Food Hydrocolloids Vol. 9no. 4pp.

291-306, 1995
Ultrastructure of low-fat ground beef patties with added whey protein
concentrate*
Salwa B.EI-Magoli, S.Larola\ and P.M.T.Hansen\,2
Department of Food Science and Technology, Faculty of Agriculture, Cairo University, Egypt and 1 Department of Food
Science and Technology, The Ohio State University, Columbus, OH 43210, USA
2To whom correspondence should be addressed
Abstract
Microstructure of gels formed at trc from different mixtures of beef myofibril protein (BMP;
6.1% protein) and whey protein (WP) were studied by transmission electron microscopy (TEM).
At a ratio of 30:10 (wlw) of whey protein concentrate (WPC; 79.5% protein) to BMP, WP formed
a network of aggregated clusters in which beef myofibril proteins were embedded. WP apparently
acts as a filler and possibly as a cementing agent for the meat pieces. At a lower ratio of 10:30 (wi
w), the WP aggregates occupied and increased the interstitial spaces between the myofibril protein
and reinforced the network. The location of WP in the interstitial spaces might explain its water
binding ability in beef patties formulated with WP and water. WP protected the beef myofibril
protein structure during heating as less disintegration in the Z-line was observed in gels with WP
compared to the control. Low-fat (10% fat) ground beef patties with added 10% water and 1-4%
whey protein concentrate (WPC), cooked to three different internal temperatures (60, 70 and
BOC), were evaluated for their cooking characteristics and examined by TEM. For all levels of
addition, WPC improved the cooking yield compared to a non-formulated control of 10% fat. Fat
retention was also improved at the highest level of WPC addition. The increased cooking yield was
shown to be caused principally by the better water retention. The textural parameters, hardness and
chewiness, were not affected by WPC addition but increased with increasing cooking temperature.
These temperature-induced changes were matched by marked changes to the ultrastructure of the
meat products.
Introduction
The US Department of Health and Human Services (1) has
issued specific nutritional guidelines to the food industry to
help the public meet established dietary goals. Consumer
trends toward reducing fat intake have increased the
demand for leaner meat products and created opportunities
for new meat product development through reformulation
using effective fat substitutes (fat mimetics, fat replacers).
However, maintenance of the customary meat flavor and
texture (2) must remain an important consideration in any
effort to reduce fat in meat products, because fat reduction
will likely decrease palatability and satisfaction (3,4),
especially when fat is reduced to 10% or lower (5).
* Presented as part of the conference entitled 'Food Hydrocolloids. Ohio
94', September 0-10.1994.
Oxford University Press
Fat replacers may be protein-based, carbohydrate-based
or fat-based constituents, where each category exhibits
different functional properties that provide advantages as
well as limitations in specific applications (6).
Iota-carrageenan in combination with water was intro-
duced (7) as a fat mimetic in the development of a
particular formulation of low-fat beef patties which has
been promoted as a commercial product (McLean De-
LuxeTM) in the US since 1990. The selection was based on
observations that the iota-carrageenan-water system pro-
vided moisture retention and improved sensory properties
of low fat 10%) ground beef. Similarly, kappa-carra-
geenan has been used to increase yield and improve texture
properties of structured beef rolls with high added water
content (8).
With respect to protein-based fat replacement, the
292 S.B.EI-Magoli, S.Larola and P.M. T.Hansen
industry relies on the ability of various proteins, such as
soy, corn and milk proteins to provide texture, mou th-feel
and water-binding. Such functionality, is mainly related to
the types of protein-protein interaction (9).
Micro-particulated proteins have been found ideal for
forming structural analogs of fat globules. Fat derives much
of its functionality in foods as an emulsified, dispersed
phase (10). Microparticulated fat substitutes achieve this
same functionality through controlled particle size reduc-
tion. The diversity of proteins with respect to their amino
acid sequence, structure, and side chain modifications,
present numerous possibilities for creating micro particles
(11). For example, it has been reported that whey proteins
(WP) can spontaneously form micro particles without any
special treatment other than heating (12) and for this
reason, WP has become a prototype for the micro-
particulation process.
Nevertheless, whey protein may be used in fat reduction
schemes without microparticulation and, in this case, the
functional properties of interest are related to the capacity
of the protein to bind water and form heat-induced gels.
This function has been described for' whey protein concen-
trate (WPC) in processed meat formulations (13). When
the functional properties of WPC, NFDM, and isolated soy
protein were compared (14), only WPC was found to be
useful in reformulation of meat emulsions.
Studies on the relationship between the microstructure
and gel strength, moisture and fat losses during cooking of
different meat batters, have assisted the industry in
improving process control and optimizing product formu-
lation (9). Light microscopy techniques as well as trans-
mission electron microscopy (TEM) and scanning electron
microscopy (SEM) have proved useful in accurately localiz-
ing the structural components within meat products (15).
It has become apparent that gel formation is of particular
importance to the textural quality of processed meats.
Globular proteins form two types of gels, depending on
how much charge is carried on the native protein. Fine-
stranded gels are formed when repulsion is large while a
network of colloidal particles is formed when the repulsion
is minimum, such as when the isoelectric point is
approached (16). However, the microstructure and physical
properties of whey protein gels are sensitive to pH,
concentration, the amount of salt addition and heating
conditions (17). The relation between structure and func-
tion is of importance in studying the basic gelation
phenomena as well as for practical aspects of food system
stability (9).
The objective of this work has been to study the
ultrastructural characteristics of composite whey protein
concentrate and beef myofibril proteins in a model system
and in real meat systems as a step towards understanding
the factors responsible for the final characteristics of low-fat
ground beef patties, formulated with WPC. Particular
attention has been given to a study of the effect of internal
cooking temperature on the ultrastructure of formulated
low-fat ground beef with added WPC and water.
Materials and methods
Gel preparation
Beef myofibril protein (BMP) was extracted at 0-4C from
freshly ground, round beef (10% fat), using a pH 7.1 buffer
of 0.1 mol/drrr' potassium chloride, 0.05 mol/dm' potassium
phosphate and 0.005 mol/dm' sodium-EDTA buffer (18).
The ground beef (100 g) was homogenized in a Waring
blender with 500 ml of the buffer for one min. Connective
tissue was removed by filtration of the homogenate through
cheesecloth. The filtrate was centrifuged at 600 g for 15 min
at 0-4C. The supernatant was discarded and the pellets
resuspended in buffer and again centrifuged. This treat-
ment was continued until the discarded supernatant was
clear. The resulting BMP pellets were collected and used as
such. Analysis of a separate preparation showed 8.7% total
solids and 6.1% protein.
Whey protein concentrate (WPC) was obtained from
New Zealand Milk Products (N.America), Inc. (San
Francisco, CA) (Alacen 878: 79.5% protein, 4.5% ash,
4.2% moisture, 4.6% fat and 6% lactose). Solutions were
prepared by adding 10, 15,20,30 and 40%g WPC and 0.5%
encapsulated salt (Morton Salt Inc., Chicago, IL) to -50 g
of water and adjusting the final weight to 100 g with water.
These WPC concentrations all contained 0.5% salt and
corresponded to WP concentrations of approximately 8, 12,
16, 24 and 32%. The ash content varied from 0.45 to
1.84%. Combined solutions of BMP and WPC were
prepared as follows. (a) 10:30 ratio: 1 g BMP + 3 g WPC +
0.05 g salt + 5.95 g water; (b) 20:20 ratio: 2 g BMP + 2 g
WPC + 0.05 g salt + 5.95 g water; (c) 30:10 ratio: 3 g BMP
+ 1 g WPC + 0.05 g salt + 5.95 g water. The estimated
protein contents of the combined solutions were 24, 17 and
10% for a, b, and c respectively.
For gel preparation, 5 ml from each sample were placed
in glass tubes, stoppered at one end, and heated at 71C for
10.5 min in a water bath. This time and temperature were
chosen to match the actual cooking time and temperature
of beef patties in the present study and was sufficient to
induce gelation in all of the samples. After heating, the
tubes were cooled to room temperature, and the gels were
removed from the tubes, cut into 1 em cubes and placed
immediately in the fixative (4% glutaraldehyde in 0.1 moll
drn' phosphate buffer pH 6.8), for TEM.
Formulation of low-fat ground beef patties
The patties were formulated using coarsely ground beef
(10% fat), 10% water, 0.5% encapsulated salt and four
different concentrations of WPC (1, 2, 3 and 4%). WPC
was first hydrated with the required amount of water to be
added to the meat, by mixing for 15 min using an electrical
stir plate. The resulting thick liquid slurry was kept
overnight at 4C, before it was added with the salt to the
meat and mixed thoroughly. The mixture was ground in
succession with 9.5 and 4.8 mm plates using a Hobart
Meat-whey protein ultrastructure 293
Cooking yield (%) =
Cooked weight
Weight of raw meat
Fat retention (%) =
Cooked weight x percent fat in cooked patties x 100
Raw weight x percent fat in raw patties
Cooking yield , corrected for the addition of WPC, was
calculated as above by subtracting from the cooked weight ,
the weight of the added WPC. Shrinkage and fat retention
values were calculated according to (2,19) as follows :
mixer. For comparison, a control was included of lean beef
(10% fat) , with only 0.5 % salt and no water added.
Quarter pound, (114 g) patties were shaped using a
household hamburger mold (Burger press, US Pat
0191367) , and quick frozen in liquid nitrogen. Each patty
was wrapped separately using pol yethylene cling film, and
every four patties were then sealed in 'Zip-Lock' pouches
and stored at -18C until further analysis.
The patties were thawed at 4C, overnight , and were
cooked to three internal temperatures (60, 71 and 80C) on
an electric household grill heated to 150C and using
variations upon a standard protocol (3 min, 2 min and 15 s
on each side). The actual cooking time varied between lO-
II minutes, depending upon the desired end-point tem-
perature. Temperatures were monitored using a hypo-
dermic probe-type thermocouple (Model HVP-2-21-1-V2-
TG-48-0ST-M Omega, Stanford, CT) connected to a data
logger recorder (21X Micrologger, Campbell Scientific
Inc ., Logan, UT) and linked to a computer (Laser
3865X/25) .
Moi sture was determined by weight loss after 12-18 h
drying in a conventional oven at 105C (20). The fat content
was determined using the Babcock method (21) , after slight
modification for meat use . Four to five grams of the sample
were placed in cream bottles with 9 ml water. A quantity of
17.5 ml sulfuric acid was then added at room temperature
to dissol ve the meat. The remainder of the procedure
followed the standard Babcock method. Texture profile
anal ysis (TPA) was conducted after cooking and cooling to
room temperature. Two whole patties were compressed (at
three different locations) each to 75% of their height , for
two cycle s using a Universal TA-XTI Texture Analyzer
(Texture Technologies Corp., Scarsdale, NY) equipped
with a 1/2 inch flat surface plunger. The machine was
programmed for a 50 kg load cell and cross head speed of
200 mm/min. Hardness and chewiness were obtained using
the available computer software .
Uncorrected cooking yield was determined relative to the
me at content :
Transmission electron microscopy
Initially, l-cm cubes from gels and meat patties were fixed
for -1 h in 4% glutaraldehyde in 0.1 mol/drrr' phosphate
buffer (pH 6.8). One millimeter cubes were then cut from
each sample and placed in fresh fixative overnight at 4C.
Samples were rinsed three times in 0.1 mol/dm" phosphate
buffer, for 45 min , post fixed at room temperature in 1%
osmium tetroxide in buffer for 90 min; then rinsed three
more times in buffer over a 90 min period and dehydrated
in graded ethanol series (50, 70,80,95, and 100%), where
each step involved two changes over 15 min. The samples
were placed in propylene oxide over a 15 min period and
then embedded in Spurr-resin and polymerized in a vacuum
oven overnight at 60C (22) . Sections (70-80 nm) , were
stained with uranyl acetate followed by Reynold's lead
citrate, and examined by a Phillips C-12 Electron Micro-
scope at 60 KV.
Results and discussion
Whey protein concentrate gel
Transmision electron microscopy of WPC gels (Figures 1
and 2) showed a distribution of aggregated protein clusters
separated by irregularly shaped void spaces, which have
also been previously recognized (23-25). The void spaces in
the network represent the compartments of water which
was removed during specimen preparation with the graded
alcohol series (26). The dimensions and extent of void
spaces are an indication of the water retained by the
system. At higher magnification (Figure IB), the clusters of
the aggregated proteins were seen to be connected by fine
threads, forming a distinct network. The presence of an
extended protein network explains the role of WP when
used as a water binder in the meat system. According to
Langton and Hermansson (16), TEM of a network of WPC
gel (12%) at pH 7.5 revealed loosely packed aggregates,
interconnected by thin threads.
In the present study, the void spaces and crevices
diminished and the structure became more compact and
dense as the concentration of WPC increased (Figure 2A
and B) . The relatively compact structure of heat induced
WP gels has been explained (16) to be related to the
presence of immunoglobulins (up to 11% in WPC) that
have an isoelectric point at a relatively high pH (6-8) and
might precipitate and , thus , contribute to a more dense
structure of WP gels.
The whe y protein started to form a gel at a concentration
of 15% WPC, corresponding to 12% WP. As the WPC
concentration increased, the micrographs (Figure 10 and
2A) showed embedded globular structures, resembling
holes or entrapped gas bubbles and varying in diameter
between 0.1 and 0.5 urn. At high magnification (Figure 2C;
112500x), a substructure was revealed, which may be
surface active protein surrounding gas bubbles. These
x 100
Shrinkage (%) =
(Raw width - cooked width) +
(Raw length - cooked length)
Raw width + Raw length
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Figure I TEM micrographs of WP gels at 71C and pH 7.1. (A and B) Fine stranded WP clusters at 15% WPC, separated by irregular shapes of void spaces .
(C and D) Aggregated clusters of WP at 30% WPC, showing the dimini shing void spaces and the compactness of the structure.
Figure 2 TEM micrographs of WP gels at 71C and pH 7.1. (A and B) Dense structure of WP at 40% WPC, showing aggregated globular particles with many rough
edges. (C) Possibly non-dissolved particle of WP.

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Figure 3 TEM micrographs of BMP (A and B) Sarcomere units of BMP before heating, showing the A-band, I-band, H-zone and Z-Iines. (C and D) Sarcomere units
after heating at 71
0C
for 10.5 min showing the disintegration of the sarcomere structure.
Meat-whey protein ultrastructure 297
particles were not detected in gels at 15% level (Figure 1A
and B).
Gels of WP, formed at 30-40% WPC (24-32% protein)
were white in color with a highly rubbery texture. Such
texture (27) may arise from the intermolecular S-S bond
resulting from an interchange reactions of SH/S-S reported
to occur at pH 7.5 over a broad concentration range. TEM
micrographs of these gels (Figure 2B) showed aggregated
globular particles with many rough edges. This structure
was not apparent in gels containing 15% Wl'C, These gels
were less rubbery, almost transparent and showed more of
a fine-stranded than aggregated structure in TEM (Figure
lB).
The results confirm that WP is capable of forming more
than one type of gel, depending on its concentration and
the condition of the system. Such observations have been
reported (25) for heat induced gels formed at 8-15% in the
pH range of 7-8. It may be noted that the temperature of
heating and pH used in the current study for gel prep-
aration were chosen to correspond to the cooking con-
ditions for the meat system.
Addition of salt (0.5%), did not affect the gel structure of
WP in this study. Sodium ions (0.25 mol/drrr') are reported
to improve the water holding capacity of WP gels (9). This
concentration is equivalent to 1.5% NaCl and thus,
considerably higher than the salt level used in this study. It
is possible that some of the variations in gel structure were
due to differences in ash levels of the WPC dispersions.
BMP gels
Immediately upon extraction BMP formed a delicate soft
gel. When heated at 71C for 10.5 min in the buffer (pH
7.1) the protein sample coagulated and formed a more
fibrous, thread-like structure. TEM micrographs of BMP
before and after heating are shown in Figure 3 (A, B, C and
D).
Before heating BMP showed a typical structure of
myofibril proteins with sarcomere length of -2 urn. The Z-
disks, A-Bands, I-bands and M-lines were identifiable at
both high and low magnifications (Figure 3A and B).
Heating of the BMP resulted in fragmentation of the Z-
disks with the loss of structure due to the coagulation of the
filaments. The structure of the myofibrils became more
amorphous and a pronounced shrinkage occurred (Figure
3C and D). The Z-disks became diffused, thickened and
disrupted. These changes are typical of those previously
reported (28-30) for the effects of heating on myofibril
proteins.
Combined gels (WP/BMP)
The distinct morphological differences between gels from
the two proteins may be used as a basis for further
comparison of the combined systems. The granular aggre-
gates of the WP are seen to persist within the complex gels.
Thus, the ultrastructure of combined gels, containing WPC/
BMP in the ratios 30:10, 20:20 and 10:30, with estimated
total protein content of 24, 17 and 10% respectively,
showed that the relative amounts of the proteins had a
determining effect on their distribution in the system
(Figure 4). At a ratio of 30:10 WPCIBMP, the WP
appeared as a continuous network embedding the BMP and
water (Figure 4A and B). It is possible that at this
concentration WP may act as a filler or as a cementing
agent.
At the 20:20 ratio (Figure 4C and D), both WP and BMP
were distributed on an equal basis, with the WP occupying
the intracellular and some of the intercellular spaces. The
effect of heating (71C for 10.5 min) on the shrinkage and
disintegration of BMP was less pronounced in the presence
of WPC, compared with the control (Figure 3C and D)
suggesting a protective action of WPC on the BMP, and
possibly explaining its role in decreasing shrinkage when
used in real meat systems.
Both the BMP fibrous network and small WP globular
aggregates were visible at the ratio of 10:30 (WPC/BMP),
with the WP clearly located in the interstitial spaces (Figure
4E and F), holding the filaments together and reinforcing
the original network of the myofibril proteins. At high
magnification (Figure 4F), WP aggregates were evenly
embedded within the BMP gel matrix. The type of gel
formed seemed similar to the 'coupled network' gel form
(31,32) possibly formed from a two-phase separated net-
work. Thus, WP might gel within the interstitial spaces of
the already formed BMP gel network. The creation of this
type of gel could have an impact on the tenderness and
juiciness of a meat product containing Wf'C. In a low-fat
meat system, the meat tends to be tough and dry.
Toughness is mainly due to drying and coagulation of
myofibrillar proteins, originating from the loss of water of
hydration surrounding thick and thin filaments. If this
water of hydration could be retained by the WP gelling
within the interstitial spaces between the myofilaments, the
meat tenderness is likely to improve.
Low-fat ground beef patties (TEM)
Specimens for TEM of the control low-fat ground beef
patties, cooked at three internal temperatures were
collected from the interior of the sample where end-point
temperatures were monitored. The micrographs revealed
pronounced changes in the myofibril patterns starting at
71C and continuing at 80C. The uncooked patties showed
well defined, closely packed and well-preserved structures
of myofibril with a sarcomere length of -2 urn. The 1-
band, A-band, H-zone and M-lines were clearly identifi-
able. Shrinkage and coagulation of the thick-band (the A-
band) began at 60C and increased significantly with the
increase in cooking temperature (Figure 5A, B, C and D).
At 80C wider endomysial spaces between the myofibrils
were evident indicating more shrinkage. Maximum shrink-
age of the endomysial connective tissue at 70C has been
previously reported (28).
298 S.B.EI-Magoli, S.Larola and P.M. T.Hansen
B 111m
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Figure 4 TEM micrographs of combined WPC/BMP gels at 71C and pH 7.1. (A and B) 30:10 WPC/BMP, arrows showing the
predominance of WP in the interacellular spaces between the BMP. (C and D) 20:20 WPC/BMP, showing equal distribution of WP in the
intracellular and intercellular spaces of BMP. (E and F) 10:30 WPClBMP, arrows showing WP occupying the interstitial spaces between
the BMP.

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Figure5 TEM micrographs of low-fat ground beef patties (0% WPC), cooked at three internal temperatures for 10.5 min (A) Before cooking; (B) cooked at 60C;
(C) cooked at 71C; (D) cooked at 80C.
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Figure 6 TEM micrographs of low-fat ground beef patties with 2% WPC, cooked at three internal temperatures for 10,5 min (A) before cooking, (B) cooked at 60C,
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Figure 7 TEM micrographs of low-fat ground beef patties with 4% WPC, cooked at three internal temperatures for 10.5 min (A) before cooking, (B) cooked at 60
DC,
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302 s. B. EI-Magoli, S. Larola and P. M. T. Hansen
Marked disintegration in the myofibril occurred at 71DC;
however, banding patterns remained visible, with some of
the Z-lines and H-zones still intact. At 80
DC
the filamentous
structural integrity was almost gone and the myofibril
showed granulation. The granular material formed at a
higher cooking temperature (90
DC)
may be related to the
partial precipitation of sarcoplasmic proteins from fluids
that collect beneath the sarcolemma during fiber shrinkage
(29).
The raw meat samples containing 2 and 4% WPC
(Figures 6 and 7) showed the same well-preserved pattern
of the myofibril structure as in the control sample (Figure
5A). The structural unit of the myofibril, the sarcomere,
was neatly organized, showing the refined distribution of
the A-band, l-band, Z-lines and M-lines.
The TEM of the low-fat ground beef patties with 2 and
4% WPC, showed that the cooking temperature had far
more impact on meat structure than the presence of WPC.
However, the structure of myofibrillar proteins seemed to
be less affected by heat in the presence of WPC at the 2 and
4% levels used than without WPC (Figures 5,6 and 7). The
myofilaments had shrunk less and have retained their Z-
lines, particularly at the 4% level. This observation suggests
that WP might form a weak gel, even when the samples are
heated to an internal temperature of 60
D
C. To reach this
point, the outer areas of the patties will obviously attain
higher temperatures with more complete gelation of WP
and stronger retention of the water around the myofila-
ments, thereby decreasing the shrinkage.
Overall, increasing cooking temperatures resulted in a
progression from a compact structure of the myofibrils at
60
DC
(Figure 6A and B) to a separation of the strands at
71DC and finally more granulation at 80
D
C (Figure 6C and
0). WP was not detected between the myofilament at the
lower cooking temperatures at the 2% WPC level; how-
ever, some of the WP aggregates were present in the
interstitial spaces between the myofibrils at the highest
cooking temperatures and at the 4% WPC level (Figure 7C
and 0). These findings support the perceived role of WP as
a water binder in the meat system, and may explain the
higher cooking yield and the lower shrinkage of the low-fat
ground beef patties with 4% WPC (Table 1).
Micrographs of samples containing 2% WPC (Figure 60)
and cooked at 80
DC,
showed a network structure of
myofibrils that seemed connected by delicate threads
resembling the WP strands in the gel system (Figure 2B)
and differed in structure from the control without WP.
However, more amorphous and granulated surfaces due to
protein coagulation were apparent at this cooking tempera-
ture than at 60 or 71DC and were also accompanied by a
Table 1 Cooking characteristics of low-fat ground beef patties (10% fat) with added whey protein concentrate, salt and water
WPC Internal Cooking yield Fat Shrinkage Texture profile analysis
(%) temperature retention (%)
eC)
(g/100 g meat) Corrected for WPC (%) Hardness Chewiness
(g/lOO g meat)
Control 60 77.6 77.6 66.1 10.5 2.79 1.22
(0%) 71 70.8 70.8 68.4 12.2 3.99 1.71
80 68.2 68.2 61.9 10.4 4.47 1.78
60 83.6 8.25 50.1 11.7 3.06 1.78
1.00 71 76.4 75.3 53.8 13.0 3.13 1.03
80 74.6 73.5 55.9 13.6 4.53 1.75
60 86.3 84.0 59.7 5.9 2.28 0.97
2.00 71 84.9 82.6 57.5 10.0 4.20 1.82
80 78.9 76.6 68.7 10.8 4.33 1.82
60 91.7 88.2 72.7 8.2 2.93 0.97
3.00 71 86.2 82.8 80.1 11.1 3.71 1.64
80 78.9 79.3 73.9 9.1 4.66 2.08
60 95.9 91.2 73.7 6.1 2.46 1.09
4.00 71 85.2 80.5 69.2 8.6 3.87 1.48
80 85.9 81.2 69.9 7.4 3.55 1.65
Standard error 1.76 1.76 4.26 1.04 0.43 0.33
All measurements are means of duplicate analyses.
Standard error values were determined from the mean square values from analysis of variance.




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8
304 S. B. EI- Magali. S. Laraia and P. M. T. Hansen
complete loss of the M-line. At 80
aC,
WP will form a more
rubbery and elastic ge l, which might the n support the
concept of it working as an agent for cementing the meat
pieces.
Water absorption and gelation phenomena are important
for the stability of ground or minced meat products (33) .
However, the TEM micrographs of the meat system
(Figure 8A and B) showed that WP may playa role as an
emulsifying agent when added to a meat system. The fat
globules were covered by the WP, with some of the WP
aggregates extending into the system, indicating an associ-
ation of protein aggregates with the fat globules. It has been
observed that globules covered with adsorbed protein
molecules may be immobilized by direct reticulation
between membrane-forming protein mo lecules and bulk
phase protein (34) .
Fat membranes are likely to be ruptured dur ing the
preparation of beef patties and upon cooking, the fat will
tend to form pools rather than retaining separate globular
structures. The micrographs in Figure 8 are selected fields
from low-fat ground beef patties formulated with 4% WPC
and cooked at 71C showing areas where fat is prominent.
Figure 8C shows a fat pool surrounded by protein, assumed
to be WP . Such fat pools were observed during examination
under the microscope in all cooked samples, with and
without WPC. However, separate fat globules were only
detected in samples with 4% Wl'C.
Cooking characteristics of low-fat ground beef patt ies
The control in these studies was a 10% beef patty with no
water and only salt added. The treatment samples were
formulated with 10% added water, salt and different WPC
levels. The results for cooking yields(Tables 1 and 2) show
that WPC addition is an effective means for retaining the
added water, because the yields gradually increased over
the control for each leve l of WPC addition. As might be
expected, yield decreased significantly as the interna l
cooking temperature was raised. However, increases in
yield due to WPC addition and losses due to heating were
independent of each other as shown by the lack of
significance of the interaction term. Some improvements in
cooking yield ma y be expected from the additional solids
provided by WPC; however, corrected yields still showed
substantial gains in cooked weight over the controls which
must be ascribed to better water or fat retention, or both.
Cooking yield has been reported to improve by reducing
the fat content of meat patties to 20% (35,36). In other
studies (4,5, 37) cooking yields did not diffe r much when the
fat levels were below 20% .
Fat retention was significantly improved (Tables 1 and 2)
at the 3 and 4% WPC addition but not at the lower levels of
addition. In fact, at the 1% WPC addition, fat retention
was adversely affected compared with the non-formulated
control. We speculate that the addition of WP to the
formulated beef patties might result in emulsification of
some of the fat which is then better retained in the WP gel.
This explanation would be consistent with the microstruc-
ture observed in Figure 8. Another possible reason for the
observed differences in fat retention for different levels of
WPC may relate to the failure of WP to gel firmly at the
lower levels of addition. The greater loss in cooking yield,
at these concentrations, implies some loss of the added
liquid, which might carry associated fat, and thus explain,
why more fat is lost from the product formulated with 1%
WPC than from the non-formulated control.
Table 2 Analysis of vari ance of cooking characteristics. Single degree of freedom comparisons between treatments and temperatures
(40)
Comparisons Cooking yield Fat Shrinkage Texture profile analysis
(%) retention (%)
(g/IOOg meat) Corrected for WPC (%) Hardness Chewiness
(g/IOO g meat)
Significance levels (P<)
Between treatments (WPC) (1%) (1%) (1%) (1%) (NS) (NS)
4 versus 3, 2, 1% WPC and control 1% 1% 5% 1% NS NS
3 versus 2, 1% WPC and control 1% 1% 1% NS NS NS
2 versus 1% WPC and control 1% 1% NS 1% NS NS
1% WPC versus control 1% 1% 1% NS NS NS
Between internal temperatures (T) (1%) (1%) (NS) (5%) (1%) (NS)
60 versus 71 and 80
aC
1% 1% NS 1% 1% 5%
71 versus 80
aC
5% 5% NS NS NS NS
Interaction (WPC x T) NS NS NS NS
Significance levels in parentheses are from two-way ANOVA; others are from single degree of freedom comparisons of the same treatments.
NS, not significant (P > 5%).
Meat-whey protein ultrastructure 305
Cooking temperature did not influence fat retention
within the temperature range studied. Therefore, the
over all losses in cooking yield observed for increasing
temperatures are principall y due to loss of water. In other
reports, cooking to well-done versus medium doneness (77
and 71"C) have been found to increase cooking losses (38).
There is evidence in the present study that fat retention is
improved when fat has been partially emulsified by WP.
Higher fat retention in low-fat meat systems has been
reported (39). These authors concluded that the denser
protein matrix of low-fat ground beef prevents fat
migration, reducing the possibility of encounter and
expansion among fat pools.
Significant changes in shrinkage values due to WPC
addition and cooking temperature were observed. Gener-
ally, WPC addition>1% reduced the degree of shrinkage,
while increasing the internal end-point temperature > 60C
increased shrinkage. The micrographs in Figures 5, 6 and 7
show features which correlate to these observations.
No significant differences due to added WPC occurred
for the textural profile traits , hardness, and chewiness,
indicating that such addition doe s not markedl y affect these
textural characteristics of meat. However, hardness in-
creased significantly as the final internal temperature was
raised above 60C. This result is understandable, since at
60C (rare to medium) less coagulation in both myofibril
proteins and endomysial connective tissue will occur. This
effect is evident in the ultrastructure micrographs (Figures
5, 6, and 7) , where changes to the myofibril become more
pronounced at 71 and 80C (medium to well-done) . Some
increase in chewiness (P > 0.05) was observed as the
cooking temperature was raised above 60Cwhich would be
consistent with the increase in hardness.
Conclusion
TEM was used to show the role of WPC as a functional
agent for formulating a low-fat ground beef system with
improved cooking yield and fat retention. Such improve-
ments may generally be rel ated to improvements in
perceived moistness and juiciness. The observations are
consistent with whey protein acting as a water binder,
cementing agent and an emulsifying agent. Results
suggested that the addition of a solution fo WPC can
enhance the cooking characteristics of low-fat ground beef
patties, depending on the concentration used. An addition
of 10 parts of water, 4 parts of WPC and 0.5 parts of
encapsulated salt to low-fat ground beef, produced im-
proved cooking yield, better fat retention and less shrink-
age compared to other concentrations tested.
Acknowledgements
Appreciation is expressed to Kathleen S. Wolken of the
OSU Microscopy and Imaging Facility for assistance with
the electron microscopy and to Dr H.W.Ockerman for
helpful advice. S.B.E.-M. acknowledges with thanks a
grant awarded to her by the Fulbright Commission. Journal
Article No. 142-94. Salaries and research support provided
by state and federal funds appropriated to the Ohio
Agricultural Research and Development Center and The
Ohio State University.
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Received on October 17, 1994, accepted on May 5, 1995