THESIS

POTENTIAL OF WHEY CHEESE YOGURT TO DECREASE

THE CHOLESTEROL LEVELS OF RATS INDUCED HIGH-
CHOLESTEROL DIET

Compiled by:
Athaya Ramadhani Janitra
NIM 3920187181454

DEPARTMENT OF PHARMACY
FACULTY OF HEALTH SCIENCE
UNIVERSITY OF DARUSSALAM GONTOR
MANTINGAN
2021
 ABSTRACT
Cholesterol is a metabolite containing sterol fat found in cell membranes and
circulated in blood plasma. Elevated cholesterol levels can lead to atherosclerosis,
coronary heart disease and so on. Some researchs showed that the consumption of
yogurt can lower the blood cholesterol content. Yogurt has a factor that can inhibit
cholesterol synthehis. Lactic acid bacteria (BAL), an alternative to decrease
cholesterol. BAL can develop faster on ingredients mixed with it, such as whey
cheese. The availability of whey cheese is abundant; utilization that has not been
maximized is the reason for the use of whey cheese as a medium to create lactic
acid in the form of fermented yogurt drinks. This study was an experimental
study with whey cheese ingredients processed into yogurt with Lactobacillus
bulgarius and Streptococus thermophyllus stater and then tested on rats produced
with a cholesterol-tingal diet with a design of 5 controls, control models that were
rats with high cholesterol, normal conductors that were healthy rats and 3
treatments of rats high in cholesterol with a yogurt induction concentration of 2
mL, 3 mL and 4 mL. The data retrieval was done by pre and post on rats blood
taken through the tail veins and measured using GCU's Easy Touch tool. The data
was analyzed using ANOVA's One-way followed by the Tukey Post Hoc test.
The results show that whey cheese yogurt has the potential as an alternative
functional drink to reduce cholesterol levels with significant reduction results.
Administering whey cheese yogurt to mice induced a high-cholesterol diet for 14
days was able to lower total cholesterol levels significantly at a dose concentration
of 4 mL / day.

Keywords: Cholesterol, Yogurt, Whey Cheese

ii
 ACKNOWLEDGEMENTS
Alhamdulillah, by raising and gratitude to Allah SWT who has bestowed
grace and guidance this thesis proposal can be completed with the title "Potential
of Whey Cheese Yogurt against The Reduction of Cholesterol Levels of Rats
Induced by High Cholesterol Diet" as one of the conditions to complete the
Bachelor Program (S1) in the Pharmacy study program of the Faculty of Health
Sciences, University of Darussalam Gontor.
This thesis proposal cannot be resolved without the support, assistance,
guidance, and advice from various parties who contributed during the preparation
of this thesis proposal. The greatest gratitude for :
1. Leader of Pondok Modern Darussalam Gontor, Al-Ustadz KH. Hasan
Abdullah Sahal., Al-Ustadz. Prof. Dr. Amal Fathullah Zarkasyi, MA., and
Al Ustadz KH. Akrim Mariyat, Dipl. A. Ed who has given me the
opportunity to study and take many life lessons at Gontor.
2. Al-Ustadz Prof. Dr. Hamid Fahmy Zarkasyi, MA., as Rector of University
of Darussalam Gontor who has instilled the Islamization of Science to his
students, so we take part in their respective fields with a solid Islamic
foundation
3. Al-Ustadz Apt. Amal Fadholah, M.Si., as Dean of the Faculty of Health
Sciences, University of Darussalam Gontor who always fights for his
students in the Faculty of Health Sciences by giving students everything
they need to get the best results.
4. Al Ustadzah Solikah Ana Estikomah, S.Si, M.Si as the main guidance
lecturer for her patience, advice, motivation, support and willingness to
guide us.
5. Al-Ustadz Rizki Awaluddin, M.Biomed as the second guidance lecturer
for his patience and willingness in guiding us, providing prayers,
motivation, support, guidance, advice, experience in the process of
completing this thesis
6. The entire range of lecturer and education personnel for their willingness
to help the final student in the success of carrying out the final task

iii
 7. Lecturer domiciled in the dormitory environment of UNIDA Gontor
Kampus Putri for prayers, motivation and life lessons that helped me to
process to become a better Muslim
Finally, the author realizes that this thesis is far from perfection. However, we
hope that this thesis can be a benefit for the people. Aamiin.

iv
 TABLE OF CONTENT

ABSTRACT............................................................................................................ii
ACKNOWLEDGEMENTS.................................................................................iv
TABLE OF CONTENT........................................................................................vi
LIST OF TABLES..............................................................................................viii
LIST OF FIGURES..............................................................................................ix
LIST OF APPENDICES.......................................................................................x
CHAPTER I............................................................................................................1
INTRODUCTION..................................................................................................1
A. Research Background................................................................................1
B. Research Question......................................................................................3
C. Research Objectives...................................................................................3
D. Research Significance................................................................................4
E. Authenticity of Research...........................................................................4
CHAPTER II..........................................................................................................7
LITERATURE REVIEW......................................................................................7
A. Theory.........................................................................................................7
1. Definition of Yogurt..................................................................................7
2. Types of Yogurt.........................................................................................8
3. Benefits of Yogurt.....................................................................................9
4. Yogurt Making Process...........................................................................10
5. Fat Content Test......................................................................................11
6. Cheese Whey............................................................................................13
7. Cholesterol...............................................................................................14
B. Theoretical Framework...........................................................................21
C. Conseptual Framework...........................................................................22
CHAPTER III......................................................................................................23
RESEARCH METHODS....................................................................................23
A. Research Type and Design......................................................................23
B. Research Location and Time...................................................................24
C. Sample and Population............................................................................24

v
 D. Research Variable....................................................................................24
E. Operational Definition Variable.............................................................24
F. Material and Instrument.........................................................................25
G. Ethical Clearance.....................................................................................25
H. Research Procedure.................................................................................26
I. Data Analysis............................................................................................28
J. Research Schedule....................................................................................29
CHAPTER IV.......................................................................................................30
RESULT AND DISCUSSION.............................................................................30
A. Making of Cheese Whey Yogurt..............................................................30
B. Fat Content of Whey Cheese Yogurt......................................................30
C. The Effect of Yogurt Whey Cheese on the Decrease of Cholesterol
Levels in Rats Levels in Rats Induced by a High Cholesterol Diet..............32
CHAPTER V........................................................................................................38
CONCLUSIONS AND SUGGESTIONS...........................................................38
A. Conclusions...............................................................................................38
B. Suggestions................................................................................................38

vi
 LIST OF TABLES
Table 1. Authenticity of Result................................................................................4
Table 2. Operational Definition Variable..............................................................24
Table 3. Research Schedule...................................................................................29
Table 4. Average Result of Yogurt Fat Content....................................................31
Table 5. Average of Result Decreased Cholesterol Levels (mg/dL).....................33
Table 6. Average Total Cholesterol Levels (Tukey)..............................................34
Table 7. Cholesterol Levels (Post Test).................................................................35
Table 8. Test of Normality Cholesterol Levels......Error! Bookmark not defined.
Table 9. Test of Homogenity Cholesterol Levels. .Error! Bookmark not defined.
Table 10. Anova Result of Delta Cholesterol........Error! Bookmark not defined.
Table 11. Tukey test of Cholesterol Levels............Error! Bookmark not defined.
Table 12. Post Hoc Pre Test...................................Error! Bookmark not defined.
Table 13. Post Hoc Post Test.................................Error! Bookmark not defined.
Table 14. Test of Normality Whey cheese Yogurt. Error! Bookmark not defined.
Table 15. Test of Homogenity Whey Cheese Yogurt..........Error! Bookmark not
defined.
Table 16. Anova of Fat Content Whey Cheese Yogurt........Error! Bookmark not
defined.
Table 17. Tukey Test Fat Content of Whey Cheese Yogurt Error! Bookmark not
defined.

vii
 LIST OF FIGURES
Figure 1. Cheese Whey (Maulana, 2018)...............................................................13
Figure 2. Theoretical Framework...........................................................................21
Figure 3. Conseptual Framework...........................................................................22
Figure 4. Measurement of Total Cholesterol Levels (mg/dL)...............................32

viii
 LIST OF ATTACHMENT
Doc 1. Fat Content Test Soxhlet Method...............Error! Bookmark not defined.
Doc 2. Drying Whey Cheese Yogurt......................Error! Bookmark not defined.
Doc 3. Aclimatitation.............................................Error! Bookmark not defined.
Doc 4. Induced High Cholesterol Food..................Error! Bookmark not defined.
Doc 6. Cheese Whey...............................................Error! Bookmark not defined.
Doc 5. Sample Treatment.......................................Error! Bookmark not defined.
Doc 7. Whey Cheese Yogurt..................................Error! Bookmark not defined.

ix
 CHAPTER I

INTRODUCTION
A. Research Background
The cheese industry in Indonesia today is quite a lot along with technological
advances and methods that accelerate the process of making cheese. But, the
process of making cheese produces a waste called whey cheese. Whey cheese is a
milk serum formed after the process of coagulation of milk in the manufacture of
cheese. Whey cheese is separated from milk curd after coagulation of milk, cream,
skim milk, or milk butter in the manufacture of cheese, casein, or similar products
primarily with rennet-type enzymes. Whey cheese is categorized as a useful waste,
but not many know the content contained in it. Therefore, it is necessary to use
optimally whey cheese, so as not to be wasted, as God almighty commands in His
word, so humans can take advantage of everything on this earth and not be a
waster.1

‫اطنْي ِ َو َكا َن الشَّْيطَا ُن لَِربِِّه َك ُف ْو ًرا‬

ِ ‫إِ َّن الْمب ِّذ ِرين َكانُوا إِخوا َن الشَّي‬
َ َ ْ ْ َ ْ َُ
The squanderers were ever brothers of the devils, and the devil was ever an
ingrate to his Lord. (QS. Al-'Isra' [17] : 27)

Whey cheese has a variety of nutrients and is beneficial for the body. The
content in whey cheese is 5% lactose, 1% protein, 0.4% fat, and some minerals.
Increasing cheese production in Indonesia means also increasing the production of
whey cheese in it. Seeing the nutrient content, especially the very high lactose in it
and not optimal utilization of whey cheese in the food field, further effort is
needed, so whey cheese waste is not just thrown away. One product that can be
processed from whey cheese with a high lactose content is in the form of
fermented drinks such as yogurt.2

1
Badan Standardisasi Nasional, “SNI 2981: 2009 Tentang Yogurt,” Jakarta: Pusat Standarisasi
Industri Departemen Perindustrian. (2009).
2
Geoffrey W. Smithers et al., “New Opportunities from the Isolation and Utilization of Whey keju
Proteins,” Journal of Dairy Science 79, no. 8 (1996).

1
 Yogurt is a fermented product produced from a process involving aerobic or
anaerobic microbial activities. Yogurt made from very beneficial bacteria is
Lactobacillus acidophilus, Lactobacillus bulgaricus and Streptococcus
thermophilous.3 Yogurt has benefits to the health of the body, among others, can
improve the process of digestion of protein and fat, triggering the secretion of
fluids needed for the digestive process such as stomach fluids, bile, and pancreas.
The benefits of yogurt are produced from bacteria contained as the basic
ingredients of making yogurt and have been proven through various research and
research conducted.4

Some researchs showed that the consumption of yogurt can lower blood
cholesterol content. Yogurt has a factor that can inhibit cholesterol synthesis, so
cholesterol shrinks and avoids the formation of blood vessel blockage
(atherosclerosis) triggers coronary heart disease. The efficacy of yogurt to lower
cholesterol levels was proven in research tried in Shiraz, Southern Iran in 2010.
They studied the consumption of yogurt with probiotics and yogurt without
probiotics in 46 respondents aged 20-67 years. As a result, giving probiotic yogurt
300 grams per day was efficient in lowering LDL and cholesterol content.
Cholesterol in the body can go down in various ways, one of which is with
bacteria contained in yogurt, namely lactic acid bacteria (BAL).5

Lactic acid bacteria (BAL) in yogurt make one of the factors of yogurt an
alternative to lower cholesterol. Lactic acid bacteria can develop more quickly on
ingredients mixed with it, such as whey cheese. Whey cheese has a high lactose
content, whereas, in the manufacture of yogurt, the presence of lactose is needed
to help the development of lactic acid bacteria. The availability of abundant whey
cheese, the utilization that has not been maximized, and aspects of area pollution
are also the reason for the use of whey cheese as a medium to create lactic acid in
the form of yogurt fermented drinks and make it one of the ideas that need to be
3
Ermina Syainah, Sari Novita, and Rusmini Yanti, “Kajian Pembuatan Yoghurt Dari Berbagai
Jenis Susu Dan Inkubasi Yang Berbeda Terhadap Mutu Dan Daya Terima,” Skala Kesehatan
(2014).
4
D. L. Hartley and G. Denariaz, “The Role of Lactic Acid Bacteria in Yogurt Fermentation,”
International Journal of Immunotherapy (1993).
5
Abdolamir Baroutkoub et al., “Effects of Probiotic Yoghurt Consumption on the Serum
Cholesterol Levels in Hypercholestromic Cases in Shiraz, Southern Iran,” Scientific Research and
Essays 5, no. 16 (2010).

2
studied whether whey cheese yogurt can be optimal for lowering cholesterol
levels.6

High cholesterol levels can be a trigger for the onset of serious diseases and
have a risk of coronary heart disease martyrs three times faster.
Hypercholesterolemia or high total cholesterol levels have a prevalence in
Indonesia in the age group of 25-34 years was 93% and increases according to age
to 15.5% in the age group of 55-64 years. Based on the results of Basic Health
Research, the proportion of the population >15 years in Indonesia with total
cholesterol levels above the normal value of 35.9%.7

Lowering cholesterol levels using yogurt or fermented beverages has many

mechanisms because the lactic acid bacteria in yogurt have the potential to lower
cholesterol levels. The lactic acid in the bacteria in yogurt causes an acidic
environment, so many other bacteria are stunted growth. Lactic acid is formed
from fermentation between milk carbohydrates (lactose) and bacteria, so it can
produce lactic acid. This is the need for this research to be done with whey cheese
ingredients as a medium for growing lactic acid bacteria and potentially to
become yogurt to lower cholesterol levels.8

B. Research Question
1. How is the quality of whey cheese yogurt reviewed from fat content?
2. Does whey cheese yogurt have the potential to be used as a functional
cholesterol-lowering drink?
3. What is the optimum concentration of whey cheese yogurt to lower
cholesterol?

C. Research Objectives
1. Knowing the quality of whey cheese yogurt reviewed from fat content

6
Rahayu Wp. Jenie Bsl, “Penanganan Limbah Industri Pangan,” Kanisius (1993).
7
Kustiasih; Sakhnan Lestari R, “The Effect Of Granting Of Aloe Vera On Blood Cholesterol
Levels Of Obese Respondents Level I,” Jurnal Proteksi Kesehatan 9, No. Vol 9 No 1 (2020): Jpk:
Jurnal Proteksi Kesehatan (2020): 30–36, Http://Jurnal.Pkr.Ac.Id/Index.Php/Jpk/Article/View/196.
8
Surya Saputra And Ani Margawati, “Pengaruh Pemberian Yoghurt Sinbiotik Tanpa Lemak
Dengan Penambahan Tepung Gembili (Dioscorea Esculenta) Terhadap Kadar Kolesterol Total
Tikus Hiperkolesterolemia,” Journal Of Nutrition College 4, No. 2 (2015): 104–109.

3
 2. Knowing the potential of whey cheese yogurt as a functional drink to
reduce cholesterol levels
3. Knowing the optimum concentration of whey cheese yogurt as a
cholesterol-lowering

D. Research Significance
1. Theoretical Benefits
The results of this research can be a reference in further research
2. Practical Benefits
The results of this study are expected to provide knowledge related to the
potential of whey cheese as yogurt and be an alternative to prevent and
reduce cholesterol sufferers.

E. Authenticity of Research
Table 1. Authenticity of Research
Research Title Design of Variable Result Different
Study in Study
Study of the Experimental Independent: The fermentation process Dependent
Potential of whey-based raised the amount of :
Whey Yogurt as yogurt whey lactic acid bacteria Cholesterol
an Ingredient by 48% and antioxidant levels
Dependent: activity by 27.7%. Whey
Natural Acne
acne Yogurt has the potential
Prevention 9
as a natural ingredient to
prevent acne.
Utilization of Experimental Independent: Caspian sea yogurt with Independe
Whey in the caspian sea the addition of whey, nt:
Manufacture of yogurt made showed that the physical, Whey
Caspian Sea from whey chemical, microbilogical cheese
Yogurt Using and organoleptic yoghurt
Lactobacillus Control: acceptance of whey that
Remoras Lactobacillu was still acceptable and Control :
Isolate And s cremonis utilized was whey with an Rat strains
Acetobacter and addition of 25% with a wistar
9
A Rahman et al., “Kajian Potensi Whey Yogurt Sebagai Bahan Alami Pencegah Jerawat (Study
of Fermented Whey keju as Natural Treatment For Acne Prevention),” Jurnal Ilmu Produksi dan
Teknologi Hasil Peternakan 02, no. 1 (2014): 238–242.

4
 Research Title Design of Variable Result Different
Study in Study
10
Orientalis Acetobacter protein content of 2.21%, induced
orientalis the amount of BAL x109c diet high in
1.41 cfu / ml, pH 4.36, cholesterol
4.55 dPAs total solids,
and viscosity 0.26.
Effect of giving Experimental Independent: Giving non-fat synbiotic Independe
non-fat Non-fat yogurt plus gembili flour nt:
synbiotic synbiotic in the P2 group as much Whey
yogurt plus yoghurt and as 3 ml can lower LDL cheese
gembili flour gembili flour cholesterol levels by yoghurt
on LDL 20.72%.
cholesterol Dependent: Dependent:
levels of Cholesterol Cholesterol
hypercholester up to LDL levels
olemia Rats 11
Control :
Hypercholes
terolemia
Rats
Effectiveness of Experimental Independent: Fermented soy milk can Independe
Lactic Acid Lactic acid lower total cholesterol nt:
Bacteria in Soy bacteria in and the accumulation of Lactic acid
Milk fermented liver triglycerides. in
Fermentation soy milk Fermented soy milk fermented
against affected reducing whey
Reduced Dependent: cholesterol in cheese
Cholesterol Cholesterol hypercholesterol patients.
12
Levels levels

10
Tabhita Larasati, Joni Kusnadi, And Endrika Widyastuti, “Pemanfaatan Whey Dalam Pembuatan
Caspian Sea Yogurt Dengan Menggunakan Isolat Lactobacillus Cremoris Dan Acetobacter
Orientalis (Whey Utilization For Making Of Caspian Sea Yogurt Using Isolate Lactobacillus
Cremoris And Acetobacter Orientalis),” Jurnal Pangan Dan Agroindustri 4, No. 1 (2016): 201–
10.
11
Surya Saputra And Ani Margawati, “Pengaruh Pemberian Yoghurt Sinbiotik Tanpa Lemak
Dengan Penambahan Tepung Gembili (Dioscorea Esculenta) Terhadap Kadar Kolesterol Total
Tikus Hiperkolesterolemia,” Journal Of Nutrition College 4, No. 2 (2015): 104–9.
12
S D Ramadhany, “Efektivitas Bakteri Asam Laktat Dalam Fermentasi Susu Kedelai Terhadap
Penurunan Kadar Kolesterol,” Jurnal Agromedicine (2015): 1–4,
Http://Juke.Kedokteran.Unila.Ac.Id/Index.Php/Agro/Article/View/1354.

5
 Research Title Design of Variable Result Different
Study in Study

Application of Experimental Independent: Lactobacillus acidophilus Independe

Lactobacillus Lactobacillu application had no nt:
Acidophilus s significant effect on the Yogurt
Against acidophilus reduction of total blood with
Decreased Total cholesterol levels after Lactobacil
Blood Dependent: hypothesis testing using lus
Cholesterol Cholesterol the One-Way ANOVA bulgarius
Levels (Mus levels and LSD Post Hoc Test, and
13
Musculus) (P>α, 0.220>0.05). Streptococ
Control : However, descriptively cus
Male mice doses of 0.5 mL and 1 mL thermophil
lowered total cholesterol us
levels (Mus musculus) to
17 mg/dL and 20 mg/dL Control :
to control. Both treatment Wistar
groups had average total male rats
cholesterol levels of
136.8 mg/dL and 133.8
mg/dL.

Uthai Sakee, “Aplikasi Lactobacillus Acidophilus Terhadap Penurunan Kadar Kolesterol Total
13

Darah Mencit (Mus Musculus),” Tetrahedron Letters 55 (2014): 3909.

6
 CHAPTER II

LITERATURE REVIEW
A. Theory
1. Definition of Yogurt
Along with the advancement of science and technology, agroindustry
activities continue to be widespread and the products produced continue to
be various. Food is a vital problem because it concerns a person's health.
To support this, nutrition is needed in the form of food or drinks that have
good nutrition and are certainly supported by proper food processing
technology.
Nutritious food is food with a certain chemical composition, so it can
function as a source of nutrients as well as have the potential to improve
health and life, and reduce the risk of chronic diseases, such as
hypercholesterolemia related to diet.14
One example of nutritious food is yogurt, which is a food product in
the form of processed milk through a fermentation process using certain
bacteria, namely, Lactobacillus bulgaricus and Streptococcus
thermophilus. The mixture of these two bacteria acts to replace lactose
(milk sugar) into lactic acid which has an impact on decreasing pH and the
occurrence of clots due to coagulation of milk protein by acid, creating a
distinctive taste because it has flavor components such as diacetyl,
acetaldehyde and carbon dioxide.15
The basic ingredients for making yogurt can come from fresh cow's
milk or goat's milk (fresh milk and pasteurized milk). Yogurt additives are
skim milk and ream to add nutritional value, sweetener, stabilizer, flavor,
and colorant to attract consumers' attention. The basic ingredients of milk
in making yogurt can also be replaced or developed into various kinds of
ingredients whose benefits are no less than cow's milk.16

14
J Silalahi, “Makanan Fungsional,” Kanisius (2006).
15
Sutrisno Maskuro, A., Juri, M., Eksperimen Pembuatan Yogurt Rasa Kenitu (Chrysophyllum
Cainito) Sebagai Peningkatan Kreasi Disertivikasi Buah Khas Jember (Jakarta, 2011).
16
Wahyudi Ahmad, Bugar Dengan Susu Fermentasi (Malang: Universitas Muhammadiyah
Malang Press, 2008).

7
 2. Types of Yogurt
There are several types of yogurt based on taste, fat content, post-
fermentation process, and method of manufacture.17

a. Yogurt based on taste

1) Plain yogurt. Plain yogurt is yogurt that is more acidic than
other yogurts. Not everyone can accept the taste of plain yogurt
because of its high sour taste. Plain yogurt takes the sour taste
of the original taste of yogurt without being mixed by anything.
2) Flavoured yogurt. Flavoured yogurt is the opposite of plain
yogurt. This type of yogurt is added with synthetic flavors and
colors such as strawberry, trambozen, orange, lychee, vanilla,
peach, cherry and honey.
3) Fruit yogurt. As the name suggests, fruit yogurt is yogurt with
fruit fillings such as sliced apples, mangoes, pineapples,
bananas. This fruit yogurt is in great demand by the public,
apart from its friendly taste and aroma, it also contains more
nutrients.
b. Yogurt based on fat content
1) Yogurt with very low fat content (-1%)
2) Yogurt with low fat content (1-2%)
3) Yogurt with medium fat content (3-4%)
4) Yogurt with hight fat content (4,5-10%)
c. Yogurt based on method manufacture
1) Strained, this method is used by removing the whey from the
milk
2) Set yogurt, a method of making yogurt that has a soft texture
and can be packed in a cardboard box
d. Yogurt based on post fermentation process
1) Consentrated yogurt. Yogurt with a total solids of about 24% or
dry yogurt with total solids of 90-94%
2) Frozen yogurt. Yogurt stored at frozen temperature.
Dwi Kusuma Surajudin, Purnomo F.R, “Yoghurt Susu Fermentasi Yang Menyehatkan,”
17

Agromedia Pustaka, Jakarta (2006).

8
 3) Dietetic yogurt. Yogurt made with lower calories and lactose
than normal yogurt, can be added with vitamins or protein.
4) Pasteurized yogurt. The pasteurization process is a process
after the incubation process aims to extend the shelf life of
yogurt

3. Benefits of Yogurt
Yogurt has many benefits in the body such as overcoming fungal and
bacterial infections18, preventing cancer19, overcoming lactose intolerant
with the content of lactic acid bacteria in yogurt which functions as a
decomposer of milk lactose into monosaccharides, namely glucose and
galactose, so milk is easily absorbed by the body and stimulates the
secretion of lactase enzymes. in the digestive tract.20 Yogurt can also
balance the digestive system by maintaining the balance of normal
intestinal flora and improving digestion.21
The decrease in cholesterol levels is influenced by the presence of
lactic acid bacterial fermentation products found in yogurt through several
mechanisms including cholesterol assimilation, bile acid deconjugation,
and the transformation of cholesterol into coprostanol.22
Cholesterol assimilation occurs through a direct mechanism by taking
cholesterol by lactic acid bacteria, which then incorporates the cholesterol
into bacterial cell membranes, causing a reduction in the amount of free
cholesterol in the body.23
The resulting lactic acid bacteria are known to secrete the enzyme Bile
Salt Hydrolase (BSH). This enzyme separates glycine and taurine resulting
18
C. Felley And Pierre Michetti, “Probiotics And Helicobacter Pylori,” Bailliere’s Best Practice
And Research In Clinical Gastroenterology, 2003.
19
Kusuma Surajudin, Purnomo F.R, “Yoghurt Susu Fermentasi Yang Menyehatkan.”
20
Dewi Rusmiati, Penyuluhan Pentingnya Konsumsi Yoghurt Dan Metode Pembuatannya Dengan
Cara Sederhana Dalam Rangka Peningkatan Derajat Kesehatan Dan Ekonomi Masyarakat Di
Kelurahan Sukaluyu Kota Bandung (Bandung, 2021).
21
Morgan E. Schafer Et Al., “Peptides And Antibiotic Therapy: Advances In Design And
Delivery,” Accounts Of Chemical Research 54, No. 10 (2021).
22
Slamet Riyanto And Hesti Muwarni R, “Yoghurt Kedelai Hitam (Black Soyghurt) Dapat
Menurunkan Kadar LDL Tikus Hiperkolesterolemia,” Jurnal Gizi Dan Dietetik Indonesia
(Indonesian Journal Of Nutrition And Dietetics) 3, No. 1 (2016).
23
Hamdan Yuwafi Naim, “Pengaruh Pemberian Yoghurt Kedelai Hitam (Black Soyghurt)
Terhadap Profil Lipid Serum,” Artikel Karya Tulis Ilmiah. Semarang: Universitas Diponegoro
Semarang (2011).

9
 in deconjugated bile salts in the form of free collision acid that is less
absorbed by the small intestine, difficult to re-absorb and recycle through
the enterohepatic cycle, so the more bile acids will be excreted through the
stool. Therefore bile acids that return to the liver become reduced. Bile
acids wasted through feces result in more cholesterol needed as precursors
to synthesize bile salts again, so cholesterol levels in the blood will
decrease and decrease partially.24
Lactic acid bacteria are also able to produce the cholesterol enzyme
reductase. Cholesterol reductase enzymes can convert cholesterol into
coprostanol, a type of sterol that cannot be absorbed by the human
intestine. In vitro studies evaluated the conversion of cholesterol to
coprostanol by Lactobacillus strains such as Lactobacillus acidophilus, L.
bulgaricus, and L. casei ATCC 393 through fluorometric analysis.
Researchers detected reductions in intra and extracellular cholesterol in
all probiotic strains tested, indicating the possibility of intra- and
extracellular cholesterol conversion into coprostanol. The concentration of
cholesterol in this medium decreases at the time of fermentation by
probiotics by increasing the concentration of coprostanol. Coprostanol is a
natural steroid produced by bacteria in the lower intestines of humans or
animals and excreted through feces. One of the functions of lactic acid
bacteria is to reduce serum cholesterol. Cholesterol in the intestines will be
converted into coprostanol, so it cannot be absorbed by the intestines and
will come out with feces.25

4. Yogurt Making Process

The manufacture of yogurt begins with basic ingredients such as
pasteurized milk at a temperature of 83-85 ° C for approximately 30
minutes. Pasteurization is done just like heating, which is reheating milk in
a heating tank that has been sterilized with hot water, then draining milk

24
Andi Nur Fadhilah, Hafsan, And Fatmawati Nur, “Penurunan Kadar Kolesterol Oleh Bakteri
Asam Laktat Asal Dangke Secara In Vitro,” Prosiding Seminar Nasional Mikrobiologi Kesehatan
Dan Lingkungan (2015).
25
M. T. Liong And N. P. Shah, “Bile Salt Deconjugation Ability, Bile Salt Hydrolase Activity
And Cholesterol Co-Precipitation Ability Of Lactobacilli Strains,” International Dairy Journal 15,
No. 4 (2005).

10
 from a homogenization device. This temperature increase can be done by
adding heat from the outside of the container. So at the time, when this
heating is done, the temperature will increase.
The milk is let stand until the temperature drops to 40°C and then
transferred into a container. Pasteurized milk in containers, inoculations
of Lactobacillus bulgarius, and Streptococcus thermophillus cultures by
2.5% each. The addition of culture is done at that temperature to make fat
and content in milk will be mixed. A decrease in the temperature of milk
in the heating tank by flowing cold water on the outside of the tank, as it is
covered with cold water. Mixing of the culture is done in the heating tank
earlier. Mixing this culture is done when the milk temperature in the tank
is cold or 45 ° C. Mixing this culture is done at cold temperatures or 45 °
C, so the culture is not damaged.26
The next step is stirred until homogeneous, then incubated at 37 ° C
until it reaches a pH of 4.5. If the Ph has reached the provisions, then
yogurt is stored in the refrigerator at a temperature of 4 ° C.
The principle of making yogurt is the fermentation of milk using the
bacteria Lactobacillus bulgaricus and Streptococcus thermophilus. Both
types of bacteria will decompose lactose (milk sugar) into lactic acid and
various components of aroma and taste. Lactobacillus bulgaricus plays a
role in aroma formation, while Streptococcus thermophiles play more of a
role in the formation of yogurt flavors. Good yogurt has a total lactic acid
of about 0.85-0.95%. While the degree of acidity (pH) that should be
achieved by yogurt is about 4.5.

5. Fat Content Test

a. Soxhlet Method
The soxhlet extraction method's fat content test has the principle of
separating fat with its solvent. The solvents used are fat solvents such
as petroleum ether, acetone, diethyl ether, N hexane and other types.
The soxhlet method requires a set of soxhletation tools consisting of
26
Friska Viviani Marpaung, “Proses Produksi Yoghurt Dan Cara Pengemasan Yoghurt Di Pt.
Bukit Baros Cempaka,” [Laporan Kerja Praktek]. Universitas Katolik Soegijapranata Semarang
(2018): 1–40.

11
 fat squash, condensers and team containers. Samples in soxhletation
need to be dried before being extracted. The purpose of drying is to
remove the water content contained in the sample, while mashed is to
facilitate dissolved compounds in solvents. After obtaining the desired
fat, then the heating method can be used to separate the solvent from
the fat. It can be used because the boiling point of the fat is higher than
27
the solvent, so, the weight of the fat extracted can be measured.

b. Gerber Method
The gerber method is used to test samples in wet form, in contrast
to the sample type in the soxhlet method. Gerber is an empirical
procedure to ascertain the value of milk fat content in units of fat per
100 ml of milk. The principle is that concentrated sulfuric acid
remodels and dissolves casein and other proteins, thus causing a loss
of fat dispersion. The separation of fat is accelerated by the addition of
amyl alcohol which will dilute the fat with the heat it causes.
Centrifugation will cause fat to accumulate in the scale part of the
butirometer. The reagent used in determining fat content by gerber
method is sulfuric acid 91-92% with colorless or lighter appearance
and clear-colored amyl alcohol.28

c. Babcock Method
Babcock has the principle of analysis of the determination of the
volume of liquid sample fat by dissolving the sample in an organic
solvent. The babcock method is used to determine the fat content of
milk. The principle of its work is to destroy the emulsion of milk fat
using H2 SO 4. This method uses centrifuges or heating, so the fat in
milk can be separated so that it can be measured in bottles that have
been calibrated bab-cock bottles.

27
Mulyo et al. Amelia, “Analisis Kadar Lemak Metode Soxhlet (AOAC 2005),” Departemen Gizi
Masyarakat, Fakultas Ekologi Manusia, IPB, 16680 Bogor, Indonesia, no. Aoac 2005 (2014).
28
Lailia Mutamimah, Sri Utami, and A. T A Sudewo, “Kajian Kadar Lemak Dan Bahan Kering Tanpa
Lemak Susu Kambing Sapera Di Cilacap Dan Bogor,” Jurnal Ilmiah Peternakan 1, no. 3 (2013).

12
 6. Whey Cheese

Figure 1. Cheese Whey (Maulana, 2018)

Whey cheese is a waste of cheese production that can be used as a
source of carbon by microbes because it contains lactose in large
quantities, so it can be used to become a fermented drink that has benefits
for the body. So far, whey cheese in the majority of factories in Indonesia
becomes waste and has not been utilized optimally and maximally, even
though in previous study there are many benefits of whey cheese waste.29
The dominant content in whey cheese is the carbohydrate component
(4.7 grams / 100 ml) and protein (0.9 grams / 100 ml) while the fat content
in whey cheese is very low (0.3 grams / 100 ml). Seeing the content that
can be beneficial for the body while in addition to the amount of whey
cheese waste that is not optimally utilized makes whey cheese a food
resource that is quite dear to waste. The number of processed foods that
should be made from milk can be replaced by whey cheese without
eliminating the benefits. One of these foods is fermented beverages.30
The benefits of whey cheese content can be processed into fermented
drinks, one of which is yogurt. Yogurt is one of the products obtained
from fermented milk or milk reconstitution using Lactobacillus bulgaricus
and Streptococcus thermophilus bacteria or other appropriate lactic acid
bacteria, with or without the addition of other foodstuffs and food
additives permitted. Functional drinks such as yogurt obtained from milk
are one of the products that can be developed from the utilization of whey
29
Benjamas Cheirsilp And Sirilaor Radchabut, “Use Of Whey Keju Lactose From Dairy Industry
For Economical Kefiran Production By Lactobacillus Kefiranofaciens In Mixed Cultures With
Yeasts,” New Biotechnology 28, No. 6 (2011).
30
Nurminabari, “Kajian Penambahan Skim Dan Santan Terhadap Karakteristik Yoghurt Dari Whey
Keju.”

13
 cheese waste. This can be a solution of whey cheese waste that has not
been fully customized can be converted into products of economic value
and higher nutrition.31
Previous research in the manufacture of Caspian sea yogurt, whey
cheese and starter concentrations had a noticeable effect on total acid, pH,
total BAL and total sugar. The interaction of the two treatments had no
real effect on protein levels, total solids and viscosity. As for the results of
the study, Caspian sea yogurt with the addition of whey cheese showed
that the acceptance of physical, chemical, microbiological and
organoleptic whey cheese that is still acceptable and utilized is whey
cheese with an addition of 25%.32
Whey cheese contains many compounds that can benefit the body, one
of which is lactoferrin which serves as an antioxidant. In addition, the
nutritional composition is quite high in whey cheese such as protein,
lactose and fat. The levels of lactose contained in whey cheese can be used
for the growth of Lactic Acid Bacteria (BAL).33
Lactic acid bacteria play an important role in the manufacture of
yogurt from whey cheese. In addition, whey cheese has the properties
needed for the health of the body. The protein contained in whey cheese is
already known to increase muscle mass and has the expertise to reduce the
risk of heart disease and cancer, and influences reducing blood pressure. In
the food industry, whey cheese protein has a position as a functional
food.34

7. Cholesterol
Cholesterol is a metabolite containing sterol fat found in cell
membranes and circulated in blood plasma. Cholesterol helps the body
produce vitamin D, several hormones, and bile acids to digest fat.
31
Badan Standardisasi Nasional, “Sni 2981: 2009 Tentang Yogurt.”
32
Larasati, Kusnadi, And Widyastuti, “Pemanfaatan Whey Keju Dalam Pembuatan Caspian Sea
Yogurt Dengan Menggunakan Isolat Lactobacillus Cremoris Dan Acetobacter Orientalis (Whey
Keju Utilization For Making Of Caspian Sea Yogurt Using Isolate Lactobacillus Cremoris And
Acetobacter Orientalis).”
33
Jane Sélia Dos Reis Coimbra And José Teixeira, Engineering Aspects Of Milk And Dairy
Products, Engineering Aspects Of Milk And Dairy Products, 2016.
34
Carlos Pereira Et Al., “Novel Functional Whey Keju-Based Drinks With Great Potential In The
Dairy Industry,” Food Technology And Biotechnology 53, No. 3 (2015).

14
 Cholesterol is one of the substances produced naturally by the liver but can
also be found in foods of animal origin, such as meat and milk.35
Cholesterol comes from the Greek Chole = bile, and Stereos = solid.
Cholesterol is a natural substance that is physically similar to fat but
swarms of steroids, like other natural compounds. Cholesterol is a
constituent of plasma membranes and plasma lipoproteins. Cholesterol is
widely distributed throughout the body's cells, especially in nerve tissue.
Based on the physical properties of chemistry, cholesterol is a sheet, a
granule with a melting point of 147.5 °C, a slightly yellow-white color,
almost odorless, oxidized by air to yellow or pale brown. Cholesterol is
insoluble in water, soluble in chloroform, acetate ether, dioxane, in
vegetable oils, quite difficult to dissolve in ethanol difficult to dissolve
slowly in ethanol 96%.36,37
Cholesterol in blood plasma combines with non-polar lipid compounds
and proteins to form lipoproteins, so they can be transported to various
tissues and organs of the body for use and storage. The three main types of
lipoproteins are low-density lipoprotein (VLDL), with a low density
(LDL) commonly referred to as bad cholesterol and high-density
lipoprotein (HDL) commonly called good cholesterol. Another type (LDL)
is triglycerides that are also produced by fats, triglycerides will produce
energy derived from carbohydrates.38
Lipid metabolism in the body occurs in the intestines and liver. Lipid
metabolism has three pathways: exogenous, endogenous and RCTP
pathways..39
1. Exogenous Metobolic Pathways
Exogenous metabolic pathways metabolize fats in the
intestines. These fats can come from food eaten or from bile

35
S. H. Cedar, “Human Biology And Health,” In Health Studies, 2015.
36
Tan Hoan Tjay and Kirana Rahardja, Obat–Obat Penting Khasiat, Penggunaan, Dan Efek–Efek
Sampingnya (Edisi VI), Jakarta: PT. Elex Media Komputindo, 2002.
37
RI Kemenkes, Farmakope Indonesia Edisi VI, Departemen Kesehatan Republik Indonesia,
2020.
38
David Savada et al., Life, The Sciene of Biology, 2011.
39
Rizka Humardewayanti Asdie, Doni Priambodo Witjaksono, and Soebagjo Loehoeri, “Buku
Ajar Ilmu Penyakit Dalam,” Interna Publishing (2017).

15
 released by the liver. The liver secretes bile that serves to emulsify
fat. Bile contains cholesterol that will be reabsorbed by the
intestines. Triglycerides and cholesterol are absorbed by intestinal
cells called enterocytes.
Triglycerides will be absorbed by the intestines in the form of
free fatty acids or free fatty acids, in enterocyte triglycerides will
be converted back into triglycerides and will combine with
apoproteins and turn into lipoproteins that are kilomicron type.
While cholesterol will be absorbed by enterocytes then converted
into cholesterol esters combine with apoprotein type Apo B and
form kilomicrons.
Kilomicrons pass through intestinal cells and into the lymph
channels then into the bloodstream. In the bloodstream,
kilomicrons again pegged triglycerides and cholesterol. Blood
vessel cells or endothelial will secrete the enzyme LPL and
hydrolyze triglycerides, so it becomes a free fatty acid and
channeled to adipose tissues, if too much it will settle in the
kilomicron and go to the heart to then become a liver tgrigliserida.
The course of the college is different from triglycerides,
cholesterol remains present in kilomicrons, unlike triglycerides.
Kilomicrons will eventually contain more cholesterol than
triglycerides because triglycerides have been channeled into
adipose tissue. Kilomicrons that have high cholesterol and
triglycerides are little called kilomicron remnants that later through
the blood vessels will go to the heart.
2. Endogenous Metabolic Pathways
Endogenous metabolic pathways metabolize fat that comes
from the liver and gets channeled into the blood vessels. Just like
the exogenous pathways that will be channeled are triglycerides
and cholesterol in the intestine. When it enters the blood vessels
triglycerides and cholesterol combines with phospholipids and
apoprotein APB100 and form VLDL (Very Low-Density

16
 Lipoprotein) lipoproteins. VLDL has higher levels of triglyceride
than cholesterol. In the blood circulation, triglycerides will be
hydrolyzed by the enzyme LPL (Lipoprotein Lipase) released by
the endothelial, hydrolyzed triglycerides from VLDL will convert
VLDL into IDL. IDL has balanced triglyceride and cholesterol
levels.
The triglycerides in IDL will be hydrolyzed by LPL for the
second time and turn into LDL. LDL has low triglyceride levels
because triglycerides have been hydrolyzed at the beginning. This
process causes cholesterol levels to tend to be higher. Cholesterol
in this case will be carried to the liver and steroidogenic tissues
that have LDL-cholesterol receptors, such as adrenaline, ovaries,
testes. On its way to hepar, LDL can undergo oxidation when
LDL is captured by the macrophage scavenger A (SR-A) respecter.
When macrophages eat too much LDL, it becomes a foam cell or
foam cell. Foam cells are the main point that causes atherosclerosis
if it settles in the blood. This oxidation can occur due to high LDL
and low HDL factors because HDL is antioxidant and can inhibit
the oxidation process by taking LDL levels and bringing them to
hepar so that atherosclerosis does not occur.
3. Reverse Cholesterol Transport (RCTP) Metabolism Pathways
The RCTP pathway is a special metabolic pathway for
metabolizing HDL. HDL is excreted by the liver and intestines into
the blood. HDL is a lipoprotein that has low cholesterol levels. The
apoproteins in HDL are APO A, C and E called nascent HDL.
HDL will approach macrophages and take the free cholesterol
stored in macrophages as found in the endogenous pathway and
then turn into mature HDL. HDL mature has high levels of free
cholesterol and will esterify and become cholesterol ester which in
its formation is assisted by LCAT (Lecithin Cholesterol
Acyltransferase).

17
 RCTP pathway, cholesterol esters undergo 2 pathways, some
of which are directly carried to the heart and captured by the
scavenger receptor B-1 (SR B-1). The second pathway is that the
cholesterol ester in HDL will exchange with triglycerides derived
from VLDL and IDL which are assisted by CETP (Cholesterol
Ester Transfer Protein), after which cholesterol will go to hepar
back.

Cholesterol biosynthesis is divided into five stages. First, acetic acid

molecules are activated into acetyl-coenzyme A. Acetyl coenzyme A
condenses with acetylase-coenzyme A into 3-hydroxy-3-methylglutaryl
coenzyme A (HMG-CoA) with the help of HMG-Co A reductase
converting HMG-CoA to mevalonate. Second, mevalonate is converted
into the basic molecule of isoprene along with the release of CO2. Third,
six isoprenoid units and two phenyl molecules hold condensation to form
compounds between the secretaries. Fourth, squalene undergoes
cyclization to form a parent steroid compound, namely lanosterol. Fifth,
cholesterol is formed from lanosterol after passing several further stages,
including through the release of three methyl groups.40.

The main lipid fraction abnormalities are a rise in total cholesterol

levels, an increase in LDL cholesterol levels, a rise in triglyceride levels,
and a decrease in HDL cholesterol levels. Hyperlipidemia is a condition
when the body experiences excess total fat content in the bloodstream.
These fats include cholesterol and triglycerides while elevated triglyceride
levels in the blood are referred to as hypertriglyceridemia. Another type is
hypercholesterolemia where there is an increase in levels (LDL) that will
inhibit its function as the main lipoprotein carrying cholesterol in the
blood. Cholesterol levels in the examination according to the type are LDL
<100 mg / dL, HDL 60 mg / dL, triglycerides <150 mg / dL and means
total cholesterol levels <200 mg / dL41.

Robert K Murray, Biokimia Harper Edisi 27, Igarss 2014, 2014.

40

41
Enrico Agabiti Rosei And Massimo Salvetti, “Management Of Hypercholesterolemia,
Appropriateness Of Therapeutic Approaches And New Drugs In Patients With High
Cardiovascular Risk,” High Blood Pressure And Cardiovascular Prevention 23, No. 3 (2016).

18
 Hypercholesterolemia is a disorder of cholesterol metabolism caused
because cholesterol levels in the blood exceed the normal limit of <200 mg
/ dL. Elevated cholesterol levels can lead to narrowing of blood vessels or
atherosclerosis of coronary heart disease, pancreatitis, diabetes mellitus,
thyroid disorders, heart disease and kidney disease. Hypercholesterolemia
is becoming one of the common health problems found in societies with
growing prevalence and its extremely dangerous complications.42
The pathophysiology of hypercholesterolemia begins with cholesterol
metabolized in the hepar, if cholesterol levels exceed normal limits as
described earlier, it will dull in the hepar. Cholesterol that enters the hepar
cannot be transported entirely by lipoproteins to the hepar from the
bloodstream, so this excess cholesterol will stick to the walls of blood
vessels and cause buildup. As a result, the walls of all elastic blood vessels
will become inelastic anymore.43
Diagnosis of hypercholesterolemia begins with a physical examination.
In some cases, xanthelasma44 appears on his eyelids. If these symptoms
have been encountered, it will be continued with sampling the patient's
direction to find out cholesterol levels significantly. When the results of
cholesterol are known, follow-up examinations such as blood sugar levels
to detect the or absence of diabetes symptoms and thyroid function tests45.
Early therapy in people with hypercholesterolemia is a non-
pharmacological therapy such as lifestyle changes consisting of weight
loss, increased physical activity, a healthy diet and exercise. A healthy
lifestyle such as not smoking can increase HDL levels and consuming
fibrous foods such as vegetables and the like can lower cholesterol levels

42
Sitti Rahma, Rosdiana Natsir, and Peter Kabo, “Pengaruh Antioksidan Madu Dorsata Dan Madu
Trigona Terhadap Penghambatan Oksidasi LDL Pada Mencit Hiperkolesterolemia,” JST
Kesehatan 4, no. 4 (2014).
43
Kathleen M. Botham and Peter A. Mayes, “Sintesis, Transpor, & Ekskresi Kolesterol,” in
Biokimia Harper, 2009.
44
Plak kekuningan akibat gumpalan lemak yang muncul di kelopak mata. Plak kekuningan ini
muncul di bagian sudut mata (canthus) dalam yang dekat dengan hidung, baik di kelopak mata atas
maupun bawah.
45
Phoebe A. Stapleton et al., “Hypercholesterolemia and Microvascular Dysfunction:
Interventional Strategies,” Journal of Inflammation, 2010.

19
 by 5-20%. If non-pharmacological therapy cannot help, then
pharmacological therapy becomes the next stage to run.46
Some classes of drugs that can be used for pharmacological therapy of
hypercholesterolemia are statins, bile acid-binding resins, niacin, derivates
of fibrotic acid, nicotinic acid, ezetimibe, omega-3 fatty acids. Statin drugs
are currently the main choice in treatment because they are generally well
tolerated by patients.47
Statin drugs inhibit the enzyme HMG-CoA reductase competitively. It
is the enzyme responsible for the conversion of HMG-CoA to mevalonate,
which is the initial pathway to cholesterol biosynthesis. Statin drugs are
generally given after dinner or before going to bed. Decreases in total and
LDL cholesterol levels occur when the drug is given at night because
cholesterol biosynthesis reaches its peak at night.48

46
Ibid.
47
Siobhra O Sullivan and Year Medicine, “Statins : A Review of Benefits And Risks .,” TSMJ 8
(2007).
48
Antonio M. Gotto, “Low High-Density Lipoprotein Cholesterol as a Risk Factor in Coronary
Heart Disease: A Working Group Report,” Circulation, 2001.

20
B. Theoretical Framework Gen

Direct Factor Gen

Free R

Faktor langsung Radika

Fatty
Hypercholesterolemia
Mak
LDL levels increase berle
Hiperkolesterolemia Obe
Cholesterol levels exceed
normal limits Obe
Str
Indirect Factor
Kadar LDL meningkat Str
Kadar kolesterol melebihi Less A
batas normal
Kurang
Ge

Other Factor JenisAk

Faktor lain U
Life

Pola

Diabetes Endothelial Cell Coro

Atherosclero
Mellitus Dysfunction Hea
sis
Diabetes Gangguan fungsi Jantu
Ateroklerosis sel endotel
Miletus koro
Whey Cheese Yogurt

Yoghurt Whey keju Lactobacillus bulgarius Streptococus thermo

Hight Lactose
Lactobacillus bulgarius Streptococus thermo
Laktosa tinggi
Lactic Acid Bacteria (BAL)
Help Growth
Bakteri Asam Laktat (BAL)
Antimutagenic, Bakterio
Cholesterol Reduction
anticarcinogenic antimicro
Digestion
Penurunan kolesterol Antimutagenik, Bakterio
antikarsinogenik antimikr
Cholesterol assimilation, Bile
Salt Deconjugation t,
Transformation of Cholesterol Decreased Cholesterol Levels
to Coprostanol

Figure 2. Theoretical Framework

21
C. Conseptual Framework Total Cholesterol
Levels in Rats
Administration of whey cheese
Figure 6. Hasil
yogurt with combined concentration
Pengukuran Kadar
in mice induced by a high
Kolesterol Total
cholesterol diet
(mg/dL)Kadar
kolesterol
Figure 5. Conseptual Framework total pada
tikus

22
 CHAPTER III

RESEARCH METHODS
A. Research Type and Design
The research was conducted experimentally with the design of pre and post
test only control group design in testing the cholesterol levels of mice. The
treatment given includes :
K model : Hypercholesterol Rats
K normal : Healthy Rats
P-1 : Induced Rats + 2 mL of cheese whey yogurt
P-2 : Induced Rats + 3 mL of cheese whey yogurt
P-3 : Induced Rats + 4 mL of cheese whey yogurt
To find out the number of repetitions in each treatment group, the Federer
formula (1956) was used, with the following calculations::49
(t-1)(n-1) ≥ 15
Description :
n : the number of each treatment
t : number of treatments
Number of repetitions : (t-1)(n-1) ≥ 15
(5-1)(n-1) ≥ 15
4n – 4 ≥ 15
4n ≥ 15 + 4
4n ≥ 19
n ≥ 4,7 = 5
To find out the dropout or risk of failure in this study, the following formula
was used50:

Drop out possibility was 10%

49
Marlina Rully Wahyuningrum And Enny Probosari, “Pengaruh Pemberian Buah Pepaya (Carica
Papaya L.) Terhadap Kadar Trigliserida Pada Tikus Sprague Dawley Dengan
Hiperkolesterolemia,” Journal Of Nutrition College 1, No. 1 (2012).
50
T. Dahriani, E. Murbawani, And B. Panunggal, “Hubungan Lingkar Leher Dan Tebal Lemak
Bawah Kulit (Skinfold) Terhadap Profil Lipid Pada Remaja,” Jurnal Kedokteran Diponegoro 5,
No. 4 (2016).

23
 n’ : Number of subjects counted

n : Minimum number of samples

f : Estimated drop out proportion

5 5
n’ : = = 5,55 = 6
1-0,1 0,9

5 x (6-5)

5x1

= 5 additional tails

B. Research Location and Time

The research was conducted at the Microbiology Laboratory and the
Laboratory of Pharmacology and Toxicology of the Faculty of Health Sciences,
University of Darussalam Gontor. The study was conducted within 3 months from
August to October 2021.

C. Sample and Population

The population used in this study was 30 Wistar strain rat test animals with 5
treatment groups with 5 rats in them. The sample used was total cholesterol levels
in the rats.

D. Research Variable
In this study used 3 types of variables, namely independet variables in the
form of whey cheese yogurt, dependent variables namely total cholesterol levels in
test animals and control variables, namely wistar strain rats

E. Operational Definition Variable

Table 2. Operational Definition Variable
N Variable Operational Measured Result
o Definition
1 Whey cheese yogurt Fermented beverage Induction
with whey cheese waste volume: 2 ml, 3
base obtained from ml and 4 ml
cheese factory in
Yogyakarta use

24
 N Variable Operational Measured Result
o Definition
bacterial starter
Lactobacillus
bulgarius and
Sterptococus
thermophillus
2 Cholesterol levels Total cholesterol levels <200 mg/dL
rise

3 Rats Rats induced with Rat strain Wistar

dietary feed high in
Age 2-3 months
coconut oil cholesterol
White color
and duck egg yolks
Male gender

F. Material and Instrument

The tools used in this study were ovens, refrigerators, large containers, stirring
rods, pH meters, rat cages, rat sonde (Wistar), microhematocrit pipettes, test
animal syringes (Syringe), Easy Touch GCU (Glucose, Cholesterol, Uric Acid)
Meter Device, dissecting set, Easy touch blood cholesterol strip, glass tool
(Pyrex), a set of Soxhleation and analytical scales.

The ingredients used in the study were whey cheese, yogurt starter,
Lactobacillus bulgarius and Sterptococus thermophillus bacteria, aquades, white
male rats, duck egg yolks, coconut oil and N-Hexan.

G. Ethical Clearance
This study with the subjects of Wistar strain rat test animals who were given
the induction treatment of dietary cholesterol feed and whey cheese yogurt with a
prescribed dose has been approved by the ethical approval submitted to the
Research Ethics Committee of Semarang State University number
389/KEPK/EC/2021.

H. Research Procedure
1. Making Whey Cheese Yogurt

25
 The manufacture of whey cheese yogurt begins with whey cheese
pasteurized at a temperature of 83-85 ° C for approximately 30 minutes.
Whey cheese was let stand until the temperature drops to 40°C and then
transferred into a container. Whey cheese that has been pasteurized in
containers, inoculations of stater culture Lactobacillus bulgarius and
Streptococcus thermophilous as much as 2.5% each. The next step was
stirred until homogeneous, then incubated at 37 ° C until it reached a pH
of 4.5. If the Ph has reached the provisions, then whey cheese yogurt was
stored in the refrigerator at 4 ° C.
2. Fat Content Testing51
Fat content testing in whey yogurt was done by the soxhlet method
with the principle of separation of solvents with samples. The sample was
in the oven at 105°C for 2 hours. Weighing was done with a sample of 4
repetitions. The drying has been perfect and the sample was weighed as
much as 2 gr at each repetition.
Timbel sample was made with filter paper and thread and naming on
each sample. The soxhletation device was arranged using n hexane solvent
and mantle heating of 26°C. Timbel was inserted and extracted for 6
hours with 5-6 cycles.
Sample back in the oven and dry for approximately 2 hours in the
oven. A decikator was used to cool the sample after it has been drained
from the oven. Reading the results by calculating the final weight on the
sample and the fat cup used. Calculation of fat levels used formula :

( CupWeight + Fat ) – Empty Cup Weight

Fat Content % = x 100%
Initial Sample Weight
3. Treatment of Animal Test
The study subjects consisted of 25 male white rats with 5 test groups,
namely the first group of control models that were fed high cholesterol
without healing, the second group of normal controls that were not given
test treatment, the third group was the treatment of 1 rat who was fed high
cholesterol and whey cheese yogurt 2 mL, the fourth group was the
51
Annytha Ina Rohi Detha et al., “Komposisi Kimiawi Dan Fraksinasi Protein Susu Kuda Sumba,”
Jurnal Veteriner September 2014 ISSN : 1411 - 8327 15, no. 4 (2014).

26
 treatment of 2 mice who were given high cholesterol feed and whey cheese
yogurt 3 mL. The fifth group was the treatment of 3 rats who were fed
high cholesterol and whey cheese yogurt 4 mL. Referring to previous
research journals conducted by James and re-researched by Riyanto that
the administration of yogurt 2-4 mL has a significant effect on reducing
cholesterol levels.
All subjects were initially identified for 7 days of standard feed and ad
libithum drinking water for test animals. Subjects were placed in a rat cage
that corresponds to the lighting and adjusting environment. Then, all
subjects were measured for total cholesterol levels before being treated.
The subjects were fed high cholesterol twice a day for 14 days except in
the normal control group. The high cholesterol feed used was a mixture of
coconut oil and duck egg yolk. Standard feeding and drinking water in rats
continue to provide the feed needed for the life of test animals.52
The manufacture of high cholesterol feed in the form of emulsion
preparations made from duck egg yolks as much as 100 grams and palm
oil 50 ml. The material was mixed and stirred using a magnetic stirrer to
get a more homogeneous result.53
High cholesterol feeding has been completed, then all the study
subjects were again tested for cholesterol levels to be sure whether the rats
had hypercholesterol with levels of <200 mg / dL. Whey cheese yogurt
testing was done by inducing preparations in mice per oral according to a
predetermined dose for 14 days. Induction of whey cheese yogurt was
done peroral using sonde to each mouse according to the dose of treatment
except on normal control.54
The last stage was testing cholesterol levels again to ascertain the
potential of whey cheese yogurt whether it can lower cholesterol levels in

52
Alodiea Yoeantafara And Santi Martini, “Pengaruh Pola Makan Terhadap Kadar Kolesterol
Total,” Media Kesehatan Masyarakat Indonesia 13, No. 4 (2017).
53
Yanita Listianasari Et Al., “Efektifitas Pemberian Jus Labu Siam (Sechium Edule) Terhadap
Profil Lipid Tikus (Rattus Novergicus) Model Hiperlipidemia,” Penelitian Gizi Dan Makanan
(The Journal Of Nutrition And Food Research) 40, No. 1 (2017).
54
Yulianto Prasetya, “Bioenkapsulasi Yoghurt Lactobacillus Acidophillus Sebagai
Antihiperkolestrolemia,” Program Studi Teknologi Laboratorium Medik (2001): 29–32.

27
 hypercholesterol rats or not. The results can be seen from the decrease of
cholesterol levels to normal.
Post-research tests animal handling was by turning them off. This
process was carried out about the ethics of handling test animals. In its
handling was used psychotropic group anesthetic namely Silazis.
However, due to the limitations of laboratory materials, inhaled anesthesia
was used with ether or chloroform. Furthermore, dislocation of test
animals as the final handling post-research. Test animals were categorized
as infectious waste and handled by the waste handling section.
4. Testing of Cholesterol Levels
Blood samples were taken in pre and post-treatment. In the blood
sample, pre-treatment was taken after a high induction of cholesterol for
14 days. The post-treatment blood sample was carried out one day after
the treatment in a state of mice was satisfied first for 12 hours. The
purpose of the rats was satisfied first, so there was no change in total
blood cholesterol levels due to food intake.
For the determination of total cholesterol levels, the Easy Touch-GCU
tool was used. Inserted cholesterol strips in the Easy Touch- GCU tool
slot. The blood samples of the mice used were taken intravenously in the
tail area until the blood came out. The blood sample was dripped in the
target strip test area until the device can read the blood sample's
cholesterol levels. Read the test results displayed on the Easy Touch-GCU
tool screen.

I. Data Analysis
The data obtained from this study was data on fat level test results and data on
test results of reduction in cholesterol levels mg / dL. Fat content test data was
calculated by the formula % fat content and presented in table form.
As for the results of the pre and post mg /dL cholesterol level test in mice after
administering whey cheese yogurt in the normality test with Shapiro wilk test if
the data was not normal then again tested with Kruskal walis test. Data that has
been declared normal based on the results of the test, in the analysis of differences
in cholesterol levels using the one way ANOVA parametric test continued the

28
post hoc test with signification indicators (P<0.05) on the comparison of pre and
post cholesterol tests and presented in graph form.

J. Research Schedule
Table 3. Research Schedule

1 2 3
No Activities
1 2 3 4 1 2 3 4 1 2 3 4
1 Preparation                        
Literature Studies                        
Preparing tools and
materials                        
Preparation of the
place                        
2 Implementation                        
Aclimatitation of
animal test                        
Making of whey
cheese yogurt
Cholesterol levels
test
Giving cholesterol
food to animal test
Induced yogurt whey
cheese to rats
Post test                        
3 Writing Result                        
Writing result and
discussion                        
Dissemination                        

29
 CHAPTER IV
RESULT AND DISCUSSION
A. Making of Whey Cheese Yogurt
Whey cheese was taken from a cheese factory domiciled in Yogyakarta. Just
like milk, whey cheese cannot stand room temperature for too long, so whey
storage should be in the refrigerator. Starters in the manufacture of yogurt was
Lactobacillus bulgarius and Streptocuccus thermophillus which will later produce
ferementation products in the form of lactic acid bacteria.
The manufacture of whey cheese yogurt begins with whey cheese and milk
heated for approximately 30 minutes until it reaches a temperature of 70 ° C.
Whey cheese and milk are let stand until the temperature drops to 40°C and is not
too hot to then transfer into a container. Whey cheese that has been left in a
container, inoculated culture stater Lactobacillus bulgarius and Streptocuccus
thermophillus as much as 2.5% each and stirred until homogeneous.
Yogurt was then incubated until it reaches a pH of 4.5 or according to the
desired acidity. If the pH has reached the provisions, then whey cheese yogurt is
stored in the refrigerator.

B. Fat Content of Whey Cheese Yogurt

The study tested fat levels on four whey yogurt formulations that had
differences in each whey content used. The method used in fat content testing is
the soxhlet method. The soxhlet method has the principle of continuous free fat
extraction using non-polar soxheletation and solvents and the presence of reverse
cooling. Sokletation can be used to test fat levels with solid or soft samples using
a relatively constant amount of solvent..55
The solvent used was n-hexane as much as approximately 500 ml in fat squash
to be boiled. The use of n-hexane solvents was because hexane is part of an
anhydrate solvent. The anhydrous solvent is a solvent that is completely water-
free so, the water-soluble materials in the preparation are not extracted and
counted as fat without reducing the activeness of the solvent.56

55
R Marjoni, “Dasar-Dasar Fitokimia,” Trans Info Media (2016).
56
Ibid.

30
 The soxhlet method is a method that using relatively fewer solvents than other
methods. In addition, the heating process in this method can be arranged simply
and has accuracy with a regular cycle. This fat test process lasts for 6 hours with
24 cycles, it is done, so the fat can be accommodated in pumpkins. Here are the
results of the calculation of the determination of the fat content of whey cheese
yogurt.

Table 4. Average Result of Yogurt Fat Content

Formulation Average of Fat Content (%)

F1 0.71a
F2 0.53a
F3 0.51a
F4 0.50a

* Different subsets show significant differences (Tukey test)

Description : F1= Cheese Whey : Milk (25% : 75%)
F2= Cheese Whey : Milk (50% : 50%)
F3= Cheese Whey : Milk (75% : 25%)
F4= Cheese Whey : Milk (100% : 0%)

The percentage of fat levels found no results in different subsets, which can be
seen from all formulations tested in the alpha subset. The results of this study
showed that the average percentage of fat levels in each formulation was not much
different, but the lowest levels were in the 4 formulation samples with a level of
0.5075%. Formulation 4 in whey yogurt preparation was a formulation with a lot
of whey cheese content of 200 mg. The difference in results shown in the above
results can be influenced by the metabolite content contained in pure cheese whey.
Formulation 4 contains more whey and less milk than other formulations.
Cow's milk commonly used for the manufacture of yogurt has high levels of
lactose and fat levels that can raise the amount of sugar and even cholesterol in the
body. When compared to the use of whey that is more dominated in the
manufacture of yogurt, it was found to be a low-fat content because whey can
balance lactose levels, protein and have a low-fat content compared to cow's milk
which is usually more dominant in yogurt mixtures.
Whey cheese has a dominant content with carbohydrate components (4.7
grams / 100 ml) and protein (0.9 grams / 100 ml) while the fat content in whey

31
cheese is very low (0.3 grams / 100 ml). In this case, whey cheese has lactose
content and is beneficial in the growth of lactic acid bacteria in yogurt
fermentation. In their infancy, bacteria will use lactose for energy and carbon
sources.57
Although the protein content in whey fluid is quite low, whey protein is easily
absorbed by the body. This high protein content is one of the low-fat factors found
in whey cheese yogurt. In the process, whey is a byproduct of the cheese-making
industry where the fat contained is already used in the manufacture of cheese and
leaves waste with low fat. Even so, the nutrients in whey cheese can still be
utilized in the manufacture of food, one of which is yogurt without or low fat.58
The fat content produced from whey cheese yogurt qualifies for lean yogurt
with a maximum fat content range of 0.5%. Different in formulation 1 which
meets the requirements in the criteria of low-fat yogurt with a range of 0.6-2.9%.
This can be a reference and supporter in the continuation of whey cheese yogurt
testing as a lowering of the cholesterol levels of mice induced high cholesterol
diet with a combination dose on the formulation of 4 whey cheese yogurt.59

C. The Effect of Yogurt Whey Cheese on the Decrease of Cholesterol Levels

in Rats Levels in Rats Induced by a High Cholesterol Diet
200 The study
180
160
was conducted
160
140
140 to test whether
120
120
100
100
whey cheese
80
80
60
yogurt had any
60
40
40
20
200
0 KM KN P1 P2 P3
KM KN P1 P2 P3
Pre Test Post Test
Pre Test Post Test

57
David Richard Hendarto et al., “Mekanisme Biokimiawi Dan Optimalisasi Lactobacillus
Bulgaricus Dan Streptococcus Thermophilus Dalam Pengolahan Yoghurt Yang Berkualitas,”
Figure 6. Measurement of Total Cholesterol Levels (mg/dL)
Jurnal Sains Dasar 8, no. 1 (2019).
58
Chirstopher Vasey, The Whey Prescription: The Healing Miracle in Milk (Inner Traditions /
Bear & Co, 2006).
59
Badan Standardisasi Nasional, “SNI 2981: 2009 Tentang Yogurt.”

32
effect on the reduction of cholesterol levels of rats induced by a high-cholesterol
diet.

Treatment KM KN P1 P2 P3
Pre Test 157.2 102.5 135.2 148.2 154
Post Test5. Average of187.2
Table 105.2 Cholesterol
Result Decreased 108.4 Levels
106.8(mg/dL)
107
∆ Total -30 -2.7 26.8 41.4 47
Reduction

Description : KM = Model Control (Cholesterol Rat Without Induction yogurt)

KN = Kontrol Negatif (Healthy Rats )
P1 = Cholesterol Rats + 2 mL of Cheese Whey Yogurt
P2 = Cholesterol Rats + 3 mL of Cheese Whey Yogurt
P3 = Cholesterol Rats + 4 mL of Cheese Whey Yogurt

The results of the study in figure 4 showed an increase in total cholesterol

levels in the pre-test results. Rats induced a high-cholesterol diet feed that was a
mixture of duck egg yolk and oil so that it was found to increase in total
cholesterol levels. Coconut oil contains 90 percent saturated fat and was not
recommended for regular use. Duck egg yolks have several deficiencies with
cholesterol levels that were twice as high as 884.00 mg / 100g compared to
chicken eggs which only amounted to 423.00 mg / 100g and quail of 844.00 mg /
100g This is in line with research conducted by the Department of Agriculture
(USDA) that 100 grams of duck eggs contain 185 calories, 13 grams of protein,
14 grams of fat, 1 gram of carbohydrates, 263 milligrams of choline, and
cholesterol which reached 884 milligrams.60

Based on induction of whey cheese yogurt there was a significant difference

between before and afterduction in the reduction of total cholesterol levels
(p<0.05). There was a decrease in total cholesterol levels in every treatment
except on model control and normal control.

Overall, giving whey cheese yogurt lowered total cholesterol levels in all
treatment groups. The average decrease was most meaningful in the P3 group
with a dose of 4 ml, this is because P3 gets the highest dose when compared to
other treatment groups. The results of this study support the results of previous
studies related to the influence of black soy yogurt on LDL levels of
60
United States Department of Agriculture Agricultural Research Service USDA, 2019.

33
hypercholesterolemic mice where the administration of yogurt in mice with a dose
variant of 2 ml, 3 ml and 4 ml was given for 21 days showed a decrease in LDL
levels of mice in the group given yogurt as much as 4 ml.61

Table 6. Average Total Cholesterol Levels (Tukey)

Treatment Average Cholesterol Decreased
(mg/dL)
KM 4.8a
KN 7.4a
P1 26.8a
P2 41.4a
P3 47a
* Different subsets show significant differences (Tukey test)
Description : KM = Model Control (Cholesterol Rat Without Induction yogurt)
KN = Kontrol Negatif (Healthy Rats )
P1 = Cholesterol Rats + 2 mL of Cheese Whey Yogurt
P2 = Cholesterol Rats + 3 mL of Cheese Whey Yogurt
P3 = Cholesterol Rats + 4 mL of Cheese Whey Yogurt

The results of the total cholesterol level test have been distributed normally
and there was a meaningful difference in the overall average with the significance
of the Anova test (p<0.05). The data was then further tested with Tukey's Hoc
Post to see the differences between treatments. The results showed each treatment
was in the same subset, alpha. This indicates that there was no meaningful
difference between each treatment even though the average result of the total
decrease gets a meaningful difference.

The average result of a decrease in total cholesterol levels does not show a
meaningful difference can be because the average result is an accumulation of the
difference between the pre test and post test values, resulting in values that were
not much different. But when viewed from pre test and post test, post test results
have a difference in treatment and control model.

Riyanto and Muwarni R, “Yoghurt Kedelai Hitam (Black Soyghurt) Dapat Menurunkan Kadar
61

LDL Tikus Hiperkolesterolemia.”

34
 Table 7. Cholesterol Levels (Post Test)
Treatment Post Test
KM 152.4b
KN 103.2a
P1 108.4a,b
P2 106.8a
P3 107a
* Different subsets show significant differences (Tukey test)
Description : KM = Model Control (Cholesterol Rat Without Induction yogurt)
KN = Kontrol Negatif (Healthy Rats )
P1 = Cholesterol Rats + 2 mL of Cheese Whey Yogurt
P2 = Cholesterol Rats + 3 mL of Cheese Whey Yogurt
P3 = Cholesterol Rats + 4 mL of Cheese Whey Yogurt

Different subsets showed significant differences. In the post test results,

namely post-induction of whey cheese yogurt there was a significant change in
cholesterol levels between treatment with the contor model (K+) except in
treatment 1. The control model was in subset 2 which means it has a subset
difference with normal control (K-), treatment 2 and treatment 3. The dose of
induction given may affect the difference in these results. In treatment 1 dose of
induction was smaller than treatment 2 and 3. This led to treatments 2 and 3
having meaningful differences in model control which meant a decrease in
cholesterol levels at doses of 3 and 4 mL had meaningful insurance results when
compared to model control.

The decrease in total cholesterol levels was influenced by the fermentation of

Lactobacillus and Streptococcus bacteria that produce the final product in the
form of lactic acid bacteria (BAL). Products fermented by BAL have a process of
reducing cholesterol that occurs in the digestive process, including through the
mechanism of cholesterol assimilation. Assimilation of cholesterol occurs directly
by the process of taking and absorption of cholesterol by bacteria that will then
integrate or merge into one on the bacterial cellular membrane, so bacteria will be
more resistant to lysis. Due to the decrease in absorption of cholesterol in the
digestive system, cholesterol levels in the blood will also decrease.62

62
Ari Yuniastuti, “Pengaruh Pemberin Susu Terfermentasi Lactobacillus Casei Strain Terhadap
Perubahan Kadar Fraksi Lipid Serum Tikus Hiperkolesterolemi,” Tesis, 2004.

35
 Several studies conducted in vitro proved the existence of cholesterol
assimilation activity by lactic acid bacteria. The fermented products of lactic acid
bacteria used are strains of the species L.acidophillus, L.bulgarius, L.gaseri, and
various other species namely Streptococcus sp, Enterococcus sp. Assimilation
activity was observed by comparing the amount of cholesterol remaining in the
inoculated media with lactic acid bacteria and control media. The results showed
that the strains tested had varying assimilation activity with a range of 8.2-23.3
μg/ml.63

Cholesterol levels can be lowered by the change in cholesterol into bile acids.
Bile acids themselves are formed from the basic synthesis of hepar cholesterol.
Before being expelled to the digestive system of the small intestine, bile acids will
be stored in the gallbladder. Bile acids have a role in emulsifying fats, producing
and preparing micelles to be ready for absorption. The total of bile acids produced
is 97% will be reabsorbed and reabsorption in the distal ileum and back again
towards the hepar. The cycle of return of bile acids to the hepar is called the
enterohepatic cycle and this cycle can occur 4-12 times/day. Meanwhile, the
remaining 3% of the total bile acid production will be removed from the body so
that cholesterol in the body is balanced. Bile acids that cannot be absorbed and
reabsorption by the body will metabolized by bacteria contained in the colon and
will experience several reactions, one of which is deconjugation. This process is
where lactic acid bacteria take a large role in the body.64

In other mechanisms, fermented products can secrete the enzyme Bile Salt
Hydrolase (BSH) in the process of decontamination of bile acids. Bile that
reaches the ileum and cecum will be deconjugated by the enzyme bile salt
hydrolase produced by these bacteria. Bile acids will dehydroxylate into
secondary bile acids where the enzyme BSH hydrolyzes the amino acids glycine
and taurine resulting in free collic acid that is less absorbed by the intestines.

63
Siti Winarti, Lilis Nuraida, and Endang Prangdimurti, “Seleksi Bakteri Asam Laktat Isolat ASI
Yang Berpotensi Menurunkan Kolesterol Secara in Vitro” (2011),
http://repository.ipb.ac.id/handle/123456789/47498.
64
Alfianti Rhamadini, “Pengaruh Pemberian Probiotik Terhadap Kadar Malondialdehid (MDA)
Ginjal Tikus Wistar (Rattus Norvegicus) Jantan Model Hiperkolesterolemia Yang Diinduksi
Mentega Putih” (Universitas Islam Indonesia, 2019),
https://dspace.uii.ac.id/handle/123456789/16389.

36
When glycine and hydrolyzed taurine eat conjugated bile acids it will not form
and break the enterohepatic cycle. Deconjugated bile salts will be less soluble and
difficult to absorb by the lumen of the small intestine, so they will be excreted
with feces. The higher the activity of the BSH enzyme in deconjugating bile acids,
the more bile acids are excreted. The body will form new bile acids and of course
will increase cholesterol catabolism to replace bile acids that are excreted and
lower the buildup of cholesterol in the body.65

Cholesterol in the body cannot fully enter the colon. Non absorbable
cholesterol is almost 200 mg per day. When it reaches the intestinal cholesterol
will be converted into coprostanol by bacteria present in the colon. When
consuming yogurt, fermented products in the form of lactic acid bacteria can
convert cholesterol into coprostanol. Coprostanol is a type of sterol that cannot be
absorbed by the small intestine and excreted through feces. This is what can be
used as one way to lower cholesterol levels in the body.66

In this study, the administration of whey cheese yogurt from all three
treatments showed a meaningful reduction in total cholesterol levels (P<0.05) in
P2 and P3. But between the two, the highest value is in P3 with a dose of 4
ml/day. This can be because the administration in treatment 3 has a meaningful
difference with positive control than the dose of other treatments. The results of
this study are in line with previous research that the administration of black yogurt
in hypercholesterolemia mice for 21 days was able to reduce LDL levels
significantly at a dose of 4 mL/day, but had no significant effect on HDL
cholesterol.67

65
Astuti, “Pengembangan Probiotik Bakteri Asam Laktat Dari Limbah Ikan Yang Berpotensi
Menurunkan Kadar Kolesterol Daging Ayam Boiler Strain Hubbart,” Jurnal Penelitian Saintek 15
(2010).
66
Rhamadini, “Pengaruh Pemberian Probiotik Terhadap Kadar Malondialdehid (MDA) Ginjal
Tikus Wistar (Rattus Norvegicus) Jantan Model Hiperkolesterolemia Yang Diinduksi Mentega
Putih.”
67
Riyanto and Muwarni R, “Yoghurt Kedelai Hitam (Black Soyghurt) Dapat Menurunkan Kadar
LDL Tikus Hiperkolesterolemia.”

37
 CHAPTER V
CONCLUSIONS AND SUGGESTIONS
A. Conclusions
Whey cheese yogurt has a low quality of fat content with an average result of
making whey cheese yogurt 0.565%. In the four experiments of formulation of
whey cheese yogurt, the formulation with the most whey content found the lowest
fat content result of 0.5075% where the level is following the criteria of low-fat
yogurt according to the Indonesian National Standard.
Whey cheese yogurt has the potential as an alternative functional drink to
lower cholesterol levels with significant reduction results. Administering whey
cheese yogurt to mice induced a high-cholesterol diet for 14 days was able to
lower total cholesterol levels significantly at a dose concentration of 4 mL/day.

B. Suggestions
Further study is needed regarding the effect of whey cheese yogurt on LDL or
HDL cholesterol levels in the body, considering that these lipoproteins are a factor
in the occurrence of advanced diseases of excessive cholesterol levels in the body.

38