients, the functional properties of the meat products, such as
sensory characteristics, must not be negatively affected, apart from
safety, microbial and physicochemical stability. Fat is a basic
ingredient of meat products with specific sensory perception in
texture and moistness, such as bite and mouthfeel (Weiss, Gibis,
Schuh, & Salminen, 2010). Additionally, fat is also a carrier com-
pound and release media for flavors (Lucca & Tepper, 1994). It is
very difficult using alternative ingredients to achieve all charac-
teristics desired, such as low calories, fat-like physical properties,
unchanging flavor, and comparable mouthfeel.
An amount of 3% fiber is required for labeling as a source of
dietary fiber in foods in Europe. A fat reduction in food can be
labeled if 30% of fat is reduced compared to the fat amount of a
standard product. The reduction of fat in a standard product by
applying only lean meat led to products that are harder to chew
(Claus, 1991). For this reason, other ingredients are required to
compensate for this lack in texture.
Low-fat beef patties were formulated with CMC or MCC at
different concentrations in water dispersion to study the impact of
CMC or MCC on the microstructural and functional characteristics of
beef patties. The higher amount of water and decreasing fat content
can also result in higher weight losses, negatively affecting the
product quality and yield. Both fibers are approved for use in foods
and do not contribute to the caloric content of foods (Holtzapple,
2003; Keeton, 1996). They are often used in combination with
other hydrocolloids (O'Connor & O'Brien, 2002). The abilities of CMC
acting as a fat replacer, emulsion stabilizer and thickener have been
explored in dairy and bakery products (Corredig, Sharafbafi, &
Kristo, 2011; O'Connor & O'Brien, 2002), as well as in low-fat meat
products, such as emulsified sausages (frankfurter-style, bologna-
style) and hamburger patties (Hoffman & Mellett, 2003; Jime
nez-
Colmenero, 1996; Sampaio, Castellucci, Pinto e Silva, & Torres,
2004; Ulu, 2005). However, the addition of CMC can result in a
decrease of firmness of the lean meat products (Ruusunen et al.,
2003). The hardness in low-fat frankfurters was also decreased
with the first bite by 16e25% (Lin, Keeton, Gilchrist, & Cross, 1988).
We hypothesized that the replacement of common ground beef
containing approximately 20% fat by a combination of ground lean
meat (10% fat) and an aqueous dispersion of cellulose fibers form-
ing soft gel particles (10 wt%) may yield a fat content reduced
product having similar sensory properties than a common beef
patty containing 20% fat. Moreover, we postulate that there may be
significant differences in ability of the two polymers to act as fat
replacers due to differences in their solubilization and interaction
behavior. Methods of analysis included confocal laser scanning
microscopy (CLSM), chemical analyses of fat and water, color
measurement, determination of weight loss and sensory evaluation
after frying. The results of these analyses assessed what impact the
addition of MCC or CMC had on the quality of fried beef patties.
237
2. Materials and methods
2.1. Materials
CMC and MCC were provided by Danisco A/S (Aarhus,
Denmark). The specifications of manufacturer are shown in Table 1.
Fluorescence stains Calcofluor White and Nile Red and acetone
were purchased from Fluka Chemie AG (Basel, Switzerland). Beef
for the preparation of ground beef was purchased from a local
retailer (MEGA, Stuttgart, Germany). Further ingredients were so-
dium chloride (Südsalz GmbH, Heilbronn, Germany) and white
pepper (Gewürzmüller, Korntal-Münchingen, Germany).
2.2. Preparation of CMC and MCC hydrocolloid dispersion
A concentrated and homogenous CMC or MCC dispersion were
prepared by mixing 5e30% wt% CMC powder and 70e95 wt% water
in a 13-L bowl chopper (Mado Garant, Maschinenfabrik Dornhan
GmbH, Dornhan, Germany) at 24 rpm with the highest revolution
speed of rotating blades (2800 rpm, 4 min). Dispersions of either
CMC or MCC (10 wt%) were added to the ground beef (90 wt%).
Sinews were roughly removed from the pure beef, it was defatted
and then coarsely minced through a 3 mm plate. Salt (1.2 g/100 g)
was added to the minced beef separately and was mixed with a
blender (Kitchen Aid, Strombeek-Bever, Belgium). The following
concentrations of MCC and CMC were used in the product before
frying: 0.5, 1.0, 2.0, and 3.0% of each powder. Controls of 100 wt%
ground beef and 90 wt% ground beef plus 10 wt% water were used.
2.3. Preparation of beef patties and frying conditions
An amount of 60 g ± 1 g of this material was formed into beef
patties with a special mold for beef patties (12 mm thick  85 mm
diameter). The patties were fried at 220  C for 2.5 min with double
contact griddle plates (Nevada, Neuma €rker, Bad Bramstedt, Ger-
many). The beef patties were laid between two sheets of aluminum
foil. Four patties were fried together with the double contact
griddle plates. For each batch, eight patties were fried on both sides
simultaneously to a core temperature of 72  C and to a surface
temperature <200  C at the end of the frying process. Surface and
core temperature were monitored during the frying process with a
temperature data logger (Almemo 8990/8, Ahlborn, Holzkirchen,
Germany).
2.4. Determination of molecular weight of CMC
The molecular weight of the CMC used was measured using a
Paar Physica MCR 300 rheometer equipped with a double coaxial
cylinder and Rheoplus/32 V3.331 operating system (Anton Paar
Germany GmbH, Ostfildern, Germany). Diluted CMC (0e0.09 wt%)
solutions (1 M NaCl) were determined by shear rate versus stress.
The slopes of the linear regression of concentration-dependent
functions (shear rate versus stress, Fig. 1A) were calculated using
Excel (Microsoft, Redmond, WA, USA). Relative, specific and
reduced viscosities were calculated to determine the intrinsic vis-
cosity [h] of the CMC sample. SigmaPlot (Systat Software Inc., San
Table 1
Specification of CMC and MCC fibers used regarding viscosity and degree of sub-
stitution ( DS) of CMC according to the manufacturer; MMW e medium molecular
weight, SPD e spray-dried.
CMC/MCC sample code Viscosity [mPa  s] Notes
®
Grindsted CMC BAK 130 Min. 1500 (1% solution) 0.55e0.75 ( DS)
Grindsted® MCC MAS 17 LS 39e130 (1.2% solution) 8.4e13.7% CMC, SPD
238 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246

400 2.5. Weight loss after frying

A 0.00 wt% Eight samples of formed raw beef patties (~60 g) were weighed.
0.01 wt% The patties were fried to a core temperature of 72  C for 2.5 min.
0.02 wt%
After 1 h, the weight of the samples was recorded again. The weight
300 0.03 wt%
loss after frying (WTLoss) was calculated as:
Shear Stress (Pa)

0.04 wt%
0.05 wt% Minitial  Mend
0.06 wt% WT Loss ½% ¼ $100 (2)
Minitial
0.07 wt%
200 0.08 wt% where Minitial is the weight of the sample prior to frying and Mend is
0.09 wt% the weight of the sample after frying.

2.6. Firmness
100
In order to imitate the inner texture of the patties, the crust was
removed and the patties were homogenized to imitate the
mouthfeel during chewing. This was simulated by an Instron uni-
versal testing machine (Model 1011, Instron Engineering Corp.,
0 Canton, MA, USA) with a 5 kN load cell operated by a control
0 200 400 600 800 1000
software (Instron Series IX V5.25, Instron GmbH, Darmstadt, Ger-
many) to determine penetration forces. The machine was equipped
Shear Rate (s -1) with a conical measurement geometry (cone diameter: 30 mm,
5000 cone height: 26 mm, cone angle: 60 ). Homogenized material was
filled into special molds with a diameter of 32 mm, and wrapped in
B Confidence interval a plastic bag to prevent drying. Each mold with material in a plastic
Preditive interval bag was then compressed under vacuum to remove any cavities.
4000 Molds with samples were stored for 1 h at 18  C to equilibrate
samples in order for them to have the same temperature prior to
Reduced Viscosity

measurements. Samples were then subjected to a compression test

and the maximum penetration force needed was recorded and then
3000
related to the sample weight (N per g material). The compressional
speed was set to 100 mm/min at a penetration depth of 85%. Four
measurements per sample were performed.
2000
2.7. Sensory test

After frying and cooling down to ambient temperature (1 h), the

1000 beef patties were stored in plastic bags at 4  C overnight. On the
next day, each patty of either CMC or MCC formulation was cut into
four roughly equal parts. Six samples were put on a plate, covered
0 with a plastic top, and reheated in the microwave for 1.5 min at
0.00 0.02 0.04 0.06 0.08 0.10 700 W. Heated samples were immediately served to 29 non-trained
sensory testers. They evaluated the samples for the attributes odor,
Concentration of CMC (wt%) flavor, texture, juiciness, and color using a hedonic scale scoring
from 0 up to 10. Sensory scores were rated using a continuous 10-
Fig. 1. Determination of molecular weight of CMC using different concentrations
cm scale for odor (0e10, too weak frying odor to burnt odor,
(0.01e0.09 wt%). (A) Shear rate (1/s) versus shear stress. (B) Intrinsic viscosity calcu-
lated from shear rate versus shear stress (y ¼ 24034x þ 1504, R2 ¼ 0.987, only values in 5 ¼ optimal), texture (0e10, too firm to too soft/pasty, 5 ¼ optimal),
the predictive interval (p ¼ 0.95) were used for the linear regression). juiciness (0e10, too dry to too moist, 5 ¼ optimal), color (0e10, very
light to very dark) and flavor (0e10, under-fried to over-fried
(burnt), 5 ¼ optimal). The panelists (n ¼ 29) evaluated the sam-
Jose, CA, USA) was used for the calculation of linear regression with ples by a mark on the scale. In addition, they had to highlight the
the confidence and predictive interval (p ¼ 0.95). Only values sample preferred. The middle of the scale (5) was the optimal and
within the predictive interval were used for the calculation of typical evaluation for each attribute each time.
intrinsic viscosity [h]. The molecular weight M is calculated by the
Mark-Houwink equation: 2.8. Determination of fat and moisture content

½h ¼ Kh $Ma
where [h] is intrinsic viscosity, M is molecular weight and K and a
are constants for a given solvent and temperature, respectively. To
calculate M from the Equation (1), K and a were taken from another
study as Kh ¼ 1.23$102 and a ¼ 0.91 (solvent 0.1 M NaCl at 25  C)
(Kulicke, Clasen, & Lohman, 2005).
The fat and moisture content of the fried patties was determined
(1) according to the instructions of the official collection of methods of
analysis (BVL, 2011).
2.9. Confocal laser scanning microscopy (CLSM)
A Nikon CLSM (Nikon D Eclipse C1, Nikon GmbH, Düsseldorf,
Germany) equipped with an argon ion single line laser (488 nm,
 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246 239

10 mW) was used for the study of the microstructure of the fried
beef patties. The crusts of the patties were cut away and samples
were taken from the inner part of different patties without crusts.
Solutions of Calcofluor White (20 mL; lEx: 440 nm, lEm:
500e520 nm) for protein staining or Nile Red (20 mL; 2 mg/10 ml
acetone; lEx: 515e530 nm, lEm: 525e605 nm) for fat staining were
added to the samples (2 mm thickness, 20 mm diameter), and
evenly distributed. Stains were used separately. The specimen was
then covered with a cover slip and fixated with adhesive tape. The
samples were kept in the dark to prevent photo-bleaching prior to
observation. Samples were excited at 488 nm and the fluorescence
emission of the samples was detected at 515/20 (Calcofluor White)
and 585/40 (Nile Red). Images of representative areas were taken
with a 4, 10 and 20, magnification objective (Plan-Apochromat
Plan Fluor 4/0.13, Plan Fluor 10/0.30, Plan Apo 20/0.75; Nikon
GmbH, Düsseldorf, Germany).
2.10. Image analysis using box counting
The microscopy images stained with Calcofluor White were
analyzed by using image analysis software ImageJ 1.47v from
Wayne Rasband (National Institutes of Health, Bethesda, MD, USA)
plus the fractal dimension tool FracLac2013Aug122 from A. Kar-
perien (Charles Sturt University, Australia). The colored images
were transformed to grayscale and then to black and white binary
images. Structure factors (Df using the box counting method) of
each binary image were determined with the box sizes automati-
cally adjusted by the software (default values were used; minimum
pixel density: 0.2; maximum: 1.0). The maximum spacing in our
method was set to 45% of the region of interest of the image, the
scan position was 12 and the box size was automatically adjusted by
using the software. A grid of known spacing (scale) was placed over
the binary image and the number of boxes that contained non-zero
pixels was counted. This process was then repeated for multiple
grids with increasing spacing. As the scale increases, the number of
boxes containing the object decreases exponentially and the
exponent is equivalent to the Df. To quantify the exponent (struc-
ture factor Df), natural logarithmic plots of the number of boxes
against scale and the slope (structure factor Df) were fitted by using
linear regression.
2.11. Statistical analysis
All measurements were repeated at least three times using
duplicate samples. Means and standard deviations were calculated
using Excel (Microsoft, Redmond, WA, USA). Statistical analysis was
carried out on measurements of firmness and weight loss using the
Statistical Analysis System (SAS 9.3, SAS Institute Inc., NC, USA). The
data was tested for assumptions, such as normal distribution, ho-
moscedasticity and one-way variance analysis using a Tukey's
studentized range test with the generalized linear model (GLM) to
determine significant differences (a ¼ 0.05) between sample and
control batches. A two-way ANOVA (SigmaPlot 12.5, Systat Soft-
ware Inc., San Jose, CA, USA) was used (a ¼ 0.05) for the structure
factor.
polymer concentrations in Fig. 1B, and was used to calculate the
molecular weight with the Mark-Houwink Equation (1). The mo-
lecular weight of the CMC used was ~390 kDa. The molecular
weight in combination with the degree of substitution (DS ¼ 0.55)
(Table 1) may have an important influence on the water-binding
capacity (Lin et al., 1988) and, thereby, on the sensory perception
of the fried beef patties. The MCC used is an additive which contains
approximately 11% CMC in addition to the MCC molecules accord-
ing to the manufacturing specification. As mixed fibers, the deter-
mination of molecular weight is not meaningful.
3.2. Weight loss after frying
The weight losses of the fried beef patties are shown in Table 2.
CMC concentrations from 2 to 3 wt% generally led to a significant
(p < 0.05) decrease in weight loss compared to the control (19.2%).
The control containing 10% water and 90% ground beef has the
highest significant weight loss (26.3% CMC batch and 24.3% MCC
batch). Addition of 3 wt% CMC reduced the weight by about 15.5%
after frying. Addition of higher viscous MCC dispersion (3%) also
showed similar weight losses (19%), but the differences were not
significantly (p > 0.05) different to the control with 100% beef, but
were significantly different to the second control containing 90%
ground beef. Generally, we found increased standard deviations in
the weight losses of the MCC samples. One reason could be the
activation process of MCC which required a high shear stress for the
MCC dispersion. It is possible that the activation is not fully
completed or the microcrystals are widely distributed in size.
Normally, approximately over 50% of the microcrystals have a
length about 0.2 mm (Lucca & Tepper, 1994). In contrast to an earlier
study on low-fat frankfurters, two different MCCs increased mois-
ture loss by 12e15% compared to the low-fat control and samples
containing CMC. In the application of CMC in emulsion-type low-fat
frankfurters, the weight losses after heating for samples containing
CMC were significantly reduced from 10 to 6% compared with the
control with substitution of fat by water (Barbut & Mittal, 1996).
Significant (p < 0.05) improvements in cooking yield (74.2%) of low-
fat patties were attributed to the water-binding ability of b-glucan
~ ero et al., 2008). These findings were comparable with results
(Pin
in our study with CMC, showing that weight loss decreased during
frying.
3.3. Moisture and fat content
The fat and moisture content of the fried beef patties are shown
in Table 3. As expected, cooked beef patties formulated with an
increasing addition of CMC fiber were significantly (p < 0.05) higher
in moisture content and lower in dry matter than the controls. The
moisture content increased from 56.5% in the control to 63.3% in the
samples with 3% CMC. The fat content decreased from 9.5% to 7.0%.
Table 2
Weight loss and texture analysis (firmness measured as penetration force) of fried
beef patties (mean ± standard deviation) containing 0e3 wt% MCC or CMC after
heating and cooling down.

MCC CMC
3. Results and discussion a a
Weight loss Force Weight lossa Forcea
(g/100 g) (N/g) (g/100 g) (N/g)
3.1. Determination of molecular weight
Control 19.2 ± 1.60a 6.3 ± 0.58a 22.0 ± 1.27a 6.1 ± 0.59a
Control 90% beef 24.3 ± 1.64b 6.4 ± 0.14a 26.3 ± 1.32b 6.4 ± 0.15a
The average molecular weight of the CMC used was determined 0.5 wt% Fiber 24.2 ± 3.04b 6.5 ± 0.45a 24.7 ± 0.88c 4.0 ± 0.99bcd
using the Mark-Houwink equation. The shear rate as a function of 1.0 wt% Fiber 22.4 ± 3.47ab 11.3 ± 0.79b 23.2 ± 1.35a 6.3 ± 0.31a
shear stress of the molecular weight of the CMC medium used is 2.0 wt% Fiber 21.3 ± 3.00a 10.7 ± 2.74b 17.9 ± 1.01d 4.5 ± 0.11b
shown in Fig. 1A. The intrinsic viscosity was determined by calcu- 3.0 wt% Fiber 19.1 ± 2.25a 10.1 ± 0.28b 15.5 ± 1.36e 2.8 ± 0.42c
a
lation of the intercept of the function of reduced viscosity versus Means with different letters are significantly different (p < 0.05).
240 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246

The main part of the reduction resulted in the lower loss of moisture significantly firmer than the control samples. As shown in Table 2,
during frying. That is also presented in the decrease of weight loss the force increased from 6.38 (control 90% beef) to 11.3 N/g of the
during frying in Table 2. Clearly, lower fat content using increased 1 wt% sample. The higher concentrations of MCC (2e3 wt%) showed
CMC concentrations compared to MCC, was found due to the lower no significant difference in firmness compared with the sample
weight loss after frying also resulting in a higher content of moisture containing 1 wt% MCC. No significant change to the control was
(Table 3). Only a slight reduction of fat content was observed using observed only when MCC was added to the formulation in a con-
increased concentrations of MCC (Table 3). Samples containing 3% centration of 0.5 wt%. These results indicate that MCC may form gel
MCC have the lowest fat content in the trial with MCC, which is a particles after cooling down to ambient temperature. CMC, present
reduction of fat of approximately 20% compared to the control with in MCC, was responsible for binding the MCC into a thick sticky
100% ground beef, but only 17% compared to the control containing paste, which accounts for the high gel strength for MCC (Correia &
90% ground beef. However, only a slight increase of water retention Mittal, 2000).
was determined in the patties containing MCC (Table 3). Similar results were reported in a recent study (Schuh et al.,
Other researchers have shown similar results (Mansour & Khalil, 2013): the firmness increased when MCC was present in the
1997; Pin~ ero et al., 2008; Schuh et al., 2013) when using CMC, MCC heated sausages after chilling. These emulsion-type sausages con-
or other fat replacers, such as oat or wheat fibers. Fat content was taining MCC showed, with the exception of 0.3 wt% MCC, signifi-
significantly (p < 0.05) lower in soluble oat fiber in beef patties cantly higher firmness than the control samples, and the batch
compared to that of the control. By contrast, an increase in the fat containing 2 wt% MCC had the highest firmness, approximately 20%
content of cooked low-fat beef patties was found. The authors higher than that of the control batch (Schuh et al., 2013). The results
concluded that oat and wheat fibers have the ability to retain fat in low-fat frankfurters showed that the addition of MCC reinforced
during heating, resulting in the lower cooking losses (Mansour & the firmness of the product in comparison to the control batch
Khalil, 1997; Pin~ ero et al., 2008). Significantly more moisture was (Barbut & Mittal, 1996; Mittal & Barbut, 1993).
retained in low-fat products containing CMC or MCC (29.5% and
31.9%, respectively) compared to the low-fat control in a study with 3.5. Sensory evaluation
low-fat sausages, but this did not indicate a significant decrease in
the cooking losses of the control samples (Mittal & Barbut, 1993). The results of the sensory evaluation of the fried beef patties
containing different cellulose fibers are presented in Fig. 2 for CMC
3.4. Texture and Fig. 3 for MCC. No significant differences were observed be-
tween the controls containing either 100% ground beef or 90%
In order to obtain a better comparison with the sensory data, we ground beef plus 10% water for any attributes in either series of
homogenized the patties before measuring the firmness, which may sensory panels. In general, low-fat beef patties containing an
show the impression of chewing and mouthfeel more. The mea- increased amount of lean meat become firmer after frying due to
surements of texture, particularly firmness, of the homogenized the additional amount of lean meat used in the formulation in
patties at ambient temperature confirmed that firmness decreased comparison to high-fat samples (Pin ~ ero et al., 2008), which nor-
with increasing concentration (>0.5 wt%) of CMC in the ground meat mally contain about 20% fat (Berry & Abraham, 1996). That is the
(Table 2). This is in agreement with previous studies (Lin et al., 1988; reason why the beef patties tested did not get the optimal scores for
Mittal & Barbut, 1993; Ruusunen et al., 2003; Schuh et al., 2013). texture and juiciness, because the fat content of the prepared
However, the addition of higher concentrations of CMC (2 or 3 wt%) patties was approximately 10% (Table 3).
significantly weakened the protein network between the fat and Fried beef patties containing soluble fibers, such as CMC, were
meat particles even more by about 50% from 6.13 N/g (control) to found to be significantly (p < 0.05) juicier than the control (Fig. 2),
2.75 N/g (3 wt% CMC). This is in agreement with findings of a recent which could be attributed to the increased moisture retention of
study (Schuh et al., 2013), which reported a decrease in firmness of the product after frying (Table 3) and lower cooking loss during
bologna-type sausages when high concentrations of CMC (3 wt%) frying (Table 2). In particular, drastic changes to softer texture with
were added to the formulation. Other authors reported a decline in increasing concentrations of CMC were observed. The patties were
firmness in the first bite, and they hypothesize that this decrease too moist and the texture of patties was significantly evaluated as
might be due to an alteration in proteineprotein or proteinewater too soft (p < 0.05). At the highest addition amount of 3 wt%, the
interactions, resulting in a decrease in gel strength of the prepared fried patties had a pasty structure and the sensory testers disliked
meat products (Lin et al., 1988). these patties. Only the fried patties containing 0.5 wt% CMC were
The addition of MCC to the formulation of patties with significantly assessed with optimal scores (approximately score 5)
increasing concentrations of MCC showed a completely different for texture compared to the controls (p < 0.05). However, the color
behavior in texture. The fried patties containing 1e3 wt% MCC were of fried patties was evaluated significantly lighter than the control
samples (p < 0.05) (Fig. 2).
These findings are in agreement with studies using b-glucans as a
Table 3
fat replacer (Desmond et al., 1998; Pin ~ ero et al., 2008). In this
Concentration of moisture and fat of beef patties (mean ± standard deviation) using
different concentrations of fibers (0e3 wt%) after frying. connection, the results of these studies showed that the b-glucans
from oat fibers increased the water-binding capacity and produced
MCC CMC
juicier low-fat beef patties. The behavior of b-glucan depends on the
Moisturea Fata Moisturea Fata origin of b-glucan (oat, barley and rye bran) (Petersson et al., 2014).
(g/100 g) (g/100 g) (g/100 g) (g/100 g)
For emulsion type sausages, addition of oat bran fibers yielded
Control uncooked 64.5 ± 0.2 11.1 ± 0.30 64.5 ± 0.2 11.1 ± 0.30 products exhibiting low process and frying losses, since these fibers
Control 56.9 ± 0.3a 10.3 ± 0.23a 56.5 ± 0.1a 9.5 ± 0.01a
contained soluble b-glucan fractions able to form gels that were
Control 90% beef 53.2 ± 0.2b 9.9 ± 0.04b 57.8 ± 0.1b 8.8 ± 0.11b
0.50% Fiber 58.7 ± 0.1c 9.4 ± 0.02c 56.5 ± 0.1c 9.4 ± 0.10a suitable as fat replacers. This is in contrast to barley fibers having
1.00% Fiber 56.2 ± 0.1d 9.3 ± 0.10c 60.6 ± 0.2d 7.6 ± 0.23c containing soluble b-glucan fractions with poor gel-forming abili-
2.00% Fiber 55.9 ± 0.4d 8.9 ± 0.04d 62.3 ± 0.3e 7.0 ± 0.21d ties. Both oat bran and barley fibers were not suitable as fat replacers
3.00% Fiber 57.9 ± 0.1e 8.2 ± 0.11e 63.3 ± 0.1f 7.0 ± 0.01d in meatballs due to them not providing a firm texture. Only rye bran
a
Means with different letters are significantly different. fibers containing high amounts of insoluble arabinoxylans and
 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246 241

Fig. 2. Sensory evaluation of fried beef patties containing 0e3% CMC using a hedonic scale for the attributes odor (0e10, too weak frying odor to burnt odor, 5 ¼ optimal), texture
(0e10, too firm to too soft/pasty, 5 ¼ optimal), juiciness (0e10, too dry to too moist, 5 ¼ optimal), color (0e10, very light to very dark), and flavor (0e10, under-fried to over-fried
(burnt), 5 ¼ optimal). (Mean ± standard deviation as grayscale interval).

cellulose providing for a firm structure was appropriate due to no
gel-forming ability for addition to meatballs thereby being accept-
able in the sensory tests (Petersson et al., 2014).
The results of the sensory evaluation for the fried patties con-
taining MCC are shown in Fig. 3. Fried patties were evaluated with
increasing concentrations of MCC as significantly softer and juicier
than the controls (p < 0.05). The patties containing MCC from 1%e
3% were scored more as typical and optimal for fried patties. The
addition of 2% MCC showed a significantly better sensory evalua-
tion regarding texture and juiciness than the controls (p < 0.05);
the panelists preferred this sample (p < 0.05) and described the
perception of mouthfeel as fat-like.
242 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246

Fig. 3. Sensory evaluation of fried beef patties containing 0e3% MCC using a hedonic scale for the attributes odor (0e10, too weak frying odor to burnt odor, 5 ¼ optimal), texture
(0e10, too firm to too soft/pasty, 5 ¼ optimal), juiciness (0e10, too dry to too moist, 5 ¼ optimal), color (0e10, very light to very dark), and flavor (0e10, under-fried to over-fried
(burnt), 5 ¼ optimal). (Mean ± standard deviation as grayscale interval).

Only a few studies described MCC as a fat replacer in sausages
(Barbut & Mittal, 1996; Mittal & Barbut, 1993; Schuh et al., 2013).
Low-fat sausages were investigated in these studies. However, the
addition of MCC resulted in products perceived to be less tender
than the high-fat product, but more tender than the reduced-fat
control product; the perception of juiciness was not affected by
the addition of MCC (Mittal & Barbut, 1993). Authors also described
that the addition of MCC did not significantly increase toughness
(Barbut & Mittal, 1996). The sensory panel did not significantly
differentiate juiciness scores between the controls and MCC prod-
ucts; the acceptability scores also indicated no difference between
the high- and low-fat MCC products (Barbut & Mittal, 1996).
 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246 243

Fig. 4. CLSM images of fried beef patties containing CMC and MCC at concentrations of 0e3 wt%, stained with the fluorescence dye Calcofluor White (lEx: 440 nm, lEm:
500e520 nm). Images were taken with an objective Plan-Apochromat Plan Fluor 4/0.13; control without hydrocolloids, 0.5e3 wt%, CMC and MCC; scale bar 500 mm.

3.6. Confocal laser scanning microscopy and structure factor of
images
Confocal laser scanning microscopy images were taken to
evaluate the changes in the structure of the meat fiber protein
network and coarsely distributed fat and meat particles (Figs. 4
and 5). Confocal laser scanning microscopy was chosen to visu-
alize major changes in the formation of the protein meat fat par-
ticle network and confirm differences in texture and sensory
evaluation, as well as water loss after heating, using hydrocolloids
in patties. This system is very prone to ionic strength, formulation,
pH, or temperature. During frying, the protein matrices in patties
formed a coarsely connected protein network between the
coarsely minced meat and fat particles incorporated. In this
connection, the myofibrillar proteins are solubilized after addition
of salt and the protein network containing the embedding parti-
cles is formed by crosslinking upon heating which enhanced the
binding and consistency (Tornberg, 2005). Thereby, a beef patty is
composed of more or less intact meat fibers as well as bundles up
to 50e70%, but they are randomly distributed compared to the
well-defined anisotropic structure of the whole muscle in light
microscopy (Tornberg, 2005). Fig. 4 shows micrographs of fried
control patties and patties containing CMC and MCC at different
polymer concentrations. The distribution of coarsely ground fat
and fat particles in a surrounding matrix of salt-solubilized pro-
teins is depicted in Fig. 5. When CMC was added to the system, the
structure of the matrix changed. The appearance of voids and the
absence of a network containing embedded particles upon use of
CMC can be seen. Typically, after salt addition, proteins are solu-
bilized thereby able to “glue” meat and fat particles in the patties
together. Upon heating a continuous particle-filled network is
thereby formed. CMC as a negatively charged polymer may
repulse soluble proteins that are also negatively charged at pH
values around 5.8. In turn, a phase separation takes place causing
the structure to become heterogenous. This phenomenon is highly
dependent on the concentration of polymer used. Changes are
visible by means of the CLSM from 1% CMC. Addition of 2 or 3 wt%
CMC led to a highly porous and uneven meat cellulose fiber
network or a matrix that contained individually distributed meat
and CMC fibers rather than a crosslinked network (Fig. 4). Meat
and cellulose fibers in the denatured protein network did not
show any connection to each other. In another study, the results
showed no alterations in network microstructure when adding
1 wt% CMC (Chattong, Apichartsrangkoon, & Bell, 2007).

Fig. 5. CLSM images of fried beef patties containing CMC and MCC at concentrations of 0 (control), 3 wt%, stained with the fluorescence dye Nile Red (lEx: 515e530 nm, lEm:
525e605 nm). Images were taken with an objective Plan-Apochromat Plan Fluor 4/0.13; scale bar 500 mm.
244 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246

Contrarily, the microstructure of MCC samples showed no

visible changes compared to the structure of the control samples.
Meat and cellulose fibers had visible connections to each other and
MCC may fill the gaps of the meat fiber network.
To determine these alterations of the microstructure in the
CSLM images, the values of a structure factor (Df) were calculated
for each magnification separately. The structure factor is an index
for characterizing fractal patterns or sets by quantifying their
complexity as a ratio of the change in detail to the change in scale.
For this approach, a model is needed to describe the high degree of
erratic behavior of surface complexity in some controlled way
(Sarka & Chaudhur, 1992).
In Fig. 6, the calculated structure factor of the images at each
magnification versus the concentration of CMC (Fig. 6A) and MCC
(Fig. 6B) are shown. The structure factors of the images with patties
containing increasing concentrations of CMC grew to higher values.
The control samples without CMC and patties containing 0.5% CMC
differed significantly from the beef patties containing the highest
CMC concentration, which had significantly higher values in
calculated structure factors of the CLSM images (p < 0.05). By
contrast, the images of fried patties containing MCC did not show
any significant difference between the calculated structure factors
of the images taken at different magnifications.
Studies of mixtures containing proteins and polysaccharides,
such as gelatin and maltodextrin, showed that phase separation
may occur even at low concentrations, and that formation of a
continuous network is highly dependent upon pH, salt and tem-
perature (Lore
n & Hermansson, 2000; Lore
n, Langton, &
Hermansson, 1999). Food hydrocolloids such as CMC may behave
in a similar way, thus leading to phase separation in coarsely
dispersed meat systems. Comparable phase separations as a result
of electrostatic repulsion between CMC and gelatin were reported
earlier (pH > pI of the protein, >0.2 M salt) (Tolstoguzov, 1991).
The distribution of fat particles in the fried meat matrix is shown
in Fig. 5. The control samples contained unevenly dispersed fat
particles throughout the meat fiber network with mean particle
sizes varying widely (Fig. 5). With the addition of CMC, the fat
particle size decreased in tendency, and the form also seems to
have a more regularly spherical shape in tendency (Fig. 5). By
contrast, added MCC show no remarked changes to the control.
However, the variations are too large due to the coarsely minced
ground beef used. Other researchers observed that fat particle size
decreased with the addition of low amounts of CMC (0.5e1 wt%) in
Thai sausages (Chattong et al., 2007). When the amount of CMC
increased to 2 wt% CMC, the particle size increased. The disruption
of the protein matrix led to aggregation of particles during heating
(Chattong et al., 2007).

3.7. Mechanistic explanation
Based on our results, both cellulose derivatives showed
completely different functionality. In Fig. 7, the schematic behavior
of the two polymers used in the fried meat matrix is shown.
Meat proteins, such as CMC, are negatively charged at pH 5.8,
which typically occurs in meat. Both polymers repulse each other.
Therefore, we suggest that at sufficiently high concentrations, CMC
(>0.5%) is thermodynamically incompatible with meat proteins
(Schuh et al., 2013), an effect that was considerably enhanced upon
frying when the proteins become increasingly hydrophobic due to
the changes in protein folding (Tornberg, 2005).
Other studies have shown that repulsion of charge between
polymers, such as proteins and polysaccharides, at pH values that are
significantly above the isoelectric point of the protein is one of the
driving forces for a possible segregative phase separation at definite
concentrations of both biopolymers. This incompatibility may be
Fig. 6. Structure factor Df (box counting method) of CLSM images after converting in
binary images with different magnifications (40, 100, 200) as a function of the
concentration of (A) CMC or (B) MCC; error bars are standard deviations.
traced back to repulsive interactions between the two polymers that
are more formed if the proteins are heated, since increasingly hy-
drophobic groups are present at the surface of proteins, thereby
increasing the tendency for proteins to aggregate, e.g. in dairy ma-
trixes (Corredig et al., 2011) or CMC, and whey proteins, which were
used as a coagulation aid (Koupantsis & Kiosseoglou, 2009). More-
over, the meat protein network, which is formed as a gel in between
the structural meat particles from heat-induced aggregated sarco-
plasmic and myofibrillar proteins being linked together, is disturbed
by using CMC in increased concentrations (>0.5%).
By contrast, MCC is an uncharged biopolymer with a mainly
crystalline structure that forms a filled particle gel network as an
inert molecule. MCC fills the gaps of the tight meat fiber network.
 M. Gibis et al. / Food Hydrocolloids 45 (2015) 236e246
Fig. 7. Suggested mechanisms of interaction of meat proteins with CMC and MCC depending on pH value and the temperatures of ~60  C and ~18  C. (Df structure factor, [ increase,
Y decrease, no change / in comparison to the control).
The uncharged MCC can be added to the ground beef and heated
without causing a disturbance of the protein network. The low
amount of CMC in the MCC powder used slightly reduced the
firmness in the sensory test at hot conditions, but, by contrast, a
filled particle gel network is formed with increasing force in texture
measurement after chilling (Fig. 7).
Both concentrations and properties of CMC and MCC as mixed
polymers influence key food attributes such as sensory perception,
texture or water-binding capacity in fried beef patties.
4. Conclusion
CMC in the concentrations (>0.5 wt%) used is not suitable as a
fat replacer, because the substance weakened the connections in-
side the protein network. Changes are visible from 1% CMC in the
CLSM images. This concentration revealed alterations in the
microstructure of fried beef patties. At higher concentrations, CMC
greatly reduced the strength of the protein network formed after
heat treatment. Sensory evaluation and texture measurements
indicated the structure disturbing effect of CMC in the meat matrix.
In the sensory test, the samples were disliked regarding the too
light color and too soft and pasty texture.
MCC is a possible fat replacer with reduction of fat of approx-
imately 50% compared to a standard product. A part of the lean
meat can also be replaced (protein replacer) in patties (10%).
Sensorially, beef patties were significantly juicier when evaluated
from 1 wt% MCC, as well as significantly softer in texture than the
controls. In addition, MCC has a fat-like mouthfeel. By contrast, the
controls were evaluated as too firm due to their high amount of
lean meat; a fact that consumers disliked. The softening impact of
CMC in texture may be interesting for producers which manu-
facture low-fat patties. Only low concentrations of CMC (0.5 wt%)
might, therefore, help to reduce this unwanted change in texture.
MCC contains approximately 11% CMC, which should be enough
245
for this softening effect. Low-fat patties with the highest concen-
tration of MCC (3 wt%) can also be labeled as a source of fiber and
containing a reduced fat content (30% fat reduction) in Europe.
Moreover, the understanding of the cause of such different
behavior and interactions between proteins and polysaccharides
should be very important for the development of new meat
products with added values relating to diet and low caloric con-
tent for the meat industry.
Acknowledgments
This research was supported by DANISCO A/S (since 2011
DuPont), Copenhagen, Denmark. The authors thank Kurt Herrmann
and Silvia Lasta for technical assistance, as well as Ann-Sophie
Ahlers, Juliane Thoma, Franziska Schneider, and Sandra Ebert for
their contributions to the project.
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