Kuwait Journal of Science 51 (2024) 100252

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Kuwait Journal of Science

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Full Length Article

Effect of chitosan concentration on hydroxyapatite/chitosan composite

synthesis using the in-situ method as a dye adsorbent
Novesar Jamarun a, * , Arika Prasejati a , Zulhadjri Zulhadjri a , Sintia Caniago a ,
Tri Yupi Amirullah a , Wulandari Wulandari a , Vivi Sisca b
a
Department of Chemistry, Faculty of Mathematics and Natural Sciences, Andalas University, Padang, Indonesia
b
Research Centre for Chemistry, National Research and Innovation Agency, B.J. Habibie Science and Techno Park, Serpong, South Tangerang, Banten, 15314, Indonesia

A R T I C L E I N F O A B S T R A C T

Keywords: Hydroxyapatite (HAp) is a versatile material with wide-ranging applications, including its utility as an adsorbent.
Hydroxyapatite-chitosan composite However, its brittleness and limited adsorption capacity pose challenges, prompting the incorporation of chi­
In-situ method tosan (CTS) fillers to enhance mechanical strength and adsorption properties. The hydroxyapatite/chitosan
Cuttlebones
(HAp/CTS) composite was synthesized via the in-situ method, utilizing natural sources such as calcium ions from
Adsorption
Rhodamine B
cuttlefish bones and chitosan extracted from shrimp shells. Various chitosan concentrations (10, 20, 30, 40, and
50 wt%) were explored in the synthesis process, with the optimal adsorption of Rhodamine B dye observed at a
30% concentration, yielding 0.1832 mg/g. Validation of the HAp/CTS composite synthesis was achieved through
X-ray diffraction, revealing the presence of CTS at a new peak at 2θ of 19.23◦ , while infrared spectroscopy
confirmed absorption bands for both HAp and CTS. Additionally, scanning electron microscopy-energy dipersive
spectroscopy (SEM-EDS) investigations revealed irregular shapes with agglomeration, resulting in a Ca/P ratio of
1.80 for the HAp/CTS composite. The HAp/CTS-30% composite demonstrated efficient Rhodamine B dye
adsorption following the Freundlich isotherm equation, pseudo-second-order adsorption kinetics, and exhibited
good reusability. These findings suggest that the HAp/CTS composite could serve as a promising, cost-effective
solution for treating industrial wastewater.

1. Introduction and low mechanical strength. Additionally, using HAp in a singular form
often requires substantial material. Innovations in the form of HAp
Hydroxyapatite (HAp) is a calcium phosphate-based biomaterial composites have emerged as a promising solution to address these
with the chemical formula Ca10(PO4)6(OH)2. The applications of HAp weaknesses process; hydroxyapatite has shortcomings, namely low
span across food additives and nutritional supplements, as well as brittleness or mechanical strength, and using hydroxyapatite in one
playing a role in the production of fertilizers, animal feed, dental im­ form can lead to large material requirements. Hydroxyapatite composite
plants, drug delivery applications, and biotechnology (Azzaoui et al., innovation is one of the solutions (Zou et al., 2023). One way to over­
2023b; Sambudi et al., 2016; Wulandari et al., 2023). The specific come these limitations is by incorporating polymers as fibers/fillers, and
structure of HAp provides ion exchange capabilities and high adsorption chitosan is one such natural polymer that can be used in composite
affinity towards various pollutants. Its crystal surface, rich in calcium fabrication (Mulyani et al., 2020). Chitosan can be integrated into
ions (Ca2+) and phosphate ions (PO3−4 ), gives it outstanding adsorption various composite materials for different purposes (Pooladi and
capacity (Amenaghawon et al., 2022; Bensalah et al., 2020). Sources of Bazargan-Lari, 2020). Chitosan is a hydrophilic and cationic polymer
hydroxyapatite can be obtained from natural materials such as snail obtained by deacetylating chitin groups in an alkaline medium. The
shell (Pooladi and Bazargan-Lari, 2023), seashells (Islami et al., 2023), adsorbent exhibits excellent performance in removing cations from
and cuttlebone (Anitta and Sekar, 2023), including mammalian teeth neutral environments due to the presence of hydroxyl and amine groups
and bones (Pooladi and Bazargan-Lari, 2023). in its structure. It can be easily extracted from various animal and plant
However, HAp has a primary drawback in its application: brittleness sources, including crab shells, snails, and the cell walls of certain fungi

* Corresponding author.
E-mail address: novesarjamarun@sci.unand.ac.id (N. Jamarun).

https://doi.org/10.1016/j.kjs.2024.100252
Received 24 October 2023; Received in revised form 25 April 2024; Accepted 21 May 2024
Available online 22 May 2024
2307-4108/© 2024 The Authors. Published by Elsevier B.V. on behalf of Kuwait University. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
N. Jamarun et al. Kuwait Journal of Science 51 (2024) 100252

and algae (Bambaeero and Bazargan-Lari, 2021). The chitosan used in solution was added to the CaO solution until the Ca/P molar ratio was
this study is derived from shrimp shells. Chitosan possesses non-toxic, 1.67. To make the chitosan solution, 50 mL of 2% acetic acid solution
biocompatible, biofunctional, and biomedical properties (Zhang et al., was used to dissolve 0.2 g of chitosan powder. Next, the chitosan solu­
2020). With hydroxyl (-OH) and amine (-NH2) groups along its polymer tion was mixed with hydroxyapatite solution while stirring. The pH was
chain, chitosan is highly effective in adsorbing colorants and metal ions adjusted to 11 by adding an ammonia solution and stirring it at 60 ◦ C for
(Hisham et al., 2021). Combining chitosan with HAp can enhance 3 h. Then, the mixture was filtered and dried at 60 ◦ C for 24 h. The same
adsorption efficiency. technique was also performed for different chitosan concentrations of
The chitosan concentration in the composite can significantly influ­ 10, 20, 30, 40, 50 wt% (Jamarun et al., 2023).
ence adsorption properties and composite structure. Therefore, it is
crucial to understand how variations in chitosan concentration can 2.5. Determination of adsorption capacity of HAp/CTS composite
affect the adsorption ability of the composite towards dyes.
The study aims to investigate how different chitosan concentrations The adsorption capacity of the HAp/CTS composite in absorbing
affect the synthesis of hydroxyapatite/chitosan composites for dye Rhodamine B dye. Firstly, 10 mL of Rhodamine B dye solution with a
adsorption using the in-situ method. It provides new insights into how concentration of 8 mg/L was adjusted to pH 3 by adding 0.1 M HCl.
chitosan concentration impacts the composite’s properties and its Subsequently, 0.1 g of HAp/CTS-10% composite was added to the
effectiveness in dye removal. Additionally, it explores the potential of Rhodamine B dye solution and stirred at 150 rpm for 120 min before
these composites as a solution for industrial dye pollution. being filtered. The absorbance of the filtrate was measured using a
UV–Vis spectrophotometer at a maximum wavelength of 553 nm. The
2. Material and methods same procedure was conducted for HAp and HAp/CTS composites with
20, 30, 40, and 50 wt% concentrations. Additionally, the adsorption
2.1. Materials capacity for each HAp/CTS composite was calculated (Xing et al., 2021).

The materials used for this research included cuttlebone (Sepia sp.), 2.6. Characterization
shrimp shells, Nitric Acid (HNO3 65%) (Merck), Diammonium Hydrogen
Phosphate ((NH4)2HPO4) (Merck), Ammonium Hydroxide (NH4OH) X-ray fluorescence (PANanalytical) was used to determine the
(Merck), Sodium Hydroxide (NaOH) (Merck), Hydrochloric Acid (HCl amount of calcium in cuttlebone powder. The type of chemical bonds
37%) (Merck), Glacial Acetic Acid (CH3COOH 100%) (Merck), most of and functional groups of the HAp/CTS composite were analyzed using
which were received in pure form. Aquadest was obtained from PT. BIO the Fourier transform infrared spectrometer (SHIMADZU) at the wave­
ANALITIKA Indonesia, Rhodamine B dye CAS 81-88-9, and Whatman number of 500–4000 cm− 1. The crystal structure of HAp/CTS compos­
filter paper number 42 (Cat No 1442090). The equipment used in this ites was analyzed using an X-ray diffractometer (PANalytical). The HAp/
research included sieve 60 mesh (ABM 230 μm) and Furnace (SCIEN­ CTS composite morphologies were observed on the surface structure
TIFIC SH-FU-5MGE). using a Scanning electron microscope-energy dispersive spectroscopy
(Hitachi Flexsem). The surface area of the HAp/CTS composite was
2.2. Preparation of CaO powder analyzed using a Surface Area Analyzer (TriStar II Plus Version 3.01).

The cuttlebones were washed, dried, and mashed. The cuttlebone 2.7. Rhodamine B adsorption isotherm
powder was calcined at 900 ◦ C for 5 h to achieve CaO powder (Charlena
and Wahyudi, 2017). Pipette 10 mL of 50 mg/L Rhodamine B dye solution, then adjust the
pH of the solution to 3. Then, the 0.1 g of HAp/CTS-30% composite was
2.3. Synthesis of chitosan from shrimp shells added into the Rhodamine B solution and stirred at 150 rpm for 120 min.
The blend was filtered, and the absorbance of the extract was checked
Shrimp shells were washed, dried, ground, and sieved. The chitosan with a UV–Vis spectrophotometer at 553 nm. The same treatment was
synthesis process consists of three steps: deproteination, demineraliza­ carried out at 100, 200, 400, 600, 800, and 1000 mg/L concentrations
tion, and deacetylation. In the initial step of the deproteination process, (Nyakairu et al., 2024).
30 g of shrimp shell powder was reacted with 1 M NaOH solution with a
powder-to-solvent ratio of 1: 5 for 20 h. After filtering, the mixture 2.8. Rhodamine B adsorption kinetics
underwent a pH neutralization process using distilled water. The results
obtained were dried at 60 ◦ C for 30 min and weighed. In the second Pipette 10 ml of 8 mg/L Rhodamine B dye solution and adjusted the
stage, the demineralization process was carried out by reacting the pH to 3 by adding 0.1 M HCl. Then, the 0.1 g HAp/CTS-30% composite
deproteinated powder with 1 M HCl for 16 h at a ratio of 1:5. The was put into the Rhodamine B solution. The mix was stirred at 150 rpm
mixture was filtered, and distilled water was used to rinse the end for 120 min. After that, the mixture was filtered, and the extract’s
product until the pH was neutral. Next, the results obtained were dried absorbance was measured using a UV–Vis spectrophotometer at a
at 60 ◦ C for 30 min. The yield of chitin obtained was then weighed. In wavelength of 553 nm. The same treatment was conducted at 30, 60, 90,
the third stage, the deacetylation process, the chitin powder was reacted 120, 150, and 180 min (Nyakairu et al., 2024).
with a 50% NaOH ratio of 1:10 at 60 ◦ C for 20 h to eliminate acetyl
groups and produce chitosan. Next, the mixture was filtered and rinsed 2.9. Regeneration
with distilled water until the pH was balanced. The formed chitosan was
then weighed after being dried for 4 h at 65 ◦ C in an oven (Hisham et al., Pipette 10 mL of the Rhodamine B dye solution into eight mg/L, put
2021). it into an Erlenmeyer, and adjust the pH of the solution to 3. Then, the
0.1 g of HAp/CTS-30% composite was added to the Rhodamine B so­
2.4. In-situ synthesis of HAp/CTS composites lution. The mixture was stirred at 150 rpm for 120 min. The mixture was
filtered, and the filtrate was analyzed with a UV–Vis spectrophotometer.
Two g of CaO powder were weighed and dissolved in 36 mL of 2 M The composite was washed with 0.1 M H2SO4 to remove the dye solu­
HNO3, filtered, and stirred for 15 min. Meanwhile, 65 mL of distilled tion, washed with distilled water, and dried. The HAp/CTS-30% com­
water was used to dissolve 2.8208 g of (NH4)2HPO4, homogenized in a posite was re-contacted with Rhodamine B solution for several
different beaker. Drop by drop, the ammonium hydrogen phosphate repetitions (cycles) (Jinendra et al., 2021).

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N. Jamarun et al. Kuwait Journal of Science 51 (2024) 100252

Table 1 Table 2
XRF result of cuttlefish bone after calcination. Effect of chitosan concentration on HAp/CTS composites with 10, 20, 30, 40,
Compounds Composition (%)
and 50 wt% concentration variations.
Concentration Observation Picture Mass
CaO 96.31
Variation (g)
Al2O3 0.33
P2O5 0.77 HAp/CTS-10% Fine powder and yellowish 3.1710
SO3 0.58 white color (+)
SrO 0.78
Other 1.24

HAp/CTS-20% Fine powder and yellowish 3.4488

3. Results and discussion white color (++)

3.1. Compositions of CaO from cuttlebone

The X-ray fluorescence (XRF) characterization of the cuttlebone is HAp/CTS-30% Fine powder and yellowish 3.5187
white color (+++)
displayed in Table 1. The percentage of CaO in the cuttlebone is 96.31%.
In addition to CaO, cuttlebone contains Al2O3, P2O5, SO3, and SrO. The
high calcium oxide content indicates that cuttlebones have the potential
as a calcium source for hydroxyapatite synthesis. The calcination pro­ HAp/CTS-40% Fine powder and yellowish 3.7801
cess of cuttlebones aims to obtain pure calcium in the form of oxide and white color (++++)
remove the organic components contained in cuttlebones. As a result of
calcination, a white, dry cuttlebone powder was obtained. The white
calcination results indicate that the process of degradation of organic
HAp/CTS-50% Fine powder and yellowish 4.5857
matter is no longer occurring (Jamarun et al., 2023). white color (+++++)
The reactions that occur in the calcination process are as follows:

CaCO3(s) + heat → CaO(s) + CO2(g)

general, the resulting HAp/CTS composite product is yellowish-white

due to the distribution of CTS in the HAp matrix; Pereira also investi­
3.2. Results of synthesis of chitosan from shrimp shells
gated the same. Following previous research, the higher the chitosan
concentration in the sample, the darker the composite’s color produced
The chitosan synthesis from shrimp shells is conducted through
(Laksono et al., 2020). The in-situ sol-gel process was used to synthesize
deproteination, demineralization, and deacetylation. With an initial
HAp/CTS composites with different chitosan concentrations, as seen in
mass of 42 g of shrimp shells, the chitosan synthesis process obtained a
Table 2.
light pink chitosan powder with a percentage yield of 61.64% and a
The mechanism of HAp/CTS composite formation is the chemical
weight of 11.7116 g. The first stage of chitosan synthesis is the depro­
interaction between the active sites of hydroxyapatite particles and the
teination process. In this deproteination reaction, the protein bonds in
hydroxyl and amino functional groups of chitosan and the dissolution of
the shrimp shells are broken, marked with a red solution, and slightly
chitosan using a 2% acetic acid solution. The function of adding acetic
thickened (Mulyani et al., 2020). Previous studies have observed that
acid is due to the presence of carboxyl groups (-COOH) in acetic acid so
the evidence of protein in the shrimp shell is released. Namely, the so­
that hydrogen bonds occur between carboxyl groups and amine groups
lution is reddish and slightly thickened. The thickening of the solution is
from chitosan. The amine group is free from chitosan and can be pro­
caused by breaking protein bonds and binding with Na+ ions in the
tonated to form a cationic amino group (-NH+ 3 ) (Szatkowski et al., 2015).
solution to form Na-proteinate, which can dissolve in the washing pro­
The reaction that occurs is:
cess (Ali et al., 2018). The second stage is the demineralization process.
The deproteinated powder was reacted with an HCl solution to remove CTS - NH2 + H+ ↔ CTS - NH+
3
the inorganic minerals in the shrimp shells. The mineral content in
shrimp shells is CaCO3 and Ca3(PO4)2. When adding HCl solution, foam After adding a chitosan solution to the hydroxyapatite solution, the
(CO2) will appear; this indicates a reaction between HCl and the min­ pH was adjusted by adding an NH4OH solution. Hydroxyapatite parti­
erals in the shrimp shell (Mulyani et al., 2020). The last step is the cles precipitate under alkaline conditions (pH ≥ 10). Hydroxyapatite
process of deacetylation. Removal of the acetyl group (-COCH3) from crystals can precipitate in the presence of chitosan solution. The formed
chitin is known as deacetylation. Chitin is immersed in a robust alkaline composite was filtered to separate the filtrate and residue. The residue
solution (NaOH) with high concentration and heat treatment for 20 h. was dried at 60 ◦ C for 24 h to eliminate humidity and protect the
Adding an alkaline solution aims to break the bond between the acetyl composite. The reaction that occurs is (Ait Said et al., 2023):
and nitrogen groups so that it turns into an amine group (-NH2) (Yanat 10Ca2+ + 6HPO2−
4 + CTS-NH3 + OH (excess) → HAp/CTS (↓) (pH ≥
+ −
and Schroën, 2021). 10)

3.3. Analysis of chitosan concentration on synthesized HAp/CTS

composites
3.4. Adsorption capacity of HAp/CTS composite
The synthesized HAp/CTS composites reveal that the final composite
is a fine powder, and the color of the composite develops yellow with Adsorption capacity states the amount of adsorbate that can accu­
increasing chitosan content. These visual results follow those reported mulate on the adsorbent surface (Jinendra et al., 2021). The adsorbent
by Trung et al. (2022), namely that hydroxyapatite is in the form of fine was HAp/CTS composite with various concentration variations of 10,
powder and changes color in the composite. Furthermore, as chitosan 20, 30, 40, and 50 wt%. Adsorption capacity of HAp and HAp/CTS 10,
concentration increases, so does the bulk of the HAp/CTS composite. In 20, 30, 40, 50% composite were 0.0624, 0.0816, 0.1025, 0.1832,

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N. Jamarun et al. Kuwait Journal of Science 51 (2024) 100252

Table 3 3.5. FTIR analysis

Adsorption capacity of HAp and HAp/CTS
composite. FTIR analysis aims to determine which molecular interactions and
Sampel qe (mg/g) functional groups of HAp, CTS, and HAp/CTS composites (Sivanandan
HAp 0.0624
and Saravanan, 2023). The spectrum of FTIR analysis results of HAp,
HAp/CTS-10% 0.0816 CTS, and HAp/CTS composites can be seen in Fig. 1. Fig. 1 (a) shows the
HAp/CTS-20% 0.1025 spectrum of the FTIR analysis results for HAp. The characteristic bands
HAp/CTS-30% 0.1832 of –OH group appear at 3277 cm− 1, and for PO3− 4 at 1023 cm− 1
HAp/CTS-40% 0.0725 − 1 − 1
(asymmetric stretching) and 599 cm and 561 cm (bending) (Rah­
HAp/CTS-50% 0.0923
mayeni et al., 2023). Fig. 1 (b) shows the spectrum of the FTIR analysis
results for CTS. The characteristic bands of O–H appear at 3433 cm− 1,
0.0725, 0.0923, respectively. The uptake of Rhodamine B dye onto N–H stretching at 3253 cm− 1, C–H asymmetric stretching at 2873 cm− 1,
HAp/CTS composite, at equilibrium qe was calculated as follows: N–H bending at 1620 cm− 1, C–H bending at 1375 cm− 1, and C–O
stretching)1008 cm− 1 (Jamarun et al., 2023). Fig. 1 (c) shows the
C0 - Ce
qe = xV (2) characteristic spectrum of the HAp/CTS composite’s FTIR analysis
W

where C0 and Ce are initial, and equilibrium concentration of Rhoda­

mine B (mg/L), W is the weight of HAp/CTS composite and V is the
volume of Rhodamine B solution.
The following adsorption capacity testing by HAp/CTS composite
can be seen in Table 3. Table 3 shows that the highest adsorption ca­
pacity value is found in the HAp/CTS-30% composite of 0.1832 mg/g. It
means every 0.1 g of composite can absorb 0.1832 mg of Rhodamine B
dye.
The HAp/CTS composite with 30% chitosan content has an optimal
absorption rate, where there is a balance between the number of
available active sites and their accessibility (interaction between
adsorbent and adsorbate). Adsorption parameters, including pH, deter­
mine the adsorption ability of an adsorbent, temperature, solution
concentration, adsorbent mass, adsorbent pore structure, adsorbent
surface area, contact time, and adsorbent particle size (Wu et al., 2020).
The HAp/CTS-30% composite will be further characterized by Fourier
transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scan­
ning electron microscopy-energy dispersive spectroscopy (SEM-EDS),
and surface area analyzer (SAA); it will also be used as an adsorbent in
Rhodamine B adsorption in this study.
Fig. 2. (a) XRD spectrum of HAp Standard, (b) pure HAp and (c) HAp/CTS-
30% composite.

Fig. 1. FTIR spectrum of (a) HAp, (b) CTS, and (c) HAp/CTS composite with various chitosan concentration.

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N. Jamarun et al. Kuwait Journal of Science 51 (2024) 100252

Fig. 3. (a) SEM image, and (b) EDS spectrum of HAp/CTS-30% composite.

results, including all HAp and CTS adsorption bands. Nevertheless, a few 3.7. Morphological analysis of HAp/CTS composite
of these distinctive peaks are slightly altered from 3277 to 3477 cm− 1
(O–H stretching), 1023 to 1014 cm− 1 (PO3− 4 asymmetric stretching), 599 SEM-EDS testing was carried out to analyze the surface morphology
to 597 cm− 1 and 561 to 557 cm− 1 (PO3− 4 bending) (Trung et al., 2022). and elemental analysis of the HAp/CTS-30% composite (Sivanandan
This shift could be attributed to the interactions between constituent and Saravanan, 2023; Vidakis et al., 2022). Fig. 3 is an SEM image of the
elements of the composite, such as hydrogen bonding between hy­ HAp/CTS-30% composite which shows that the composite consists of an
droxyapatite and chitosan (Szatkowski et al., 2015). irregular agglomeration of spherical-like particles (Wulandari et al.,
2023). Agglomeration tends to be found in HAp crystals. It can occur due
3.6. XRD analysis to the reaction between Ca2+ and PO3− 4 . Fig. 3 (b) displayed the EDS
spectrum of HAp/CTS-30%. According to the EDS spectrum, the
The XRD of HAp and HAp/CTS-30% composite are shown in Fig. 2. HAp/CTS-30% composite had a Ca/P ratio of 1.80, revealing the exis­
Fig. 2(b) displayed the diffraction pattern of pure HAp. The structure of tence of P, O, C, N, and Ca elements. According to earlier studies, HAp
HAp is hexagonal, which agrees with the HAp diffraction standard (ICSD could form at Ca/P ratios of 1.2–2.0 (Charlena and Wahyudi, 2017).
#157481) (Zhou et al., 2023; Ilnicka et al., 2023). The characteristic
peak for HAp is at 2θ with a value of 25.9◦ ; 31.8◦ ; 32.9◦ ; and 39.9◦ ; 3.8. Surface area analyzer analysis
46.7◦ ; 48◦ ; 49.6◦ ; 50.5◦ ; and 53.2◦ . Fig. 2(c) shows a new peak of chi­
tosan in the HAp/CTS-30% composite diffraction pattern at 2θ 19.23◦ . Surface area analyzer analysis was conducted to determine the sur­
According to previous research, it was explained that chitosan has a face area of the HAp/CTS-30% composite using the BET method (Bra­
distinctive peak at 2θ 10◦ –20◦ . These peaks will shift slightly depending nauer-Emmett-Teller) (Gao et al., 2017). This method uses
on the degree of deacetylation of the chitosan (Mulyani et al., 2020). adsorption/desorption of N2 gas to determine the surface area of the
In addition, the crystallite size of HAp and HAp/CTS-30% composites composite. The surface area of the HAp/CTS-30% composite was
were determined using the Scherrer equation, 61.1824 m2/g. Previous researchers found that the surface area of the
kλ hydroxyapatite-chitosan composite was 41.0 m2/g (Szatkowski et al.,
D= (1) 2015).
β Cos θ

where k is the Scherrer constant (0.89), β is the Full-Width radiant at half 3.9. Rhodamine B adsorption isotherm analysis
maximum (FWHM) of the diffraction line, λ is the wavelength of the Cu-
Kα radiation (1.5406 Å), and θ is the Bragg diffraction (Bazargan-Lari The adsorption isotherm is known as an equilibrium relationship
et al., 2011a). The crystallite size of HAp and HAp/CTS-30% composites between the concentration of adsorbate in the solution and the con­
were 21.49 nm and 19.63 nm, respectively. The widening of the hy­ centration of adsorbent particles (Azzaoui et al., 2023a, 2023b). In this
droxyapatite mineral diffraction peaks in the HAp/CTS-30% composite investigation, we employ two widely recognized adsorption isotherm
exhibited a decrease in the crystallinity and size of the apatite crystals models: the Langmuir and Freundlich isotherms. Equation (3) represents
(Sun et al., 2022). The distinctive peaks of the mineral and the polymer the Langmuir isotherm:
matrix broaden and weaken following the formation of the composite,
reflecting the bonding of the two phases (Szatkowski et al., 2015). The 1 1 1 1
= + (3)
characteristic peak of HAp and CTS on the diffractogram indicated that qe KL qm Ce qm
the HAp/CTS-30% composite had been successfully synthesized.
In Equation (3), the variables are defined as follows: Ce (mg/L) repre­
sents the equilibrium concentration of Rhodamine B, KL (L/mg) is the

Fig. 4. Linear graph of Langmuir isotherm (a) Freundlich isotherm (b).

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N. Jamarun et al. Kuwait Journal of Science 51 (2024) 100252

Table 4 findings.
Parameters of Rhodamine B dye adsorption isotherms on the HAp/CTS-30%
composite. ln (qe - qt) = ln qe – k1 t (5)
Isotherm Parameter Value t 1 t
= + (6)
Langmuir qm 0.0560 qt k2 (qe )2 qe
KL 0.0429
R2 0.5970 where qt (mg. g− 1) is the adsorption capacity at contact time t, qe (mg.
Freundlich KF 0.5526 g− 1) is the equilibrium adsorption capacity, and k1 and k2 are the first-
1/n 0.0045 and second-order kinetic rate constant. The slope and intercept were
R2 0.9082 leveraged to determine the k1 and k2 (Bazargan-Lari et al., 2011b;
Pooladi and Bazargan-Lari, 2023). The kinetic curves of the
Langmuir constant associated with adsorption energy, qe (mg/g) is the HAp/CTS-30% composite, displayed in Fig. 5, are pseudo-first-order and
pseudo-second-order, respectively. A pseudo-second-order kinetic
equilibrium adsorption capacity, and qm (mg/g) is the maximum
adsorption capacity. This equation further states that by examining the model described the Rhodamine B adsorption kinetics using the
HAp/CTS-30% composite. It is indicated by the coefficient of determi­
slope, which is 1/(KL.qm), and the intercept, which is 1/qm, of the linear
relationship, one can calculate the values of qm and KL (Bazargan-Lari nation (R2) value, more significant than the first-order pseudo kinetics
model, which equals 0.8913. Besides, the theoretical sorption value (qe)
et al., 2011a; Lakrat et al., 2023). The Freundlich isotherm is repre­
is 0.1368 mg/g, which are close to the experimental quantities (qe. exp)
sented by Equation (4):
0.1843 mg/g. It may be inferred that Rhodamine B adsorption kinetics
log qe = log KF + (1/n) log Ce (4) were pseudo-second-order and that chemical interactions occurred
during the adsorption of Rhodamine B dyes with HAp/CTS-30% com­
where KF (mg/g)(L/mg)1/n is the Freundlich constant of the relative posite (Jinendra et al., 2021). Thus, it implies that chemical adsorption
capacity value of the adsorbent, and 1/n is the heterogeneity on the is the process by which the adsorbates are adsorbed onto the synthesized
adsorbent surface. According to Eq. (4), KF and n can be determined composites.
from the slope 1/n and intercept log KF. The graph of Langmuir and The formation of a monolayer by chemical bonding in HAp/CTS
Freundlich isotherms shown in Fig. 4. The adsorption results of composite-based adsorbents may restrict the rate of the adsorption
Rhodamine B using HAp/CTS-30% composite showed that the Langmuir process (Amalina et al., 2022). The parameters of the adsorption kinetics
isotherm obtained an R2 value of 0.5970 and a Freundlich isotherm of model of Rhodamine B dye by HAp/CTS-30% composite are shown in
0.9082. Because the R2 value obtained by the Freundlich isotherm is Table 5. Table 5 contains detailed information from Fig. 5.
higher than the Langmuir isotherm, this announces that the Rhodamine
B adsorption process pursues the Freundlich isotherm equation. One of
the most used adsorption equilibrium models is the Freundlich isotherm 3.11. Regeneration analysis
(Sen Gupta and Bhattacharyya, 2014).
The adsorption of Rhodamine B with HAp/CTS-30% composite is Regeneration analysis is an essential aspect of adsorbents. Regener­
likely to adopt the Freundlich isotherm, where the adsorption takes ation is necessary for adsorbent applications because it can reduce costs
place by physisorption by forming a multilayer. The physisorption and improve process efficiency. The desorption agent for the HAp/CTS-
process enables bonds to develop with the adsorbent and between dye 30% composite utilizes an H2SO4 solution (Trung et al., 2022). The
molecules in solution. These two bindings are weak because Van der investigation of the regeneration of the HAp/CTS-30% composite in
Waals forces bind each other. A multilayer adsorption process is absorbing Rhodamine B dye is depicted in Fig. 6. The HAp/CTS-30%
generated because the adsorbate can move freely. The detail parameters composite has the potential to absorb Rhodamine B up to the second
of Rhodamine B dye adsorption isotherms on the HAp/CTS-30% com­ cycle by 38.01% and 14.48%. The decrease in adsorption percentage is
posite are shown in Table 4 (Table 4 contains detailed information from due to the increasing number of cycles, and the dye molecules will
Fig. 4).
Table 5
Parameters of the adsorption kinetics model of Rhodamine B dye by HAp/CTS-
3.10. Analysis of Rhodamine B adsorption kinetics
30% composite.
qe.exp Pseudo first order Pseudo second order
The study of adsorption kinetics is to ascertain the rate at which the
(mg/g)
adsorbent absorbs against the adsorbate (Wu et al., 2020). To check the qe (mg/ k1 R2 qe (mg/ k2 (g/mg. R2
adsorption capacity of HAp/CTS-30% composite for Rhodamine B, g) (minute− 1) g) minute)

Equations (5) and (6) illustrate how the pseudo-first-order and 0.1843 0.2291 0.0063 0.1099 0.1368 0.469 0.8913
pseudo-second-order models were enforced based on the experimental

Fig. 5. Linear graph of pseudo first order (a) and (b) pseudo second order kinetic models on Rhodamine B dye adsorption by HAp/CTS-30% composite.

6
N. Jamarun et al. Kuwait Journal of Science 51 (2024) 100252

Investigation, Funding acquisition, Conceptualization. Arika Prasejati:

Writing – original draft, Visualization, Software, Methodology, Investi­
gation, Formal analysis, Data curation. Zulhadjri Zulhadjri: Writing –
review & editing, Supervision. Sintia Caniago: Writing – review &
editing. Tri Yupi Amirullah: Writing – review & editing. Wulandari
Wulandari: Writing – review & editing. Vivi Sisca: Writing – review &
editing.

Declaration of competing interest

The authors declare that they have no known competing financial

interests or personal relationships that could have appeared to influence
the work reported in this paper.

References

Ait Said, H., Mabroum, H., Lahcini, M., Oudadesse, H., Barroug, A., Ben Youcef, H.,
Noukrati, H., 2023. Manufacturing methods, properties, and potential applications
in bone tissue regeneration of hydroxyapatite-chitosan biocomposites: a review. Int.
J. Biol. Macromol. 243, 1–27. https://doi.org/10.1016/j.ijbiomac.2023.125150.
Ali, M.E.A., Aboelfadl, M.M.S., Selim, A.M., Khalil, H.F., Elkady, G.M., 2018. Chitosan
Fig. 6. Rhodamine B dye adsorption cycle by HAp/CTS-30% composite. nanoparticles extracted from shrimp shells, application for removal of Fe(II) and Mn
(II) from aqueous phases. Separ. Sci. Technol. 1–12. https://doi.org/10.1080/
01496395.2018.1489845.
Table 6 Amalina, F., Abd Razak, A.S., Krishnan, S., Zularisam, A.W., Nasrullah, M., 2022.
A review of eco-sustainable techniques for the removal of Rhodamine B dye utilizing
Comparison of Rhodamine B dye adsorption capacity with other adsorbents biomass residue adsorbents. Phys. Chem. Earth 128, 1–11. https://doi.org/10.1016/
reported in the open study the literature. j.pce.2022.103267.
Amenaghawon, A.N., Anyalewechi, C.L., Darmokoesoemo, H., Kusuma, H.S., 2022.
Adsorbent pH qe (mg/ References
Hydroxyapatite-based adsorbents: applications in sequestering heavy metals and
g)
dyes. J. Environ. Manag. 302, 1–19. https://doi.org/10.1016/j.
Cassava slag biochar 4 105.6 Wu et al. (2020) jenvman.2021.113989.
RGO-Ni composite 8 65.31 Liu et al. (2022) Anitta, S., Sekar, C., 2023. Voltammetric determination of paracetamol and ciprofloxacin
Chitosan/Ag-hydroxyapatite 7 40.11 Li et al. (2018) in the presence of vitamin C using cuttlefish bone-derived hydroxyapatite sub-
nanocomposite beads microparticles as electrode material. Results Chem 5, 1–11. https://doi.org/
10.1016/j.rechem.2023.100816.
Walnut shell-derived N-doped activated 7.2 39.91 Husain et al.
Azzaoui, K., Aaddouz, M., Akartasse, N., Mejdoubi, E., Jodeh, S., Hammouti, B.,
carbon (2024)
Taleb, M., Es-Sehli, S., Berisha, A., Rhazi, L., Lamhamdi, A., Algarra, M., 2023a.
HAp/CTS composite 3 0.1832 Current study
Synthesis of β-Tricalcium phosphate/PEG 6000 composite by novel dissolution/
precipitation method: optimization of the adsorption process using a factorial
design—DFT and molecular dynamic. Arabian J. Sci. Eng. https://doi.org/10.1007/
saturate the active sites on the adsorbent. Additionally, the adsorbent s13369-023-08390-8.
degrades at very high pH levels, and obstruction occurs in its pores and Azzaoui, K., Jodeh, S., Mejdoubi, E., Hammouti, B., Taleb, M., Ennabety, G., Berisha, A.,
Aaddouz, M., Youssouf, M.H., Shityakov, S., Sabbahi, R., Algarra, M., 2023b.
active sites (Hevira et al., 2020) Therefore, the performance of the Synthesis of hydroxyapatite/polyethylene glycol 6000 composites by novel
HAp/CTS-30% composite still needs to be improved for repeated use. dissolution/precipitation method: optimization of the adsorption process using a
Table 6 presents the adsorption capacity of Rhodamine B dye for factorial design: DFT and molecular dynamic. BMC Chem 17, 1–15. https://doi.org/
10.1186/s13065-023-01061-7.
various adsorbents. The pH parameter is also presented because it is an Bambaeero, A., Bazargan-Lari, R., 2021. Simultaneous removal of copper and zinc ions
important parameter that influences adsorption capacity (Hevira et al., by low cost natural snail shell/hydroxyapatite/chitosan composite. Chin. J. Chem.
2020). The HAp/CTS composite has good absorption. However, the Eng. 33, 221–230. https://doi.org/10.1016/j.cjche.2020.07.066.
Bazargan-Lari, R., Bahrololoom, M.E., Nemati, A., 2011a. Sorption behavior of Zn (II)
amount is relatively lower than that of adsorbents made from
ions by low cost and biological natural hydroxyapatite/chitosan composite from
biomass-based materials. This research can also reduce pollution prob­ industrial waste water. J. Food Agric. Environ. 9, 892–897.
lems and utilize solid waste. Bazargan-Lari, R., Bahrololoom, M.E., Nemati, A., Salehi, Z., 2011b. Adsorption of Cu (II)
ions from industrial wastewater on natural hydroxyapatite extracted from bone ash.
J. Food Agric. Environ. 9, 652–657.
4. Conclusion Bensalah, H., Younssi, S.A., Ouammou, M., Gurlo, A., Bekheet, M.F., 2020. Azo dye
adsorption on an industrial waste-transformed hydroxyapatite adsorbent: kinetics,
isotherms, mechanism and regeneration studies. J. Environ. Chem. Eng. 8, 1–10.
HAp/CTS composites produced from cuttlebone and shrimp shells https://doi.org/10.1016/j.jece.2020.103807.
were properly manufactured utilizing the in-situ sol-gel technique. The Charlena, Bikharudin A., Wahyudi, S.T.Erizal, 2017. Synthesis and characterization of
variation of chitosan concentration in the HAp/CTS composite synthesis hydroxyapatite-collagen-chitosan (Ha/Col/Chi) composite by using Ex-situ wet
precipitation method. Rasayan J. Chem. 10, 766–770. https://doi.org/10.7324/
can affect the composite’s final product. The higher the concentration of
RJC.2017.1031768.
chitosan, the more yellow the color of the resulting composite. HAp/CTS Gao, Z., Zhang, Y., Song, N., Li, X., 2017. Biomass-derived renewable carbon materials
composite with 30% chitosan concentration has optimum results for electrochemical energy storage. Mater. Res. Lett. 5, 69–88. https://doi.org/
10.1080/21663831.2016.1250834.
compared to other concentrations. HAp/CTS composite use in adsorbing
Hevira, L., Zilfa, Rahmayeni, Ighalo, J.O., Zein, R., 2020. Biosorption of indigo carmine
Rhodamine B dye follows the Freundlich isotherm model, which shows from aqueous solution by Terminalia Catappa shell. J. Environ. Chem. Eng. 8, 1–11.
the formation of multilayer layers by following the second-order pseudo https://doi.org/10.1016/j.jece.2020.104290.
kinetics model. The HAp/CTS composite’s reusability occurred twice Hisham, F., Maziati Akmal, M.H., Ahmad, F.B., Ahmad, K., 2021. Facile extraction of
chitin and chitosan from shrimp shell. Mater. Today Proc. 42, 2369–2373. https://
during the cycle, so the HAp/CTS composite can be used as an adsorbent doi.org/10.1016/j.matpr.2020.12.329.
to reduce Rhodamine B dye waste in aqueous solutions. Husain, A., Ansari, K., Mahajan, D.K., Kandasamy, M., Ansari, M.N.M., Giri, J., Al-
Lohedan, H.A., 2024. Harnessing sustainable N-doped activated carbon from walnut
shells for advanced all-solid-state supercapacitors and targeted Rhodamine B dye
CRediT authorship contribution statement adsorption. J. Sci. Adv. Mater. Devices 9, 100699. https://doi.org/10.1016/j.
jsamd.2024.100699.
Novesar Jamarun: Writing – review & editing Writing – review &
editing, Validation, Supervision, Resources, Project administration,

7
N. Jamarun et al. Kuwait Journal of Science 51 (2024) 100252

Ilnicka, A., Skorupska, M., Szkoda, M., Zarach, Z., Lukaszewicz, J.P., 2023. N-doped delivery, and bioactivity. RSC Adv. 6, 43041–43048. https://doi.org/10.1039/
carbon materials as electrodes for highly stable supercapacitors. Mater. Res. Lett. 11, c6ra03147a.
213–221. https://doi.org/10.1080/21663831.2022.2139163. Sen Gupta, S., Bhattacharyya, K.G., 2014. Adsorption of metal ions by clays and
Islami, D.M., Wulandari, Jamarun, Syukri, N., Sisca, V., 2023. In-situ Hydrothermal inorganic solids. RSC Adv. 4, 28537–28586. https://doi.org/10.1039/c4ra03673e.
method for graphene oxide/hydroxyapatite synthesis from scallop shells. Rasayan J. Sivanandan, T., Saravanan, S., 2023. Effects of calcination temperatures on structural,
Chem. 16, 456–462. https://doi.org/10.31788/RJC.2023.1618169. functional, morphological, and magnetic properties of strontium ferrite (SrFe2O4)
Jamarun, N., Amelia, D.Rahmayeni, Septiani, U., Sisca, V., 2023. The effect of nanoparticles. Kuwait J. Sci. 50, 271–275. https://doi.org/10.1016/j.
temperature on the synthesis and characterization of hydroxyapatite-polyethylene kjs.2023.01.011.
glycol composites by in-situ process. Hybrid Adv 2, 1–7. https://doi.org/10.1016/j. Sun, T., Wang, Hongze, Gao, Z., Wu, Y., Wang, M., Jin, X., Alex Leung, C.L., Lee, P.D.,
hybadv.2023.100031. Fu, Y., Wang, Haowei, 2022. The role of in-situ nano-TiB2 particles in improving the
Jinendra, U., Bilehal, D., Nagabhushana, B.M., Kumar, A.P., 2021. Adsorptive removal of printability of noncastable 2024Al alloy. Mater. Res. Lett. 10, 656–665. https://doi.
Rhodamine B dye from aqueous solution by using graphene–based nickel org/10.1080/21663831.2022.2080514.
nanocomposite. Heliyon 7, e06851. https://doi.org/10.1016/j.heliyon.2021. Szatkowski, T., Kołodziejczak-Radzimska, A., Zdarta, J., Szwarc-Rzepka, K., Paukszta, D.,
e06851, 1–9. Wysokowski, M., Ehrlich, H., Jesionowski, T., 2015. Synthesis and characterization
Lakrat, M., Jodati, H., Miloud, E., Evis, Z., 2023. Synthesis and characterization of pure of hydroxyapatite/chitosan composites. Physicochem. Probl. Miner. Process. 51,
and Mg , Cu , Ag , and Sr doped calcium-deficient hydroxyapatite from brushite as 575–585. https://doi.org/10.5277/ppmp150217.
precursor using the dissolution-precipitation method. Powder Technol. 413, 118026 Trung, T.S., Minh, N.C., Cuong, H.N., Phuong, P.T.D., Dat, P.A., Nam, P.V., Hoa, N. Van,
https://doi.org/10.1016/j.powtec.2022.118026. 2022. Valorization of fish and shrimp wastes to nano-hydroxyapatite/chitosan
Laksono, A.D., Amatosa Jr., T.A., Sitorus, H.P.O., Asih, W.P.K., Sulistijono, 2020. Study biocomposite for wastewater treatment. J. Sci. Adv. Mater. Devices 7, 1–9. https://
on antibacterial of chitosan/hydroxyapatite doped magnesium composite as a doi.org/10.1016/j.jsamd.2022.100485.
material for bone graft applications. Makara J. Technol. 23, 119–125. https://doi. Vidakis, N., Petousis, M., Velidakis, E., Tzounis, L., Mountakis, N., Boura, O.,
org/10.7454/mst.v23i3.3755. Grammatikos, S.A., 2022. Multi-functional polyamide 12 (PA12)/multiwall carbon
Li, L., Iqbal, J., Zhu, Y., Zhang, P., Chen, W., Bhatnagar, A., Du, Y., 2018. Chitosan/Ag- nanotube 3D printed nanocomposites with enhanced mechanical and electrical
hydroxyapatite nanocomposite beads as a potential adsorbent for the efficient properties. Adv. Compos. Mater. 31, 630–654. https://doi.org/10.1080/
removal of toxic aquatic pollutants. Int. J. Biol. Macromol. 120, 1752–1759. https:// 09243046.2022.2076019.
doi.org/10.1016/j.ijbiomac.2018.09.190. Wu, J., Yang, J., Huang, G., Xu, C., Lin, B., 2020. Hydrothermal carbonization synthesis
Liu, X., Guo, Y., Zhang, C., Huang, X., Ma, K., Zhang, Y., 2022. Preparation of graphene of cassava slag biochar with excellent adsorption performance for rhodamine B.
oxide/4A molecular sieve composite and evaluation of adsorption performance for J. Clean. Prod. 251, 1–10. https://doi.org/10.1016/j.jclepro.2019.119717.
Rhodamine B. Sep. Purif. Technol. 286, 120400 https://doi.org/10.1016/j. Wulandari, W., Islami, D.M., Wellia, D.V., Emriadi, E., Sisca, V., Jamarun, N., 2023. The
seppur.2021.120400. effect of alginate concentration on crystallinity, morphology, and thermal stability
Mulyani, R., Mulyadi, D., Yusuf, N., 2020. Chitosan membrane from shrimp shells properties of hydroxyapatite/alginate composite. Polymers 15. https://doi.org/
(Panaeus Modonon) as an antibacterial food. J. Phys. Conf. Ser. 1477, 1–6. https:// 10.3390/polym15030614.
doi.org/10.1088/1742-6596/1477/7/072006. Xing, W., Liu, Q., Wang, J., Xia, S., Ma, L., Lu, R., Zhang, Y., Huang, Y., Wu, G., 2021.
Nyakairu, G.W.A., Kapanga, P.M., Ntale, M., Lusamba, S.N., Tshimanga, R.M., High selectivity and reusability of biomass-based adsorbent for chloramphenicol
Ammari, A., Shehu, Z., 2024. Synthesis, characterization and application of Zeolite/ removal. Nanomaterials 11, 1–13. https://doi.org/10.3390/nano11112950.
Bi2O3 nanocomposite in removal of Rhodamine B dye from wastewater. Clean. Yanat, M., Schroën, K., 2021. Preparation methods and applications of chitosan
Water(Washingt.) 1, 100004. https://doi.org/10.1016/j.clwat.2024.100004. nanoparticles; with an outlook toward reinforcement of biodegradable packaging.
Pooladi, A., Bazargan-Lari, R., 2023. Adsorption of zinc and copper ions simultaneously React. Funct. Polym. 161 https://doi.org/10.1016/j.reactfunctpolym.2021.104849.
on a low-cost natural chitosan/hydroxyapatite/snail shell/nano-magnetite Zhang, S., Li, Jianyong, Li, Jianfeng, Du, N., Li, D., Li, F., Man, J., 2020. Application
composite. Cellulose 30, 5687–5705. https://doi.org/10.1007/s10570-023-05219-3. status and technical analysis of chitosan-based medical dressings: a review. RSC Adv.
Pooladi, A., Bazargan-Lari, R., 2020. Simultaneous removal of copper and zinc ions by 10, 34308–34322. https://doi.org/10.1039/d0ra05692h.
Chitosan/Hydroxyapatite/nano-Magnetite composite. J. Mater. Res. Technol. 9, Zhou, X., Qiao, S., Yue, N., Zhang, W., Zheng, W., 2023. Soft X-ray emission spectroscopy
14841–14852. https://doi.org/10.1016/j.jmrt.2020.10.057. finds plenty of room in exploring lithium-ion batteries. Mater. Res. Lett. 11,
Rahmayeni, Wendari, T.P., Ramadani, S., Stiadi, Y., Sofyan, N., Zulhadjri, 2023. 239–249. https://doi.org/10.1080/21663831.2022.2141586.
CuFe2O4/hydroxyapatite magnetic nanocomposite synthesized using pensi clam Zou, B., Wang, L., Zhang, Y., Liu, Y., Ouyang, Q., Jin, S., Zhang, D., Yan, W., Li, Z., 2023.
shells as a source of calcium for degradation of dye and anti-bacterial applications. Enhanced strength and ductility of metal composites with intragranularly dispersed
Case Stud. Chem. Environ. Eng. 8, 100482 https://doi.org/10.1016/j. reinforcements by additive manufacturing. Mater. Res. Lett. 11, 360–366. https://
cscee.2023.100482. doi.org/10.1080/21663831.2022.2153630.
Sambudi, N.S., Cho, S., Cho, K., 2016. Porous hollow hydroxyapatite microspheres
synthesized by spray pyrolysis using a microalga template: preparation, drug