CHAPTER I: Background of the Study

Osteoporosis affects the bones, making them thin and fragile and putting them at increased

risk of fracture. This condition is more common among the elderly women, and osteoporotic

fracture can result in patients becoming bed-ridden. According to WHO an estimated 9.0 million

fractures were listed in the year 2010 worldwide, and the prevalence in South East Asia accounted

for 15.3% of the world occurrence.

According to a 2011 survey carried out a Filipino orthopedic hospital, 62.9% of patients

aged 50 years and over who had been admitted for fracture suffered from osteoporosis. In addition,

the reported calcium intake per person per day in the Philippines, according to the Sixth National

Nutrition and Health Survey in the year 2009, was 440 mg, which was only 57% of the

recommended nutritional intake for Filipinos.

As this problem arises the UNILAB Phil. Conducted a study and came a with a product

named Calciumade, this product contains Calcium, Vitamin D, Magnesium, Zinc and Manganese

which are nutrients known to help protect against osteoporosis and promote bone health. But as

because of poverty some people can’t afford to buy these medicine.

An eggshell is the outer covering of a hard-shelled egg and some form of eggs with soft outer

coats. Eggshells are a rich source of calcium carbonate, they also have a little strontium and some

other bone-protective nutrients. But there is little evidence that getting calcium carbonate from

eggshells offers any significant advantage over other sources. According to NCBI study, the

natural calcium in eggshells can have positive effects on bone density and can help prevent and

treat osteoporosis.
 According to International Journal of Food Sciences and Nutrition, eggshells are great

source of natural calcium. Half an eggshell can provide daily calcium intake. The researcher wants

to conduct or make a study wherein eggshells can be utilized to form capsule supplement. The

study was entitled “Utilization of eggshells as a capsule supplement for osteoporosis.”

STATEMENT of the PROBLEM

This study aims to introduce an eggshell as a potential calcium capsule supplement for

osteoporosis. Specifically, it aims to answer the following:

1. What are the components/nutrition in an eggshell which can be used to prevent

osteoporosis?

2. What is the microbial analysis of the eggshell capsule in terms of:

a) Molds;

b) Coliforms;

c) And, Yeast.

3. What is the effectivity of the capsule as to be tested in Albino rats?

4. What is the moisture analysis of the capsule?

5. What is the dissolution time of the capsule?

6. What is the shelf life of the capsule?

Hypothesis
 Based from the given problems or objectives the following hypothesis are formulated:

1. The component present in eggshells that can prevent osteoporosis are calcium carbonate

and strontium.

2. The microbial analysis of the capsules is negative in terms of:

a. Molds;

b. Coliforms; and

c. Yeast.

3. The moisture content of the capsule about 13-16%.

4. The capsule will be effective as tested in rats.

5. The possible dissolution time of the capsule is approximately 44 mins.

6. The possible shelf life of the capsule is 3-4 years.

Significance of the study

 Community- this study helps the community by producing a cheaper and easier to produce

products.

 Eggshells - this study can help to lower the number of eggshells disposed.

 Osteoporosis patient’s- this study helps those people specifically who ar prone to

osteoporosis
Scope and Delimitations

This study aims to determine that eggshells can be utilize as a capsule supplement for

osteoporosis. The gathering of eggshells and other materials will be conduct at the hometown of

the researcher at Tayug, Pangasinan. The formulation of capsule will be conducted on the

Pharmaceutical Building at Virgen Milagrosa University Foundation. The testing on albino rats

will be conducted also at Virgen Milagrosa University Foundation and continously observed day

by day. For the testing of microbial analysis the researcher will make a sample and give it to the

laboratory at Virgen Milagrosa University Foundation. The dissolution time testing of the capsule

will be conducted at Tayug National High School assisted by his research adviser. And the capsule

will be monitored everyday by the researcher for its shelf life observation. The gathered results

and data were tallied to be computed by the researcher or statistician.

Definition of terms

Albino rats- rats with white fur and pink eyes used extensively in laboratory experiments.

Calcium- is a soft white element which helps to maintain a good and healthy bones.

Capsule- a small case or container, especially a roun or cylindrical one.

Dissolution- become or cause to become incorporated into a liquid so as to form a solution.

Eggshell – the thin hard outer layer of an egg, especially a hen’s egg.

Microbial analysis- is the use of biological, biochemical, molecular or chemical methods for

detection, identification or enumeration of microorganisms in a material.

Strontium- is a silvery metal found naturally as a non-radioactive element. About 99% of the

strontium in the human body is concentrated in the bones. Strontium can also be found in dietary

supplements for building bones .

Supplement- something that completes or enhances something else when added to it.

Utilization- the action of making practical and effective use of something.

CHAPTER II

Related Literatures

Figure 1: Eggshell

The key to eating egg shells and other calcium rich foods is moderation. Egg shells are

extremely high in calcium so it’s definitely not recommended to start eating egg shells as if your

life depends on it because that obviously won’t do you any good. Too much calcium can actually

be harmful to your body.

Calcium Intake. The recommended amounts of calcium for adults are as follows: For pre-

menopausal women 25-50 years old and post-menopausal women on estrogen replacement

therapy: 1,000-1,200 milligrams of calcium per day. 1,500 milligrams of calcium per day is

recommended for pregnant or lactating women. Half an eggshell may provide enough calcium to

meet the daily requirements for adults, which is 1,000 mg per day (2, 4). Summary Eggshells are

commonly used as a calcium supplement. Just half an eggshell may provide enough calcium to

meet the average daily requirements of an adult.

Retrieved from: https://www.healthline.com/nutrition/eggshells-benefits-and-risks

https://www.spine-health.com/conditions/osteoporosis/calcium-and-vitamin-d-

requirements

CALCIUM

Calcium is a chemical element that is essential for living organisms, including humans. It

is the most abundant mineral in the body and vital for good health. We need to consume a certain

amount of calcium to build and maintain strong bones and healthy communication between the

brain and other parts of the body. Calcium is found naturally in many foods; it is also added to

certain products, and supplements are available.

Around 99 percent of the calcium in the human body is found in the bones and teeth; it is

essential for the development, growth, and maintenance of bone. Calcium continues strengthening

the bones of humans until they reach the age of 20-25 when bone density is highest. After that age,

bone density declines, but calcium continues to help maintain bones and slow down bone density

loss, which is a natural part of the aging process. People who do not consume enough calcium

before the age of 20-25 have a considerably higher risk of developing brittle bone disease

or osteoporosis later in life; this is because calcium is drawn from the bones as a reserve.

OSTEOPOROSIS

Osteoporosis affects men and women of all races. But white and Asian women —

especially older women who are past menopause — are at highest risk. Medications, healthy diet

and weight-bearing exercise can help prevent bone loss or strengthen already weak bones.
 Retrieved from: ,https://doi.org/10.1093/ajcn/64.1.71

Symptoms

There typically are no symptoms in the early stages of bone loss. But once your bones have been

weakened by osteoporosis, you may have signs and symptoms that include:

 Back pain, caused by a fractured or collapsed vertebra

 Loss of height over time

 A stooped posture

 A bone fracture that occurs much more easily than expected

You may want to talk to your doctor about osteoporosis if you went through early menopause or

took corticosteroids for several months at a time, or if either of your parents had hip fractures.
 Causes

Osteoporosis weakens bone

Your bones are in a constant state of renewal — new bone is made and old bone is broken

down. When you're young, your body makes new bone faster than it breaks down old bone and

your bone mass increases. Most people reach their peak bone mass by their early 20s. As people

age, bone mass is lost faster than it's created.

 How likely you are to develop osteoporosis depends partly on how much bone mass you

attained in your youth. The higher your peak bone mass, the more bone you have "in the bank"

and the less likely you are to develop osteoporosis as you age.

Risk factors

A number of factors can increase the likelihood that you'll develop osteoporosis — including your

age, race, lifestyle choices, and medical conditions and treatments.

Unchangeable risks

Some risk factors for osteoporosis are out of your control, including:

 Your sex. Women are much more likely to develop osteoporosis than are men.

 Age. The older you get, the greater your risk of osteoporosis.

 Race. You're at greatest risk of osteoporosis if you're white or of Asian descent.

 Family history. Having a parent or sibling with osteoporosis puts you at greater risk,

especially if your mother or father experienced a hip fracture.

 Body frame size. Men and women who have small body frames tend to have a higher risk

because they may have less bone mass to draw from as they age.

Hormone levels

Osteoporosis is more common in people who have too much or too little of certain hormones in

their bodies. Examples include:

  Sex hormones. Lowered sex hormone levels tend to weaken bone. The reduction of estrogen

levels in women at menopause is one of the strongest risk factors for developing

osteoporosis. Men experience a gradual reduction in testosterone levels as they age.

Treatments for prostate cancer that

 reduce testosterone levels in men and treatments for breast cancer that reduce estrogen levels

in women are likely to accelerate bone loss.

 Thyroid problems. Too much thyroid hormone can cause bone loss. This can occur if your

thyroid is overactive or if you take too much thyroid hormone medication to treat an

underactive thyroid.

 Other glands. Osteoporosis has also been associated with overactive parathyroid and

adrenal glands.

Dietary factors

Osteoporosis is more likely to occur in people who have:

 Low calcium intake. A lifelong lack of calcium plays a role in the development of

osteoporosis. Low calcium intake contributes to diminished bone density, early bone loss

and an increased risk of fractures.

 Eating disorders. Severely restricting food intake and being underweight weakens bone in

both men and women.

  Osteoporosis causes bones to become weak and brittle — so brittle that a fall or even mild

stresses such as bending over or coughing can cause a fracture. Osteoporosis-related

fractures most commonly occur in the hip, wrist or spine.

 Bone is living tissue that is constantly being broken down and replaced. Osteoporosis

occurs when the creation of new bone doesn't keep up with the removal of old bone.

Retrieved from: https://www.mayoclinic.org/diseases-conditions/osteoporosis/symptoms-

causes/syc-20351968

RELATED STUIES

Eggshells as natural calcium carbonate source in combination with hyaluronan as beneficial

additives for bone graft materials, an in vitro study

Jörg Neunzehn, Thomas Szuwart, and Hans-Peter Wiesmann

Abstract

In bone metabolism and the formation especially in bone substitution, calcium as basic

module is of high importance. Different studies have shown that the use of eggshells as a bone

substitute material is a promising and inexpensive alternative. In this in vitro study, the effects of

eggshell granulate and calcium carbonate towards primary bovine osteoblasts were investigated.

Hyaluronan (HA) was used as artificial extracellular matrix (ECM) for the used cells to facilitate

proliferation and differentiation and to mimic the physiological requirements given by the egg in

vivo.
Chicken eggshell as suitable calcium source at home

Lucas R. Brun,Maela Lupo,Damián A. Delorenzi,Verónica E. Di Loreto &Alfredo Rigalli

Abstract

Aim: Taken into consideration that the deficiency of calcium (Ca) in the diet is a common

problem, the aim of this work was to study the chicken eggshell as Ca source at home. It was

evaluated: (1) different mechanisms to process eggshells and find an easy way to determine the

required amount of Ca at home and; (2) the flavor and the texture for eggshell fortified

food. Methods: Chemical and mechanical methods of eggshell processing were evaluated.

Changes in flavor and texture were evaluated in volunteers coordinated by a professional

chef. Results: A single eggshell contains 2.07 ± 0.18 g of Ca; therefore half an eggshell could

provide the amount of Ca needed by adult human beings per day. The best way to use chicken

eggshell as Ca dietary supplement is powdered to add to bread, pizza or spaghetti as there were

small changes in texture and no changes in flavor.

Keywords: Calcium, calcium dietary supplement, eggshell

Effects of supplementation of the diets with calcium and calcium-rich foods on bone density

of elderly females with osteoporosis.

C J Lee G S Lawler G H Johnson

Abstract

Diets of 20 elderly females with osteoporosis were supplemented daily with 2.25 oz of

processed cheese, and 3 capsules of dicalcium phosphate + vitamin D containing 350 mg Ca, 270

mg P, and 399 IU

vitamin D, for a 6-month period. The bone density of the 3–2 phalanx was measured

densitometrically from the hand x-ray taken with a standard aluminum stepwedge. Fasting blood

and urine were analyzed at the beginning and end of the experimental period. The mean bone

density increased (p < 0.05). There were 11 subjects with increased bone density, three with no

changes and six with reductions in bone density. Calcium, P, and alkaline phosphatase in serum

and Ca/creatinine and P/creatinine in urine were unchanged. The changes in bone density were

correlated to the changes in body weight (r = 0.6529). The results suggested that, even with a mean

age of 70 yr, some elderly persons can benefit from supplementary Ca and Ca-rich foods to

improve bone density.

Keywords: Bone density, Ca intake, osteoporosis

Eggshell calcium in the prevention and treatment of osteoporosis.

(PMID:15018022)

Stancíková M ,
Masaryk P ,

Svík K ,

Istok R

Abstract

In this paper the most significant biological and clinical aspects of a biopreparation made

of chicken eggshells are reviewed. Eggshell powder is a natural source of calcium and other

elements (e.g. strontium and fluorine) which may have a positive effect on bone metabolism.

Experimental and clinical studies performed to date have shown a number of positive properties

of eggshell powder, such as antirachitic effects in rats and humans. A positive effect was observed

on bone density in animal models of postmenopausal osteoporosis in ovariectomized female rats.

In vitro eggshell powder stimulates chondrocyte differentiation and cartilage growth. Clinical

studies in postmenopausal women and women with senile osteoporosis showed that eggshell

powder reduces pain and osteoresorption and increases mobility and bone density or arrests its

loss. The bioavailability of calcium from this source, as tested in piglets, was similar or better than

that of food grade purified calcium carbonate. Clinical and experimental studies showed that

eggshell powder has positive effects on bone and cartilage and that it is suitable in the prevention

and treatment of osteoporosis.

A follow-up study on the effects of calcium-supplement withdrawal and puberty on bone

acquisition of children

W T Lee S S Leung D M Leung J C Cheng

ABSTRACT

Recent calcium supplementation trials in children have confirmed a positive but moderate

effect of calcium intake on bone mineral accretion. However, the lasting effect of a higher bone

mineral mass after calcium-supplement withdrawal is not known. This is an 18-mo follow-up study

conducted after an 18-mo controlled calcium supplementation trial to study the persistent effect of

higher bone mineral mass in children. Radial bone mineral mass was determined by single-photon

absorptiometry; lumbar spine and femoral neck bone mineral mass were evaluated by dual-energy

X-ray absorptiometry in 84 healthy Hong Kong children at age 8.5 y and these evaluations were

repeated at age 10 y. Pubertal status was determined by Tanner staging. At the end of the follow-

up, the differences in percentage gains in lumbar spine bone mineral content (12.1 +/- 8.2%

compared with 14.9 +/- 10.05%, P = 0.24) and lumbar spine area (8.6 +/- 5.1% compared with 9.4

+/- 5.5%, P = 0.47) between the study and control groups disappeared. Dietary calcium intakes

during follow-up were similar for the two groups (555 and 640 mg/d, P = 0.23). In multiple-

regression analyses, pubertal status was the strongest correlate of bone acquisition and linear

growth in the study period. In conclusion, higher percentage gains in bone mineral mass in

childhood by calcium supplementation for 18 mo were reversible. Our study showed that the

benefits of calcium supplementation disappear after treatment is withdrawn. Longer-term calcium

trials are necessary to determine whether peak bone mass can be modified through sustained

supplementation so that appropriate calcium intakes can be determined.

Reducing fracture risk with calcium and vitamin D

Paul Lips

Roger Bouillon

Natasja M. Van Schoor

Dirk Vanderschueren

Abstract

Studies of vitamin D and calcium for fracture prevention have produced inconsistent

results, as a result of different vitamin D status and calcium intake at baseline, different doses and

poor to adequate compliance. This study tries to define the types of patients, both at risk of

osteoporosis and with established disease, who may benefit from calcium and vitamin D

supplementation. The importance of adequate compliance in these individuals is also discussed.

Calcium and vitamin D therapy has been recommended for older persons, either frail and

institutionalized or independent, with key risk factors including decreased bone mineral density

(BMD), osteoporotic fractures, increased bone remodelling as a result of secondary

hyperparathyroidism and increased propensity to falls. In addition, treatment of osteoporosis with

a bisphosphonate was less effective in patients with vitamin D deficiency. Calcium and vitamin D

supplementation is a key component of prevention and treatment of osteoporosis unless calcium

intake and vitamin D status are optimal. For primary disease prevention, supplementation should

be targeted to those with dietary insufficiencies. Several serum 25‐hydroxyvitamin D (25(OH)D)

cut‐offs have been proposed to define vitamin D insufficiency (as opposed to adequate vitamin D
status), ranging from 30 to 100 nmol/l. Based on the relationship between serum 25(OH)D,

BMD, bone turnover, lower extremity function and falls, we suggest that 50 nmol/l is the

appropriate serum 25(OH)D threshold to define vitamin D insufficiency. Supplementation should

therefore generally aim to increase 25(OH)D levels within the 50–75 nmol/l range. This level

can be achieved with a dose of 800 IU/day vitamin D, the dose that was used in successfull

fracture prevention studies to date; a randomized clinical trial assessing whether higher vitamin D

doses achieve a greater reduction of fracture incidence would be of considerable interest. As

calcium balance is not only affected by vitamin D status but also by calcium intake,

recommendations for adequate calcium intake should also be met. The findings of community‐

based clinical trials with vitamin D and calcium supplementation in which compliance was

moderate or less have often been negative, whereas studies in institutionalized patients in whom

medication administration was supervised ensuring adequate compliance demonstrated significant

benefits.

CHAPTER III

METHODOLOGY
Research Design
REPLICATIONS
TREATMENTS
1 2 3

T1
x x x

T2
x x x

T3
x x x

LEGEND: Where X serves as a substitute for the destined result.

Methods:
I.GATHERING OF ALL THE MATERIALS
The researcher prepare all the materials. The egg shells will be gather at the hometown
for the researcher.
II.CLEANING OF EGGSHELLS
The gathered eggshells will be clean using water.
III. BOILING OF EGGSHELLS
The eggshells will be put in a casserole and boil in a 100 C boiling point of the water to
get rid of the salmonella.
IV. TESTING FOR SALMONELLA
To test for its salmonella content the eggshells will be put in an empty and clean
container, and will be submitted to the VMUF.
V. GRINDING OF EGGSHELLS
The eggshells will be grind or pound using mortar and vessels or by a coffee grinder.
VI. FORMULATION OF CAPSULES
The researcher will formulate the capsule at VMUF and measured its weight.
VII.LABORATORY ANALYSIS
1. MICROBIAL ANALYSIS.
The researcher will submit the finish product to the VMUF to test its microbial
analysis.
2. CALCIUM DETERMINATION
The researcher submit the capsule to determine its calcium content at the VMUF.
X. PROXIMATE ANALYSIS OF MOISTURE
To test the proximate analysis of moisture the researcher put a (kg) of cocoa nibs in a
clean container and will submit it to the VMUF for the laboratory testing.
IX. DISSOLUTION TESTING
The researcher will conduct the dissolution testing of the capsule at TNHS with the
assistance of his research adviser.
X.TESTING of TREATMENTS
The researcher will conduct and test the treatments (1,2, and 3) at VMUF.
XI.SHELF LIFE OBSERVATION
The researcher will continue to observe the shelf life observation of the capsule.

XII.GATHERING OF THE RESULTS

The researcher gather all the results and ready for the analyzation.

MATERIALS

For Boiling

Eggshells. Casserole, burner, thermometer and thick gloves. Laboratory gown, facial mask,

and gloves.

For Grinding

Cleaned eggshells, mortar and vessels, coffee grinder. Gloves and facial mask for sanitary

purposes.

For Moisturize Analysis

Powdered eggshells. Container for storing the product that was submitted to the laboratory

for analysis.

For production of Capsule

Powdered eggshells for the content of the capsule. Capsule shell. Weighing scale for

measuring the total mg of the capsules. Laboratory gown, facial mask, and gloves for sanitary

purposes.

For Calcium Determination

 The formulated eggshell capsule. Laboratory gown, facial mask, and gloves for sanitary

purposes. Weighing scale, tupperware and the equipment for calcium determination.

For microbial analysis

Laboratory gown, facial mask and gloves for sanitary purposes. Eggshell capsule. Petri

plates, cotton swabs, nutrient agar, distilled water, difibrinated blood, and incubator.

For dissolution testing

Lab. gown, facial mask and gloves. Six capsules and distilled water.

For experimental animals

Rat cage. Albino rats supplied with food and water.

For testing of the treatments

Experimental rats. Thick gloves and mask. Calciumade, eggshell capsule and syringe for

the treatments.

FLOWCHART

Gathering of all the materials

Cleaning of eggshells
 Boiling of eggshells

Grinding of eggshells

Formulation of capsules

Laboratory
Analysis

Calcium Microbial
Determination Analysis

Proximate moisture analysis

Dissolution testing

Shelf life determination

Data gathering and analysis