1. Skim Latihan Akademik Bumiputera (SLAB): KURSUS PENYELIDIKAN 2006
Bilik Mesyuarat Fakulti, FKKKSA, 07 July 2006
“CHARACTERIZATION OF Hibiscus rosa-sinensis
Linn LEAVES AQUEOUS EXTRACT ON THE
ANTIPYRETIC ACTIVITY”
SAIFUL IRWAN BIN ZUBAIRI
RESEARCH OFFICER (QA41)
CHEMICAL ENGINEERING PILOT PLANT (CEPP)
UNIVERSITI TEKNOLOGI MALAYSIA
UTM SKUDAI, JOHOR
 e-mail: saiful-z@utm.my
Hibiscus rosa-sinensis leaves

2. INTRODUCTION
 Fever or pyrexia is an adaptive mechanism of the body to react against pathogens
 The core body temperature is elevated to levels which denatures, weaken or kill the pathogen
 Unfortunately, there are circumstances when fever accelerates to temperature where the body is
at risk of devastating its own cells and should be brought under control with suppressive medication
TYPES OF FEVER
Pyrexia can be classed as low grade (38 °C to 39 °C), moderate (39 °C to 40 °C) or high grade
(more than 40 °C) depending on how much the body temperature has deviated from normal
FEVER BENEFICIAL
 Fever may be of benefit and is part of the body's response to a disease
 However, if the fever goes above 42 °C (107.6 °F), then it may cause significant tissue damage and
would most certainly be harmful. This is termed as hyperpyrexia
NORMAL TREATMENT
 Drugs that reduce fever are known as antipyretics
 Common antipyretics are acetaminophen, also called paracetamol and Non Steroidal Anti-
inflammatory Drug (NSAIDs) such as ibuprofen
 A popular household remedy is soaking a cloth in cold water and placing it on the patient's forehead

3. WHY CHOOSE THE CONVENTIONAL WAY TO THREAT FEVER?
 Nowadays, the concern is on the modern antipyretic drugs which associated with the
hepatotoxicity problems
 For that reason, Hibiscus rosa-sinensis Linn leaves is used traditionally as an antipyretic
remedy (Edward, 1999)
 This is because, the aqueous extract of Hibiscus rosa-sinensis Linn leaves are comprised
with β-sitosterol, taraxeryl acetate, flavonoids compound and high in calcium and
phosphorus content (Duke and Ayensu, 1995)
 These bio-active compounds, minerals and vitamins that available in the extracts are
assumed to lower down the temperature and acts as a coolant to the fever patient
 Since there is not scientific research on this particular plant species as regards to the
antipyretic activity, this research could be a new BREAKTHROUGH or ALTERNATIVE
REMEDY to the chemically synthesized ANTIPYRETIC DRUG in which posse major side
effects especially associated with hepatic organ
 Hence, the synergistic of these compounds could be contributes to the antipyretic activity

4. WHAT IS β-SITOSTEROL?
 β-sitosterol is a major bio-active compound of phytosterol group in the higher plants
 β-sitosterol, with an empirical of C29H50O, is a sterol compounds with a molecular weight
of 414 g/mol
 β-sitosterol is not synthesized endogenously in mammals with minimal intestinal absorption
 β-sitosterol has been shown to have anti-inflammatory and antipyretic properties and a
study on animal model have found that antipyretic effects of β-sitosterol was comparable
to that of aspirin (Gupta et al., 1980)
 Moreover, there have been numerous clinical and animal studies that have established the
benefits of β-sitosterol such as immune system support (especially during stress)-relieves
allergies-reduces cancer risk (prostate, breast and colon)-analgesic properties-relieves
symptoms of enlarge prostate Molecular structure of β-sitosterol

5. POSTULATE
An aqueous extract of Hibiscus rosa-sinensis is used as an acute
dermal and acute oral to treat the elevation of body temperature
above normal temperature (i.e.: 98.4 °F or 37 °C)
PROBLEM STATEMENT/RESEARCH PROBLEM/QUESTION
HOW does β-sitosterol, flavonoids compound, minerals and vitamins in
the aqueous extract of Hibiscus rosa-sinensis affect the body immune
system simultaneously alleviate the elevation of body temperature?
HYPOTHESIS
β-sitosterol, flavonoids compound, minerals and vitamins in the aqueous
extract of Hibiscus rosa-sinensis acts as an antipyretic, antioxidant and
enhancer of body immune system by producing a SUFFICIENT amount of
white blood cells (leukocytes: WBC count/mm3) to fight the infections of
pyrogens simultaneously alleviate the elevation of body temperature

6. RESEARCH OBJECTIVE
To investigate the efficacy of bio-active constituents, trace minerals and vitamins in the
aqueous extract of Hibiscus rosa-sinensis Linn leaves on the antipyretic activity
RESEARCH SCOPES
 Literature review on the current development of conventional (ethno-botanical studies) and
un-conventional antipyretic drugs
 To identify and analyze the major bio-active constituents, trace minerals and vitamins in
the extract of Hibiscus rosa-sinensis
 To study the effect of major bio-active constituents in the extract of Hibiscus rosa-sinensis
on the antioxidant and antimicrobial activity
 To study the effect of major bio-active constituents in the extract of Hibiscus rosa-sinensis
on the white blood cells count (WBC count/mm3) and antipyretic activity
 To study the correlation between the antipyretic activity with the major bio-active
constituents in the extract of Hibiscus rosa-sinensis and white blood cells count
(WBC count/mm3)
 Thesis write-up and publications

7. RESEARCH PLANNING AND EXECUTION
Research scope/semester S1 S2 S3 S4 S5 S6
1. Literature review (conventional and unconventional antipyretic drugs)
2. To identify and analyze the major bio-active constituents, trace
minerals and vitamins
3. To study the effect of major bio-active constituents on the antioxidant
and antibacterial activity
4. To study the effect of major bio-active constituents on the white
blood cells count (WBC count/mm3) and antipyretic activity
5. To study the correlation between the antipyretic activity with the
major bio-active constituents and white blood cells count
(WBC count/mm3)
6. Thesis write-up and publications     
 Publications: International Scientific Journal

8. PROCESS FLOW AND RESEARCH METHODOLOGY
(A) Pretreatment of the Hibiscus fresh leaves:
Forced with air oven at 30 0C (3 hours) and ground into fine powder (particles size of 0.5 mm ∅)

(B) Extraction of the Hibiscus ground dried leaves:
Microscale batch solid-liquid Pyrex extractor: ζ = 5.0 mlg-1, t = 40 0C for 24 hours
HPLC: Bio-active compounds 
and flavonoids content (C) Physico-chemical characterization:
Relative density (20d20) and refractive index (20nD)
 Lyophilization process
(D) Nutritional composition and proximate analysis: (LIQUID EXTRACT)
a) Identification of trace minerals: Induced Coupled Plasma-mass spectrometry (ICP-MS)
b) Proximate analysis (Ash, Fat, Carbohydrate and Vitamin A, B, C: AOAC Method)

(E) Antioxidant test (LIQUID EXTRACT):
Radical scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) method

(F) Microbiology test - Agar dilution method (SOLID EXTRACT):
Fungal colony of Cladosporium sphaerospermum

(G) Antipyretic test (LIQUID AND SOLID EXTRACT):
(1) An ED50 value calculated as the dose of Hibiscus extract solution required to decrease pyresis by 50 %
(2) Acute Dermal Heat Transfer Assessment (Q = Joules) - Newtonian method/Cooling method

(H) Blood cells count (count/mm3):
Complete blood count (RBC + WBC) and leukocytes content

9. EXPECTED RESULTS
 Mechanism of β-sitosterol, flavonoids compound, trace minerals and vitamins in the aqueous
extract of Hibiscus rosa-sinensis Linn leaves on the antibody immunization system, antioxidant,
antimicrobial and antipyretic activity will be observed significantly due to its well known TRADITIONAL
USAGE to treat fever and as a food supplementary
 β-sitosterol is expected to be the major contribution of all constituents in the extract due to the fact that
plant sterol appears to modulate the immune function by controlling the production of inflammatory
cytokines that relieve the pain activity
 This phytosterol compound is also expected to produce high inhibitory on the targeted
microorganisms given that phytochemicals which consist with antioxidant and anti-inflammatory
properties will also produced a significant effect on the antimicrobial activity
 Whilst flavonoids compound (e.g.: kaempferol, rutin, catechin, luteolin and quercetin) are
expected to yield significantly in the aqueous extract in which these compounds will scavenged the
pre-radical molecules (oxidation agent) that usually found abundantly in the body system during the
demotion of body immunization system
 The macro mineral (e.g.: calcium, potassium, phosphorus and sodium) are expected to increase
the hormones activity to the hypothalamus system function by regulating the high body temperature
to normal temperature (negative feed back mechanism)
 As for the pyrexia effect of the Hibiscus extract, it is expected to produce high effective dose (ED 50)
values to decrease the pyrexia level of rat induced fever (↑ ED50; ↓ pyrexia level)
 The higher effective dose (ED50) indicates the higher of changes in mean dorsal temperature lowering
produced by the Hibiscus extracts treatment versus the non treatment animal of an induced fever

10. CONCLUSION AND RECOMMENDATION
 From an extensive literature reviews study, understanding the mechanism of
plant sterol, flavonoids compound, minerals and vitamins that existed
abundantly in the particular plants or herbs as a potential antipyretic drug and
body immunization system enhancer is an important innovative
breakthrough and alternative to allopathic remedy (e.g.: tylenol, aspirin,
acetaminophen or paracetamol) in the future to come
 The study can be a viable references and the configuration of the research
can be a useful tools for developing on other potential plant that posses a
specific therapeutic action
**THE END**

11. THANK YOU FOR YOUR KIND
ATTENTION
QUESTION AND ANSWER SESSION