DOI: 10.22270/jmpas.V10I6.1608 ISSN NO. 2320–7418
Journal of medical pharmaceutical and allied sciences, Volume 10 - Issue 6, 1608, November - December 2021, Page – 3874 - 3877 3874




Research articles
Phytochemical properties of methanol and ethyl acetate extracts of cananga odorata flowers and their
pharmacological activities

Sarah Niaci
1
, Rosnani Nasution
1
*, Nurdin Saidi
1
, Muhammad Bahi
1
, Marianne Marianne
2


1.
Universitas Syiah Kuala, Banda Aceh, Indonesia
2.
Sumatera Utara University, Medan, Indonesia
ABSTRACT
This work aimed to investigate the phytochemical contents of methanol as well as ethyl acetate extracts from Cananga odorata flower and
their activities against a skin acne bacterium - Propionibacterium acnes. Herein, methanol extract of C. odorata was sequentially partitioned using n-
hexane and ethyl acetate as solvents. Each extract was studied for its antibacterial activity against P. acnes bacteria. Antibacterial testing was
conducted based on disc diffusion assay technique, where the discs were dripped with several variants of concentrations of C. odorata flower extract
ranging from 0.5, 2, 4, 6, and 8%. The results showed that the ethyl acetate extract had a higher inhibition zone than the methanol extract. At a
concentration of 0.5% ethyl acetate extract the diameter of the inhibition zone formed was 7.3 mm and at a concentration of 8% it was 14 mm. While
the methanol extract at a concentration of 0.5% did not show antibacterial effect, but was active at a concentration of 8% with the inhibition diameter
reaching 8.3 mm. The effect of extract concentration on the diameter of the inhibition zone is concentration-dependent. In conclusion, the methanol
and ethyl acetate extracts of C. odorata flowers had weak inhibitory activities against P. acnes.
Keywords: Cananga odorata, biological activity, bacterial inhibition, Propionibacterium acnes, medicinal plant.
Received - 20-09-2021, Reviewed - 02/10/2021, Revised/ Accepted- 26/10/2021
Correspondence: Rosnani Nasution*  [email protected]
Universitas Syiah Kuala, Banda Aceh, Indonesia


INTRODUCTION
Skin diseases such as acne are sometimes considered
unimportant, even trivial when compared to other organ diseases.
[1]
.
The cause of acne could be stem from 4 factors that may influence
each other, namely changes in follicular keratinization,
Propionibacterium acnes colonies, increased sebum production and
inflammation.
[2,3]
. P. acnes bacteria could produce several
inflammatory substances that induce acne development
[4]
. In general,
acne appears due to the presence of P. acnes bacteria on the facial
skin
[5]
. These bacteria will enter the pores of the skin that are
clogged with dust and eventually cause acne. Before the bacteria
growth becoming uncontrollable on the facial skin, prevention and
treatment should be carried out.
The preventive treatment could be conducted either by
applying healthy diet (i.e. consuming low-fat foods) or by using
medicines. Generally, the commercial drugs use a lot of chemicals
that contain sulfur and other astringent elements that potentially cause
side effects on the skin
[2]
. Hence, the employment of natural
ingredients in the treatment is required, where plant extracts have
been reported to be capable of preventing the development of acne
associated with P. acnes bacteria.
[6, 7]
. The utilization of natural

compounds deriving from plants as curative agents are welcomed in
general medical practices.
[8, 9]
.
One of the medicinal plants that can be used to manufacture
natural medicines and cosmetics is Cananga odorata – a member of
family Annonaceae. Its flower has several properties including as a
medicine for skin diseases, asthma, mosquito repellent, antibacterial,
antioxidant and antidepressant
[10]
. C. odorata flowers contain
flavonoids and saponins which are antibacterial and anti-
inflammatory. Flavonoids are the most dominants in phenolic
compounds possessing effective properties to hamper the growth of
microorganisms
[8,11]
. Based on previous reports, the bark of the plant
C. odorata has antifungal, antibacterial, and cytotoxic activities
[12]
.
The stem bark of C. odorata has high bacteriostatic activity towards
P. acnes bacteria
[13]
. Furthermore, C. odorata leaves contain
saponins, flavonoids, and polyphenols which are thought to be useful
as antimicrobials
[14]
. Lastly, C. odorata flower oil has antimicrobial,
antioxidant, antifungal and antidepressant activities
[15]
.
Several other studies related to C. odorata flowers have
been carried out, including the extraction of C. odorata flowers as
aromatherapy liquid soap
[16]
, C. odorata oil extraction for insect

DOI: 10.22270/jmpas.V10I6.1608 ISSN NO. 2320–7418
Journal of medical pharmaceutical and allied sciences, Volume 10 - Issue 6, 1608, November - December 2021, Page – 3874 - 3877 3875
repellent lotion
[17]
, and C. odorata flower essential oil extraction for
perfume and mosquito repellent lotion
[18]
. The antibacterial activity
test of C. odorata flowers against the growth of Staphylococcus
aureus bacteria has also been reported. However, so far, researches on
C. odorata flowers against P. acnes bacteria is still relatively scarce.
This is the novelty of our study, where we investigated the biological
activities of extracts from C. odorata against P. acnes bacteria. It is
considered strategic since C. odorata flowers have high antibacterial
activity, which allows them to be used as natural medicines and
cosmetics
[19]
.
MATERIAL AND METHOD
Material and Bioindicator
In this study, the apparatus used included glass utensils
such as maceration containers, separating funnels, glass funnels,
beakers, Erlenmeyer, ose needles, micro pipettes, gauze, discs, filter
paper and other supporting tools, then a rotary evaporator (Buchi R-
100), distillation apparatus (Ruchi), Ultraviolet spectrophotometer
(AE-S60-2UP UV), incubator (Memmert), analytical balance
(CPA22AS Sartorius), oven (Memmert), Laminar flow (Esco).
As for the materials, this work required n-hexane, ethyl
acetate and methanol solvents; the solvents used for extraction were
technical solvents. Phytochemical reagents, Dragendorff (Bi (NO3)3),
Mayer (potassium tetra iodo mercurate), Wagner (I2 in KI),
Liebermann-Burchard (glacial acetic acid-H2SO4(P)). Mueller Hinton
Agar (MHA) and Nutrient Broth (NB) media. Otherwise stated, all
chemicals were procured from Merck (Selangor, Malaysia) and
analytical grade. Drugs used as positive control were purchased from
PT. Kimia Farma (Jakarta, Indonesia).
C. odorata flower samples were collected on Friday,
August 23
rd
, 2019 from Ie Masen, Banda Aceh, Aceh, Indonesia. The
determination of C. odorata plant has been carried out at the
Herbarium Laboratory of the Department of Biology, Universities
Syiah Kuala University.
The bioindicator used was the bacterium P. acnes (ATCC
27853) obtained from the Microbiology Laboratory, Sumatera Utara
University, Medan, Indonesia.
Extraction of C. odorata
Collected fresh C. odorata flowers weighing 5 kg was
cleaned using running water and dried at room temperature. Dried C.
odorata flowers (1.5 kg) were cut into small pieces and macerated
using methanol for 1x24 hours repeatedly for 3 days. Thereafter, the
sample was filtered to obtain the filtrate, and followed by evaporation
in a rotary evaporator to obtain methanol extract. The methanol
extract was partitioned with increasing polarity employing n-hexane
and ethyl acetate, where each soluble was labeled as n-hexane extract
and ethyl acetate extract. Afterward, each extract was investigated for
its antibacterial activity against P. acnes. The overall process of the
extraction could be seen in the schematic diagram presented below
(Figure 1). Qualitative phytochemical screenings were conducted
according to the previous reports.
[20,21]
.
Figure 1: Schematic diagram of C. odorata flowers extraction

Antibacterial Test of C. odorata Flower Extracts against P. acnes
Antibacterial testing of C. odorata flower extract was
conducted through disc diffusion assay, as suggested by published
work
[22]
. Bacterial suspension was made by adding 1-2 oses of P.
acnes (ATCC 27853) bacteria into NB liquid medium after which it
was incubated for 24 hours with 100 rpm shaking using a shaker. The
test bacteria suspension was equalized with the turbidity of the
McFarland solution 0.5-0.8.
A total of 20 μL bacterial suspension was dripped onto
MHA media which had been priorly solidified in a petri dish, then
leveled using a spreader. The entire plate was allowed to stand for a
while for the bacteria to reach their logarithmic phase. On the agar
media, a 6 mm diameter disc paper was placed on which the test
solution was dripped with a concentration variant of 0.5, 2, 4, 6, and
8% (w/v) of obtained extracts from C. odorata flower, the solvent of
each extract as a negative control and a solution of tetracycline
antibiotics. with a concentration of 100 g/mL as a positive control,
each was dripped on a different paper disc of 30 μL. Furthermore, the
petri dishes were incubated at 37ºC for 18 hours.

RESULT AND DISCUSSION
Phytochemical Profile of The Extract
Phytochemical tests were done to evaluate the
phytoconstituents of C. odorata flowers. The secondary metabolites
tested included alkaloids, terpenoids, steroids, flavonoids, saponins
and phenols. According to Table 1, it could be seen the methanol
extract of C. odorata flower contained secondary metabolites of
alkaloids, terpenoids, saponins and phenols. Meanwhile, extract
partitioned with ethyl acetate was found to contain terpenoids,
saponins, and phenols. Alkaloids and steroids were mostly present
in a form of plant salts, thus becomes more soluble in methanol
extract that has higher polarity. This is the reason why alkaloids and
steroids were not found in ethyl acetate sample. Based on previous
reports, C. odorata flowers contained phytocompounds such as
flavonoids, saponins, along with volatile oil components of

DOI: 10.22270/jmpas.V10I6.1608 ISSN NO. 2320–7418
Journal of medical pharmaceutical and allied sciences, Volume 10 - Issue 6, 1608, November - December 2021, Page – 3874 - 3877 3876
monoterpene and polyphenol compounds
[23]
.
Table 1. Phytochemical screening of the extracts from C. odorata flowers
Secondary metabolites Methanol Ethyl acetate
Alkaloids
Dragendorff + -
Mayer - -
Wagner - -
Steroids + -
Terpenoids + +
Flavonoids - -
Saponins + +
Phenols + +
(+) = detected; (-) = not detected
The results of phytochemical testing carried out in this
study are in line with several previous reports
[16,24-26]
. In accordance
with previous reports that the bark of C. odorata contains
flavonoids, tannins and contains essential oils such as linalool and
eugenol
[24]
. C. odorata leaves are reported to contain flavonoid
compounds, saponins and polyphenols.
[16,24]
. Meanwhile, C.
odorata flowers have been reported to contain essential oils such as
α-caryophyllene, caryophyllene, benzyl benzoate, germacrene D, as
well as α-linalool,
[25]
. Furthermore, it was also reported that the
content of flavonoid compounds, tannins, saponins, and steroids in
C. odorata flowers was also reported.
[26]
.
Antibacterial Activities of The Extract
Antibacterial activity test of C. odorata flower extract
against P. acnes bacteria was carried out to evaluate the inhibitory
effect of the extract against the tested bacteria. The selection of this
test bioindicator is because P. acnes is one of the causes of human
acne
[5]
. The bacterial test method was conducted by the disc
diffusion assay by placing the disc that had been dripped with C.
odorata flower extracts of several concentrations on the test
medium. The testing process was repeated three times. The results
of the inhibition zone of the flower extract of C. odorata against P.
acnes bacteria can be seen in Figure 2, and the results of these
investigations can be seen in the following Table 2. As for the n-
hexane extract, the results would be reported separately.
Figure 2: Inhibition zone to assess the antibacterial activities of C. odorata
flower extracts; (a) ethyl acetate and (b) methanol

Overall, our findings suggest the ethyl acetate extract
showed greater inhibitory activity than the methanol extract. Ethyl
acetate extract with a concentration of 0.5% showed an inhibition
zone diameter of 7.3 mm and at a concentration of 8% it was 14
mm. While the methanol extract at a concentration of 0.5% did not
show an inhibition zone, but the largest concentration showed an
inhibition zone diameter of 8.3 mm.
Table 2. Inhibition zone of methanol and ethyl acetate extracts from C.
odorata flowers against P. acnes
Samples
Inhibition zone (mm)
0.50% 2% 4% 6% 8%
Ethyl acetate
extract
7.33 ±
0.57
8.33 ±
0.57
8.67 ±
0.57
11 ± 1
14 ±
1
Methanol
extract
0 ± 0
6.3 ±
0.57
7±0
7.3 ±
0.57
8.3 ±
0.57
Tetracycline 20 ± 1
Solvent 0 ± 0

Tetracycline is an antibiotic used as a positive control
which is one of the antibiotics that can treat various types of
bacterial infections, one of which is P. acnes. Positive control
produces an inhibition zone of 20-22 mm. The negative control used
was the solvent of each extract, which did not show the formation of
an inhibition zone, this proves that the solvent used to extract the
sample of C. odorata flower had no effect on the growth of P. acnes
bacteria. The antimicrobial activity was categorized as strong
having inhibition zone diameter > 20 mm, moderate (16 - 20 mm),
weak (10-15 mm), very weak < 10 mm
[27]
. Therefore, in this study,
the ethyl acetate extract with a concentration of 0.5-8% had a weak
zone of inhibition category, while the methanol extract had a very
weak zone of inhibition. Differences in the antibacterial inhibition
are due to the presence of different phytocompounds drawn into
each extract sample. Weaker inhibition activity produced by
methanol extract was ascribed to the presence of inert and inactive
compounds, leading to the reduction of the concentration of the
active compounds. Consequently, ethyl acetate extract containing
less impurities (after the partitioning) would yield higher
antibacterial activity. Nonetheless, the antibacterial activities could
be further enhanced using topical deliver strategies including masks
prepared using various biopolymers
[28-32]
and lotions
[30]
.

CONCLUSION
Phytochemical screening revealed that methanolic extracts
from C. odorata flowers contained alkaloids, terpenoids, saponins,
and phenols. As for the ethyl acetate extract, it contained terpenoids,
saponins, and phenols. The P. acnes inhibitions by the methanol and
ethyl acetate extracts were very weak and weak, respectively.

REFERENCE
1. Collier C N, Harper J C, Cantrell W C, Wang W, Foster K W,
Elewski B E, 2008. The prevalence of acne in adults 20 years
and older, J Am Acad Dermatol, 58, 56-59.
2. Chularojanamontri L, Tuchinda P, Kulthanan K, Pongparit K,
2014. Moisturizers for Acne, what are their Constituents?. J Clin
Aesthet Dermatol, 7, 36-44.
3. Ahmad T, Dhama K, Tiwari R, Chaicumpa W, Hui J, 2021.
Bibliometric analysis of the top 100 most cited studies in
apolipoprotein E (ApoE) research Narra J, 1, e2.
4. Fisk W A, Lev-Tov H A, Sivamani R K, 2014. Botanical and
phytochemical therapy of acne, a systematic review, Phytother
Res, 28, 1137-1152.
5. Dessinioti C, Katsambas A D, 2010. The role of
Propionibacterium acnes in acne pathogenesis, facts and

DOI: 10.22270/jmpas.V10I6.1608 ISSN NO. 2320–7418
Journal of medical pharmaceutical and allied sciences, Volume 10 - Issue 6, 1608, November - December 2021, Page – 3874 - 3877 3877
controversies, Clin Dermatol, 28, 2-7.
6. Hamdy A A, Kassem H A, Awad G E A, El-Kady S M, Benito
M T, Doyagüez E G, 2017. In-vitro evaluation of certain
Egyptian traditional medicinal plants against Propionibacterium
acnes, S Afr J Bot, 109, 90-5.
7. Alnabati N A, Al Hejin A M, Noor S O, Ahmed M M M, Abu-
Zeid M, Mleeh N T, 2021. The antibacterial activity of four
Saudi medicinal plants against clinical isolates of
Propionibacterium acnes, Biotechnol Equip, 35, 415-424.
8. Hasballah K, Sarong M, Rusly R, Fitria H, Maida D R,
Iqhrammullah M, 2021. Antiproliferative Activity of
Triterpenoid and Steroid Compounds from Ethyl Acetate Extract
of Calotropis gigantea Root Bark against P388 Murine
Leukemia Cell Lines, Scientia Pharmaceutical, 89 (2), 21.
9. Vimaladevi K, Selladurai M, Poonkodi K, Prabhu V, Mini R,
Manoj Kumar B, 2021. Chemical Composition of Essential Oil
of Cananga odorata (Lam.) Hook F & Thomson Leaves and Its
Biological Activities, J Essent Oil-Bear Plants, 24, 596-602.
10. Juca M M, Cysne Filho F M S, de Almeida J C, Mesquita D D
S, Barriga J R M, Dias K C F, 2020. Flavonoids biological
activities and therapeutic potential, Nat Prod Res, 34, 692-705.
11. Rahman M M, Lopa S S, Sadik G, Harun O R, Islam R,
Khondkar P, 2005. Antibacterial and cytotoxic compounds from
the bark of Cananga odorata, Fitoterapia, 76, 758-761.
12. Kusuma I W, Murdiyanto, Arung E T, Syafrizal, Kim Y U,
2014. Antimicrobial and antioxidant properties of medicinal
plants used by the Bentian tribe from Indonesia, Food Sci Hum
Wellness, 3, 191-196.
13. Kuspradini H, Putri AS, Sukaton E, Mitsunaga T, 2016.
Bioactivity of Essential Oils from Leaves of Dryobalanops
Lanceolata, Cinnamomum Burmannii, Cananga Odorata,
Scorodo carpus Borneensis Agric Sci Procedia, 9, 411-418.
14. Yunita D, Aisyah Y, Maulidya R, 2019. Quality Characteristics
and Antibacterial Activity of Transparent Solid Soap with
Addition of Cananga Oil (Cananga odorata), In Proceedings of
the 2
nd
International Conference of Essential Oils, 1, 112-118.
15. Tan L T, Lee L H, Yin W F, Chan C K, Abdul Kadir H, Chan K
G, 2015. Traditional Uses, Phytochemistry, and Bioactivities of
Cananga odorata (Ylang-Ylang), Evid Based Complement
Alternat Med, 2015, 896314.
16. Soonwera M, Phasomkusolsil S, 2015. Efficacy of Thai herbal
essential oils as green repellent against mosquito vectors, Acta
Trop, 142, 127-130.
17. Pavela R, Maggi F, Giordani C, Cappellacci L, Petrelli R,
Canale A, 2020. Insecticidal activity of two essential oils used in
perfumery (ylang Ylang & frankincense), Nat Prod Res, 35, 1-7.
18. Thielmann J, Muranyi P, Kazman P, 2019. Screening essential
oils for their antimicrobial activities against the foodborne
pathogenic bacteria Escherichia coli and Staphylococcus aureus,
Heliyon, 5, e01860.
19. Sembiring E N, Elya B, Sauriasari R, 2017. Phytochemical
Screening, Total Flavonoid and Total Phenolic Content and
Antioxidant Activity of Different Parts of Caesalpinia bonduc
(L) Roxb, Pharmacognosy Journal, 10, 123-127.
20. Ali M, Obaid R, Obaid R, 2019. Antibacterial Activity for Acne
Treatment through Medicinal Plants Extracts Novel Alternative
Therapies for Acne, Journal of Pure and Applied Microbiology,
13, 1245-1250.
21. Sacchetti G, Maietti S, Muzzoli M, Scaglianti M, Manfredini S,
Radice M, 2005. Comparative evaluation of 11 essential oils of
different origin as functional antioxidants, antiradicals and
antimicrobials in foods, Food Chemistry, 91, 621-632.
22. Pujiarti R T B, Wisowati, Kasmudjo K, Sunarta S, 2015.
Kualitas, Komposisi Kimia, dan Aktivitas Antioksidan Minyak
Kenanga (Cananga odorata), Jurnal Ilmu Kesehatan, 9, 1-4.
23. Putri A R Y, Putri A M, Muham A O, Anggraini S, Maisarmah
S, 2020. Analisis Kualitatif Kandungan Bunga Kenanga
(Cananga odorata) Secara Fitokimia dengan Menggunakan
Pelarut Etanol, Journal of Research and Education Chemistry, 2,
43-48.
24. Morales G, Sierra P, Mancilla A, Paredes A, Loyola L A, 2003.
Secondary metabolites from four medicinal plants from
Northern Chile antimicrobial activity & bio toxicity against
artemia salina, Jou. of the Chilean Chem. Soc., 48, 13-18.
25. Iqhrammullah M, Suyanto H, Rahmi, Pardede M, Karnadi I,
Kurniawan KH, Chiari W, Abdulmadjid SN, 2021. Cellulose
acetate-polyurethane film adsorbent with analyte enrichment for
in-situ detection and analysis of aqueous Pb using Laser-Induced
Breakdown Spectroscopy (LIBS), Environ Nanotechnol Monit
Manag, 6, e04590.
26. Safitri E, Humaira H, Murniana M, Nazaruddin N,
Iqhrammullah M, Md Sani N D, 2021. Optical pH Sensor Based
on Immobilization Anthocyanin from Dioscorea alata L, onto
Polyelectrolyte Complex Pectin-Chitosan Membrane for a
Determination Method of Salivary pH Polymers, 13, 1276.
27. Nazaruddin N, Afifah N, Bahi M, Susilawati S, Sani N D M,
Esmaeili C, 2021. A simple optical pH sensor based on pectin
and Ruellia tuberosa L-derived anthocyanin for fish freshness
monitoring F1000Research, 10, 422.
28. Fathana H, Iqhrammullah M, Rahmi R, Adlim M, Lubis S,
2021. Tofu wastewater-derived amino acids identification using
LC-MS/MS and their uses in the modification of chitosan/TiO2
film composite Chem Data Collect, 35, 100754.
29. Rahmi, Lubis S, Az-Zahra N, Puspita K, Iqhrammullah M, 2021.
Synergetic photocatalytic and adsorptive removals of metanil
yellow using TiO 2/grass-derived cellulose/chitosan
(TiO2/GC/CH) film composite, Int J Eng, 34(8), 184-191.
30. Arifin B, Nasution R, Desrianti N, Marianne M, Helwati H,
2019. Antimicrobial Activity of Hand Lotion of Flower
Mimusops elengi, Open Access Maced J Med Sci, 7, 3748-3756.
31. Rahmad R, Earlia N, Nabila C, Inayati I, Amin M, Prakoeswa C
R S, 2019. Antibacterial cream formulation of ethanolic Pliek U
extracts and ethanolic residue hexane Pliek U extracts against
Staphylococcus aureus IOP Conference Series Materials Science
and Engineering, 523, 012011.
32. Sufriadi E, Meilina H, Munawar AA, Muhammad S, Idroes R,
2021. Identification of β-Caryophyllene (BCP) in Aceh
patchouli essential oil (PEO) using gas chromatography-mass
pectrophotometry (GC-MS) IOP Conference Series Earth and
Environmental Science, 667, 012032.

How to cite this article
Sarah Niaci, Rosnani Nasution, Nurdin Saidi, Muhammad
Bahi, Marianne,

2021. Cananga odorata, biological activity,
bacterial inhibition, Propionibacterium acnes, medicinal plant,
Jour. of Med. P’ceutical & Allied. Sci. V 10 - I 6, 1608, P-
3874 - 3877. doi: 10.22270/jmpas.V10I6.1608