TUGAS MATA KULIAH

PUBLIKASI DAN PENULISAN ILMIAH

Dosen Pengampu :
Drs. Agustinus Yuswanto, SU., PdD, Apt

dikerjakan oleh,

Diyan Ajeng Rossetyowati

16/405843/00141

PROGRAM STUDI S3 ILMU FARMASI

FAKULTAS FARMASI
UNIVERSITAS GADJAH MADA YOGYAKARTA
 Courses: Publication and Methodological Research
Task: Reviewing Journal

Name: Diyan Ajeng R, MPh., Apt

Reviewing results:

1. Does the paper contain new information?

Yes, it does
Research about Pacar air all this time concern to antibacterial activity,
immunomodulatory activity by testing imunosupresant from Pacar air
still rarely performed so this research has novelty value information.
But there is no contribute new information like as method that
researcher has used, and therere no significant data from the results,
but the researches claimed that Pacar Air (Impatiens balsamina Linn.)
have immunosuppressive activity

2. Does the title of manuscript covers its content?

Yes, it does
The tittle of manuscript covers did comply with the content of course
from thats study, but its to long and make do not show a clear title.
The title is a little more to report the contents of the journal

3. Is the summary appropiate?

No, isnt
There isnt summary from this study,but abstrack from this study isnt
complete.Some content like as the aim and background from this
study arent mentioned clearly. The method, tool and materials that
used are not listed. Technique in this research not focus and how
cultivate analysis done arent listed too. Abstracts should summarize
the objectives, methods, results and conclusions.

4. Is the aim of the work clearly established?

yes, it is
The aim from this study not clearly established. The aim in a research
must be spesific, concept and variable must be clearly, with accurate
measurement. The purpose of research it was clear that to observe
the immunomodulatory activity of dichloromethane fraction of Pacar air
plant (Impatiens balsamina L.) and to determine the effective dose.

5. Are the methods adequate?

No, they arent. The methods werent complete to describe about the
equipment and materials used. There are no information about the
average body of the animals/ rats used, no specifications reagents, no
step to determining dose from plant extracts, not listed antibody level.
 Experiment to immunomodulator needs to be added effect of extract
and fractions on in vitro phagocytosis by human PMN cells.
Observation and measurement B lymphocytes and T lymphocytes to
look immune response specific not explained in this research.
Immunostimulant activity, can be observed with increased immune
cells which correlated with spleen weight.

6. Are the results reliable?

No, they arent.
Many things related to incomplete outlined the results obtained. In
addition there is some incongruity between the description of the
results obtained by the methods used. This research still needed an
percentage of specific leukocyte cell that eosinophils, neutrophils,
lymphocytes and monocytes to support the discussion. The data were
collected in the table/ drawings must be complete, also narrative text
and presented in a form easily understood. Every important
relationship between variables in the experiment and any correlation
between variables is not obvious.

7. Are discussion and conclusions well documented and scientifically

coherent?
No, they arent
The results from analysis arent clearly. There arent normality and
homogenity test, also the hypothesis. Identification of test compounds
must be discussed and continued with a discussion antigen and dose
of plant extracts. Any data presented should be relates back to the
question stated in the introduction.

Recommendation:
The recommendation based on the review to seven points above is
receiving the manuscript with revisions, like as complementary methods of
research and discussion that is more structured and appropriate review .
Immunomodulatory Activity of Pacar Air (Impatiens balsamina Linn.)
Dichloromethane Fraction with Haemagglutination of Titer
Antibodies and Delay Type Hypersensitivity (DTH) Methods in Mice
(Mus musculus)
ABSTRACT
Imunomodulatory activity of dichloromethane fraction was studied
by haemaglutination of titer antibodies and delay type hypersensitivity
(DTH) methods in Balb/C mice. Each method was used 5 groups of mice
which are normal control group (CMC-Na 1% p.o), positive control group
of immunostimulant (levamisole 50 mg/kg bw, p.o), positive control group
of immunosuppressant (methylprednisolone 15 mg/kg bw, i.p), and
dichloromethane fraction groups doses of 125 mg/kg bw (DF I) and 250
mg/kg bw (DF II) orally.Based on the study, both doses of
dichloromethane fraction have a potention to suppress humoral and
celullar specific immune system. It showed lower humoral antibody (HA)
titre value than normal and control group (p<0.05) at doses of 125 mg/kg
bw.Primary and secondary titer antibody value were (5,094 0,5) and
(5,937 0,5) respectively. It also showed lower percentage of right foot
thickness in DTH method at 48 hours after induction measurement
compared with normal and control groups (p <0.05) atboth doses
groups.Based on this study, dose of 125 mg /kg bwof Pacar Air (Impatens
balsamina Linn.) dichloromethane fraction was effective dose in
imunosupressant activiy on humoral and selular spesific immune system.
Keywords :Delayed Type Hypersensitivity, Haemaglutination Titer
Antibodies, Immunomodulatory, Impatiens balsamina Linn.
INTRODUCTION
Immunomodulators are substances that can modulate the immune system.
Many synthetic drugs are efficacious as immunomodulators cause side
effects, such as urticaria, agranulocytosis, toxic to the liver, and others.1
Therefore, research must be done to prove the immunomodulatory activity
of extracts and isolated plants are expected to have less side
effects.Family Balsaminaceae is one group of plants that are the main
source of naftaquinon.2 Other studies have shown that the compound is
1.4 naphtoquinon was a significant inhibitor of the lipopolysaccharide
stimulation of macrophages through immunosuppressive activity with
inhibition percentage of 97.4 to 98.9% , The same study also showed that
1.4 naphtoquinon can show promising activity in the treatment of breast
cancer and lung cancer through immunomodulatory activity and anti-
proliferative activity.3Other study showed that ethanolicextract of Impatiens
balsamina with doses of 250 and 500 mg/kg bw have immunosuppressive
activity. It also had lower activity in Adjuvant Induced Arthritis (AIA) animal
model, best percent inhibition is shown by Ethanolic Extract of Impatiens
balsamina dose 500 mg/kg bw. Ethanolic Extract of Impatiens balsamina
also has the potential to be developed as an alternative medicine for the
treatment of rheumatoid arthritis.4The purpose of this study is to observe
the imunomodulator activity of dichloromethane fraction of pacar air
(Impatiens Balsamina L.) and to determine its effectivedose.

MATERIAL AND METHODS

Animal Used
Balb/C albino mice (Approx 20 to 25 gm) were procured from
Gadjah Mada University, Yogyakarta.
Plants Preparation
Stems and leaves of Pacar Air (Impatiens balsamina Linn.) were
used in ratio of 3:1. Samples were collected from Pontianak, West
Kalimantan and determinated at Biology Laboratory of Mathematics and
Natural Sciences, University of Tanjungpura, Pontianak.The collected
sampleswere washed, air dried, shaped into small shape and extracted
with ethanol 96% as the solvent with maceration and then fractinated with
dichloromethane by separated funnel. Extract and fraction were
concentrated by vacuum distillation. The yield of the resulting extract and
dichloromethane fraction are 12,95% and 2,46%. Dichloromethane
fraction were suspended in CMC-Na 1% to prepare suitable dosage forms
on the experiment.
Experimental Design
Animals were divided into different groups each containing 7 animals.
Group I - Normal Control, 1% CMC-Na suspension
Group II - Standard, Levamisole, 50 mg/kg b.w, p.o
Group III - Standard, Methylprednisolone, 15 mg/kg bw, i.p
Group IV, V- DF 125, and 250 mg/kg b.w, p.o
Antigen
Sheep Red Blood Cells (SRBC) was collected from Biofarma,
Bandung, Indonesia. SRBC was washed 3-4 times with large quantity of
sterile and pyrogen free saline
Delay Type Hipersensitivity
Antigen Challenge
th
On 0 day, all groups were sensitized by 0.1 ml/10 g bw of SRBC,
th
i.p. On 6 day prior to injection, right hind footpad thickness was measured
with digital calipers. Then animals were challenged by injecting 1% SRBC
th th
(0,05 ml) into the right hind footpad. On 7 and 8 day footpad thickness
wasmeasured. Difference between prior and post challenge footpad
thickness was reported as DTH response.
Titer Antibody Response to SRBC
Experimental design was done same as mentioned in Delayed
th
typehypersensitivity model. On 5 day before challenge, blood was
withdrawn from tail vein of each animal. Blood was centrifuged, and serum
was separated. Serial two fold dilutions were made i.e. 50 l of serum was
st
added to 1 well of 96-well micro titre plate containing 50 l phosphate
buffer saline. To this 1% SRBC (50 l) dissolved in phosphate buffer
st nd
saline was mixed. From 1 well 50 l of dilutedserum was added to 2 well
containing 50 l phosphate buffer saline and 50l 1% SRBC.
th
Suchdilutions were done till 12 well. Plates were incubated at 37C for 1
hr. Highest dilution that has shown visible agglutination was considered as
haemaglutination antibody titre5,6.
Statistical Analysis
Values were expressed as mean standard deviation (SD), n=5
mice in each group. Statistical analysis was performed with one way
analysis of variance (ANOVA). All analysis and comparasion were
evaluated at 95% level (p<0,05) was considered statistically significant.
RESULT AND DISCUSSION
Phytochemical Assay
Phytochemical screening showed that dichloromethane fraction of
Impatiens balsamina had more secondary metabolite compounds.
Dichloromethane fraction revealed of the following classes of chemical
compounds : polyphenols, tanninscatechin, flavonoid, steroid/triterpenoid
and kuinon.

Titer antibody response to SRBC

Titre antibody method shown humoral immunity response activity by
B lymphocyte. B lymphocytes undergo proliferation and differentiation into
plasma cells produce antibody. The primary antibody response that was
characterized by the appearance of IgM formed in 5-10 days after the
entry of antigen (SRBC) for first time. At the second time of antigen
exposure, secondary antibody response occurs that marked the
emergence of IgG reached the peak on day 10 to day 14. 7,8

Fig. 1 Titre Score Comparison of dichloromethane fraction of Impatiens

balsamina (Linn.) with control groups

Dichloromethane fraction dose of 125 mg / kg bw (DF I) has the

value of primary and secondary antibody titers lower than the normal
control group, and showed a significant difference (p <0.05) and it wasnt
showed significant difference (p> 0.05) on methylprednisolone as positive
control group immunosuppressants. From these results can be expected
that DF I group has immunosuppressive activity similar to
methylprednisolone in this test.
Delayed Type Hypersensitivity
Delayed type hypersensitivity reaction occurs more than 24 hours
after contact with the antigen and usually lasts for several days. Delayed
type hypersensitivity reactions involving Th cell activity. The increased
activity of Th cells will increase the activity of macrophages thus increasing
the inflammatory reaction in the test is characterized by swelling of the feet
around the place after antigen injection.9
 F
ig. 2 Activity of dichloromethane fraction of Impatiens balsamina(Linn.)on
delay type hypersensitivity
Based on the study, dichloromethane fraction doses of 125 (DF I)
and 250 mg / kg bw (DF II) showed lower percentage of footpad thickness
than the normal control group with a significant difference (p <0.05). The
average percent change in thickness of paws of mice in the both doses
also showed a smaller value than methylprednisolone with a significant
difference (p> 0.5). It shows that both doses of dichloromethane fraction
have greater potential immunosuppressants compared with
methylprednisolone as positive control group immunosuppressants.
The results showed that the phytochemical screening of
dichloromethane fraction of Pacar Air containing quinone compounds.
Compounds of quinone class that suspected having immunosuppressive
activity is naftaquinoncompound. Family Balsaminaceae is one of groups
of plants as main source of naftaquinon.10
CONCLUSION
Dichloromethane fraction of Impatiens balsamina Linn. suppress
both cellular and humoral spesific immune systems based on the test by
delayed type hypersensitivity and hemaglutination antibody titre method
with dose of 125 mg / bw (DF I) as the effective dose.
Acknowledgement
Acknowledgements submitted to the research grant from Directorate
General of Higher Education, Indonesia (DIRJEN DIKTI) through
Decentralization on funding research grants awarded (HIBAH DOSEN
PEMULA) in 2015.
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