Indonesian Impression Journal (JII)
Vol. 4, No. 2, February 2025
p-ISSN: 2828-1284 e-ISSN: 2810-062x
Website: https: //rivierapublishing.id/JII/ index.php / jii /index
Doi: 10.58344/jii.v4i2.6305 1159
IN VITRO EVALUATION OF THE ANTI-INFLAMMATORY POTENTIAL OF
ETHANOL EXTRACT FROM PINK CHRYSANTHEMUM
(CHRYSANTHEMUM MORIFOLIUM)
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali3, Fona Dwiana
Hermina Budiarso4, Billy Johnson Kepel5
Sam Ratulangi University, Indonesia
gloriabaya011@student.unsrat.ac.id1, aldakussoy@yahoo.com2, fatimawali@unsrat.ac.id3,
fonabudiarso@unsrat.ac.id4, billy.kepel@unsrat.ac.id5
Keywords
Abstract
anti-inflammatory, pink
chrysanthemum, ethanol
extract, in vitro
Inflammation is the body's normal response to injury. However, acute
and chronic inflammatory reactions can lead to various diseases.
Nonsteroidal anti-inflammatory drugs (NSAIDs) have been proven
effective as anti-inflammatory agents, but excessive and long-term use
of NSAIDs can cause various side effects. Pink chrysanthemum flowers
(Chrysanthemum morifolium) are one of many medicinal plants used as
traditional medicine due to their content of compounds such as
flavonoids, triterpenoids, and steroids, which have anti-inflammatory
properties and the potential to serve as alternatives to NSAIDs. To
identify active compounds and analyze the anti-inflammatory activity of
ethanol extract from pink chrysanthemum flowers (Chrysanthemum
morifolium). This study is an experimental laboratory research. The
extraction method used was Microwave-Assisted Extraction (MAE).
The anti-inflammatory activity of the ethanol extract of pink
chrysanthemum flowers was measured using the red blood cell
membrane stabilization method. The tests were performed using a UV-
Vis spectrophotometer at extract concentrations of 50 ppm, 100 ppm,
200 ppm, 400 ppm, and 800 ppm. Pink chrysanthemum flowers contain
several phytochemical compounds, including alkaloids, flavonoids,
tannins, triterpenoids/steroids, and phenolics. The anti-inflammatory
activity measurement showed that higher extract concentrations resulted
in lower absorbance values and increased stabilization percentages,
indicating anti-inflammatory activity that protects red blood cell
membranes from damage. The ethanol extract of pink chrysanthemum
flowers contains several phytochemical compounds with potential anti-
inflammatory activity. The extract at a concentration of 800 ppm
demonstrated the highest membrane stabilization ability, with a
stabilization rate of 59.459%.
Corresponding Author: Gloria Prisca Susana Angelique Baya
E-mail: gloriabaya011@student.unsrat.ac.id
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1160
INTRODUCTION
When an injury occurs, the body releases several substances in response. These
substances include histamine, prostaglandins, cytokines, bradykinin, chemokines, leukotrienes
and serotonin (Suryandari et al., 2021). The release of substances such as histamine results in
dilation of blood vessels and increased permeability of the capillary walls so that phagocyte
cells (leukocytes) can move towards the area to destroy substances that are considered harmful
(phagocytosis) (Luthfianto et al., 2023). Excessive phagocytosis process can worsen
inflammation. It is characterized by redness, swelling, heat, pain, and loss of tissue function
(Ross et al., 2021). Inflammation is divided into two types, namely acute inflammation which
is usually followed by immediate recovery, and there is also inflammation that develops into
chronic when the causative agent is settled (Akrom, 2021).
Basically, inflammatory reactions are normal. But in fact, acute and chronic
inflammatory reactions can cause various diseases. One example is rheumatoid arthritis (Arida
et al., 2018). This disease has a reported prevalence rate of 20 percent of the world's population
according to the World Health Organization (WHO) and also reaches 7.30% in Indonesia
(Prince et al., 2015). In addition, there is another example, namely gout arthritis which is an
inflammatory arthritis. In fact, in North Sulawesi itself, especially North Minahasa, has a
prevalence rate of 29.2% (Hendayana, 2023). This prevalence also increases with age and
unhealthy lifestyle which is directly proportional to the thick customs of Minahasa such as
drinking alcoholic beverages and foods high in purines (red meat and certain seafood). Due to
the high prevalence of diseases caused by inflammatory reactions, the use of anti-inflammatory
drugs has also increased.
Administration of OAINS (nonsteroidal anti-inflammatory drugs) has been shown to be
effective as an anti-inflammatory. However, excessive and long-term use of OAINS can cause
various side effects such as cardiovascular tract disorders and kidney disorders (Arfania et al.,
2023). This is one of the factors to consider when switching to traditional medicine. This is
also related to Indonesia's nature, which is very rich in various plants that are useful as
medicinal plants. Of the 30,000 types of plants in Indonesia, there are approximately 7,000
types of plants that are considered efficacious as medicinal ingredients. In Indonesia, this herbal
treatment method has existed for generations in accordance with the prevailing customs in the
community (Adiyasa & Meiyanti, 2021).
Pink chrysanthemum (Chrysanthemum morifolium) is one of the many medicinal plants
that are used as herbal medicine by the Indonesian people (Prabawati et al., 2021). This flower
contains several compounds such as flavonoids, triterpenoids and steroids that are efficacious
as anti-inflammatory for the body. Wrong
One chemical compound in pink chrysanthemums that is beneficial to the body is
flavonoids that have anti-inflammatory, antioxidant, antiallergic, and antiviral properties (Yuan
et al., 2020). In addition, there are also triterpenoids that act as anti-inflammatory. This
triterpenoid functions to prevent the enzyme cyclooxygenase, which converts arachidonic acid
into prostaglandins (inflammatory mediators) (Wahdaniah et al., 2023).
Although many studies have been conducted on various anti-inflammatory properties of
medicinal plants, special studies on the effectiveness of pink chrysanthemum flower extract
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1161
(Chrysanthemum morifolium) by testing anti-inflammatory activity using red blood cell
stabilization method and extraction method by Microwave Assisted Extraction (MAE) method
are still very limited. This research is expected to fill this gap and make new contributions in
the field of herbal pharmacology, especially related to the potential of pink chrysanthemum
flowers (Chrysanthemum morifolium) as a potential source of anti-inflammatory compounds.
RESEARCH METHOD
This study uses laboratory experimental methods, which aims to test the anti-
inflammatory activity of ethanol extract of pink chrysanthemum flowers (Chrysanthemum
morifolium) in vitro. Study uses a quantitative approach to analyze anti-inflammatory activity
based on data from spectrophotometric testing.
Population and Sample
1. Population: Pink chrysanthemum plant (Chrysanthemum morifolium).
2. Sample: Pink chrysanthemums obtained from Kakaskasen Tiga, Tomohon City, North
Sulawesi Province, with a geographical location of 770 meters above sea level.
Data Collection Techniques
Data is obtained through a series of laboratory tests which include:
1. Phytochemical screening for identification of active compounds (flavonoids, tannins,
alkaloids, etc.).
2. Anti-inflammatory activity testing using erythrocyte membrane stabilization method with
UV-Vis spectrophotometry.
Data Analysis Techniques
1. The data from the anti-inflammatory activity test results were analyzed using SPSS
statistical software.
2. The normality test was carried out using Shapiro-Wilk, and the homogeneity test was
carried out using the Levene test.
3. One-way variance analysis (ANOVA) is used to identify significant differences between
groups.
4. If a significant difference is found, continue with the smallest real difference test (Tukey).
RESULTS AND DISCUSSION
Pink Chrysanthemum Flower Extraction Results
Pink chrysanthemum leaf simplicia powder of 98 grams was extracted by the Microwave
Assisted Extraction method which uses 96% ethanol solvent with a ratio of 1:10 between
simplicia powder and ethanol solvent. A total of 50 grams of simplicia powder is added with
500 ml of 96% ethanol solvent and then extracted using a microwave with a power of 700 watts
for 1-6 minutes. The extraction solution is irradiated at regular intervals with a 1-minute
radiation cycle and 2-minute turned off to keep the temperature no more than 80◦C. After
irradiation, the solution is left at room temperature and filtered using filter paper until it
produces a pink chrysanthemum filtrate and concentrated using a rotary vacuum evaporator.
Once the filtrate has thickened, it is placed in an oven at 46◦C. From the evaporation process,
14,805 gr of thick extract was obtained.
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1162
%Rendement=
=
Based on the calculation above, the percentage value of the pink chrysanthemum extract
was obtained of 29.61% which shows a good yield value. Because the higher the yield value,
the higher the compound of interest and the more extracts produced.
Phytochemical Screening Results
The results of phytochemical screening of pink chrysanthemums from ethanol extract
showed the presence of active compounds such as phenols, flavonoids, tannins,
steroids/triterpenoids, and alkaloids so that they have great potential to have anti-inflammatory
activity. The results of the phytochemical screening examination can be seen in the following
Table 1:
Table 1. Phytochemical Screening Results
No.
Examination
Reagents
Result
Information
1.
Alkaloids
Wagner
+
No brown deposits
formed
Mayer
-
White deposits form
Dragendorff
+
Orange deposits are
formed
2.
Flavonoids
Hcl and Mg
+
Embossed dark red
color
3.
Saponins
Aquades
-
No stable foam formation
4.
Tannins
FeCl3
+
There is a discoloration
to greenish-brown
5.
Triterpenoids/Steroids
Concentrated
H2SO4
+
There is a discoloration
become red-purple
6.
Phenolic
FeCl3
+
There is a discoloration
to blackish green
Information:
+ : contains a group of compounds
- : does not contain a group of compounds
Anti-Inflammatory Activity Test Results of Pink Chrysanthemum Ethanol Extract with
Red Blood Cell Membrane Stabilization Method
The anti-inflammatory activity of pink chrysanthemum ethanol extract was tested with various
concentration variations, namely 50 ppm, 100 ppm, 200 ppm, 400 ppm, and 800 ppm using the
red blood cell membrane stabilization method. This measurement was carried out by three
repetitions using a UV-Vis spectrophotometer with a wavelength of 576 nm. Stabilization of
red blood cell membranes has been used as a method to determine anti-inflammatory activity
in vitro.
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1163
Table 2. Results of Measurement of Absorbance Value of Pink Chrysanthemum Extract by Red
Blood Cell Membrane Stabilization Method
Sample
Concentration
(ppm)
Absorbance
Average
%Stabilization
Pink Chrysanthemum
Flower Extract
50 ppm
0,749
0,747
15,8784
0,752
0,74
100 ppm
0,464
0,484
45,4204
0,451
0,539
200 ppm
0,46
0,456
48,6111
0,456
0,453
400 ppm
0,458
0,432
51,3514
0,419
0,419
800 ppm
0,361
0,36
59,4595
0,361
0,358
Based on the measurement results, it can be seen that the greater the concentration of the
test solution, the lower the absorbance value and the higher the percent of stabilization which
indicates the presence of anti-inflammatory activity that is able to protect the red blood cell
membrane from damage.
Results of IC50 Analysis of Pink Chrysanthemum Flower Extract (Red Blood Cell
Membrane Stabilization Method)
1. IC50 Pink Chrysanthemum Flower Extract
The results of the anti-inflammatory activity test of ethanol extract of pink
chrysanthemum flowers with the red blood cell membrane stabilization method obtained the
absorbance value used to obtain the percent stabilization value. The linear regression equation
is obtained by plotting the concentration of the test solution with percent stability, where the
concentration is as the abscess and the percent absorbance is the ordinate. The equation
obtained is y = 0.0395x + 31.944 with a correlation coefficient value of 0.527.
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1164
Figure 1. Graph of Anti-Inflammatory Activity Test Results of Pink Chrysanthemum Flower Extract
In the linear regression equation y = ax + b, y is the % of anti-inflammatory activity, a is
the slope setting, b is the intercept setting, and x is the concentration of the test solution. The
IC50 value of pink chrysanthemum flower extract was obtained through a linear regression
equation, with the y variable being 50 and the x variable being the IC50 value which is
interpreted as the sum of the concentration of the solution that evaluates and compares the
effectiveness of an extract in inhibiting hemolysis. Anti-inflammatory activity is categorized
as very active if the IC₅₀ value is less than 50 μg/mL, active if it is in the range of 50-100
μg/mL, medium if it is in the range of 101-250 μg/mL, weak if it is in the range of 251-500
μg/mL, and inactive if the IC₅₀ value is greater than 500 μg/mL.
The following is the calculation of the IC50 value of ethanol extract of pink chrysanthemums:
y = ax + b
y = 0.0396x + 31.898
50 = 0.0396x + 31,898
x = (50-31,898)/0.0396
x = 457.121212 ppm
Based on these calculations, the IC50 value of ethanol extract of pink chrysanthemums
of 457.121212 ppm is included in the weak category.
2. IC50 Na Diclofenak
Figure 2. Graph of Anti-Inflammatory Activity Test Results of Diclofenac Sodium Solution
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1165
The equation obtained is y = 0.062x + 44.477 with a correlation coefficient value of
0.7923. The following is the calculation of the IC50 value of diclofenac sodium solution:
y = ax + b
50 = 0.0865x + 25,352
x = (50-25.352)/0.0865
x = 284.947977
From the two experiments, quite good results were obtained, with an increase in the
stabilization percentage as the concentration of the test solution increased. The IC50 value
obtained for chrysanthemums is 5. Pink is 457,121, while for positive control (sodium
diclofenac) is 284,947. These results show that both test solutions have potential anti-
inflammatory activity.
Results of Statistical Data Analysis
Table 3. Kruskal-Wallis Test Results
Solution
Concentration
(ppm)
p-
value
Interpretation
Pink chrysanthemum flower
ethanol extract
50
0,001
There are differences between
groups
100
200
400
800
Diclofenac Sodium
50
100
200
400
800
Table 3 shows the results of the Kruskal-Wallis test on pink chrysanthemum flower
extract and diclofenac sodium at five different concentrations. The results of the analysis
showed a p-value of 0.001, which indicated that there was a significant difference between
pink chrysanthemum flower extract and diclofenac sodium in all concentrations tested.
Table 4. Difference in Test Group Values
BP50
BP100
BP200
BP400
BP800
KP50
KP100
KP200
KP400
KP800
BP50
0,138
0,086
0,070
0,003
0,676
0,404
0,012
0,001
0,000
BP100
0,138
0,817
0,745
0,150
0,286
0,516
0,307
0,063
0,023
BP200
0,086
0,817
0,926
0,228
0,194
0,378
0,430
0,104
0,041
BP400
0,070
0,745
0,926
0,265
0,164
0,330
0,486
0,126
0,051
BP800
0,003
0,150
0,228
0,265
0,012
0,037
0,676
0,676
0,404
KP50
0,676
0,286
0,194
0,164
0,012
0,676
0,037
0,003
0,001
KP100
0,404
0,516
0,378
0,330
0,037
0,676
0,095
0,012
0,003
KP200
0,012
0,307
0,430
0,486
0,676
0,037
0,095
0,404
0,210
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1166
Table 4 shows the results of the Post Hoc Test using the Dunn's Test to find out the groups
that have significant differences. The group that had the most difference from the other group
was the 800 ppm positive control. This group has significant differences with the other 5 groups.
The 800 ppm positive control was different from the group with the extract group at 50 ppm,
100 ppm, 200 ppm and with the positive control group at 50 ppm and 100 ppm. The group that
had the least difference from the other groups was the 100 ppm and 200 ppm extract groups.
These two groups have significant differences with 1 other group. The two groups of 100 ppm
and 200 ppm extracts both had differences with the positive control group at 800 ppm.
Discussion
Pink chrysanthemums are taken as samples and go through a series of processes, from
drying to extraction, to producing a viscous extract that will be tested for its anti-inflammatory
activity. This extraction process aims to separate the compounds contained in the plant from
the mixture or simplisia. The extraction method chosen is the Microwave Assisted Extraction
method because it has the ability to speed up the extraction process so that it takes a shorter
time to get optimal results when compared to conventional methods. This extraction method
produces good extract quality even though it has a simpler and easier process (Damayanti et
al., 2021).
In the extraction process, a thick extract of 14.805 grams was obtained with a yield of
29.61%. The greater the yield value, the better the value of the extract produced. Higher yield
values indicate that more dissolved compounds are successfully extracted. High yields
generally reflect the success of extraction methods in obtaining active ingredients efficiently
(Yulianti et al., 2021). Many factors affect the yield of a viscous extract, including the type of
solvent, extraction time, temperature, particle size of the material, type and source of the
material, extraction method, and concentration of solvent and material (Senduk et al., 2020).
Based on the results of phytochemical screening, it was obtained that pink
chrysanthemum flower ethanol extract contains active compounds such as alkaloids,
flavonoids, tannins, steroids/triterpenoids, and phenolates where these compounds are able to
act as anti-inflammatory. This result is in line with research conducted by Belhachat et al.,
(2023) that the results of phytochemical screening of chrysanthemum flower extract contain
alkaloid compounds, flavonoids, triterpenoids, and tannins. Pink chrysanthemum ethanol
extract has anti-inflammatory activity because there are compounds that play a role in reducing
and relieving inflammation by inhibiting inflammatory mediators (Liu et al., 2018).
The phenol test showed positive results because a blackish-green color was formed.
Phenols are one of the main compounds that act as anti-inflammatory. Phenols make an
important contribution to anti-inflammatory activity through inflammatory mediator
inhibition mechanisms and antioxidant activity that donates hydrogen atoms, so that free
radicals are reduced to a more stable form. In other words, it can reduce inflammation caused
by oxidative stress (Lumintang et al., 2022).
KP400
0,001
0,063
0,104
0,126
0,676
0,003
0,012
0,404
0,676
KP800
0,000
0,023
0,041
0,051
0,404
0,001
0,003
0,210
0,676
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1167
The flavonoid test showed positive results because there was a brick red/dark red color.
One of the functions of flavonoids is to help regulate the immune response by suppressing
excessive activity in immune cells such as macrophages and T lymphocytes (Fitri & Putra,
2021). The tannin test showed a positive result because there was a greenish-brown color.
Tannin compounds have an OH group whose hydrogen atoms can be donated to free radicals
so that they become more stable forms thereby reducing oxidative stress that contributes to
the inflammatory process (Novika et al., 2021).
The steroid and triterpenoid tests showed positive results because there was a purplish-
red color. Steroids/triterpenoids can inhibit the production of inflammatory mediators such as
TNF-α and IL-6. These compounds can lower levels of pro-inflammatory cytokines in
experimental models, thereby helping to relieve inflammatory symptoms (Rezeki et al., 2023).
The alkaloid test showed positive results because there were orange deposits in the
Dragendorff reagent and brown precipitates in the Wagner reagent. However, in the Mayer
reagent there is no white deposit. Alkaloids can modulate immune system activity, lowering
hyperactive responses that trigger inflammation (Anggarani et al., 2023).
The anti-inflammatory activity test uses the red blood cell membrane stabilization
method. In the red blood cell membrane stabilization method, it is used to evaluate anti-
inflammatory activity in vitro because the red blood cell membrane has a structural similarity
to the lysosomal membrane. Lysosomal membranes play an important role in inflammation,
and their stabilization can prevent the release of enzymes from within the lysosomes during
the inflammatory process. The rupture of the lysosomal membrane will release the enzyme
phospholipase, which then converts the phospholipid into arachidonic acid. This acid further
triggers the formation of prostaglandins, which act as mediators in the inflammatory process
(Wahdaniah et al., 2023). The release of this enzyme, which often occurs due to neutrophil
activation, can cause tissue damage associated with acute and chronic inflammation (Saepudin
et al., 2024).
In the anti-inflammatory test, five main solutions are used, namely isosaline solution,
hyposaline, phosphate buffer, red blood cell suspension, and test solution. The results show
that the negative control absorbance value is 0.888. Meanwhile, the positive control
absorbance for concentrations of 50 to 800 ppm consecutively was 0.721; 0,667; 0,388; 0,215;
and 0.133. For the test solution with pink chrysanthemum flower extract, the absorbance value
obtained consecutively was 0.747; 0,484; 0,456; 0,432; and 0.36. These results show that the
absorbance value of the test solution is quite good, with a consistent increase in effectiveness
along with increasing concentration.
The anti-inflammatory activity of the ethanol extract of pink chrysanthemum flowers
was shown in the decrease in absorbance value in the test solution mixture. A decrease in
absorbance signals the occurrence of lower hemolysis, which reflects a higher anti-
inflammatory activity of the sample (Wasiaturrahmah & Amalia, 2023). The absorbance of the
solution was measured at a wavelength of 576 nm to detect hemoglobin released by lysis of
red blood cells. Diclofenac sodium is used as a positive control because it is a nonsteroidal
anti-inflammatory drug (NSAID) that works by inhibiting prostgland synthesis (Dr . Apt .
Diana Laila Ramatillah, 2020). This mechanism involves inhibiting the enzyme
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1168
cyclooxygenase (COX) responsible for the synthesis of prostaglandins, compounds that play
a role in the inflammatory process. In addition, diclofenac sodium was chosen because of its
widespread use as an anti-inflammatory agent and its easy availability (Rodwell et al., 2015).
The results showed that the ethanol extract of pink chrysanthemum flowers at a
concentration of 800 ppm had the highest ability in stabilizing red blood cell membranes with
a stabilization rate of 59.459%. In contrast, the lowest concentration, which is 50 ppm, only
produces a stabilization of 15.878%. These findings indicate that the increase in extract
concentration is directly proportional to the ability of red blood cell membranes to stabilize.
The higher the concentration, the greater the protection it provides against the red blood cell
membrane, which also reflects the greater potential for anti-inflammatory activity. This
analysis is also supported by statistical testing. The analyzed data included the stability of red
blood cell membranes in diclofenac sodium solution and pink chrysanthemum ethanol extract
with concentrations of 50, 100, 200, 400, and 800 ppm.
After obtaining absorbance data from UV-Vis spectrophotometry, calculations were
carried out to obtain the stabilization percentage (%stability) and IC50 value. For positive
control (sodium diclofenac), the percentage stabilization of concentrations of 50 to 800 ppm
consecutively was 18.768%; 24,811%; 56,230%; 75,712%; and 84.984%. As for pink
chrysanthemum flower extract, the value of the consecutive stabilization percentage at the
same concentration was 15.878%; 45,420%; 48,611%; 51,351%; and 59.459%. This result is
considered good if the % stabilization value of the pink chrysanthemum flower extract
approaches or even exceeds the value produced by the positive control, in this case sodium
diclofenac, which is used as the standard anti-inflammatory drug. This indicates the potential
for competitive anti-inflammatory activity of pink chrysanthemum flower extract.
After obtaining the stabilization %, the IC50 value was calculated to determine the
stabilization ability of the membrane against lysis at a concentration of 50%. The results
showed that the IC50 value for the positive control (sodium diclofenac) was 284.947977,
while for the pink chrysanthemum flower extract was 457.121212. The IC50 value of pink
chrysanthemum flower extract indicates the presence of anti-inflammatory activity. However,
this difference in IC50 values indicates that the anti-inflammatory activity of pink
chrysanthemums is weaker compared to the positive control of sodium dichlorofenac.
The results of the analysis from SPSS using the normality test showed that the data was
not normally distributed, with a significance value of 3 out of 5 concentrations greater than
0.05. Based on these results, the next step is to use the Kruskal-Wallis non-parametric test,
because the data distribution is abnormal. The Kruskal-Wallis test produced an asymptotic
significance value of 0.011, which indicates a significant difference between the
concentrations tested. Therefore, this test can be continued with the Post-Hoc test using the
Dunn-Bonferroni method.
The results of the Post-Hoc test showed a significance value of less than 0.05, which
indicates that there is a significant difference between the concentrations tested. Thus this
indicates that there is a significant difference between the concentrations of each test solution.
Gloria Prisca Susana Angelique Baya1, Aaltje Ellen Manampiring2, Fatimawali Fatimawali3, Fona
Dwiana Hermina Budiarso4, Billy Johnson Kepel5
In Vitro Evaluation of the Anti-Inflammatory Potential of Ethanol Extract from Pink Chrysanthemum
(Chrysanthemum morifolium)
Journal of Indonesian Impressions (JII) Vol. 4, No. 2, February 2025 1169
CONCLUSION
This study shows that the ethanol extract of pink chrysanthemum flower
(Chrysanthemum morifolium) contains a variety of active compounds, including alkaloids,
flavonoids, tannins, steroids/triterpenoids, and phenolates, which have the potential as anti-
inflammatory agents. Based on testing using the erythrocyte membrane stabilization method,
the anti-inflammatory activity of the extract increased along with the increase in concentration.
The highest concentration tested, which was 800 ppm, showed the highest level of red blood
cell membrane stabilization of 59.459%. However, the IC50 value obtained at 457.12 ppm
indicates that the ethanol extract of pink chrysanthemum flower has anti-inflammatory activity
in the weak category. These findings indicate that pink chrysanthemums may be an alternative
source of natural anti-inflammatory compounds, although more research is needed to improve
their effectiveness and optimize their potency compared to positive controls such as sodium
diclofenac.
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