Applied Physics A (2025) 131:637

https://doi.org/10.1007/s00339-025-08744-z

Synthesis and optimization of GSH-capped Cu-In-Zn-S quantum Dots

for biomedical applications
Swati Sharma1 · Shikshita Jain1 · S. K. Tripathi1

Received: 18 February 2025 / Accepted: 24 June 2025

© The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature 2025

Abstract
The present research work reports the synthesis of novel GSH-capped Cu-In-Zn-S (CIZS) quantum dots (QDs), with
excellent electronic and optical properties, making them suitable for biomedical applications. The CIZS QDs are syn-
thesized via hydrothermal technique by optimizing the Zn precursor ratio, pH value, reaction time, and temperature.
The photoluminescence (PL) emission peak at 720 nm increases initially with the Zn molar ratio due to reduced surface
defects but subsequently declines due to lattice strain-induced non-radiative relaxations. The pH variation from 5.5 to 11.5
significantly influences PL intensity, with lower pH leading to particle aggregation and reduced emission, while basic pH
enhances PL efficiency. The reaction time (5–8 h) and temperature (80–160 °C), also impact PL intensity. Below 120 °C,
weak emission is observed due to inactive S²⁻ ions, followed by an increase due to radiative recombination. However,
beyond 140 °C, PL intensity decreases due to thiol hydrolysis. The direct band gap is also tuned for different pH values
using UV-visible (UV-vis) spectra, ranging from 1.96 to 3.86 eV with varying diameters. The optimal luminescence is
achieved at a Cu: In: Zn: S ratio of 1:4:12:2, synthesized at 140 °C for 6 h with a pH value of 10.5. Fourier-transform
infrared spectroscopy (FTIR) confirmed effective GSH capping via hydrogen bonding. Structural analysis revealed a
tetragonal phase with a crystallite size of ~ 1 nm, a spherical morphology (3.49 nm diameter), and a hydrodynamic size
of 93.5 nm. The Zeta potential results demonstrate that synthesised CIZS QDs as stable and efficient nanomaterials for
biomedical applications.

Keywords CIZS QDs · Photoluminesence · Hydrothermal synthesis · Biocompatible

1 Introduction [14], is constrained by the heavy metal toxicity associated

with Cd [15]. Consequently, there is a growing interest in
The quantum dots (QDs) are nanoscale semiconductor par- heavy-metal-free QDs as alternatives [16]. Notably, copper
ticles that exhibit quantum mechanical properties [1–6]. (Cu) based QDs have become a focal point of research due
Their size-dependent electrical and optical characteristics to their reduced toxicity this includes binary, ternary, and
make them useful for a variety of applications, especially quaternary QDs [17]. The photoluminescence intensity of
in the biomedical domain where they can be employed in quaternary nanoparticles (NPs) is higher than binary and
therapeutic actions [7], drug administration [8], biosens- ternary NPs because the photoluminescence (PL) of quater-
ing, and bio-imaging [9]. While traditional Cd-based QDs nary NPs can be adjusted by controlling the diffusion of zinc
such as Cadmium Selenide (CdSe) [10] and Cadmium Tel- (Zn) into ternary NPs to create quaternary NPs. This behav-
luride (CdTe) [11], offer tunable spectra across the visible iour is explained by the fact that adding Zn initially removes
region but their applicability in research, particularly in non-radiative surface states, thus increasing the density of
sensitive sensors [12], bio-imaging [13], and drug delivery radiative channels within the NPs [18]. Consequently, there
is a growing interest in the synthesis of quaternary CIZS
QDs.
Several synthesis routes of NPs such as solvothermal,
S. K. Tripathi
[email protected] microwave irradiation, hot injection, and thermal deposi-
tion are reported earlier but in all these synthesis methods
1
Department of Physics, Centre of Advanced Study in the solvents used are in the organic phase, which requires
Physics, Panjab University, Chandigarh 160014, India

13
637 Page 2 of 12 S. Sharma et al.

high temperature and an inert atmosphere which results CIZS QDs (Fig. 1) can act as a highly promising candidate
in the formation of hydrophobic NPs [19, 20]. To make for several biomedical applications, including drug delivery,
them hydrophilic, surface modifications are done. For sur- bio-imaging, and bio-sensing.
face modifications, several strategies are put forward one The regulation of pH is also crucial as it influences
of which is ligand exchange or conjugation of NPs surface the particle size and PL emission processes of inorganic
by some biomolecules but these results in poor biocompat- NPs which is essential for biomedical applications. So, it
ibility, lower photoluminescence, low chemical stability, is important to study the synthesis of NPs in different pH
increased NPs size, and have high bio-toxicity [21–23]. One media [31]. In this article, the study of variation in optical
method that is commonly used to synthesize hydrophilic studies, the change in size of NPs, and their energy band
NPs is the hydrothermal method to obtain stable, uniform, gaps as the pH of the solution changes. It is noted that as the
good quality, and highly luminescent NPs. Other than this, pH of the solution increases, the energy band gap of CIZS
the microwave irradiation method can be used but it does QDs increases and the size of the QDs decreases, because
not yield good-quality NPs [24–26]. Considering these at low pH the agglomeration of QDs starts. Therefore, for
challenges, there is a compelling need to yield hydrophilic all studies or measurements the pH value 10.5 is considered
and good-quality NPs. Xie et al. [27] reported the synthesis because at this pH the QDs are small with highest PL, also
of quaternary CIZS QDs with exceptional luminescence, for medical point of view the smaller NPs are best because
boasting a quantum yield (QY) exceeding 70%, all achieved they have a better chance of entering the membrane of any
without the need for coating a wide band gap shell. They cell as compared to larger NPs and provide a high surface-
successfully achieved tuneable emission across the visible to-volume ratio due to which their reactivity increases [32].
to near-infrared (NIR) spectrum by changing the reaction Determining the stability of NPs is necessary for their
temperature and the ratio of precursors, enabling the control biomedical applications because NPs possess high surface
of both particle size and composition. energy due to their small size therefore it is a tedious task to
Now, the main objective of our present research work is prepare stable NPs. The prepared CIZS QDs in the present
to overcome all the above-mentioned problems, for which work are in an aqueous medium. Therefore, it is essential to
our strategy is to introduce an environment-friendly, facile, calculate their hydrodynamic size and stability in solution.
and biocompatible method. This manuscript reports the syn- This article reports the results of zeta potential (ZP) and
thesis of GSH-capped CIZS QDs via hydrothermal method dynamic light scattering (DLS) measurements to evaluate
and optimization of various parameters such as the effect of the stability and hydrodynamic size of the NPs. As far as we
the precursor’s ratio of Zn, reaction temperature, time, and are aware, this is the first report that includes these measure-
pH of the solution. Here GSH is used as a stabilizing as well ments for GSH-capped CIZS QDs [33].
as capping agent because GSH coating not only facilitates
the aqueous dispersal of CIZS QDs but also establishes a
biocompatible surface, enabling seamless integration into 2 Experimental section
biological systems [28]. To the best of our knowledge,
there are only a few studies [29, 30] that report the effect 2.1 Materials and methods
of all four parameters simultaneously on the synthesis of
CIZS QDs. The best results at which the highest intensity Cupric chloride dihydrate (CuCl2·2H2O, CAS number:
is achieved are with a Cu: In: Zn: S molar ratio of 1:4:12:2, 10125-13-0), and sodium hydroxide (NaOH, CAS number:
Cu: GSH ratio of 1:36, at 140 °C for 6 h, and a pH value 1310-73-2) are purchased from SRL and NICE. Zinc ace-
of 10.5. For the synthesis of hydrophilic CIZS QDs, this is tate dihydrate (Zn (CH3CO2)2·2H2O, CAS number: 5970-
the shortest time and lowest temperature ever reported. This 45-6) is purchased from Qualigiens. Indium (III) chloride
distinctive amalgamation of inorganic core GSH-capped (InCl3, CAS number: 10025-82-8) and sodium sulphide
(Na2S, CAS number: 1313-82-2) are from Avra and Fisher
Scientific. Glutathione (GSH, CAS number: 70-18-8) is
purchased from CDH. All the chemicals employed are of
analytical-grade quality and no additional purification is
done. In all experiments, deionized (DI) water is utilized.

2.2 Characterization details

High-resolution Transmission electron microscopy

(HRTEM) images are captured using the JEM 2100
Fig. 1 Shows the diagram of capping of GSH with CIZS QDs

13
Synthesis and optimization of GSH-capped Cu-In-Zn-S quantum Dots for biomedical applications Page 3 of 12 637

plus electron microscope operating at 200 kV to achieve The hydrothermal approach is used for the synthesis of
enhanced image resolution. For HRTEM sample prepara- hydrophilic GSH-capped CIZS QDs in an aqueous solution.
tion, a solution of NPs is dropped onto an ultrathin carbon- In a standard procedure, firstly CuCl2·2H2O (2 µM), InCl3
coated 200-mesh copper grid, then the samples are allowed (8 µM), and Zn (CH3CO2)2·2H2O (different molar con-
to dry. The grid is dried naturally under ambient conditions centrations of Zn are added from 0.016 µM to 0.028 µM)
without the use of any nitrogen stream and specified dry- are added in 62 mL of deionized (DI) water and after that,
ing equipment. Based on the DLS principle, the hydrody- GSH (4.7µM) is added dropwise to the solution. After that,
namic size of synthesized NPs is determined using Zetasizer freshly prepared NaOH solution (2 mol L−1) is added to the
(Nano ZS, Malvern Instruments, Malvern, UK). On an mixture solution to adjust the pH at different values vary-
“X’PERT PRO TW 3050 Diffractometer”, a powder X-ray ing from 5.5 to 11.5 with continuous stirring. The stirring
diffraction (XRD) pattern is recorded using a Cu-Kα beam continues for 15 min, followed by the addition of 10 µM
with a wavelength of 1.54 Å as the incoming X-ray source, of Na2S to the solution, after addition, the solution colour
covering a scan range of 5° to 70° for the scattering angle changes to light brown. The concentration of Zinc is varied
(2θ). ZP is used to characterize the produced NPs using to obtain the best result, and at 1:4:12:5 precursor ratios for
the Delsa Nano C particle analyzer (Beckman Coulter) to Cu: In: Zn: S best results are obtained. All these experimen-
investigate the surface charge. Pressed KBr pellets are used tal steps are carried out at room temperature.
in Fourier transform infrared (FT-IR) analysis performed After that, the resulting solution is transferred to an 80
on a Perkin Elmer (Spectrum 400-FTIR) spectrometer. mL stainless steel autoclave lined with Teflon. Then auto-
LABINDIA spectrophotometer is used to record absorption clave is placed in oven which have to maintained at tem-
spectra of wavelengths between 250 and 750 nm. A Shi- peratures (80 °C, 100 °C, 120 °C, 140 °C, and 160 °C) for
madzu Spectro fluorophotometer (RF-5301 PC) is used to different time durations (5, 6, and 8 h) to obtain the most
measure fluorescence emission spectra with a 1 cm path- fluorescent QDs, following that, a natural cooling process
length quartz cuvette. At room temperature, both absorption brings the autoclave down to room temperature. When the
and fluorescence spectra are recorded. The pH of aqueous reaction is run for 6 h at 140 °C, the best results are pro-
is determined using an electronic pH meter (Max Electron- duced. The molar concentration of Zn is optimised first then
ics, India). To ensure accuracy, the pH meter was calibrated pH and then the time and temperature of the reaction are
using standard buffer solutions at pH 4.0 and 7.0 before each optimised simultaneously to get the best results.
measurement. These are the different techniques to analyze
the optical, morphological, and chemical studies of GSH-
capped CIZS QDs. 3 Results and discussions

2.3 Synthesis of CIZS QDs 3.1 Effect of change in molar ratio of Zn in CIZS QDs

The systematic diagram of the synthesis of CIZS QDs is Zn-incorporated CIS QDs are produced through hydro-
represented in Fig. 2. thermal methods with GSH acting as the stabilizing agent.
Fig. 2 Representing the system-
atic diagram of the synthesis
of CIZS QDs via hydrothermal
method by optimizing them for
different parameters such as Zn
molar ratio, reaction time and
temperature and pH values

13
 637 Page 4 of 12 S. Sharma et al.

Following Lewis’s electronic theory of acid and base, S2− ratio in specific reactions, typically, incorporating Zn into
functions as a soft base, easily engaging with soft acids like Cu-based ternary NPs. The photoluminescence of the NPs
Zn2+ and Cu2+, and GSH can act as both an acid and a base. can be modulated by precisely managing the Zn diffusion
GSH has multiple functional groups, including a thiol (-SH) into CIS, culminating in the formation of quaternary CIZS
group, a carboxyl (-COOH) group, and an amino (-NH2) as shown in Fig. 3(b). Initially, when Zn is added the PL
group, allowing it to participate in acid-base reactions. intensity increases with the increase in the concentration
However, in an aqueous solution, GSH typically acts as a of Zn, the increased PL intensity could be explained by the
weak acid due to its ability to donate protons from the car- easy passivation of Zn in the crystal or in the defects on the
boxyl and thiol groups to yield a stable product [34, 35]. surface of QDs, which makes it easier to block the non-radi-
The chalcopyrite structure typically contains copper (Cu) ative recombination route [38, 39]. As Zn is added further,
and indium (In) atoms in a solid solution. To maintain elec- the PL intensity decreases because of the increasing non-
trical charge balance, Zn atoms can replace these atoms radiative relaxations, which are connected to the intrinsic
in the structure. Specifically, two Zn atoms can randomly flaws and are most likely to increase in the lattice’s strain
eliminate one Cu atom and one In atom to satisfy the charge [37]. Ionic size mismatches and compositional gradients
requirements for Zn²⁺, Cu⁺, and In³⁺ ions. When we write a during synthesis are some of the factors that cause lattice
formula like ZnCuInS, it means that there is a probability strain. Because Zn²⁺ has a smaller ionic radius than Cu⁺ and
that any site normally occupied by Cu or In in the crystal lat- In³⁺, adding more Zn to CIZS QDs may result in compres-
tice could instead be occupied by a Zn atom, reflecting the sive strain. Defects like dislocations or vacancies may occur
random substitution that occurs within the structure [36]. as a result of the crystal structure being distorted by this
Figure 3 shows the (a) absorption and (b) PL spectra of strain. By serving as non-radiative recombination centers,
CIZS QDs at different concentrations of Zn. The absorption these flaws allow excited electrons to recombine without
spectra of CIZS QDs at various Zn concentrations are the emitting photons, which lowers the PL intensity [37].
same as that of Cu-based ternary and quaternary QDs which
means the synthesis of CIZS QDs starts even at the lowest 3.2 Influence of reaction temperature and time
concentration of Zn as shown in Fig. 3 (a) [35].
In the general mechanism of photoluminescence emis- Commencing the discussion on optimization, we delve into
sion, the electron is stimulated to go from the valence band the kinetics and thermodynamics of the synthesis methodol-
to the conduction band leaving behind the hole. This elec- ogy. This method exhibits remarkable potential for achiev-
tron comes back in two ways either through electron-hole ing elevated productivity and ensuring reproducibility by
pair recombination which is known as exciton or another strategically fine-tuning the reaction temperature and time.
possibility they get trapped in surface defects. From sur- The typical PL and absorption spectra of CIZS QDs with
face defects, they will also be released again in two ways: varying reaction times and temperatures are displayed in
radiative decay which results in emission and the other is Fig. 4. Compared to binary II-VI QDs, the UV-Vis spectra
non-radiative decay which results in heat only [26]. In CIZS of CIZS QDs do not exhibit a conspicuous exciton absorp-
QDs, the origin of their PL emissions can be traced because tion peak. No recognizable exciton fingerprints in absorp-
of carriers recombination confined within donor and accep- tion spectra may be explained by the uneven composition
tor levels, which arise from various defects within CIZS distribution, a wide range of sizes, and unique electri-
NPs including S and Cu vacancies, as well as interstitial cal characteristics of CIZS QDs [40, 41]. The synergistic
atoms [37]. Cu-based QDs offer greater stoichiometric flex- effect of the uneven composition distribution and the wide
ibility. This might be achieved by modifying the precursors size dispersion among several CIZS QDs results in the
Fig. 3 Shows the (a) absorption
and (b) PL spectra of CIZS QDs
at different concentrations of Zn
from 16 µM to 28 µM

13
Synthesis and optimization of GSH-capped Cu-In-Zn-S quantum Dots for biomedical applications Page 5 of 12 637

structureless absorption spectra that are characteristically in this area (~ 570 nm) is primarily present in hydrother-
detected in quaternary compounds [42, 43]. Thus, it can be mally synthesized mercaptopropionic acid or glutathione
deduced that due to the above reasons the ternary and qua- capped CIZS QDs which are not much studied, this band
ternary Cu-based I-III-VI QDs are characterized by poorly may have originated from the band-edge transition or InxSy
defined excitonic peaks in the absorption spectra as shown species emission which are related to defects [46, 47]. CIZS
in Fig. 4 [43]. The PL properties of CIZS QDs are also QDs emissions are observed in the deep red area or extend
dependent on several parameters, including differences in to the 700–800 nm (near-infrared region) is associated with
synthesis method, size of NPs, and chemical composition. the donor-acceptor pair (DAP) transition, in which the
Two peaks are visible in the CIZS QDs luminescence spec- acceptor state is Cu and Zn vacancy, and the donor state is
tra, one at about 570 nm in the visible range and the other either sulfur vacancy or copper Indium substitution (InCu).
at about 720 nm in the red-NIR when NPs are excited by From these possibilities, the band-edge PL emission does
480 nm wavelength. These peaks are similar to those found not take place, but rather the radiative recombination of
in Cu-doped II-VI and I-III-VI QDs [44, 45]. The PL band

Fig. 4 Shows the absorption ((a),

(c), and (e)) and PL ((b), (d) and
(f)) spectra of CIZS QDs at dif-
ferent times and temperatures, for
(a) and (b) time is 5 h, for (c) and
(d) time is 6 h, and for (e) and (f)
time is 8 h

13
 637 Page 6 of 12 S. Sharma et al.

donor-acceptor pairings, which are inherent defect-related 3.3 Influence of reaction pH

states [42, 48].
No shift at the beginning of absorption is observed in the The study of optical characteristics at various pH values
temporal evolution of the absorption spectra, indicating that is relevant since pH is a crucial parameter in the research
the CIZS QDs formulated in 5 h. In contrast, the CIZS QDs of biological fluids and the alterations in particle size and
PL intensity rises with reaction time, reaching a maximum shape are also linked to the surface changes caused by varia-
at 6 h at 140 °C, and then progressively decreases for higher tions in the medium’s pH [51]. Furthermore, GSH-capped
temperatures and higher time (8 h at different temperatures) CIZS exhibits pH-responsive behaviors in solution due to
as given in Fig. 4. Notably, almost no shift is observed in its functionalization with pH-responsive groups like carbox-
the wavelength peak of the CIZS QDs produced under vari- ylic acid [52]. Figure 5 (a) and (b) show the UV-Vis absorp-
ous reaction times, further demonstrating that the CIZS QDs tion and PL spectra of CIZS QDs at different pH values.
formation had already initiated in 5 h, remaining consistent The UV-Vis spectra of CIZS QDs show no strong peak in
with the absorption spectra. The increase in PL intensity the region (250–750 nm), but only a change in intensity of
may result from enhanced ligand passivation of surface QDs at different pH (5.5–11.5) is observed, as illustrated in
defects. In contrast, the decrease could be attributed to thiol Fig. 5.
hydrolysis, which reduces ligand availability, leading to The PL spectra as shown in Fig. 5(b) demonstrated a dis-
system instability and an excess of surface defects during cernible pattern of initially increasing and then decreasing
rapid QD growth, thereby facilitating non-radiative decay intensity as the pH value increased from 5.5 to 11.5. Nota-
[29, 42, 49]. bly, the peak PL intensity is most prominent at a pH value
At every fixed time when temperature is varied the absor- of 10.5. This peak could be attributed to enhanced binding
bance spectra is same having no peak, but the PL intensity forces between the cations and GSH present at this specific
show variation, first it increases and then decreases without pH level [42].
changing in the position of peak, the PL intensity is low Additionally, at low pH values, the COO‾ group of GSH
when the temperature is less than or equal to 120 °C. The undergoes protonation. This reduces the electrostatic repul-
reason for this is that S2− and metal ion reaction activity is sion among QDs, causing the QDs to agglomerate. This
insufficient to support CIZS QD development. It is unable agglomeration is evident in the absorption and photolumi-
to participate in the development of QDs in aqueous solu- nescence spectra (Fig. 5 (a) and (b)), showing increased
tion below 120 °C [50]. The synthesis of QDs can be done particle size and reduced or minimal emission in the acidic
more quickly at higher temperatures. So, as the temperature medium. At higher pH, GSH has a COO‾ group. This
increases the PL intensity starts increasing due to complete increases cluster distribution and colloid formation stabil-
or proper formation of CIZS QDs but when the temperature ity, reducing the tendency for particle aggregation due to
is increased further for a fixed time the PL intensity starts the cluster’s complete charging, which maximizes repulsive
decreasing because the rupture in the bonds of QDs starts electrostatic interactions. Small spherical NPs are formed
and the QDs become unstable. As seen in Fig. 4, the QDs at elevated pH, and also NPs formed are higher in quantity
exhibited optimal PL intensity after a 6-hour reaction at 140 [48]. The changes in pH during synthesis impact QD nucle-
°C. Beyond this duration or at higher temperatures, the PL ation and growth. Deprotonation of ligands like glutathione
intensity decreases [48, 49]. Consequently, 140 °C and 6 h improves their binding to the QD surface at higher pH val-
are determined to be the ideal temperature and time for the ues, which may lessen surface imperfections and increase
formation of CIZS QDs. PL intensity. On the other hand, hydroxide species or other
byproducts may arise due to an overly high pH, adding
Fig. 5 Shows the effect of differ-
ent pH values (5.5–11.5) on (a)
UV-Visible and (b) PL spectra of
CIZS QDs

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Synthesis and optimization of GSH-capped Cu-In-Zn-S quantum Dots for biomedical applications Page 7 of 12 637

strain and flaws that promote non-radiative recombination is given by R. The energy band gap exhibits a rise from
[48]. 1.95 ± 0.01 eV to 2.76 ± 0.01 eV with an increase in pH from
Through UV–Vis spectra analysis, the energy band gap 5.5 to 11.5, as depicted in Fig. 6 (a)-(d). The alterations in
of CIZS QDs is determined by using Tauc plots [48] and particle size are calculated using the effective mass approxi-
presented in Table 1. The band gap is calculated using the mation (EMA) equation as given above and variation shown
Tauc relation given below. in Table 1, as the pH value increases the size of the par-
ticle decreases. In an acidic medium, the NPs are not sta-
A(hν − Eg )
n
α = ble therefore, aggregation occurs because a more rapid ion
hν release rate is observed. Conversely, ions are released under
control in a basic media through regulated nucleation and
The absorption coefficient (α), Planck’s constant (h), and growth processes [56]. This is also the main reason behind
the optical bandgap (Eg) are used to determine the bandgap conducting the synthesis at pH 10.5, therefor PL intensity is
of a material, with the value of 𝑛 depending on the type of maximum at this pH value [57].
electronic transition: 𝑛=1/2, 3/2 for allowed and forbidden On optimising the four different parameters using Absorp-
direct transitions and 2,3 for allowed and forbidden indirect tion and PL spectroscopy the best results are obtained at a
transitions. Here, 𝑛=1/2 is used for allowed direct transi- molar ratio of Cu: In: Zn: S of 1:4:12:2, at 140 °C for 6 h,
tions, as indicated by the flat regions in the plots, which are and a pH value of 10.5. So, from here, the best sample is
characteristic of direct allowed transitions [53]. used for further characterisations.
A straight line can be fitted to the linear segment of a
plot of (𝛼ℎ𝜈)2 vs. ℎ𝜈 in order to extrapolate the band gap. 3.4 X-ray diffraction
This line can then be extended to cross the energy axis
(ℎ𝜈). These plots, known as Tauc plots, display both linear The XRD pattern of GSH-capped CIZS QDs is displayed in
and non-linear portions. The optical curve’s linear portion, Fig. 7. Four prominent diffraction peaks are found at 27.5°,
which is primarily found on the higher energy side of the 32.3°, 47.5°, and 55.6° corresponding to the planes (111),
Tauc plot, relates to the material’s basic absorption, whereas (200), (220), and (312) respectively [53]. These peaks indi-
the non-linear portion is caused by residual absorption cate the formation of a tetragonal CIZS alloyed structure
involving defect states. The optical bandgap of CIZS QDs which is consistent with previous reports [58]. In contrast to
is calculated by extrapolating the linear portion [54]. After the (1 1 1), (2 2 0), and (3 1 1) peaks of ZnS, the four char-
calculating the band gap value for different pH values, the acteristic peaks of CIZS exhibited a leftward shift. Nota-
effective mass approximation (EMA) equation, which is bly, all four characteristic peaks are situated between the
provided as follows, has been used to determine the CIZS distinct diffraction peaks of CIS and ZnS with JCPDS card
QDs average size, as given in Table 1. number (JCPDS 65-1572) and (JCPDS 65-9585), signifying
( ) the Zn2+ ions incorporation and the resultant formation of a
h2 1 1 1.8e2 CIZS alloyed structure [37, 38].
∆ E (R) = Eg (R) + + ∗ −
8R2 m∗e me 4π Rϵ α ϵ 0 The average crystallite size (D) of a material can be cal-
culated using the Debye-Scherrer equation [59]:
Where E(R) and Eg(R) are the band gap of the NPs and
bulk (1.77 eV for CIZS QDs) [55], me* and mh* represent Kλ
D=
the effective mass of an electron and a hole. Planck’s con- β cosθ
stant, represented by h (6.626*10−34 Js) and the NPs radius
where D is the average crystallite size, λ is incident X-ray
Table 1 The variation of the energy gap of CIZS QDs at different pH wavelength (1.54 Å for Cu Kα radiation), β is the full width
values varies from 5.5 to 11.5
at half maximum (FWHM) of the diffraction peak, θ is the
S.No. pH Value Energy Gap (± 0.01 eV) Diameter
of CIZS QDs of CIZS Bragg angle, K is the form factor, approximately around
QDs (nm) 0.91. The crystallite size For CIZS QDs is calculated using
1. 5.5 1.95 eV 6.66 ± 0.32 the Debye-Scherrer equation, which is approximately 1 nm,
2. 6.5 2.23 eV 4.16 ± 0.08 less than the size determined by TEM images. Also, the
3. 7.4 2.24 eV 4.10 ± 0.15 crystallite size is always less than the particle size, which
4. 8.5 2.30 eV 3.86 ± 0.23 verifies the result. Also, the XRD structure determines the
5. 9.5 2.39 eV 3.58 ± 0.19 tetragonal lattice parameter values a and c from the equation
6. 10.5 2.53 eV 3.22 ± 0.48 given below [60].
7. 11.5 2.76 eV 2.82 ± 0.14

13
 637 Page 8 of 12 S. Sharma et al.

Fig. 6 Shows the effect of dif-

ferent pH values from acidic to
basic (a) pH 5.5, 6.5, (b) pH 7.4,
8.5, (c) pH 9.5, 10.5, and (d) pH
11.5 on the bandgap of CIZS
QDs

Fig. 7 Shows the XRD spectra of

GSH-capped CIZS QDs synthe-
sized at a molar ratio of Cu: In:
Zn: S of 1:4:12:2, at 140 °C for
6 h, and 10.5 pH

13
Synthesis and optimization of GSH-capped Cu-In-Zn-S quantum Dots for biomedical applications Page 9 of 12 637

1 h2 + k 2 l2 presence of secondary phases of CuₓSγ, which could arise

= +
d2 a2 c2 from slight stoichiometry errors during synthesis [61].

In this context, the interplanar distance is denoted by d, the 3.5 FTIR spectroscopy
lattice constants or unit cell parameters by a and c, and the
Miller indices by h, k, and l. Bragg’s law is used to calculate Figure 8 shows the FT-IR spectra of the pure GSH and CIZS
d’s value. The lattice parameters a and c are then determined QDs. The FT-IR spectra of both GSH and CIZS QDs are
using these d values. For the planes (111), (200), (220), and obtained at room temperature and under normal conditions.
(312), the d values are determined to be 3.22 Å, 2.76 Å, 1.91 It tells about the type of bonding formed between various
Å, and 1.65 Å. The corresponding lattice parameters a and precursors used during the synthesis process.
c are calculated to be 5.40 Å and 10.2 Å, respectively. The The strongly centred signals at 672 cm−1 and 1650 cm−1
peaks at 27.5°, 32.3°, 47.5°, and 55.6° exhibit a slight incli- indicate that the C‒H bend and C = O stretching originated
nation towards greater angles in contrast to the chalcopyrite from the carboxylic acid group of pure GSH [62, 63],
CuInS₂ (JCPDS 65-1572). This shift is attributed to the drop but after capping with CIZS QDs these peaks are shifted
in the lattice constant caused by introducing smaller radius to 700 cm−1 and 1642 cm−1 as shown in the above figure.
Zn²⁺ ions into the crystal lattice, suggesting the formation These findings showed that the–COO group of GSH is fur-
of an alloy structure Cu–In–Zn–S [42]. Additionally, very ther functionalized with CIZS QDs [64]. The FTIR peak at
small peaks found at different positions may indicate the 3461 cm−1 in GSH-capped CIZS QDs is attributed to the–
OH (water) stretching vibration [65]. The peaks located at

Fig. 8 Shows the FT-IR spectra

of pure GSH (upper) and GSH-
capped CIZS QDs (lower)

13
 637 Page 10 of 12 S. Sharma et al.

Fig. 9 Shows the (a) HRTEM

images, insets display a magni-
fied view of the sample (CIZS
QDs outlined with blue circle)
and (b) size distribution histo-
gram of CIZS QDs obtained by
taking 80 QDs

Fig. 10 Shows the (a) DLS

(hydrodynamic size, encompass-
ing the core, surface ligands,
and solvation layer), and (b) ZP
measurements of GSH-capped
CIZS QDs

2136 cm−1, 1125 cm−1, and 1009 cm−1 in pure GSH rep- analyzing HRTEM images using the open-source software
resent the ‒C = C‒ stretch and ‒C‒O bond [66]. The − SH ImageJ by taking the data of 80 QDs.
stretching and N − H stretching might be attributed to the
absorption bands at 2524 cm−1 and 3436 cm−1, respectively 3.7 DLS and ZP measurements
in pure GSH [42].
The GSH is integrated into the surface of CIZS QDs, as ZP and DLS are great resources for assessing the dispers-
evidenced by the removal of the two peaks at 2524 cm−1 ibility and stability of colloids. Also, to regulate the ligand
and 3436 cm−1, which are attributed to the ‒SH stretching chemistry and performance of bioconjugation NPs in bio-
vibration and the ‒NH deformation vibration. Conversely, logical imaging and drug delivery, evaluation of the hydro-
the lack of ‒SH and ‒NH may suggest that the GSH is effec- dynamic size and surface charge of NPs is necessary [68].
tively conjugated on CIZS QDs. This clearly illustrates the Figure 10 (a) and (b) show the DLS and ZP of GSH-
creation of a covalent link between the metal atoms of CIZS capped CIZS QDs obtained at pH value of 10.5. at basic
QDs and the thiol group on the surface of GSH, with the pH value deprotonation of surface functional groups like
hydrophilic carboxylate group-oriented outward [39, 42]. carboxylates and thiols increases electrostatic repulsion
between particles which reduces aggregation and improves
3.6 HRTEM of CIZS QDs dispersion stability. The average size of CIZS NPs obtained
from DLS is 93.5 nm which is appropriate for use in bio-
The HRTEM visuals of GSH-capped CIZS QDs obtained medical applications especially in drug delivery [69]. The
under ideal experimental settings are displayed in Fig. 9 (a). diameter determined through DLS tends to be larger than
The inset image in Fig. 9 (a) shows the magnified version of those obtained from HRTEM as shown in Fig. 9 (a). This
HRTEM images of CIZS QDs [42, 48, 67]. It is clear from discrepancy arises from the different measurement tech-
the images that CIZS QDs have a well-dispersed, nearly niques employed and the distinct particle size aspects they
spherical form ranging in size from 2 nm to 6 nm. The assess because DLS analyses samples in a solvated state,
size distribution histograms are formed by using HRTEM while TEM operates on dried samples in ultrahigh vacuum
images (as given in Fig. 9(b)) and the mean size of CIZS conditions. Additionally, the two techniques differ fun-
QDs obtained is 3.49 nm. The standard error comes out to damentally: HRTEM relies on a number-based approach,
± 0.42 nm. The particle size distribution is calculated by while DLS is intensity-based [70]. The five runs are carried

13
Synthesis and optimization of GSH-capped Cu-In-Zn-S quantum Dots for biomedical applications Page 11 of 12 637

alization. S.K. Tripathi: Writing– review & editing, Supervision and

out in a row with the same setup to guarantee the accuracy Conceptualization.
and repeatability of our measurements. A mean hydrody-
namic diameter of 93.5 nm with a standard deviation of Data availability The data supporting these findings is part of an ongo-
± 2.1 nm was obtained from the results. ing study and cannot be shared at this time.
In general, NPs with a zeta potential between − 10 and
+ 10 mV are considered as neutral. From ZP measurement, Declarations
it is observed that the ZP value obtained is nearly (~ −6.3
Conflict of interest According to the authors, no known conflicting fi-
mV), as shown in Fig. 10 (b). Additionally, the standard nancial interests or personal relationships appear to have influenced
deviation for the zeta potential is ± 1.3 mV. It would be easy any material presented in this paper.
to reduce the strong interaction between NPs and serum pro-
teins in the circulatory system when the NPs are neutral and
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