The Effects of Cheese Substitutions on Macaroni and Cheese Product Outcomes

By: Rawan Abdalla and Laura Johnston

Abstract:
Hypercholesterolemia is a condition characterized by high serum cholesterol levels. The
consumption of saturated fats in the diet raises serum cholesterol levels, increasing the risk of
developing cardiovascular disease. The product in this experiment was macaroni and cheese, a
dish typically high in saturated fat content. The main ingredient, sharp cheddar cheese, was
substituted for three different alternatives: reduced fat cheddar cheese, part skim low moisture
mozzarella cheese, and nutritional yeast. These are more favorable substitutions for individuals
with hypercholesterolemia because of the reduced saturated content compared to the control
recipe. For the objective tests, recipe variations with reduced saturated fat content will be more
viscous, less tender, but have similar masses when sampled and evaluated. For the sensory
evaluation, judges will find the variations with reduced saturated fat to be have reduced
tenderness, creaminess, moisture, and buttery aroma; however, there will still be a higher overall
liking for the recipe variations. The objective tests did not yield statistically significant results.
The sensory evaluation did yield some statistically significant results. The nutritional yeast
recipe was significantly different from the control recipe when it came to average rating of
moisture, buttery aroma, and overall liking. Because fat contributes richness, tenderness, and
satiety factors to food, evidence agrees with the fact the nutritional yeast recipe was not well
perceived by sensory judges. The results have implications for those adjusting traditionally
saturated fat dense recipes in order to improve the nutritional value of the recipe. While a simple
one ingredient substitution may seem appropriate, the balance of the ingredients in the recipe
must be considered in order to create an appealing and tasty recipe.

Introduction:
​The clinical issue addressed in this experiment is hypercholesterolemia.
Hypercholesterolemia is characterized by high cholesterol levels in the blood. This can be caused
by an unhealthy diet or a hereditary condition known as familial hypercholesterolemia. Acquired
hypercholesterolemia is attributed to the consumption of a diet high in saturated and trans fats

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(Kelly 2010). In 2015, the FDA ruled trans fats unsafe to consume due to their extremely
negative effect on cardiovascular health (FDA 2018). However, saturated fats are still recognized
as safe. Both saturated and trans fats raise serum LDL cholesterol levels. Elevated LDL
cholesterol levels and high serum total cholesterol are major risk factors for the development of
cardiovascular disease, the leading cause of death in the country (“Hypercholesterolemia” 2018).
Currently, it is estimated that over 34 million adults have high cholesterol levels
(“Hypercholesterolemia” 2018). Reducing dietary saturated fat intake and eliminating trans fat
intake is proven to lower serum cholesterol levels (Hooper et al. 2015). Studies have shown
individuals who consume diets lower in saturated fats and higher in unsaturated fats tend to have
healthier lipid profiles, including lower serum cholesterol levels (Dietary Guidelines Advisory
Committee​ ​2015).
The product created in this experiment was macaroni and cheese. Sharp cheddar cheese
was used as the control ingredient. The variable ingredients were reduced fat cheddar cheese,
part skim low moisture mozzarella cheese, and nutritional yeast. These three products were
chosen as alternatives to the control due to their varying saturated fat content levels, all of which
are below the saturated fat content level of the control. One ¼ cup serving of regular cheddar
cheese has 6 grams of saturated fat on average. In comparison, one ¼ cup of reduced fat cheddar
cheese has 4 grams of saturated fat on average, one ¼ cup serving of part skim low moisture
mozzarella cheese has an average of 3.5 grams of saturated fat, and a 1.5 tablespoon serving of
nutritional yeast has no saturated fat content. Ideally, these products would be more favorable
substitutions to control saturated fat in a recipe designed to be more suitable for individuals with
hypercholesterolemia.
The control, sharp cheddar cheese, is one of the more popular cheese choices in the
country. It is produced by separating the curds and whey of whole milk using the enzyme rennet.
Then, the curds are cut, cooked and drained. This is followed by a special process unique to
cheddar cheese, called cheddaring. The curds are kneaded with salt, cut, stacked and turned to
allow the remaining whey to drain out. Then, the cheddar is allowed to age. The stronger the
cheddar flavor, the longer the aging period (Early 2000).​ ​The first variable, reduced fat cheddar
cheese, is modified from regular cheddar by removing some fat from the milk before the cheese

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is produced (Nelson 2004). The reduction in the fat content results in increased viscosity and
melting time, and a decrease in the flowability and stretchability of the cheese when it is baked
(Guinee 2000).​ ​However, the intensity of the cheddar flavor should be unaffected, since it resides
in the water soluble portion of the cheese, which is not modified (Nelson 2004). The second
variable, part skim low moisture mozzarella cheese, is made with a lower fat content than regular
mozzarella. Though this is desirable for health, research shows that this fat reduction results in
an increase in firmness and a decrease in the amount of free oils for baking and flowability,
which may be undesirable in palatability (Guinee 2000). The third variable, nutritional yeast, is a
flaky yellow powder created from the deactivated yeast ​Saccharomyces cerevisiae​. It has a nutty,
cheesy, creamy flavor similar to parmesan cheese, and is thus typically used by vegans as a
cheese substitute (Dubey 2010). Nutritional yeast is rich in protein and B vitamins, and tends to
be low in fat and sodium (​Harusekwi 2014​). Therefore, it may be used as a healthier, animal-free
alternative to cheese in a wide variety of recipes.
The purpose of the experiment was to evaluate the effects of cheese substitutions on the
palatability and overall liking of a macaroni and cheese product. Three objective measures were
tested; viscosity (relating to the cheese sauce), mass, and tenderness. Additionally, six subjective
measures were evaluated by a panel of judges. These measures were color, texture, moistness,
creaminess, buttery aroma, and overall liking.​ ​For the objective tests, recipe variations with
reduced saturated fat content will be more viscous, less tender, but have similar masses when
sampled and evaluated. For the sensory evaluation, judges will find the variations with reduced
saturated fat to be have reduced tenderness, creaminess, moisture, and buttery aroma; however,
there will still be a higher overall liking for the recipe variations.

Materials and Methods:

The materials and equipment used for the experiment were one large pot, four saucepans,
four measuring spoons, four dry measuring cups, four liquid measuring cups, four mixing bowls,
four whisks, four pyrex baking dishes, one colander, one microwave, one timer, eight copies of
sensory ballots, one scale, one line spread test tool, 32 sample cups, one black marker, eight
plastic spoons, eight small white trays, and two pairs of small lab gloves. The ingredients used
for the experiment are listed in Table 1 below.

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Table 1: Ingredients Used in the Experiment, Specific Amounts, and Their Sources

Ingredient Amount Used Source

2% Milk 4 cups total Wakefern Food Corp

1 cup per recipe Keasbey, NJ

Salted Butter 4 tbsp total Wakefern Food Corp

1 tbsp per recipe Keasbey, NJ

Shredded Sharp Cheddar 5 oz Wakefern Food Corp

Cheese Keasbey, NJ

Shredded Reduced Fat 5 oz Wakefern Food Corp

Cheddar Cheese Keasbey, NJ

Shredded Part Skim Low 5 oz Wakefern Food Corp

Moisture Mozzarella Cheese Keasbey, NJ

Nutritional Yeast 1.25 oz Lesaffre Food Corp

Milwaukee, WI

Garlic Powder 1 oz Wakefern Food Corp

Elizabeth, NJ

All Purpose Flour 4 tbsp total Wakefern Food Corp

1 tbsp per recipe Keasbey, NJ

Salt 1 tsp total Wakefern Food Corp

¼ tsp per recipe Keasbey, NJ

Ground Black Pepper 1 tsp total Mccormick & Company, Inc.

¼ tsp per recipe Hunt Valley, MD

Elbow Macaroni 32 oz total Riviana Foods Inc.

8 oz each recipe Houston, TX

Cooking Spray Enough to coat the bottom of Wakefern Food Corp

4 baking dishes Keasbey, NJ

The first step of the procedure was preheating the oven to 400°F. Then, in a large pot, 8
quarts of water were measured using a liquid measuring cup and brought to a boil. After the
water boiled, 32 ounces of pasta were added to the water and cooked for 8 minutes. The pasta

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was then drained into a colander placed in the sink and set aside. Eight (8) fl oz. of milk were
measured using a glass liquid measuring cup and microwaved for 1 ½ minutes on high, then set
aside. One (1) tablespoon of butter was placed into a saucepan and melted over medium-low
heat. One (1) tablespoon of flour was then whisked with the butter until smooth, then whisked
for another minute. The milk was then slowly poured into the saucepan and the mixture was
whisked constantly for 5 minutes until it thickened. Then, ¼ teaspoon of salt and ¼ teaspoon of
black pepper were measured with measuring spoons and whisked into the mixture along with 4
ounces of sharp cheddar cheese, which was measured using a bowl placed on a scale.
One-quarter (¼) of the cooked pasta was then added to the saucepan and folded in the sauced
until fully coated. One baking dish was sprayed lightly with cooking spray. The mixture was
then poured into the baking dish. One (1) oz. of sharp cheddar cheese was then sprinkled on top
of the mixture. The product was placed in the oven and baked for 20 minutes.
For the variations using reduced fat cheddar cheese and part skim low moisture
mozzarella cheese, the procedure was repeated, substituting sharp cheddar cheese for reduced fat
cheddar cheese and part skim low moisture mozzarella cheese of the same quantities,
respectively. For the variation using nutritional yeast, sharp cheddar cheese was substituted for
2.5 ounces of nutritional yeast and 1 ounce of garlic powder.
For the objective tests, viscosity of the cheese sauce was measured using a line spread
test. A metal cylinder was placed in the center of a line spread test board. The cylinder was filled
to the brim with the first sample. The cylinder was then lifted vertically. Measurements at each
corner of the test were taken after 30 seconds had passed. The average of the four readings was
calculated and recorded. The test procedure was repeated two more times per sample, three times
total for each of the four products.
Mass was tested by weighing two tablespoons of the first sample in a small bowl on a
scale. The weight was recorded in grams. This test was repeated two more times per sample,
three times total for each of the four products.
Tenderness was tested by measuring 2 oz. of the first sample using a small bowl on a
scale. The entire sample was placed in the mouth of one researcher. She chewed the sample and

5
counted the number of chews required in order to be able to swallow the sample. This test was
repeated two more times per sample, three times total for each of the four products.
The subjective tests were prepared by labeling 32 sample testing cups using a black
marker. Eight cups were labeled “118”, five cups were labeled “304”, eight cups were labeled
“708”, and eight cups were labeled “828”. Two ounces of the sharp cheddar cheese product were
scooped into each of the eight cups labeled “118”. Two ounces of the reduced fat cheddar cheese
product were scooped into each of the eight cups labeled “304”. Two ounces of the part skim low
moisture mozzarella cheese product were scooped into each of the eight cups labeled “708”. Two
ounces of the nutritional yeast product were scooped into each of the eight cups labeled “828”.
One sample testing cup of each product was placed in a small white tray. The tray of products
and one sensory ballot were placed next to each other in a separate room. This step was repeated
seven more times, thus preparing a total of eight subjective test panels. Judges were allowed into
the room to evaluate the samples. After the judges completed their subjective evaluations, the
sensory ballots were collected and evaluated. Statistical analysis was done on the results
obtained.

Results:
Objective Results
To evaluate the recipe variations, line spread, mass, and number of chews were chosen as
objective tests. Each test was performed in triplicate, and the average calculated. Refer to Figure
1- Figure 3 for average objective test results with standard deviation error. Refer to Appendix C
for calculations of objective results.

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Figure 1: Average Line Spread Score for Mac and Cheese Variations (n=3)
The average line spread score for sharp cheddar recipe (control recipe #118) was 1.92 with a std
dev. of +/- 1.01. The average line spread score for the reduced fat cheddar recipe (#304) was
0.42 with a std dev. of +/- 0.14. The average line spread score for the part skim low moisture
mozzarella recipe (#708) was 2.58 with a std dev. of +/- 0.80. The average line spread score for
the nutritional yeast recipe (#828) was 1.67 with a std dev. of +/- 0.38. With significance
established at p-value <0.05, the recipe variations were not significantly different from the
control recipe with a p-value = 0.12 for reduced fat cheddar, p-value = 0.46 for part skim low
moisture mozzarella, and p-value = 0.58 for nutritional yeast.

Figure 2: Average Mass in Grams for Mac and Cheese Variations (n=3)
The average mass in grams for the sharp cheddar recipe was 36g with a std dev. of +/- 2.65g. The
average mass for the reduced fat cheddar recipe was 39.67g with a std dev. of +/- 2.08g. The
average mass for the part skim low moisture mozzarella recipe was 40.67g with a std dev. of +/-

7
1.53g. The average mass for the nutritional yeast was 38.67g with a std dev. of +/- 4.16g. With
significance established at p-value <0.05, the recipe variations were not significantly different
from the control recipe with a p-value = 0.29 for reduced fat cheddar, p-value = 0.08 for part
skim low moisture mozzarella, and p-value = 0.56 for nutritional yeast. While the part skim low
moisture mozzarella was not significant at a p-value = 0.08, it can be said the p-value is trending
towards significance.

Figure 3: Average Number of Chews for Mac and Cheese Variations (n=3)
The average number of chews for the sharp cheddar recipe was 10.67 chews with a std dev. of
+/- 1.15 chews. The average number of chews the reduced fat cheddar recipe was 9.33 chews
with a std dev. of +/- 0.58 chews. The average number of chews for the part skim low moisture
mozzarella recipe was 12.67 chews with a std dev. of +/- 1.53 chews. The average number of
chews for the nutritional yeast recipe was 11.33 chews with a std dev. of +/- 1.15 chews. With
significance established at p-value <0.05, the recipe variations were not significantly different
from the control recipe with a p-value = 0.27 for reduced fat cheddar, p-value = 0.18 for part
skim low moisture mozzarella, and p-value = 0.42 for nutritional yeast.

Subjective Results
A sensory ballot was created to evaluate attributes of each recipe variation. Orange color was
selected to evaluate appearance, chewiness for texture, moistness for mouthfeel, creaminess for
mouthfeel, buttery aroma for aroma, and finally overall liking. A total of 8 judges participated in

8
the sensory analysis (n=8) and the average rating was calculated along with standard deviation
and p-value using a student t-test. Statistical significance was established with p< 0.05. Refer to
Figure 4- Figure 9 for average objective test results with standard deviation error. Refer to
Appendix D for calculations of subjective results.

Figure 4: Average Rating for Orange Color of Mac and Cheese Variations on a Scale
of 1 to 5 (1 being the least, 5 being the most) (n=8)
With regards to orange color, the sharp cheddar recipe had an average rating of 4.13 and a std
dev. of +/- 0.83. The reduced fat cheddar recipe had an average rating of 3.38 and a std dev. of
+/- 0.74. The part skim low moisture mozzarella recipe had an average rating of 1.00 and a std
dev. of +/- 0. The nutritional yeast recipe had an average rating of 3.00 and a std dev. of +/-
1.77. With significance established at p-value <0.05, the reduced fat cheddar recipe was trending
on statistically significant at p-value = 0.05. The part skim low moisture mozzarella recipe was
statistically significant at p-value= 0.00. The nutritional yeast recipe was not statistically
significant with a p-value= 0.14.

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Figure 5: Average Rating for Chewiness of Mac and Cheese Variations on a Scale of 1 to 5 (1
being the least, 5 being the most) (n=8)
With regards to chewiness, the sharp cheddar recipe had an average rating of 3.88 and a std dev.
of +/- 0.83. The reduced fat cheddar recipe had an average rating of 4.13 and a std dev. of +/-
0.64. The part skim low moisture mozzarella recipe had an average rating of 3.63 and a std dev.
of +/- 1.41. The nutritional yeast recipe had an average rating of 4.00 and a std dev. of +/- 0.68.
With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe with a p-value = 0.17 for reduced fat cheddar, p-value = 0.56 for
part skim low moisture mozzarella, and p-value = 0.68 for nutritional yeast.

Figure 6: Average Rating for Moisture of Mac and Cheese Variations on a Scale of 1 to 5 (1
being the least, 5 being the most) (n=8)

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With regards to moisture, the sharp cheddar recipe had an average rating of 3.75 and a std dev. of
+/- 1.39. The reduced fat cheddar recipe had an average rating of 3.75 and a std dev. of +/- 1.16.
The part skim low moisture mozzarella recipe had an average rating of 3.13 and a std dev. of +/-
0.99. The nutritional yeast recipe had an average rating of 2.50 and a std dev. of +/- 1.31. With
significance established at p-value <0.05, the nutritional yeast recipe was statistically significant
at p-value= 0.03. The reduced fat cheddar recipe and the part skim low moisture mozzarella
recipes were not statistically significant with a p-value= 1.00 and p-value = 0.18 respectively.

Figure 7: Average Rating for Creaminess of Mac and Cheese Variations on a Scale of 1 to 5
(1 being the least, 5 being the most) (n=8)
With regards to creaminess, the sharp cheddar recipe had an average rating of 3.75 and a std dev.
of +/- 1.16. The reduced fat cheddar recipe had an average rating of 4.00 and a std dev. of +/-
1.07. The part skim low moisture mozzarella recipe had an average rating of 2.75 and a std dev.
of +/- 1.16. The nutritional yeast recipe had an average rating of 2.50 and a std dev. of +/- 1.20.
With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe with a p-value = 0.65 for reduced fat cheddar, p-value = 0.10 for
part skim low moisture mozzarella, and p-value = 0.13 for nutritional yeast.

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Figure 8: Average Rating for Buttery Aroma of Mac and Cheese Variations on a Scale of 1 to
5 (1 being the least, 5 being the most) (n=8)
With regards to buttery aroma , the sharp cheddar recipe had an average rating of 3.38 and a std
dev. of +/- 1.30. The reduced fat cheddar recipe had an average rating of 3.50 and a std dev. of
+/- 1.20. The part skim low moisture mozzarella recipe had an average rating of 3.25 and a std
dev. of +/- 1.67. The nutritional yeast recipe had an average rating of 1.50 and a std dev. of +/-
1.07. With significance established at p-value <0.05, the nutritional yeast recipe was statistically
significant at p-value= 0.04. The reduced fat cheddar recipe and the part skim low moisture
mozzarella recipes were not statistically significant with a p-value= 0.87 and p-value = 0.85
respectively.

Figure 9: Average Rating for Overall Liking of Mac and Cheese Variations on a Scale of 1 to
5 (1 being the least, 5 being the most) (n=8)

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 With regards to overall liking, the sharp cheddar recipe had an average rating of 4.13 and a std
dev. of +/- 0.64. The reduced fat cheddar recipe had an average rating of 4.00 and a std dev. of
+/- 1.07. The part skim low moisture mozzarella recipe had an average rating of 3.38 and a std
dev. of +/- 0.92. The nutritional yeast recipe had an average rating of 2.00 and a std dev. of +/-
1.07. With significance established at p-value <0.05, the nutritional yeast recipe was statistically
significant at p-value= 0.00. The reduced fat cheddar recipe and the part skim low moisture
mozzarella recipes were not statistically significant with a p-value= 0.76 and p-value = 0.14
respectively.

Nutrition Facts Labels

In Table 2, below, are some of the highlighted differences among the created nutrition facts
labels for each recipe variation. Total fat, saturated fat, cholesterol, and sodium when consumed
in excess can be risk factors for increasing LDL cholesterol as well as other heart health issues.
Vitamin A and calcium are found in dairy products, and because the fat content of the cheese
was changing, the change in those nutrients was noted. Refer to Appendix E for complete
nutrition facts labels.

Table 2: Nutrition Facts Label Selected Nutrients

Nutrients Sharp Cheddar RF Cheddar PSLM Mozzarella Nutritional Yeast

Recipe Recipe Recipe Recipe

Calories 160 calories 150 calories 150 calories 140 calories

Total Fat (g) 7g 6g 5g 3g

Saturated Fat (g) 4.5g 3.5g 3g 1.5g

Cholesterol (mg) 25mg 15mg 15mg 5mg

Sodium (mg) 190mg 230mg 190mg 115mg

Vitamin A (%) 6% 6% 4% 2%

Calcium (%) 15% 15% 15% 4%

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Discussion:
It was hypothesized reducing the saturated fat content would lead to a more viscous, less
tender product that is similar in mass. Studies have shown saturated fats increase the levels of
LDL cholesterol in the blood, which is a factor in the development of hypercholesterolemia.
Macaroni and cheese is a classic American comfort food; however, the basic ingredients are
cheese, butter, and milk which equates to a high amount of saturated fat. In order to create a
recipe which is lower in saturated fat, and potentially a better choice for those with
hypercholesterolemia, the cheddar cheese in a control recipe was substituted with three
variations. The control cheese was sharp cheddar cheese and the variations were reduced fat
sharp cheddar, part skim low moisture mozzarella, and nutritional yeast.
The objective measures looked to evaluate changes in viscosity, tenderness, and mass of
the recipes when substituting sharp cheddar cheese for ingredients increasingly lower in
saturated fat content. Fat contributes to characteristics of food, such as aroma, richness, texture,
and satiety when eating. For the line spread scores none of the measures were statistically
significant; therefore, it cannot concluded replacing the control cheese with a lower saturated fat
option alters the viscosity of the baked cheese sauce. For the mass in grams, the part skim low
moisture mozzarella cheese was trending towards significance with a p-value = 0.08. The other
recipes were not statistically significant; therefore, it cannot be concluded replacing the control
cheese with a lower saturated fat option alters the weight of the mac and cheese. For the number
of chews, none of the measures were statistically significant; therefore, it cannot be concluded
replacing the control cheese with a lower saturated fat option alters the tenderness of the mac and
cheese. Refer to Appendix C for detailed results of the objective tests.
Based on background research, a significant difference was expected for line spread and
number of chews because fat content decreases viscosity and increases tenderness. Contrary to
the hypothesis, none of the results were statistically significant, with only the mass of the part
skim low moisture mozzarella trending on significance (p-value = 0.08). The reduction of
saturated fat content should have resulted in a more viscous cheese sauce; however, the line
spread measures did not show those results. It was noted the part skim low moisture mozzarella
recipe did not hold together in the mouth, potentially increasing the number of chews because the

14
components moved around the mouth more freely. As fat contributes to tenderness, it was
surprising the reduced fat cheddar recipe had the lowest average number of chews. It was
expected the control recipe with full fat would have the highest average number of chews.
For the sensory evaluation, it was hypothesized judges will find the variations with
reduced saturated fat to be have reduced tenderness, creaminess, moisture, and buttery aroma;
however, there will still be a higher overall liking for the recipe variations. The subjective
measures looked to evaluate the appearance, texture, tenderness, aroma, and overall liking of the
macaroni and cheese for the control and recipe variations. A two oz. sample was placed into
plastic containers marked 118 (control) 304 (reduced fat cheddar) 708 (part skim low moisture
mozzarella) and 828 (nutritional yeast). A total of 8 (n=8) judges completed sensory ballots.
The part skim low moisture mozzarella recipe was statistically different from the control
recipe ( p=0.00); however, the other recipes did not display a clear significant difference. The
difference in color is simply because the part skim low moisture mozzarella is white and did not
display any orange color. A surprising result from the orange appearance of the recipe was how
the average rating of the reduced fat cheddar recipes was almost 1 point lower than the control
recipe. The actual color of the cheese before cooking was not different, and it is unknown why
the color difference was perceived by judges. As fat content contributes to tenderness, it was
expected chewiness would increase as saturated fat content decreased; however, the results
differed. The part skim low moisture mozzarella had the lowest average rating for chewiness
despite its lower fat content compared to the control. This could be attributed to the reduced
hold noted before with the objective measurement for number of chews. For chewiness, none of
the recipes were statistically significant with regards to the control recipe; therefore, it cannot be
concluded texture was affected by replacing the cheese in the recipes. For moisture, the
nutritional yeast was significantly different from the control with a p-value of 0.03. This was
expected as it had the lowest saturated fat content of the recipe variations. None of the recipes
were statistically significant with regards to a difference in creaminess; therefore, it cannot be
concluded replacing the control cheese altered the creaminess of the macaroni and cheese. This
is surprising considering judges noticed a difference in moisture, but not creaminess. The part
skim low moisture mozzarella recipe and nutritional yeast recipe were on the cusp of being

15
considered for trending on significance, and with repeated sensory evaluation may have been
found to be significantly different. For buttery aroma, the p-value for the nutritional yeast recipe
was statistically significant with a p-value of 0.04. The other recipes were not statistically
significant. It can be concluded the nutritional yeast recipe is significantly different from the
control recipe with regards to buttery aroma. For the overall liking of the recipe, the p-value for
the nutritional yeast recipe was statistically significant with p-value = 0.00. The other recipes
were not statistically significant from the control recipe. It can be concluded the nutritional yeast
is significantly different from the control recipe with regards to overall liking. Refer to
Appendix D for detailed results of the subjective tests.
Potential sources of error did exist in objective measures of this experiment. The line
spread was performed after baking the recipes with a short rest period. This rest period
introduced some error as the mac and cheese cooled and lost some fluidity. To overcome this,
samples were reheated in a microwave prior to testing. The cooling and reheating of the samples
in the microwave could have altered the viscosity. When reading the line spread, some
individual noodles fell farther from the rest of the sample, and those noodles were excluded
when reading the linespread. With regards to the mass, it was noted the scoops of macaroni and
cheese were not uniform, with some consisting of more noodles or more cheese sauce. This had
implications for both mass readings and number of chews, as uniformity is important for both
measures.
If the experiment were to be repeated, a number of procedures would be changed. For the
purpose of measuring viscosity, the cheese sauce would be measured separate from the noodles
in order to get a more accurate reading. Additionally, the line spread reading would be taken
immediately after preparation to avoid issues with cooling and reheating. For the mass, the
entire mass of the final dish would be taken rather than an individual sample. This eliminates the
issue of uniformity with a two tablespoon sample consisting of different ingredient ratios.
By replacing the sharp cheddar cheese in the control recipe with options lower in
saturated fat content, the saturated fat content was effectively lowered. Looking at the nutrition
facts labels, the total fat, saturated fat, cholesterol, and sodium all reduce in the recipe variations.
For the reduced fat cheddar recipe, there is an increase in sodium, most likely related to the

16
processing of low fat products that occurs to preserve quality when fat is removed. With the
nutritional yeast there is the largest decrease in nutrients; however, this is because it is compared
to dairy products that naturally contain more of these nutrients. There were observable changes
in the variations; however, only a couple results from the sensory ballots were statistically
significant. It can be concluded replacing the control cheese with part skim low moisture
mozzarella significantly altered the perception of color for the macaroni and cheese. It can also
be concluded replacing the control cheese with nutritional yeast significantly altered the
perception of moisture, buttery aroma, and overall liking of the macaroni and cheese. Based on
these results, replacing sharp cheddar cheese may produce a mac and cheese lower in saturated
fat; however, it may not be as liked by a consumer because of the loss of moisture and richness
supplied by the original fat content. As stated before, removing the saturated fat from a recipe
may improve the nutritional value of the recipe; however, it can remove the taste and appeal
from the dish. Balance should be considered when altering recipes higher in fat, as more than one
ingredient may need to be substituted to maintain similar textures and flavors supplied by fat.

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Appendix Table of Contents

Appendix A: Macaroni and Cheese Recipe with Variations

Appendix B: Sensory Ballot
Appendix C: Objective Test Averages, Std Dev, and T-Test Calculations
Appendix D: Objective Test Averages, Std Dev, and T-Test Calculations
Appendix E: Nutrition Facts Labels

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 Appendix A: Macaroni and Cheese Recipe with Variations

Recipe: Macaroni and Cheese

Chefs: Laura Johnston and Rawan Abdalla

Yield: 22.5 oz ​ Portion Size: ​2.5 oz.

AMT UNIT INGREDIENTS PROCEDURES

*Preheat oven to 400°F

8 oz Elbow Macaroni Bring 4 quarts of water to a boil, add 32 oz of pasta, cook,

drain, and set aside.

1 tbsp Unsalted Butter In a saucepan, melt 1 tbsp butter over medium to high heat

1 tbsp AP Flour Add 1 tbsp of flour to melted butter, whisk continuously for
1-2 minutes

8 fl oz 2% milk Microwave 1.5 cups of 2% milk on high for 2 minutes, slowly

pour into roux in saucepan while whisking. Continue whisking
for 5 minutes or until thickened.

5 oz Sharp Cheddar Cheese Shred 5 oz of sharp cheddar cheese and add to saucepan,
whisking until smooth.

0.25 tsp Ground Black Pepper Add 1/4 tsp of salt and pepper to cheese mixture. Remove
from heat and fold into 8 oz of elbow macaroni

0.25 tsp Salt

Spray glass baking dish with cooking spray, add mac and
cheese, and sprinkle remaining 1 oz of sharp cheddar
cheese. Bake for 20 minutes.

5 oz Reduced Fat Cheddar *Use in place of sharp cheddar cheese for variation 1
Cheese

5 oz Part Skim Low Moisture *Use in place of sharp cheddar cheese for variation 2
Mozzarella

2.5 oz Nutritional Yeast *Use in place of sharp cheddar cheese for variation 3

0.25 tsp Garlic Powder

19
Appendix B: Sensory Ballot

Laura Johnston and Rawan Abdalla

Section 1, Monday, Station 4

INSTRUCTIONS:

Hold the sample in your mouth for about 5 seconds.

Be sure to bring sample into contact with all tasting areas.
Rinse your mouth with room temperature water in between each sample
Wait at least 10 seconds in between each sample
Re-tasting each sample is permitted once before proceeding to the next sample.

20
 Please Rate from 1 (LEAST) to 5 (MOST)

SAMPLE 118

APPEARANCE (COLOR)
How ORANGE is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
TEXTURE
How CHEWY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
MOUTHFEEL
How MOIST is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
How CREAMY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
AROMA
How BUTTERY does the sample smell?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
OVERALL LIKING
▢ ▢ ▢ ▢ ▢
1 2 3 4 5

Comments:

21
 Please Rate from 1 (LEAST) to 5 (MOST)

SAMPLE 304

APPEARANCE (COLOR)
How ORANGE is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
TEXTURE
How CHEWY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
MOUTHFEEL
How MOIST is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
How CREAMY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
AROMA
How BUTTERY does the sample smell?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
OVERALL LIKING
▢ ▢ ▢ ▢ ▢
1 2 3 4 5

Comments:

22
 Please Rate from 1 (LEAST) to 5 (MOST)

SAMPLE 828

APPEARANCE (COLOR)
How ORANGE is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
TEXTURE
How CHEWY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
MOUTHFEEL
How MOIST is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
How CREAMY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
AROMA
How BUTTERY does the sample smell?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
OVERALL LIKING
▢ ▢ ▢ ▢ ▢
1 2 3 4 5

Comments:

23
 Please Rate from 1 (LEAST) to 5 (MOST)

SAMPLE 708

APPEARANCE (COLOR)
How ORANGE is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
TEXTURE
How CHEWY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
MOUTHFEEL
How MOIST is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
How CREAMY is the sample?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
AROMA
How BUTTERY does the sample smell?
▢ ▢ ▢ ▢ ▢
1 2 3 4 5
OVERALL LIKING
▢ ▢ ▢ ▢ ▢
1 2 3 4 5

Comments:

24
Appendix C: Objective Test Averages, Std Dev, and T-Test Calculations

LINE SPREAD Sharp Cheddar RF Cheddar PSLM Mozzarella Nutritional Yeast

Measurement 1 1.75 0.25 3.5 1.75

Measurement 2 3 0.5 2.25 2

Measurement 3 1 0.5 2 1.25

Average 1.92 0.42 2.58 1.67

Std. Dev 1.01 0.14 0.80 0.38

t-Tests 0.12 0.46 0.58

MASS (grams) Sharp Cheddar RF Cheddar PSLM Mozzarella Nutritional Yeast

Measurement 1 35 39 42 42

Measurement 2 34 42 39 40

Measurement 3 39 38 41 34

Average 36.00 39.67 40.67 38.67

Std. Dev 2.65 2.08 1.53 4.16

t-Tests 0.29 0.08 0.56

# OF CHEWS Sharp Cheddar RF Cheddar PSLM Mozzarella Nutritional Yeast

Measurement 1 10 10 11 10

Measurement 2 12 9 13 12

Measurement 3 10 9 14 12

Average 10.67 9.33 12.67 11.33

Std. Dev 1.15 0.58 1.53 1.15

t-Tests 0.27 0.18 0.42

25
Appendix D: Subjective Test Averages, Std Dev, and T-Test Calculations

Orange (color) Sharp Cheddar RF Cheddar (304) PSLM Nutritional

(118) Mozzarella (708) Yeast (828)

Judge 1 3 2 1 2

Judge 2 5 4 1 5

Judge 3 5 3 1 5

Judge 4 4 4 1 3

Judge 5 3 4 1 5

Judge 6 4 3 1 2

Judge 7 5 4 1 1

Judge 8 4 3 1 1

ave 4.13 3.38 1.00 3.00

std dev 0.83 0.74 0.00 1.77

t-Tests 0.05 0.00 0.14

Chewy (texture) Sharp Cheddar RF Cheddar (304) PSLM Nutritional

(118) Mozzarella (708) Yeast (828)

Judge 1 4 4 4 4

Judge 2 4 4 2 5

Judge 3 4 5 5 5

Judge 4 4 4 4 5

Judge 5 4 4 2 4

Judge 6 4 4 5 3

Judge 7 5 5 5 5

Judge 8 2 3 2 1

26
 ave 3.88 4.13 3.63 4.00

std dev 0.83 0.64 1.41 1.41

t-Tests 0.17 0.56 0.68

Moist (mouthfeel) Sharp Cheddar RF Cheddar (304) PSLM Nutritional

(118) Mozzarella (708) Yeast (828)

Judge 1 5 5 3 4

Judge 2 4 3 3 3

Judge 3 1 3 2 1

Judge 4 3 2 2 1

Judge 5 3 5 4 3

Judge 6 5 3 5 1

Judge 7 5 5 3 4

Judge 8 4 4 3 3

ave 3.75 3.75 3.13 2.50

std dev 1.39 1.16 0.99 1.31

t-Tests 1.00 0.18 0.03

Creamy Sharp Cheddar RF Cheddar (304) PSLM Nutritional

(mouthfeel) (118) Mozzarella (708) Yeast (828)

Judge 1 2 4 3 5

Judge 2 5 4 1 2

Judge 3 2 4 2 2

Judge 4 4 2 2 2

27
 Judge 5 4 5 3 3

Judge 6 5 5 5 2

Judge 7 4 5 3 3

Judge 8 4 3 3 1

ave 3.75 4.00 2.75 2.50

std dev 1.16 1.07 1.16 1.20

t-Tests 0.65 0.10 0.13

Buttery (Aroma) Sharp Cheddar RF Cheddar (304) PSLM Nutritional Yeast

(118) Mozzarella (828)
(708)

Judge 1 4 3 5 2

Judge 2 1 5 1 4

Judge 3 4 2 4 1

Judge 4 4 2 1 1

Judge 5 4 4 2 1

Judge 6 3 3 5 1

Judge 7 5 5 4 1

Judge 8 2 4 4 1

ave 3.38 3.50 3.25 1.50

std dev 1.30 1.20 1.67 1.07

t-Tests 0.87 0.85 0.04

28
Overall Liking Sharp Cheddar RF Cheddar (304) PSLM Nutritional Yeast
(118) Mozzarella (828)
(708)

Judge 1 4 4 3 3

Judge 2 5 4 3 2

Judge 3 3 4 4 2

Judge 4 4 2 2 2

Judge 5 4 5 4 4

Judge 6 4 3 5 1

Judge 7 5 5 3 1

Judge 8 4 5 3 1

ave 4.13 4.00 3.38 2.00

std dev 0.64 1.07 0.92 1.07

t-Tests 0.76 0.14 0.00

29
Appendix E: Nutrition Facts Labels for the Recipe Variations

30
 References:

Dietary Guidelines Advisory Committee. “Scientific Report of the 2015 Dietary Guidelines
Advisory Committee: Advisory Report to the Secretary of Health and Human Services and the
Secretary of Agriculture.” ​U.S. Department of Agriculture,​ Agricultural Research Service,
2015.

Dubey, R. C., et al. ​Industrial Exploitation of Microorganisms.​ I.K. International Publishing

House, 2010.

Early, Ralph. ​Technology of Dairy Products​. John Wiley, 2000.

FDA. “Trans Fat.” ​US Food and Drug Administration​, Center for Food Safety and Applied
Nutrition, 2018, ​www.fda.gov/food/ucm292278.htm​.

Guinee, Timothy P, et al. “The Effect of Fat Content on the Rheology, Microstructure and
Heat-Induced Functional Characteristics of Cheddar Cheese.” ​International Dairy Journal​,
vol. 10, no. 4, 5 Apr. 2000, pp. 277–288., doi:10.1016/s0958-6946(00)00048-0.

Harusekwi, Serere Julien, et al. “Development of High Protein Content Homemade Bread by
Nutritional Yeast Fortification for Disadvantaged Communities.” ​International Journal of
Nutrition and Food Sciences,​ vol. 3, no. 3, 20 May 2014, pp. 194–198.,
doi:10.11648/j.ijnfs.20140303.20.

Hooper, L., et al. “Reduction in Saturated Fat Intake for Cardiovascular Disease.” ​Cochrane
Database of Systematic Reviews,​ no. 6, 2015, doi:10.1002/14651858.cd011737.

“Hypercholesterolemia.” ​U.S. National Library of Medicine,​ National Institutes of Health, 4 Dec.

2018, ghr.nlm.nih.gov/condition/hypercholesterolemia.

Kelly, Robert B. “Diet and Exercise in the Management of Hyperlipidemia.” ​American Family
Physician,​ vol. 81, no. 9, 1 May 2010, pp. 1097–1104.

Nelson, B.K., and D.M. Barbano. “Reduced-Fat Cheddar Cheese Manufactured Using a Novel
Fat Removal Process.” ​Journal of Dairy Science,​ vol. 87, no. 4, 2004, pp. 841–853.,
doi:10.3168/jds.s0022-0302(04)73228-2.

31
Extra Credit Assignment

32
Table 1: Average Line Spread Score, Std Dev, and T-test Calculations with Nutritional Yeast as
Control
LINE SPREAD Nutritional Yeast Sharp Cheddar RF Cheddar PSLM
Mozzarella

Measurement 1 1.75 1.75 0.25 3.5

Measurement 2 2 3 0.5 2.25

Measurement 3 1.25 1 0.5 2

Mean 1.67 1.92 0.42 2.58

Std. Dev 0.38 1.01 0.14 0.80

t-Tests 0.58 0.04 0.17

With significance established at p-value <0.05, the part skim low moisture mozzarella recipe was
statistically significant with a p-value =0.04. The other recipe variations were not significantly
different from the control recipe

Table 2: Average Mass (g), Std Dev, and T-test Calculations with Nutritional Yeast as Control
MASS Nutritional Yeast Sharp Cheddar RF Cheddar PSLM
Mozzarella

Measurement 1 42 35 39 42

Measurement 2 40 34 42 39

Measurement 3 34 39 38 41

Mean 38.67 36.00 39.67 40.67

Std. Dev 4.16 2.65 2.08 1.53

t-Tests 0.56 0.68 0.51

With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe

33
Table 3: Average Number of Chews, Std Dev, and T-test Calculations with Nutritional Yeast as
Control
# OF CHEWS Nutritional Yeast Sharp Cheddar RF Cheddar PSLM
Mozzarella

Measurement 1 10 10 10 11

Measurement 2 12 12 9 13

Measurement 3 12 10 9 14

Mean 11.33 10.67 9.33 12.67

Std. Dev 1.15 1.15 0.58 1.53

t-Tests 0.42 0.18 0.06

With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe. However, with a p-value =0.06, the part skim low moisture
mozzarella is trending towards significance.

Table 4: Average Line Spread Score, Std Dev, and T-test Calculations with PSLM Mozzarella
Recipe as Control
LINE SPREAD PSLM Mozzarella Nutritional Yeast Sharp Cheddar RF Cheddar

Measurement 1 3.5 1.75 1.75 0.25

Measurement 2 2.25 2 3 0.5

Measurement 3 2 1.25 1 0.5

Mean 2.58 1.67 1.92 0.42

Std. Dev 0.80 0.38 1.01 0.14

t-Tests 0.17 0.46 0.06

With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe. However, with a p-value =0.06, the reduced fat cheddar recipe
is trending towards significance.

34
Table 5: Average Mass (g), Std Dev, and T-test Calculations with PSLM Mozzarella Recipe as
Control
MASS PSLM Mozzarella Nutritional Yeast Sharp Cheddar RF Cheddar

Measurement 1 42 42 35 39

Measurement 2 39 40 34 42

Measurement 3 41 34 39 38

Mean 40.67 38.67 36.00 39.67

Std. Dev 1.53 4.16 2.65 2.08

t-Tests 0.51 0.08 0.67

With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe. However, with a p-value =0.08, the sharp cheddar recipe is
trending towards significance.

Table 6: Average Number of Chews, Std Dev, and T-test Calculations with PSLM Mozzarella
Recipe as Control
# OF CHEWS PSLM Mozzarella Nutritional Yeast Sharp Cheddar RF Cheddar

Measurement 1 11 10 10 10

Measurement 2 13 12 12 9

Measurement 3 14 12 10 9

Mean 12.67 11.33 10.67 9.33

Std. Dev 1.53 1.15 1.15 0.58

t-Tests 0.06 0.18 0.11

With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe. However, with a p-value =0.06, the nutritional yeast recipe is
trending towards significance.

35
Table 7: Average Line Spread Score, Std Dev, and T-test Calculations with RF Cheddar Recipe as
Control
LINE SPREAD RF Cheddar PSLM Mozzarella Nutritional Yeast Sharp Cheddar

Measurement 1 0.25 3.5 1.75 1.75

Measurement 2 0.5 2.25 2 3

Measurement 3 0.5 2 1.25 1

Mean 0.42 2.58 1.67 1.92

Std. Dev 0.14 0.80 0.38 1.01

t-Tests 0.06 0.04 0.12

With significance established at p-value <0.05, the part skim low moisture mozzarella recipe was
statistically significant with a p-value =0.04. With a p-value =0.08, the part skim low moisture
recipe is trending towards significance. The other recipe variations were not significantly
different from the control recipe

Table 8: Average Mass (g), Std Dev, and T-test Calculations with RF Cheddar Recipe as Control
MASS RF Cheddar PSLM Mozzarella Nutritional Yeast Sharp Cheddar

Measurement 1 39 42 42 35

Measurement 2 42 39 40 34

Measurement 3 38 41 34 39

Mean 39.67 40.67 38.67 36.00

Std. Dev 2.08 1.53 4.16 2.65

t-Tests 0.67 0.68 0.29

With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe

36
Table 9: Average Number of Chews, Std Dev, and T-test Calculations with RF Cheddar Recipe as
Control
# OF CHEWS RF Cheddar PSLM Mozzarella Nutritional Yeast Sharp Cheddar

Measurement 1 10 11 10 10

Measurement 2 9 13 12 12

Measurement 3 9 14 12 10

Mean 9.33 12.67 11.33 10.67

Std. Dev 0.58 1.53 1.15 1.15

t-Tests 0.11 0.18 0.27

With significance established at p-value <0.05, the recipe variations were not significantly
different from the control recipe

37