Article

Nanoparticles Formulated from Young Areca Nut Extract Utilizing Sodium

Alginate as a Polymer and Calcium Chloride (CaCl₂)

Indis Hilwa Rohmahdana¹, Indri Maharini^2*, Eka Novita Br Purba³, Indah Salsabilah⁴, Aulia

Hafizhah Nugroho⁵, Yulwilla Virginia⁶

^1,2,3,4,5,6Department of Pharmacy, Universitas Jambi, Jambi, 36361, Indonesia

Abstract

Nanoparticles are an innovative formulation designed to enhance the bioavailability of drugs with poor absorption

while allowing for a more targeted release of active compounds to minimize the risk of side effects. This study aims

to develop nanoparticle formulations of young areca nut extract. The ionic gelation method, utilizing 0.02% CaCl2

and 0.1% sodium alginate, was employed in the preparation process. The three formulas were developed with

different concentrations and volumes of extract. The evaluation of nanoparticles included phytochemical screening,

particle size analysis (PSA), zeta potential, % transmittance, and FTIR for functional group identification. The

characterization results of the nanoparticles from young areca nut seed ethanol extract showed that formulas F1,

F2, and F3 had particle sizes of 84.267±1.250 nm, 97.367±1.079 nm, and 82.333±0.723 nm, respectively. The

polydispersity index values ranged from 0.254±0.046 to 0.325±0.02, suggesting good particle distribution. The zeta

potential values, all below -30 mV, indicate the stability of the colloidal suspension system. FTIR analysis showed that

the young areca nut seed extract nanoparticles in all formulas contained functional groups such as alcohol, alkene,

and amide.

Keywords: CaCl₂, nanoparticles, sodium alginate, young areca nut

Graphical Abstract

*

Corresponding author

Email addresses: indri.maharini@unja.ac.id

DOI: https://doi.org/10.22437/chp.v8i2.38128

Received October 30^th2024; Accepted December 26^th2024; Available online December 31^st2024

(https://creativecommons.org/licenses/by/4.0 )

101

Chempublish Journal, 8(2) 2024, 101-108

Introduction

sodium alginate and calcium chloride can form

nanoparticle preparations with good physical

characteristics [7][8][9].

One of the leading and largest commodities in

Jambi province is areca nut. Areca nut, also

known as Areca catechu L., is a flowering plant

that belongs to the Arecaceae family and is still

one of the palmae family plants. Traditionally,

areca nut seeds have been used to manufacture

food, beverages, medicines, natural dyes, and

cosmetics. Areca seeds are known for their

therapeutic properties in health. They are used in

the treatment of various conditions such as

malaria, diarrhoea, vaginal discharge, skin

wounds, and intestinal worms. Additionally,

areca seeds are also believed to help strengthen

teeth and gums [1] [2].

Material and Methods

Materials and Instrumentations

Young areca nut (Areca catechu L.) seeds were

obtained from the Betara Sub-district, West

Tanjung Jabung Regency, Jambi Province, ethanol

p.a, ethanol 96%, deionized water, Na alginate,

CaCl2, chloroform, FeCl₃, HCl, Mg and Meyer

reaction. The instrumentation used in this

research is magnetic stirrer (DLAB), blender

(Philips), hot plate (Cimarec+), micropipette, petri

dish, stirring rod, analytical balance (Pioneer™),

FTIR (Perkin Elmer), Particle Size Analyzer (Horiba

Scientific SZ-100), Vacuum rotary evaporator

(Büchi Rotavapor R-114®), oven (Memmert), digital

pH meter (Hanna), glass tools (Pyrex).

According to research data conducted by

Fredison et al. [3], ethanol extracts from young

areca nut seeds have been shown to contain

various

bioactive

compounds,

including

alkaloids, flavonoids, tannins, saponins, and

polyphenols, all of which possess antibacterial

properties. These compounds effectively inhibit

the growth of bacteria responsible for conditions

like canker sores, particularly Staphylococcus

aureus, with inhibition zones ranging from

moderate to vigorous (6-21 mm). In addition to

their antibacterial activity, young areca nut seeds

Methods

Preparation and Extraction. The young areca nut

seeds were freshly collected, ensuring they were

unripe, intact, and green in color, with the stalk

still attached to the base of the fruit. The seeds

were separated from the fruit's skin and sliced

into thin pieces. These slices were dried in an

oven at 50°C. Once dried, the material was

ground into powder using a blender and sieved.

Subsequently, 600 grams of the powdered

material were macerated with 96% ethanol at a

1:10 ratio for 5 days. The resulting filtrate was

then concentrated using a rotary evaporator to

obtain a thick ethanol extract of the young areca

nut seeds.

demonstrate

antimutagenic

antiseptics.

potential

agents,

as

antioxidants,

astringents,

and

Developing

a

formulation in the form of

nanoparticles can enhance the effectiveness of

the delivery system for young areca nut seed

extract as an antibacterial agent. Nanoparticles

are an innovative preparation with the main

advantage of increasing the poor bioavailability

of drugs and releasing active substances that are

more targeted to reduce the risk of side effects.

In this case, Nanotechnology has a fast and

precise drug delivery system because it has a

molecular size of <1000 nm [4][5][6].

Phytochemical Screening. Young areca nut extract

was identified by conducting phytochemical

screening,

including

identifying

alkaloids,

tannins, flavonoids, saponins and polyphenols.

The Preparation of young areca nut extract

solutions with 1, 3 and 5% concentrations. Weighed

1, 3, and 5 grams of young areca nut. Dissolve

each extract with 5 ml of ethanol p.a., and add

distilled water to a volume of 100.0 ml.

This study aims to formulate young areca nut

seed extract as nanoparticles using the polymer

sodium alginate (Na alginate) and calcium

chloride (CaCl₂) as a cross-linking agent. Sodium

alginate is a natural polymer and biocompatible

and biodegradable material, making it safe for

pharmaceutical formulations. The interaction of

The preparation of nanoparticles of young areca nut

extract. Young areca nut extract nanoparticles

102

Chempublish Journal, 8(2) 2024, 101-108

were prepared by mixing 0.1% Na Alginate with a

magnetic stirrer for 30 minutes. Then, 0.02%

CaCl₂was gradually added drop by drop while

stirring with the magnetic stirrer. The colloid

solution formed was sonicated for 1 hour. The

formula of nanoparticles is shown in Table 1.

than -30 mV and more significant than +30 mV

have higher stability [7].

Functional group analysis with FTIR. The young

areca nut nanoparticle extract solution was

placed on the sample holder and then analyzed

with an infrared spectrophotometer.

Table 1. Nanoparticle preparation formulation of

young areca nut seed extract

Results and Discussions

% Yield

Material

Formula (mL)

FI

1

FII

3

FIII

5

In this stage, young areca nut seeds are oven-

dried at 50°C. After drying, the simplisia is sorted

and pulverized using a blender. The resulting

powder from the young areca nut seeds is then

extracted using the maceration method with 96%

ethanol. The extract is evaporated using a rotary

evaporator to obtain a concentrated extract,

yielding 35.86%. The yield value can be

influenced by various factors, including the

solvent's polarity, the particle size of the

simplisia, solvent concentration, and soaking

time. Polar active compounds are more soluble

in polar solvents, while nonpolar compounds

dissolve better in nonpolar solvents. Choosing a

solvent with the appropriate polarity for the

Young areca nut seed

extract (1% w/v)

Sodium alginate (0.1% w/v)

CaCl₂(0.02% w/v)

5

25

5

25

5

25

Nanoparticle Characteristics of Young Areca Nut

Extract. The characteristics of nanoparticles can

be evaluated by transmittance test, particle size

distribution, polydispersity index test, zeta

potential test, and Functional group analysis with

FTIR.

Transmittance Test. The transmittance test was

performed using a UV-Vis spectrophotometer

with a maximum wavelength of 650 nm, and a

blank aquadeion was used as the reference[10].

A higher transmittance value indicates that the

particle size is decreasing [7].

active

compounds

improves

extraction

efficiency. Smaller particle sizes of the simplisia

increase the surface area in contact with the

solvent, enhancing interaction and yield. The

proper solvent concentration improves the

solubility of active compounds, thus increasing

yield. Adequate soaking time allows the solvent

to interact with the active compounds, optimizing

the yield fully. The success of separating active

compounds during extraction depends on the

polarity or solubility differences between the

active compounds and the solvent. In conclusion,

optimizing factors like solvent polarity, particle

size, solvent concentration, and soaking time is

crucial for achieving the best extraction yield [13].

Particle Size Distribution Test. Particle size

determination was carried out using the Particle

Size Analyser (PSA). This test is carried out by

measuring particles within ± 15 minutes [4].

Polydispersity Index test. The polydispersity index

describes the homogeneity of the colloidal

solution. The polydispersity index has a range of

values from 0 to 1. Values near 0 indicate a

homogeneous dispersal, while values greater

than 0.5 indicate high heterogeneity [7]. The

results of the polydispersity index can meet the

range of a good polydispersity index with a value

range of 0.1-0.6 [8].

Table 2. Rendemen Extract Results

Extract

Simplisia

Weight (g)

Extract Yield (%)

Weight

(g)

Zeta Potential test. Zeta potential was measured

using a zeta sizer. This test is used to characterize

the surface charge properties of nanoparticles.

Nanoparticles with zeta potential values smaller

Young

600 g

215.20 g

35.86%

Areca Nut

Extract

103

Chempublish Journal, 8(2) 2024, 101-108

Secondary Metabolites

The results of the phytochemical screening Table

3 showed that the ethanol extract of young areca

nut seeds contains alkaloids, tannins, flavonoids,

saponins and polyphenols. From these results, it

can be concluded that young areca nut seeds

have the potential to have the ability as an

antioxidant, antimutagenic, astringent, antiseptic

and antibacterial[3][14][15].

Young areca nut extracts were identified by

conducting

phytochemical

screening.

The

content of secondary metabolites found in young

areca nut seeds is listed in Table 3.

Table 3. Phytochemical Screening Results

Secondary

Metaboles

Alkaloids

Tannins

Flavonoids

Saponins

Methods

Results

Nanoparticle Characteristics of Young Areca Nut

Extract.

Mayer reaction

Chloroform+FeCl₃

HCl+Mg

+

The characteristics of nanoparticles can be

evaluated

by

testing

the

percentage

Chloroform+FeCl₃

transmittance value, particle size distribution,

polydispersity index test, zeta potential test, and

Functional group analysis with FTIR.

Polyphenols Chloroform+FeCl₃

Description: (+): Positive; (-) : Negative

Table 1. Results of Transmittance Test of Nanoparticles

Formulation

%Transmittance

Average±SD

Replication I

Replication II

Replication III

F1

91.50%

93.80%

90.70%

92±1.61

F2

F3

85.80%

76.50%

82.00%

75.10%

74.40%

67.20%

80.73±5.80

72.93±5.01

The characterization of % transmittance was

carried out to observe nanoparticle formation

indirectly. In general, nanoparticles have a cloudy

appearance. Based on the transmittance values

in Table 4, % transmittance values smaller than

100% indicate the formation of an opaque

solution due to ionic interactions between

Alginate and CaCl₂. When the calcium chloride

solution interacts with the sodium alginate

solution, bonds form between the Ca²⁺ ions from

calcium chloride and the carboxylate groups of

sodium alginate, resulting in a polyelectrolyte

complex due to opposing electrostatic forces,

forming an egg-box structure. The Ca²⁺ ions will

bind with the guluronate acid carboxyl groups of

alginate [10,16,17].

it can be observed that higher concentrations of

young areca seed extract in the nanoparticle

formulation result in lower % transmittance

values. However, this does not align with the

particle sizes observed. Formula 3, despite

having a higher extract concentration, shows the

smallest particle size, which could be attributed

to the effect of sonication. On the other hand,

Formula 2 has the largest particle size, likely due

to impurities in the young areca nut extract

solution.

The polydispersity index represents the particle

size distribution. A lower polydispersity index

value indicates a more stable formulation, as

higher values suggest that the particles are

uneven in size, which can lead to quicker

flocculation of the formula. The polydispersity

index is close to 0.01, indicating that the formed

dispersion system tends to be stable for a long

time [11], [12]. The results of the polydispersity

The particle size test was analyzed using a

particle size analyzer. The evaluation aims to

determine the particle size formed, indicated by

the z average value. All of the formulation meets

the nanoparticle size requirements as they fall

within the 1-1000 nm range (Table 5). Formula 3

exhibits the smallest particle size. When

compared to the % transmittance data in Table 4,

index can meet the range of

a

good

polydispersity index, with values ranging from

0.254 – 0325 (Table 5 and Figure 1). The

polydispersity index value for Formula 2 is higher

104

Chempublish Journal, 8(2) 2024, 101-108

than that of the other formulas, suggesting that

its particle size distribution is more varied

compared to the other two formulas.

forces within the nanoparticle dispersion help

prevent aggregation. Conversely, a low zeta

potential leads to attractive forces, which may

cause the dispersion to break or flocculate.

Nanoparticles with a zeta potential greater than

+30 mV or less than -30 mV are typically regarded

as stable colloidal suspension systems. Based on

the measurement results (Table 5 and Figure 2),

the zeta potential value ranges from -31.033 to -

34.733 mV, indicating that the sample is stable

[18][19].

Another parameter that was analyzed is zeta

potential, which is crucial for predicting the

stability of colloidal dispersions. Zeta potential

refers to the potential difference between the

phase boundaries of solids and liquids, indicating

the electric charge of particles suspended in a

liquid. A high zeta potential suggests a stable

formulation, as the repulsive and stabilizing

Table 2. Results of Z-Avarage, PI and Zeta Potential

Formulation

Z-Average

Polydispersity Index

Zeta Potential

F1

84.267±1.250

0.258±0.091

-34.733 ±0.208

-32.400±0.436

-31.033±1.106

F2

F3

97.367±1.079

82.333±0.723

0.325 ± 0.02

0.254±0.046

Figure 1. The particle size distribution of nanoparticles of young Areca nut. (a) Formula 1, (b) Formula 2,

(c) Formula 3.

105

Chempublish Journal, 8(2) 2024, 101-108

Figure 2. Zeta potential of nanoparticle young areca nut. (a) Formula 1, (b) Formula 2, (c) Formula 3.

Fourier transform infrared (FTIR) spectroscopy is

an analytical technique that utilizes molecular

vibrational spectra in the infrared range to

characterize nanoparticles' chemical properties

and molecular structure. In this case, FTIR can

provide information related to chemical bonding,

surface functionality, and the presence of certain

compounds in nanoparticle preparations. Some

aspects that can be characterized using FTIR for

nanoparticles are compound identification,

chemical bond analysis, surface functionality,

compound purity, chemical reactions and

synthesis, and phase change monitoring [20].

This study used an FTIR (Fourier Transform

Infrared) spectrophotometer to identify the

functional groups contained in the ethanol

extract of young areca nut seeds with their

bioactive content. The following are the results of

the FTIR test analysis data in Figure 3.

The functional groups observed in the 3200-3600

wavelength

hydroxyl groups (O-H). In the 1600-1680

wavelength range, the functional groups

range

correspond

to

alcohol

identified are alkenes with double C bonds (C=C),

while at wavelengths between 1550-1640,

amines and amides (N-H) are detected (Fig. 1).

These results indicate that the young areca nut

extract nanoparticles contain alcohol, alkene,

and amide groups. Flavonoids and alkaloids,

secondary metabolite compounds, can form in

the presence of O-H, N-H, and C=C functional

group bonds. Flavonoids typically contain

structures with double bonds and hydroxyl

groups, whereas alkaloids may include amine (N-

H) groups and various other structures. Both

flavonoids and alkaloids are known for their

significant biological activities and are commonly

found in a variety of plants [21].

Figure 3. IR spectrum of nanoparticle formulas

106

Chempublish Journal, 8(2) 2024, 101-108

Conclusion

https://epublikasi.pertanian.go.id/berkala/

wartabun/article/view/3460

The nanoparticle formulation of young Areca nut

extract can be prepared using 0.1% sodium

alginate as the polymer and 0.02% calcium

chloride as a crosslinking agent. Formula 3

produced the smallest particle size, measuring

82.333 ± 0.723 nm, with a polydispersity index of

0.254 ± 0.046, indicating uniform particle size

distribution, and was found to be stable.

[2]

L. Nurdiana and R. N. Hardiyanti. (2022).

“Konsumsi Biji Pinang dengan Kejadian

Keputihan pada Wanita Usia Subur di

Wilayah Kerja Puskesmas Rum Kota Tidore

Kepulauan.”

Media

Publikasi

Promosi

Kesehatan Indonesia (MPPKI). 5 12:. 1626–

1632. 10.56338/mppki.v5i12.2892.

Fredison, R. Triyadi, M. Iqbal, D. A. Ramdini,

and Suharmanto. (2023). “Kajian Potensi

Biji Pinang (Areca catechu L.) sebagai

Antibakteri.” JK Unila. 7 1:. 51–59.

[3]

[4]

Acknowledgement

S. Nabillah, Noval, and N. Hidayah. (2022).

“Formulasi dan Evaluasi Nano Hidrogel

This research is supported by the Directorate of

Learning and Student Affairs (Belmawa) of the

Ministry of Education, Culture, Research and

Technology (Kemdikbudristek). We sincerely

thank the Directorate of Learning and Student

Affairs (Belmawa) and Jambi University for

providing support and assistance through

resources, facilities, and technical support during

this research process. Thank you to all team

members and parties who have helped make this

activity possible. Without the support of various

parties, this research would not have been

realized.

Ekstrak

Daun

Serunai

(Chromolaena

odorata L.) dengan Variasi Konsentrasi

Polimer Carbopol 980.” Jurnal Ilmiah Ibnu

Sina (JIIS): Ilmu Farmasi dan Kesehatan. 7 2:.

340–349. 10.36387/jiis.v7i2.995.

Z. Hami. (2021). “A Brief Review on

Advantages of Nano-based Drug Delivery

Systems.” Annals of Military and Health

[5]

Sciences

Research.

19

1:.

1–6.

10.5812/amh.112274.

[6]

[7]

M. Ozdal and S. Gurkok. (2022). “Recent

advances in nanoparticles as antibacterial

agent.” ADMET and DMPK. 10 2:. 115–129.

10.5599/admet.1172.

A. Ngafif, E. D. Ikasari, and L. W. Ariani.

(2020). “Optimasi Natrium Alginat Dan

Kalsium Klorida (Cacl2) Sebagai Agen

Sambung Silang Nanopartikel Ekstrak

Etanol Daun Katuk (Sauropus Androgynus

(L.) Merr).” Berkala Ilmiah Mahasiswa

Farmasi Indonesia (BIMFI). 7 2:. 13–23.

10.48177/bimfi.v7i2.33.

Author Contributions

Conceptualization, I. H. R. and I. M.; Methodology,

I. H. R. and I. M.; Software, I. S. and A. H. N.;

Validation, I. M.; Formal Analysis, Y. V. and A. H.

N.; Investigation, I. H. R.; Resources, I. H. R., I. S.

and E. N. B. P.; Data Curation, I. M.; Writing –

Original Draft Preparation, I. H. R.; Writing –

Review & Editing, I. S., I. M. and E. N. B. P.;

Visualization, I. S.; Supervision, I. M.; Project

Administration, I. H. R., I. S. and E. N. B. P.

[8]

[9]

S. Kumar and D. C. Bhatt. (2021). “Influence

of Sodium Alginate and Calcium Chloride

on the Characteristics of Isoniazid Loaded

Nanoparticles.”

Research

Journal

of

Conflic of Interest

Pharmacy and Technology. 14 1:. 389–396.

10.5958/0974-360X.2021.00071.8.

N. Wathoni, Y. Herdiana, C. Suhandi, A. F. A.

Mohammed, A. El-Rayyes, and A. C. Narsa.

The authors declare no conflict of interest

(2024).

Nanoparticles for Antibacterial Agents

Delivery.” International Journal of

5021–5044.

“Chitosan/Alginate-Based

References

[1]

K. Suheiti, H. Ardi, I. W. Putri, and D.

Medionovianto. (2023). “Panen Dan

Pascapanen Pinang Betara.” Warta BSIP

Nanomedicine.

19.

10.2147/IJN.S469572.

Perkebunan.

Available:

1

3:. 16–20.

[Online].

[10] W. C. G. Budastra, W. Hajrin, and D. G.

107

Chempublish Journal, 8(2) 2024, 101-108

Wirasisya. (2022). “Pengaruh Kecepatan

Pengadukan terhadap Karakteristik

Nanopartikel Sari Buah Juwet (Syzygium

cumini L.).” Unram Medical Journal. 11 3:.

1000–1006. 10.29303/jk.v11i3.4742.

[18] Yunida, M. T. Kamaluddin, Theodorus, and

S. Mangunsong. (2021). “Formulasi dan

Karakterisasi Nanopartikel Kafein Hasil

Isolasi dari Biji Kopi Robusta.” Jurnal

Mandala Pharmacon Indonesia. 7 1:. 47–59.

10.35311/jmpi.v7i1.68.

[11] I. Maharini, F. Fitrianingsih, and D. T. Utami.

(2023). “Green synthesis nanosilver using

[19] M. K. Waqas, S. Safdar, M. Buabeid, A.

Ashames, M. Akhtar, and G. Murtaza.

dadap

serep

(Erythrina

Subumbrans

(Hassk.) Merr) stem extract Green.” Journal

of Pharmaceutical and Sciences. 1:. 255–259.

10.36490/journal-jps.com.v6i5-si.410.

(2022).

“Alginate-coated

chitosan

nanoparticles for pH-dependent release of

tamoxifen citrate.” Journal of Experimental

Nanoscience.

[12] N. Auliasari, R. F. Azzahra, and N. Aji. (2024).

17

1:.

522–534.

“Volume 29 No 2 , Agustus 2024.” Formulasi

10.1080/17458080.2022.2112919.

Dan

Karakteristik

α-Mangostin

Dengan

[20] O. Bouzekri et al. (2023). “Green Synthesis

of Nickel and Copper Nanoparticles Doped

with Silver from Hammada scoparia Leaf

Extract and Evaluation of Their Potential to

Inhibit Microorganisms and to Remove

Variasi Kitosan-TPP Sebagai Polimer. 29 2:.

168–180.

[13] D. L. Y. Handoyo. (2020). “The Influence Of

Maseration Time (Immeration) On The

Vocity Of Birthleaf Extract (Piper Betle).”

Dyes

Sustainability

from

Aqueous

(Switzerland)

Solutions.”

15 2:.

Jurnal Farmasi Tinctura.

2

1:. 34–41.

.

10.35316/tinctura.v2i1.1546.

10.3390/su15021541.

[14] S. C. Rangani, R. A. U. J. Marapana, G. S. A.

Senanayake, P. R. D. Perera, M. M.

Pathmalal, and H. K. Amarasinghe. (2023).

[21] A. Kresnamurti, F. Izazi, and D. Camelia.

(2022). “Standarisasi dan Analisis Ftir

Ekstrak Etanol 70% Bulu Babi (Echinometra

mathaei) dari Sabang, Nanggroe Aceh

Darussalam.” Farmasainsꢀ: Jurnal Ilmiah

“Correlation

analysis

of

phenolic

compounds, antioxidant potential, oxygen

radical scavenging capacity, and alkaloid

content in ripe and unripe Areca catechu

from major cultivation areas in Sri Lanka.”

Ilmu

Kefarmasian.

9

1:.

1–8.

10.22236/farmasains.v9i1.7200.

Applied Food Research.

3

2:. 1–11.

10.1016/j.afres.2023.100361.

[15] H. Sun, W. Yu, H. Li, X. Hu, and X. Wang.

(2024). “Bioactive Components of Areca

Nut: An Overview of Their Positive Impacts

Targeting Different Organs.” Nutrients. 16

5:. 1–23. 10.3390/nu16050695.

[16] P. Maharani, E. Ikasari, U. Purwanto, and I.

Bagiana. (2022). “Optimasi Na-Alginat dan

Ca-Klorida Pada Nanopartikel Ekstrak

Terpurifikasi Fukoidan Dari Rumput Laut

Cokelat (Sargassum Polycystum).” Jurnal

Farmasi Medica/Pharmacy Medical Journal

(PMJ).

5

2:.

38–45.

10.35799/pmj.v5i2.45100.

[17] C. Bennacef, S. Desobry, J. Jasniewski, S.

Leclerc, L. Probst, and S. Desobry-Banon.

(2023). “Influence of Alginate Properties

and Calcium Chloride Concentration on

Alginate Bead Reticulation and Size: A

Phenomenological Approach.” Polymers. 15

20:. 1–15. 10.3390/polym15204163.

108