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Chapter 1
Introduction
living. A healthy body allows an individual to live an active life and ward
be able to perform new skills, develop ideas and help their communities.
Philippines, over 2.7 million people experiences hunger but do not eat
while 1.5 million have gone through the day without eating. Decreased
K.1995).
Appetite is a term widely used to refer to the desire to eat food. There
is one of the most common diseases that may contribute to low appetite.
effect of some diseases, and a side effect of some drugs. By increasing our
healthy habits, we can decrease our illnesses that are caused by obesity
and malnutrition. There are a several factors that may cause decreased in
appetite. Such factors may differ with age and underlying health
appetite and weight gain with unintentional weight loss or poor food
intake. One of the specific concerns in elderly adults is low appetite. There
dental care, bowel regularity, and basic hygiene (Suzanne Falck, 2018).
wise use of medications and avoiding vices are some of the causes
(Godwin, 2018).
From the beginning up to now, nature has a lot to offer. Nature has
been a source of medicinal agents for years and huge numbers of modern
drugs have been derived from natural sources. Plant samples are widely
constituents can be derived from any part of plant like bark, leaves,
flowers, roots, fruits, and seeds. Medicinal plants play a major role in
human health care. Most of the world population relies on the use of
Blessed thistle, wormwood, gentian and bitter boost are some of the herbal
plants used for appetite enhancing and even indigestion. In addition, some
grows like grass, its underground stem or rhizome is succulent and yellow
on the inside, the leaves are grass-like, about 20 to 60 cms long and green
Laguna. This plant is also found in the province of Benguet and Bontoc
mostly in swamps. Acorus calamus leaves are well known in its alkaloidal
activity of its roots alone and with the present studies about the chemical
constituents of the leaves of the Sweet flag, the researchers have decided
to make use of the leaves of the Sweet flag, scientifically known as Acorus
calamus from the family Acoraceae, as the source of Methanolic extract for
is most commonly found in some muddy areas and along river side.The
Acoraceae family formerly Arum family – Araceae. The said plant is one of
the most common and most frequently used herbal plant among Chinese
and Indians. In other countries, Sweet flag was often added to wines. It
has been medicinally used for various ailments and conditions. The
medicines. This herb is generally used from the Ancient and Vedic periods
normalizing the appetite. Significant weight loss can occur and long-term
nutrition may be affected, leading to weakness and fatigue that can not
only inhibit daily activities but can also lead to malnutrition, consequently
formulated to help the body promote appetite and weight gain with
about by the phytoconstituents of its rhizome as well as its leaves. For this
reason, the student researchers were able to study further the appetite
Conceptual Framework
calamus leaves for appetite enhancer property. Scientific study was made
compound and volatile oils. But most of the biological actions of sweet flag
7
with design that will satisfy all the parameters set to measure the appetite
used for witchcraft. Further, in some folkloric they are used to stimulate
appetite. The researchers deepen its underlying effect which was being
leaves extract?
enhacer?
5. What is the appropriate dosage form for the Acorus calamus leaves
The study would like to prove that the crude extract from Acorus
Nutrition is vital for our body and all of its systems to function
well. Having a good nutrition can help reduce the risk of some illnesses
and diseases. Nowadays, a condition that can change the level of appetite
further in children the loss of appetite is very common, they tend to focus
themselves more of playing with toys, gadgets, playing with their friends
outside that they almost forget about eating nutritious food 3 times a day,
however they spend their eating time buying junk foods. According to
cost medications, a lot of people are forced to used herbal medications for
to the health and a cause of mortality and morbidity not only in our
country but also in the whole world. Regular exercise, regulating the food
we eat and appetite enhancers are some best solutions to address this
researchers are looking forward to the success of the study which will be
property of the crude extract from the leaves of sweet flag from the family
flavoniods test. The Acorus calamus extract was also subjected to toxicity
form was done yet it was not able to have its efficacy testing and quality
controls.
Definition of Terms
For better understanding of the study, the following terms have been
identified:
11
weight. The illness occurs in both sexes but mainly in women. Mostly, it
nutrition, 2003)
management reasons.
Sprague Dawley rats. It has long and narrow head, longer tail which may
Swiss mice. Common laboratory mice which are smaller than the
Chapter 2
interest to play activities, have a low sense of well-being and are less able
psychological factors, determines one’s hunger, the desire to eat and fed.
loss of appetite especially when the child is not feeling well for any reason.
(Khandke, 2011)
the patient. Anabolic steroids have been used showing some positive
which lead to dying and follows strict diet especially avoiding foods with
high calorie content. The top candidate for this kind of eating disorder are
Appetite
There are numerous herbs and herbal formulations which are widely
was used to enhance appetite, good quality of food intake and make the
children feel the desire to eat. More of these, the study found to be safe
and do not possess any adverse events. Results were obtained from the
decreased sense of well-being and are less able to cope with disease-related
determines one’s hunger, desire to eat and satiety. It is quite normal for a
the child is not feeling well for any reason. Hence, it is difficult to
term for this is anorexia. A wide variety of conditions can cause your
illnesses.
Appetite Enhancer
appetite and weight gain with unintentional weight loss or poor food
16
experiencing. Although many agents have been evaluated for their use as
are megestrol, oxandrolone, and dronabinol. They are indicated for the
treatment for anorexia and to promote weight gain for some patients. The
the appetite stimulants may be used for the treatment for anorexia.
(Archer, 2014)
Acorus calamus
a sweet, tangerine like scent when cut. There are a lot of therapeutic claims
(Mukherjee, et. al 2007).In most experimental studies, the parts used are
the leaves, rhizomes and stem of the plant. Phytochemical studies have
extracts of A. calamus. The findings of other studies show that the major
than beta and alpha asarone, hydrocarbon compounds are also present in
in the leaves and rhizomes of a plant sample had been already proven.
experiments that can be conducted which has more activities that shows
the pharmaceutical and also in bioethical scale for its great potency and
such plant with distinctive solvent or extract. This herb is widely used from
the Ancient and Vedic periods for the reason of its power of rejuvenation
Chandra, 2017).
Plants are being used to cure various diseases. Active constituents have
been isolated and studies from these plants to yield treatment. Most of the
drugs that are available in the market today were originally made from
Acoruscalamus was also known to many early American settlers and used
intellectual and mental health for the elderly (Hou and Jin, 2008). It is also
affected, leading to weakness and fatigue that can not only inhibit daily
risk of morbidity and mortality. Children with intestinal failure, those who
have undergone a transplant, and those who have cancer or cystic fibrosis
to the present. According on some literatures there are many plants that
centimeters wide, with waved margins and stout midribs, base of leaf
moist places with running water, on boulders, etc., at low and medium
The parts used in most of the experimental studies are the leaves,
are mostly used. Decoction of dried roots and leaves to a pint of boiling
furthermore it can be used for scenting snuff and for flavoring alcoholic
sub-temperate regions of Eurasia and the Americas. The plant has a rich
ethnobotanical history dating back possibly to the time of Moses in the Old
Testament of the Bible and in early Greek and Roman medicine. Sweet
flag, known to be indigenous to India that has been valued for its rhizome
and fragrant oils which have been used medicinally, in alcoholic beverages,
terpenes, a few commonly occurring steroids and xanthones had also been
namely herbal multiaction skin gel, which was found to be effective against
rhizome for calamus oil are widely used now in pharmaceuticals and
Thus, it is a common item in the international drug market and the raw
new drugs from plants related to many disorders. A large number of drugs
of an Acorus calamus. The rhizome, roots and leaves of the plant have
Acorus calamus (Sweet flag) has a history of use and has many
Chinese medicine, etc. for the treatment of various aliments like nervous
calamus has been used since ancient times, many of its uses are yet to be
While rhizomes of Acorus calamus, having high ADH and low LDH
(100 and 200 mg/kg) as well as saponins (10 mg/kg) isolated from the
as by other cultures, and many of these uses continue to this day (Motley,
26
This herb is generally used from the Ancient and Vedic periods due to its
system. Acorus calamus has many wide varieties which were used in
a medicinal plant dates back to Egypt, Greek and Roman times. Sweet flag
was distributed from its native range by rhizomes through trade and
of Acorus calamus plant are alpha and beta-asarone. The plant has also
are reported to have psychoactive effects. This has been attributed to the
smooth muscle tissue, and the oil has been found to induce spasmolytic
medicine by Chinese, Indians, American Indians and others, and are still
properties were attributed to the rhizome and it was used in North America
medicinal aid for stomach complaints and colic in children. Since ancient
times it has been reputed for its stimulant digestive virtues. In India, the
New Guinea, the leaves of sweet flag are taken as a tonic, and chewed to
relieve toothache. The crushed rhizome is rubbed into the hair to kill lice.
In Brunei, sweet flag is used to treat gastritis and diarrhea and also as a
cabinet. Historically, it has been used all over the world for many different
rejuvenator for the brain and nervous system, and as a remedy for
and getting the stomach churning and ready for food. This is what it does
infusion can be made by soaking it at the top of a closed jar of cold water
overnight. Hot water infusions and alcoholic tinctures are dreadful, will
Sweet flag was included in many of the early herbals and has a rich
history in the Chinese and Indian cultures. Few plants have gained such
much attention as they are well tested for their efficacy and are generally
31
biological activities of sweet flag provides detailed evidence for the use of
biological activities of sweet flag, it seems that there are still a large
isoasarone, methyl isoeugenol and safrol are also responsible for medicinal
activity but most of the biological actions of sweet flag have been attributed
(Imam, 2013).
32
system going, and that makes the person feel hungry. Acorus calamus is
also known for its calming and centering effect that is why it can be both
stimulant and sedative, it puts your energy into balance and gets you
equal as a treatment for panic and anxiety attacks. Have the person chew
on Sweet flag and breathe deeply, fully and slowly and the anxiety and
panic will fade. Incidentally, its antianxiety effects are the reason why it
was used for quitting smoking, not because it causes a 'distaste for
Tobacco'.
Research findings suggest that sweet flag may have applications for
several ailments for which it has a historical record of use. It may still have
which seem to be free from this component offers new opportunities for
more extensive use in the future after selection. Sweet flag might have good
For ages, Acorus calamus rhizome has been used for the treatment
polyploidy, and the composition of the essential oil obtained from the plant
effects of Acorus calamus rhizome and its constituents α.-asarone and β.-
CNS active herb from Ayurvedic tradition and requires further research to
et.al., 2008)
of works and losing up energy, people must not forget to have proper diet
for the body most especially to those who struggled in such loss of appetite
and weak food intake. This study parallel with the literatures stated, the
researchers will easily find its way to develop the underlying component of
Chapter 3
plant sample used to determine the appetite enhancer property from the
extract of sweet flags leaves (Acorus calamus L.) on Sprague Dawley rats.
selectively collected from San Antonio, Nueva Ecija, Philippines, during the
Industry in San Andres St, Malate, Manila for validation and identified as
Bulacan.
Methods of Research
35
calamus).
The leaves were collected and washed multiple times with water and
air dried, cut into small pieces and powdered using mechanical blender.
hexane, methanol, ethyl acetate and 50% methanol for 24 hours. Then
filtered and evaporated into dryness over rotavap. The collected residue
evaluation.
36
Serial Extraction
n-Hexane Ethyl acetate Methanol 50% Methanol
Organoleptic Glycosides
Flavonoids
Saponins
Turpenoids
Tannins
Alkaloids
Only that extract that gave the most number of positive results in Flavonoids test
Biological testing
Efficacy
37
Figure 1
A small amount of the crude extract of sweet flag was observed for the
Phytochemical Screening
subjected in a water bath for 15 minutes. Observe for any color changes.
color.
A few drops of lead acetate were added to the test solution. The
Libermann-Burchard test:
and cool. With the test tube tilted, concentrated sulfuric acid is added the
39
by the side of the test tube, shows brown ring at the junction of two layer
and the upper layer turns green which shows the presence of sterols and
Salkowski’s test
With the crude extract treated chloroform and with several drops of
concentrated sulfuric acid and mixed, shaken well and allow to stand for
some time. The indication that there is presence of sterol will appear red
color in the lower layer and formation of yellow colored lower layer indicate
Molisch test:
In a test tube treat the 2ml solution with several drops of alcoholic
junction.
Fehling’s test:
40
mixed together and added several drops of sample and boiled, indication
precipitate.
Foam test:
The extract was diluted with distilled water and for 15 minutes
Borntrager’s test:
To the 3ml test solution, diluted sulphuric acid was added and
for 5 minutes was boiled and then filtered. After, equal volume of benzene
was added to the cold filtrate and mixed well. The organic layer was
layer.
acid and a drop of 5% ferric chloride were added. With the test tube tilted,
carefully add 0.5 ml of concentrated sulphuric acid by the side of the test
Mayer’s reagent
Then the filtrate solution will be treated with several drops of mayer’s
42
alkaloids.
Dragendorff’s reagent
Then the filtrate solution will be t reated with several drops of dragendorff’s
presence of alkaloids.
Wagner’s reagent
Then the filtrate solution will be treated with several drops of wagner’s
Toxicity Testing
Animals should be fasted prior to dosing (e.g. with the rat, food but not
water should be withheld over-night; with the mouse, food but not water
43
should be withheld for 3-4 hours). Following the period of fasting, the
animals should be weighed and the test substance administered. After the
substance has been administered, food may be withheld for a further 3-4
changes should be calculated and recorded. At the end of the test surviving
Efficacy Testing
rats from food and water for 24 hours before starting the experiment.
The rats are randomly divided and selected into 5 groups: group 1
mg/kg test sample group 4 treated with extract containing 100 mg/kg test
sample and group 5 treated with extract containing 150 mg/kg test sample
The rats in the vehicle group received distilled water and 200g food
every day and those in the untreated group received no intervention. The
hours. The weight comparison will be based on the initial testing up to the
ingredient.
Statistical Treatment
The statistical method used in the research was the bartlett’s test
for equal variances. All the raw data used in the research was presented
for the analysis of significant differences within the group of rats during
Chapter 4
This chapter dealt with the results obtained from the preliminary
collected.
Physicochemical Evaluation
Organoleptic Test
All the extracts of Acorus calamus have the same physical properties
Table 1
Organoleptic Test
Table 2
Methanol
Preliminary Screening
expected result of pink scarlet odor, Ferric chloride test with the expected
Alkaline test with the expected result of yellow solution to colorless and
Lead Acetate test with the expected result of yellow precipitate formation.
50% methanol was used as the solvent for the flavonoidal extraction
phytochemical screening.
The dried Lubigan leaves were extracted using 50% methanol as the
Limit Test
methanolic extract. All the mice survived the limit test. Data obtained from
Efficacy Testing
baseline of each rats and controlling the diet. The study used Sprague
49
dawley rats as test animals with either male or female sex. The food
were weighed before it was given and collected the remaining food after
8 hours then weighed again. The rats were given the corresponding
dose of drugs and extracts that was computed. Each rats were
The data obtained from getting the baseline and controlling the
diet for testing the appetite enhancer effect of the Flavonoid extract can
WEIGHT INCREASE IN
PERCENTAGE
150mg/kg 9.91
100mg/kg 9.3
50mg/kg 5.67
NEGATIVE
0.87
POSITIVE 5.13
Figure 2
Statistical Treatment
and 150mg/kg compared to the positive control (Heraclene Forte 2 mg) but
rats therefore the researchers conclude that the said concentration was
Dosage Formulation
The dried leaves ware macerated in 50% methanol for 48 hours and
prepared simple syrup through agitation with the aid of heat with the
following ingredients such as sucrose (80 g) and distilled water (qs.ad). The
removed from the heat, the sucrose was added gradually with occasional
51
Acorus calamus was mixed with the simple syrup until dissolved.
Chapter 5
chapter.
Summary of Findings
Based on the following results of the study, the summary of the findings
solvent which has the highest percentage yield was methanol but
for toxicity testing. The rats were weighed and their toxic dose was
food consumption for a week, the biological testing was done the
conducted. The rats were observed for 2 consecutive weeks and was
weight again. The most effective dose for the appetite enhancing
improved.
5. The use of liquid formulation is the most common due to its ease
prepared syrup dosage form. It was done by agitation with the use
formation that does not appear during the process and shelf life. The
Conclusions
All doses administered to the rats are effective, yet the most effective
Recommendations
the lubigan plants for the determination of other therapeutic activity that
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Emanuele, M., (2016), Anorexia of Aging: Risk Factors, Consequences, and
Potential Treatments. Nutrients,8(6), 1-10.
Glanz, K., & Rimer, B. K., (1995), Theory at a Glance: A Guide for Health Promotion
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Greta, W., Maria, R., Zoe, R., (2012), Contribution of social and family factors in anorexia
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Janet, T., & June, A., (2013), Anorexia Nervosa. A Recovery Guide for Sufferers, Families
and Friends, Second edition.
Katherine L. Godwin, MD., (2018), Eating Disorders Can Mask Malnutrition. 1(3), 37-41.
Kho, S.L., Chan, S.W., (2016), Antioxidant and antibacterial activity of Acorus calamus.
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59
APPENDIX D
APPENDIX E
Results
Dose Compuations
POSITIVE CONTROL
3 mg
61
50 mg/kg :
100 mg/kg :
BODY WEIGHT DOSE
150 mg/kg :
control
control
=66.77g
64
04,2018 administration
Dec 11,2018
POSTIVE POSTIVE
1.
NEGATIVE NEGATIVE
50mg/kg 50mg/kg
100mg/kg 100mg/kg
150mg/kg 150mg/kg
control
Control
=70.94
=79.50
67
Control
Control
=82.00g
Dec 11,2018
POSTIVE POSTIVE
NEGATIVE NEGATIVE
2).276g 2).276g
3).261.5g 3).263g
4).248g 4).250g
5).275.g 5).279g
50mg/kg 50mg/kg
100mg/kg 100mg/kg
150mg/kg 150mg/kg
PERSONAL INFORMATION
Gender:Female
Birthdate: June 22, 1998
Birthplace: Nueva ecija
Age: 20 years old
Civil Status: Single
Nationality: Filipino
Religion: Roman Catholic
Language: English and Tagalog
EDUCATIONAL ATTAINMENT
Tertiary Education: Bachelor of Science in Pharmacy
Centro Escolar University – Malolos
Km. 44, Macarthur Highway, City of Malolos,
Bulacan
2015 – 2019
Secondary Education: Bethel Christian Academy
Plaridel, Bulacan
2011 – 2015
Elementary Education: Lord’s Shepherd Academy
Tabang Guiguinto, Bulacan
INTERNSHIP TRAINING
Community Internship: 2016 - Mercury Drug Corporation
(Puregold Guiguinto Branch)
Hospital Internship: 2017- Allied Care Experts Medical Center
(Baliuag, Bulacan)
Manufacturing Internship: 2018 – MDB Pharma Trading
(Paombong, Bulacan)
Major Internship: 2018 – La Consolacion University General Hospital
(Malolos City, Bulacan)
72
Jonna B. Esguerra
#0322 Pungo, Calumpit, Bulacan
esguerra.jonna17@gmail.com
+639056963220
PERSONAL INFORMATION
Gender: Female
Birthdate: June 17, 1999
Birthplace: Calumpit, Bulacan
Age: 20 years old
Civil Status: Single
Nationality: Filipino
Religion: Roman Catholic
Language: English and Tagalog
EDUCATIONAL ATTAINMENT
Tertiary Education: Bachelor of Science in Pharmacy
Centro Escolar University – Malolos
Km. 44, Macarthur Highway, City of Malolos,
Bulacan
2015 – 2019
Secondary Education: Mary Chiles College of Arts and Sciences
Pulilan, Bulacan
2011 - 2015
Elementary Education: Pungo Elementary School
Calumpit Bulacan
2008 - 2011
INTERNSHIP TRAINING
Community Internship: 2016 - Mercury Drug Corporation
(Calumpit, Bulacan)
Hospital Internship: 2017 - Sacred Heart Hospital of Malolos Inc.
(Malolos City, Bulacan)
Manufacturing Internship: 2018 – Primecare Lifesciences Laboratories Corp.
(Hagonoy, Bulacan)
Major Internship: 2018 - Sacred Heart Hospital of Malolos Inc.
(Malolos City, Bulacan)
73
PERSONAL INFORMATION
Gender: Female
Birthdate: December 22, 1998
Birthplace: Bocaue, Bulacan
Age: 20 years old
Civil Status: Single
Nationality: Filipino
Religion: Roman Catholic
Language: English and Tagalog
EDUCATIONAL ATTAINMENT
Tertiary Education: Bachelor of Science in Pharmacy
Centro Escolar University – Malolos
Km. 44, Macarthur Highway, City of Malolos,
Bulacan
2015 – 2019
Secondary Education: St. Paul College of Bocaue
Turo, Bocaue Bulacan
2011 - 2015
Elementary Education: St. Paul College of Bocaue
Igulot, Bocaue Bulacan
2008 - 2011
INTERNSHIP TRAINING
Community Internship: 2016 - Mercury Drug Corporation
(Bocaue, Bulacan)
Hospital Internship: 2017- Our Lady of Lourdes Hospital
(Sta. Mesa, Manila)
Manufacturing Internship: 2018 - Morishita- Seggs Pharmaceuticals Inc.
(Sta. Maria, Bulacan)
Major Internship: 2018 – Dr. Yanga Hospital Inc.
(Bocaue, Bulacan)
74
Pamela P. Baluyut
#118 Sta. Rita Macabebe, Pampanga
pamela.baluyut00@gmail.com
+639268017690
PERSONAL INFORMATION
Gender: Female
Birthdate: September 15, 1999
Birthplace: Apalit, Pampanga
Age: 19 years old
Civil Status: Single
Nationality: Filipino
Religion: Roman Catholic
Language: English, Tagalog, Kapampangan
EDUCATIONAL ATTAINMENT
Tertiary Education: Bachelor of Science in Pharmacy
Centro Escolar University – Malolos
Km. 44, Macarthur Highway, City of Malolos,
Bulacan
2015 – 2019
Secondary Education: Colegio De San Lorenzo
Macabebe, Pampanga
2011 - 2015
Elementary Education: Jesus Mary Joseph Montessori School
Macabebe, Pampanga
2008 – 2011
INTERNSHIP TRAINING
Community Internship: 2016 - Mercury Drug Corporation
(Macabebe, Pampanga)
Hospital Internship: 2017- Pampanga Premier Medical Center Inc.
(Sampaloc, Apalit Pampanga)
Manufacturing Internship: 2018 –MDB Pharma Trading
(Paombong, Bulacan)
Major Internship: 2018 - Mercury Drug Corporation
(Macabebe, Pampanga)
75
Christian A. Domingo
Esguerra St. Poblacion, Pulilan Bulacan
christian.domingo29@gmail.com
+639174533608
PERSONAL INFORMATION
Gender: Male
Birthdate: February 22, 1997
Birthplace: Pulilan, Bulacan
Age: 22 years old
Civil Status: Single
Nationality: Filipino
Religion: Born again Christian
Language: English and Tagalog
EDUCATIONAL ATTAINMENT
Tertiary Education: Bachelor of Science in Pharmacy
Centro Escolar University – Malolos
Km. 44, Macarthur Highway, City of Malolos,
Bulacan
2014 – 2019
Secondary Education: Virgilio V. Dionisio Memoria High School
Pulilan, Bulacan
2010 - 2014
Elementary Education: Balatong Elementary School
Brgy. Balatong Pulilan, Bulacan
2004 - 2010
INTERNSHIP TRAINING
Community Internship: 2015 - South Star Drug
(Robinsons Pulilan, Bulacan)
Hospital Internship: 2016- Our Lady of Mercy General Hospital
(Longos Pulilan, Bulacan)
Manufacturing Internship: 2018 –RoyMed Pharmaceuticals
(Kalayaan village, San fernando Pampanga)
Major Internship: 2019 – Polymedic Hospital
(Guiguinto Bulacan)
76