1. 1
What’s Your Beef? The Effects of Breadcrumb Substitutions for Ground Beef in Hamburgers
Related to Lysinuric Protein Intolerance
A Research Paper
Submitted to Jodie Seybold, MS, RD, LDN
In Partial Fulfillment of the
Requirements for
FDNT 362 Experimental Foods
Maria Wendt
Indiana University of Pennsylvania
December 2, 2014

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Abstract
Lysinuric protein intolerance [LPI] has required a diet low in protein. Thus, reducing amino acid
intake has been essential to living a healthy, normal lifestyle. A common food consumed high in
protein used was 80/20 ground beef for hamburgers. Throughout the course of the experiment,
subjective and objective data were collected and used for later results. The substitution of
breadcrumbs for ground beef to reduce protein content contributed to specific changes of
dependent variables in texture, color, and taste, along with density and tenderness. The
independent variables that caused these changes were 0% breadcrumbs, 25% breadcrumbs, 50%
breadcrumbs, and 75% breadcrumbs. There were few alternatives for those with LPI, but this
experiment has proven more, depending on the desired outcome of the consumer.
Acknowledgements
I would like to thank Mrs. Seybold for contributing incredible organization and simplicity to this
paper, and always willing and enthusiastic to lend a helping hand. Thank you to my mother for
helping me pick a disease outside of the box and live up to the challenge. Thank you to the
students at our lab island for letting me borrow necessary extra materials for my experiment.
Thank you to Kate Numer for always offering advice and peer reviewing this paper over and
over. Finally, thank you to the sensory panelists for going through this experimentation carefully
and producing good outcomes when using SPSS. Together we made this possible!
Introduction
In today’s society, it seemed all was about gaining muscle and losing fat. But for some, too much
protein can be fatal. Experimentation with food has given dietitians a deeper knowledge of the
specific ingredients needed to be put in, or taken out of a food or diet in order to compensate for
a medical diagnosis. The focus of this study was to draw attention to lysinuric protein intolerance

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[LPI], alter a high protein recipe, and find substitutions for the protein in order for these patients
to enjoy a common, easily cooked meal. LPI was defined as faulty carrying of diamino acids,
specifically being arginine, lysine, and orthinine, in an autosomal recessive pattern.1-3 The signs
and symptoms of LPI were evident after children were taken off breast feeding.4 LPI was
referred to as a multi-organ disease, that had serious complications and has been known to be
fatal.2,3 Diagnosis was said to be time consuming, as there were many listed misdiagnoses.3,4
Treatments have previously included low-protein diets, reducing risks of complications such as
hyperammonia, or a citrulline supplementation.3,4
The recipe experimented on was hamburgers. The independent variables used were 0%
breadcrumbs, 25% breadcrumbs, 50% breadcrumbs, and 75% breadcrumbs. The dependent
variables were volume and tenderness that were tested by objective evaluations with machines
such as the volumeter and penetrometer, and texture, color, and taste, which were measured
through sensory evaluation with the senses.5,6 In this study, different amounts of water were used
which acted as bound water to hold the ingredients together.7 The breadcrumbs came from
generic whole grain bread crumbled in a food processor. These breadcrumbs made the
hamburgers dense, also leaving patients fuller longer due to the whole grain content and fiber.8-10
The addition of breadcrumbs also contributed to the increasing softness as more were added and
ground beef was taken away.11
A portion of ground beef was substituted with breadcrumbs to reduce the overall protein
content of a hamburger. The ground beef was 80/20 to slightly reduce the protein content by
raising the fat percentage. It remained important, however, that patients with LPI received
enough protein in their diet, through the hamburgers, for example, as ground beef in a previous
study has been paired with better nutrient consumption and an overall improved diet.12 In fact,

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McNeill stated that, “beef…and beef mixed dishes rank ninth among the top 10 sources of
energy in the US diet, and hamburgers rank 12th”12, which gave more reason as to why
experimentation was done on this food item. It has been hard to limit beef consumption, and data
has proven that 96% of American adults have testified to consuming beef.12
In this study, each burger was cooked to 165oF for 15 seconds to avoid foodborne illness,
such as E. coli.13 Heating between 131-176oF broke down protein and non-enzymatic browning,
also known as a Maillard reaction, occurred as the heat was continually applied.14 Meat has been
found to be made up of 20% protein, therefore, by reducing the proportion of meat with a greater
proportion of breadcrumbs differing per variable, the protein content was significantly reduced.13
Restaurateurs have been known to decrease cooking time on burgers when using extreme
temperatures with pan frying due to possible overcooking on the outside to try and reduce a
drastic texture change, which in turn not only increased the acceptability of a product, but also
the risk of food borne illness.13
Research questions to be answered from this study were: Which hamburger will be more
tender: the one made with 25% breadcrumbs or the one with 50% breadcrumbs? Which
hamburger will be more tender: the one made with 50% breadcrumbs or the one with 75%
breadcrumbs? Which hamburger will have more volume: the one with 25% breadcrumbs or the
one with 75% breadcrumbs? Will the panelists find that the burger made with 75% breadcrumbs
to be more airy in texture than the one with 25% breadcrumbs? Will the color of the burger made
with 25% breadcrumbs be more dark brown than the one made with 75% breadcrumbs? Will the
burger made with 25% breadcrumbs be saltier than the one with 50% breadcrumbs? and Will the
burger made with 25% breadcrumbs be saltier than the one with 75% breadcrumbs? The
hypotheses were as follows: By substituting 75% breadcrumbs for ground beef in hamburgers,

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the saltiness will significantly change. By substituting 50%-75% breadcrumbs for ground beef in
hamburgers, the volume will significantly change. By substituting 75% breadcrumbs for ground
beef in hamburgers, the tenderness will significantly change. The purpose of this experiment was
to determine the effect of decreased protein with the addition of breadcrumbs on volume,
tenderness, color, taste, and texture in hamburgers.
Methodology
The recipe used was Hamburgers found on page 205 in The Good Housekeeping Illustrated
Cookbook.1 The recipe was changed from English to metric measurements using the USDA
Handbook 8, as seen in Table 1.
Table 1. Nutrition Information for Each Recipe Variation*
Control 100%
ground beef
75% ground
beef
50% ground beef 25% ground beef
Calories 138.75kcal 122.75kcal 149.25kcal 129.25kcal
Fat (g) 8.72g 8.84g 9.2g 5.32g
Saturated Fat
(g)
6.64g 5.06g 3.5g 1.9g
Cholesterol
(mg)
44g 33g 22g 11g
Total CHO
(g)
0.56g 5.42g 10.3g 15.12g
Dietary Fiber
(g)
0.1g 0.4g 0.7g 1.00g
Sugars (g) 0.21g 0.63g 1.05g 1.45g
Protein (g) 13.6g 5.03g 5.93g 4.8g
Zinc 3.32ug 1.00ug 1.22ug 0.81g
B12 1.41ug 0.61ug 0.63ug 0.36g

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Each ingredient, including the variables with breadcrumbs, were entered manually into the
USDA Handbook 8 with respective serving sizes. Then, each was converted from English to
metric measurements. The serving size, calories, protein, total lipid, saturated fat, cholesterol,
carbohydrate by difference, fiber, sugars, and a specific vitamin and mineral were listed and
recorded for data for each recipe.
Control Recipe- 100% Ground Beef
Each day before cooking time, the ingredients were pre-measured and placed aside until ready to
use. When ready to pre weigh, 12 half cup plastic cups with lids were obtained, taken to lab
station 12, and hands were washed. Next, salt, pepper and onion were gathered and taken to the
station. The top-loading electronic balance was retrieved out of the cabinet and placed on the
counter. Before that, a cutting board and 8 inch knife were obtained and the onion was then
minced. Once cut, the balance was turned on, one plastic cup was placed on top, and it was
TARED. Then, the minced onion was added into the cup using hands until it read 20grams [g].
Lids were put on each container and set aside. Another plastic container was placed on the
balance and salt was shaken from the container directly into the plastic cup until it read 6g.
Again, a plastic cup was placed on the balance and pepper was shaken directly into container
until it read 0.58g. Next,80-20 ground beef was taken from the refrigerator and placed at the
station. Once the pack was opened, a bowl was placed on the balance, and weighed 226.67g for
the control recipe. The measurement was set aside, and the same was done for the three variables
of ground beef which were 170g, 113g, and 56.67g. Plastic wrap was placeded over the four
bowls and put aside until ready for mixing. Next, the whole grain bread (generic brand) was
taken out of the fridge and placed at the station. The food processor was taken out of storage and
plugged into the nearest outlet. The bread was ripped and fed into it one slice at a time, usually

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between two to three slices per blend before the machine became too full. A new bowl was
placed on the balance, and breadcrumbs were added until it read 56.67g, 113.67g, and 170g for
the three variable recipes. It was put aside until ready for later use. The cooking process began
by washing hands thoroughly to avoid any transferring of bacteria to the food. Next, the PAM
cooking spray and a large skillet were obtained. The plastic wrap from bowls with ground beef
were removed and thrown away. The control recipe began by using a large mixing bowl and
placing one container of onion, salt, and pepper into it, and also 226.67g of ground beef. The
ingredients were mashed together by hand about 20 times. Once well mixed together, the beef
was divided into four patties, each about one inch thick. Next, the skillet was placed over the
stove and warmed to medium heat. Also, the oven was preheated to 2000F to keep burgers warm
for serving. Then, the PAM cooking oil was generously applied to the skillet for about ten
seconds. Once warmed, all of the patties were placed in the skillet and the lid placed over them.
While waiting for the burgers to cook, a cookie sheet was pulled out to place the cooked burgers
on. The burgers were cooked two minutes on each side, flipped with a rubber spatula, then
checked every one and a half minutes with the temperature gauge until they reached 1650F. The
burgers were then removed from the skillet and placed on the baking sheet in an orderly fashion
that distinguished between recipes, and placed in warmed oven. A new skillet was placed to
warm while the other was removed for washing. These two skillets were used for every other
recipe.
Independent Variable- 25% Breadcrumbs
All of the same procedures were followed in the control recipe, but 170g ground beef and 56.67g
breadcrumbs were used. It took 20 hand squeezes to mash the ingredients together. The cooking
time was slightly shorter than the control as noted by when it reached 1650F.

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Independent Variable- 50% Breadcrumbs
All of the same procedures were followed in the control recipe, but 113g of ground beef and
113.67g of breadcrumbs were used. An additional ¼ cup of water was added to avoid dryness.
This took more hand squeezes to mash ingredients together, upwards of 30 to 50.
Independent Variable- 75% Breadcrumbs
All of the same procedures were followed in the control recipe, but 56.67g ground beef and 170g
of breadcrumbs were used. An additional ½ cup of water was added to avoid dryness. This recipe
took 50 hand squeezes.
Subjective Methods
A score card was created with sensory objectives on it. This included texture, with a range from
1-5: 1 being dense, 3 semi-dense, and 5 airy; color, with 1 as light brown, 3 brown, and 5 dark
brown; and lastly taste with 1 as salty, 3 semi-salty, and 5 very salty. Four numbers were made
up, each with three digits to correspond with each recipe. The numbers used for this experiment
were 464, 772, 213, and 117. The sensory score card can be seen in Figure 1. These will be the
same throughout, but vary as to which recipe they were assigned. Five copies of the score card
were made each time the experiment was done, and placed in each panelist’s seat when done
cooking. The seats were side by side, but blocked off from view of others by a divider, so it was
a blind taste test. Next, five paper plates were obtained and were each divided into four quadrants
with a sharpie and marked each at the top with the four different numbers stated earlier. Each
week the numbers would change to which variable it corresponded to. The plates were placed in
blocked off judging seats along with five score cards. The score cards were collected by the

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researcher when completed. The sensory panelists went through a 35 minute training session for
this process.
Figure 1. Sensory Scorecard
Characteristic 772 464 117 213
Texture a
Color b
Taste c
a Texture
1________ 2_______ __3_____ ____4________ _5
Dense Semi dense Airy
b Color
1________ 2_______ __3_____ ____4________ _5
Light brown Brown Dark brown
c Taste
1________ 2_______ __3_____ ____4________ _5
Not salty Semi salty Very salty
Objective Methods
The burgers were tightly wrapped with plastic wrap and taken to the volumeter for objective
testing. Each burger was placed on a top-loading electronic balance and the gram weight of each
was recorded. The volumeter was calibrated by pulling the metal slip out until the rape seeds
stopped flowing down The number was recorded at the line where the seeds ended. The
volumeter was turned 1800 to return seeds to the top. When they stopped flowing, the metal slip

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was pushed back in, and the volumeter was turned right side up. The bottom was then unlatched
and the control burger was placed inside, then locked again. The metal slip was pulled out until
the seeds stopped flowing and the number at which they stopped was again recorded. The
volumeter was inversed 1800 until the seeds stopped flowing, returning the metal slip when done,
and turning right side up. The bottom was unlatched and the burger removed. It was then
replaced with variable one. Repeat these steps for each burger and record all results. In
calculating density, the formula used was D=m(g)/V(cm3). Volume represented by cm3 was
determined by subtracting the reading with the burger in the volumeter from the calibration
(sample reading – calibration = V(cm3) ). The products of the formula for each variable were
averaged over the three weeks of experimentation.
The second objective test was the penetrometer to test for tenderness. First, the plastic wrap was
removed from the burgers. Then, the top of each burger was cut off with a butter knife in order to
test the inside of each variable for tenderness. The control recipe was placed on the block and the
weight lowered with the crank on the right so it was slightly touching the burger. Next, the
stopwatch clicked and the small handle on the penetrometer was squeezed simultaneously for
one minute. Once the minute was up, the handle was released and the timer stopped. Afterwards,
one finger was used to tap the loose metal slip at the top of the penetrometer until the handle in
the scale stopped moving Results were recorded as 1/10 millimeters. To return the scale back to
zero, the handle was squeezed down again, and the metal rod was gently pulled up on. Repeat
this process for the three remaining variables.

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SPSS
SPSS was used to analyze data, specifically with One-way ANOVA and the Tukey test. These
were calculated to test judging and objective testing accuracy. They gave week by week averages
for all of the data recorded throughout the three weeks of experimentation. The results will now
be interpreted.
Results and Discussion
Statistical analysis tests were used to test judging accuracy along with subjective and objective
measurements taken during the three weeks of experimentation. Explanations were given using
these results of the Tukey test and One Way ANOVA, and represented the significances between
variables designated by p-values. A significant difference in sensory and objective measurements
was noted as a p-value less than 0.05 (p< 0.05).
Subjective Evaluation
The judges were almost perfectly accurate each week with the subjective measures of texture
(p=0.927), color (p=0.998), and taste (p=0.926).
Texture
Through further testing, it was found that 100% ground beef had a significant difference in
texture compared to 50% ground beef (p=0.000) and 25% ground beef (p=0.000), but not a
significant difference compared to 75% ground beef (p=0.590).. A previously mentioned
research question stated, will the panelists find that the burger made with 75% breadcrumbs to be
more airy in texture than the one with 25% breadcrumbs? It was found that the hamburgers made
with 75% ground beef had a considerable difference in texture compared to 50% ground beef

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(p=0.040) and 25% ground beef (p=0.001). For texture, the burger made with 100% ground beef
was the obvious choice when looking for a dense hamburger. However, when looking for a more
airy burger, the one made with 25% ground beef would be preferable. This was due to the
breadcrumbs contributing to an increased softness compared to the 100% ground beef burger.11
Color
When compared to the control, the burgers made with 75% , 50%, and 25% all had an equal
significant difference in color (p=0.000). A research question beforehand asking whether the
color of the burger made with 25% breadcrumbs would be more dark brown than the one made
with 75% breadcrumbs, was answered with a very significant color change (p=0.000). This color
change was due to the lack of ground beef available to produce a Maillard reaction. The burger
made with 25% breadcrumbs was the best variable when compared with the control when
looking for minimal color change in this recipe modification.
Taste
There was not a significant difference when comparing taste with the control to the burger made
with 75% ground beef (p=0.974) or made with 50% ground beef (p=0.358). However, the
control compared to the burger made with 25% ground beef (p=0.000) did have a significant
difference in taste. One hypothesis that was raised before this study was proven, which stated
that by adding 75% breadcrumbs for ground beef in hamburgers, the saltiness would
significantly change. The panelists tasted decreased levels of sodium with a comparable
difference from the control (p=0.000). This was due to the lack of fatty meat containing extra
sodium aside from the 6g already added from the recipe. A research question previously
mentioned asked will the burger made with 25% breadcrumbs be saltier than the one with 50%

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breadcrumbs? Again, there was no significant difference in the saltiness between the burger
made with 25% and 50% breadcrumbs (p=0.189). Another question raised before this study was
will the burger made with 25% breadcrumbs be saltier than the one with 75% breadcrumbs?
There was a significant difference in the saltiness of the burger made with 25% and 75%
breadcrumbs (p=0.000). This was due to the lack of salt from less beef in the burger made with
75% breadcrumbs. The burgers made with 75% (p=0.974) and 50% (p=0.189) ground beef had
no significant difference in taste, which makes them good alternatives for a person with LPI.

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Table 2. Table of Means* for Dependent Variables: Sensory Evaluation
Dependent
Variable
Condition Mean P-significance
Texture
Control 1.466 ----------
75% ground beef 2.398 0.59
50% ground beef 4.066 .000
25%ground beef 4 .000
75%-50% ---------- .040
75%-25% ---------- .001
Color
Control 5 ----------
75% ground beef 3.334 .000
50% ground beef 2.47 .000
25%ground beef 1.532 .000
100%-75% ground beef ---------- .000
100%-50% ground beef ---------- .000
100%-25% ground beef --------- .000
75%-50% ---------- .011
75%-25% ---------- .000
50%-25% ---------- .006
Taste
Control 3.932 ------------
75% ground beef 4.066 .974
50% ground beef 3.398 .358
25%ground beef 2.2 .000
75%-25% ---------- .000
50%-25% ---------- .007
*SPSS was used to create averages from three consecutive weeks of data for each sensory evaluation. Significance considered as p<.05.
Control is being compared to each of the three variables.

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Objective Evaluation
The second hypothesis, which stated that by substituting 50%-75% breadcrumbs for ground beef
in hamburgers, the volume would significantly change, was not proven correct. There was not a
significant change in volume when 50% breadcrumbs (p=0.58) and 75% breadcrumbs (p=0.297)
were substituted for ground beef, therefore, the null hypothesis failed to be rejected. The research
question that coincided with this was which hamburger will have more volume: the one with
25% breadcrumbs or the one with 75% breadcrumbs? There was not a significant difference
between the volume of burgers made with 25% and 75% breadcrumbs (p=0.526). The last
hypothesis, which stated by substituting 75% breadcrumbs for ground beef, the tenderness will
significantly change, was also not proven to be true (p=0.865). The null hypothesis failed to be
rejected. A foregoing question asked was which hamburger will be more tender: the one made
with 25% breadcrumbs or the one with 50% breadcrumbs? This research showed that there was
no significant difference in tenderness between 25% and 50% breadcrumbs (p=0.960) and,
therefore, made burgers cooked with 25%-50% breadcrumbs a good alternative. The second
question asked when comparing tenderness was which hamburger will be more tender: the one
made with 50% breadcrumbs or the one with 75% breadcrumbs? The answer was the same, there
was no significant difference between the tenderness of burgers made with 50% and 75%
breadcrumbs (p=0.588), and would also be a good alternative recipe. Figure 2 demonstrates how
close the volume (density) and tenderness were when calculating objective evaluation averages.

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Limitations
Limitations of this study were lack of participants, lack of availability of equipment, time
constraints, a budget, and the ovens were outdated which either ran too high, too low, or just
right. Experimenters and panelists were also in the same room which could have swayed their
sense of taste with other aromas in the air.
Conclusion
The greatest change in texture was between the control and 50%breadcrumbs (p=0.000) and 75%
breadcrumbs (p=0.000). The color had significant changes when all variables were compared to
the control, all having the same significance (p=0.000). Lastly, when comparing the control’s
taste with the variables, the burger made with 75% breadcrumbs (p=0.000) was the only burger
to have a significant difference in taste. When comparing objective measures, there was no
significant change in density between the control and 25% breadcrumbs (p=0.958), 50%
0
5
10
15
20
25
30
Control 75% beef 50% beef 25% beef
Figure 2. Objective Evaluation Averages
Density
Tenderness

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breadcrumbs (p=0.580), and 75% breadcrumbs (p=0.297). In addition, tenderness did not have a
significant difference from the control to 25% breadcrumbs (p=1.000), 50% breadcrumbs
(p=0.611), and 75% breadcrumbs (p=0.865). Patients with LPI can use these results to alter this
hamburger recipe depending on their desired outcomes. Specifically, for a burger low in protein,
the burger made with 75% breadcrumbs would be the best option containing 4.8g of protein
compared to 13.6g with the control. No research has been conducted testing the effects of
substituting a percentage of breadcrumbs for ground beef in hamburgers related to those with
LPI. More research needs to be done on how to modify recipes for those in need of a low protein
diet, while also maintaining the quality of a product, and also on breadcrumbs. From the
foregoing it has been made clear that the choice of an alternative recipe lies in the hands of the
consumer, deciding what he or she desires as a final product. The goal of this experiment was to
find a recipe that could be used by patients with LPI to help them enjoy a normal lifestyle while
eating the same, common foods as everyone else, and that alternative was reached.

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