RESEARCH ARTICLES

Green synthesis, characterization and

applications of TiO2 nanoparticles using
aqueous extract of Erythrina variegata leaves
Jayaram Mari Selvi1, Mariappan Murugalakshmi1,*, Ponnusamy Sami2,
Mariappan Gnanaprakash3 and R. Thanalakshmi1
1
PG & Research Department of Chemistry, The Standard Fireworks Rajaratnam College for Women, Sivakasi 626 123, India
2
PG & Research Department of Chemistry, Virudhunagar Hindu Nadar’s Senthikumara Nadar (VHNSN) College, Virudhunagar 626 001, India
3
Mepco School of Management Studies, Mepco Schlenk Engineering College (Autonomous), Sivakasi 626 001, India

electrical, optical, magnetic, catalytic, biomedical and anti-

Green synthesis is a simple, non-toxic, economical and
eco-friendly approach for the synthesis of nanoparticles bacterial activities which cannot be achieved by their bulk
(NPs). The implementation of new technologies has led counterparts1–9. Several traditional synthetic methods have
to the new area of nano revolution which unfolds the been developed by various research groups to synthesize
role of plants in bio- and green synthesis of nanomate- NPs using physico-chemical methods10,11.
rials. The plant extracts employed are neem, lemon NPs are defined as a cluster of atoms between 1 and
grass, aloe vera, Indian gooseberry, etc., focusing on 100 nm in size that behave like a whole unit with respect
the green chemistry principles. In the present work, to all their properties12. NPs are one of the most im-
NPs of titanium dioxide (TiO2) were synthesized using portant gifts of science in the modern era. Nanotechnology
an aqueous extract of Erythrina variegata leaves as a mainly deals with the synthesis of NPs of variable size,
capping agent. The leaf extract was utilized as a re- shape, chemical composition and their potential use for the
ducing agent for the conversion of metal precursors
benefit of humanity13. The preparation of NPs can be
into metal-oxide NPs. E. variegata-mediated TiO2 NPs
were characterized by UV–Vis absorption spectrosco- done under three main conditions: (i) choice of environ-
py, Fourier transform infrared spectroscopy, X-ray ment-friendly solvent medium, (ii) reducing agent and
diffraction (XRD), energy-dispersive X-ray spectro- (iii) a non-toxic material for their stabilization14. NPs are
scopy and morphological studies were conducted by the building blocks of the next generation of technology
scanning electron microscopy. The UV–Vis absorption with applications in several other fields15.
spectrum showed an absorption band at 317.6 nm, Nanotechnology is an important branch of science. It
which supports the formation of TiO2 NPs. The optical deals with the synthesis and development of various types
band-gap energy was determined to be 2.35 eV. Fur- of NPs in sizes ranging from 1 to 100 nm (ref. 16). It has
ther characterization by XRD supported the crystal- been noted that the physical and chemical properties of
linity and purity of the synthesized TiO2 NPs. These any material change when its size decreases to nano-
NPs may have effective dye degradation ability. The
scale17,18. The characteristics depend on certain traits such
green-synthesized TiO2 NPs exhibited interesting photo-
catalytic efficacy on methylene blue dye under UV as size, particle distribution, morphology and high sur-
irradiation (using a multi-lamp photo reactor) and anti- face/volume ratio19. Metal-oxide NPs have gained attention
bacterial activity against pathogenic organisms like for their potential applications in optoelectronics, nanodevi-
Streptococcus, Staphylococcus, Escherichia coli and ces, nanoelectronics, nanosensors, information storage
Pseudomonas aeruginosa. and catalysis20. Various metal-oxide NPs have been im-
plemented in a wide range of applications. They can be
used as catalysts in reduction, oxidation electrocatalysis,
Keywords: Antibacterial activity, Erythrina variegata, photocatalysis and gas-phase reactions21.
green synthesis, nanoparticles, photocatalytic efficacy, tita- Green synthesis is eco-friendly and manufactures stable
nium dioxide. and multiuse nanomaterials which involves on the action
of biomolecules and works as a reducing and capping
IN the last few years, nanoparticles (NPs) have gained agent without toxicity, and supports the ease in manufac-
importance in the scientific field due to their typical size, turing. The physical and chemical techniques are cost-
shape, surface area and technological applications like effective and they involve the usage of instruments or
chemical agents. Another limitation is low stability, conta-
mination with residuals of agents and less compatibility in
*For correspondence. (e-mail: m_murugalakshmi@[Link]) the pharmaceutical and cosmetics industry22,23. Moreover,

CURRENT SCIENCE, VOL. 123, NO. 1, 10 JULY 2022 59

RESEARCH ARTICLES
plant leaf extracts are the best candidates for the synthesis Chemicals
of NPs.
In the present study, the leaf extract of Erythrina varie- Titanium tetraisopropoxide (TTIP; Sigma Aldrich, AR-
gata belonging to the family Fabaceae, commonly known grade, purity >97%) was purchased from the National
as ‘Kalyana murungai’, was chosen. It has high medicinal Scientific Company in Villapuram, Madurai, Tamil Nadu,
value and properties. E. variegata possesses distinctive India.
biological actions, including antibacterial/dental caries
prevention, CNS effects, cardiovascular effects, antioxidant,
Glassware
smooth muscle relaxant, analgesic and anti-inflammatory,
calcium homeostasis, trypsin/proteinase inhibitors and
Glassware such as conical flask, measuring cylinder,
cytotoxicity24. Its leaves have metal and salt-reducing
beaker, watch glass, magnetic stirrer, funnel, standard mea-
properties. E. variegata has been used in traditional medi-
suring flask, round-bottom flask, condenser and Soxhlet
cine as a nervine sedative, febrifuge, anti-asthmatic and
apparatus were used.
antiepileptic. The leaves are used to treat fever, inflamma-
tion and joint pain. The juice of the leaves is used to treat
earache, toothache, constipation, cough, and to stimulate Plant extract preparation
lactation and menstruation25. The bark is used to treat fe-
ver, liver problems and rheumatism. E. variegata leaves were washed several times with water
E. variegata contains several phenolic metabolites, to remove dust particles. The leaves were dried in the
such as peterocarpans, isoflavones, flavanones and chal- shade at room temperature for ten days. The dried leaves
cones, some of which display antiplasmodial, antimyco- were ground to a fine powder. The aqueous extract was
bacterial and cytotoxic activity against various cancer cell prepared using the Soxhlet extraction method. In the
lines. It also contains alkaloids like N-norprotosinomenine Soxhlet apparatus, solvent (150 ml of distilled water) was
(I), dimethyltryptophan, hyparphorine and sterols like added to a round-bottom flask, which was attached to a
campesterol, β-sitosterol and β-amyrin. The isoflavones Soxhlet extractor and condenser on an isomantle. Then
indicanines D and E, genistein, wighteone, alpinum iso- 15 g of dried leaf powder of the plant material was loaded
flavones, dimethyl alpinum isofavone, 8-prenyl erythrinin into the thimble placed inside the Soxhlet extractor. The
C and erysenegalensein E – erythrinassinate B, apigenin, side arm was lagged with glass wool. The solvent was
genkwanin, isovitexin, swertisin, saponarin, 5-O-gluco- heated using the isomantle and began to evaporate at
sylswertisin and 5-O-glucosylisoswertisin. Glucoside 100°C, moving through the apparatus to the condenser.
swertiamarin, a triterpene betulin have also been isolated. The condensate drips into the reservoir containing the
The alcohol insoluble portion of the unsaponifiable matter thimble. Once the level of solvent reached the siphon, it
has yielded n-hexosamol, heptacosine, nonacosane and was poured in dropped into the flask and the cycle started
non-saponifiable matter of the petroleum ether extract has again. The process should run for a total of 6 h. After 6 h
yielded myristic, stearic and oleic acids26–28. of continuous extraction, the fine extract containing vari-
In this study, E. variegata leaf aqueous extract was used ous natural compounds was filtered using Whatmann No.
for the synthesis of TiO2 NPs to evaluate the reduction 1 filter paper and stored safely for further use.
power of this extract as a green route of TiO2 NPs pro-
duction. The effect of extract volume was studied using
Synthesis of TiO2 NPs
UV-Vis spectroscopy, Fourier transform infrared spectro-
scopy (FTIR), X-ray diffraction (XRD), scanning electron
TTIP (1 N) was prepared using distilled water and a col-
microscopy (SEM) and energy-dispersive X-ray spectro-
ourless solution was obtained. Next, 20 ml of E. variega-
scopy (EDAX). Next, its application in photocatalytic
ta extract was added to 0.1 N TTIP solution. The colour
degradation of methylene blue dye and antibacterial activity
of the mixture changed from pale yellow immediately and
action on Streptococcus, Staphylococcus, E. coli and Pseu-
slowly turned to sandal after 6 h of continuous stirring.
domonas aeruginosa was analysed.
This colour change is due to the formation of TiO2 NPs.
The synthesized NPs were separated by centrifuging and
Materials and methods dried in a hot air oven at 100°C for 6 h. The NPs were
characterized by UV-Vis spectrum, FTIR, XRD, SEM and
Plant material EDAX studies (Figure 1).
Leaves of E. variegata were collected from the garden of
The Standard Fireworks Rajaratnam College for Women, Characterization of TiO2 NPs
Sivakasi, Virudhunagar district, Tamil Nadu, India and dried
in the shade. These were then powdered and stored in an The colour change in the reaction mixture was visually
air-tight container at room temperature until further use. examined. The absorption maxima in UV–Vis (Shimadzu
60 CURRENT SCIENCE, VOL. 123, NO. 1, 10 JULY 2022
 RESEARCH ARTICLES

Figure 1. Schematic view of titanium dioxide (TiO2) nanoparticles (NPs) synthesis using Erythrina variegata leaves.

3H2O, MW = 319.85 g/mol) solution was performed using

green-synthesized TiO2 NPs in UV light (multi-lamp pho-
to reactor with light source @ 365 nm). In this experiment,
a suspension was prepared by adding 0.02 mg of TiO2
NPs to 100 ml of 10 ppm methylene blue solution. Then
the mixture was stirred for about 30 min in darkness to
ensure the constant equilibrium of TiO2 NPs in the dye so-
lution. Degradation was visually detected by a gradual
change in the colour of the dye solution from deep blue to
colourless. Simultaneously, the absorbance of dye solution
Figure 2. Colour change indicating the TiO2 NPs formation. before and after degradation by TiO2 NPs at regular time
intervals of exposure to UV light was measured.
UV-2450) double-beam spectrophotometer in the range Antibacterial activity of TiO2 NPs
between 200 and 900 nm using distilled water as a reference.
The FTIR spectrum of TiO2 NPs was recorded (Shimadzu The antibacterial activity of E. variegata leaf extract-medi-
IR-Affinity-1 FTIR spectrophotometer, The Standard Fire- ated TiO2 NPs was examined using broth dilution and
works Rajaratnam College for Women, Sivakasi, Tamil disc-diffusion method against pathogenic organisms like
Nadu, India). The functional groups present in the sam- Streptococcus, Staphylococcus, E. coli and P. aeruginosa.
ples were identified using FTIR spectral data. The sample The results were predicted based on the zone of inhibition.
was prepared as 0.25 mm thick KBr pellets (1 mg in
100 mg KBr). The spectrum was recorded between 4000 Results and discussion
and 400 cm–1 nm for 70 scans. XRD patterns were ob-
tained for the powder samples (Bruker D8 advance ECO Visual observations
X-ray diffractometer, Kalasalingam University, Krishnan-
koil, Tamil Nadu, India). Phase purity and size were de- This is the primary test for checking the formation of TiO2
termined by XRD analysis. The average size of TiO2 NPs NPs. The sequential colour change indicates the for-
was calculated using the Debye–Scherrer equation. The mation of TiO2 NPs by the plant material. Leaf extract
morphology and chemical composition of the synthesized was added to the TTIP solution. Within 2 min, the colour
NPs were examined by EDAX (6490 LA) at an accelera- changed from pale yellow to sandal. After incubation, the
tion voltage of 20 kV. colour changed from pale yellow to sandal, indicating the
formation of TiO2 NPs (Figure 2).
Photocatalytic activity of TiO2 NPs
UV-Vis spectral studies
The photocatalytic degradation of methylene blue (chemi-
cal name (3,7 bis (dimethylamino) phenazathionium chlo- The UV–Vis spectra of the green-synthesized TiO2 NPs
ride tetramethylthionine chloride), MF = C16H18ClN3S, were recorded using a double-beam spectrophotometer
CURRENT SCIENCE, VOL. 123, NO. 1, 10 JULY 2022 61
RESEARCH ARTICLES

Figure 3. a, UV–visible spectrum of aqueous extract of Erythrina variegata leaves. b, Synthesized TiO2 NPs.

The value of Eg was 2.35 eV for 317.6 nm TiO2 NPs.

The decrease in Eg with increasing thickness may be due
to the possibility of structural defects, which in their rule
lead to the formation of donor levels within the energy gap.
These results were identical to those of Jagpreet et al.29.

FTIR spectral studies

FTIR spectral analysis was carried out to identify the

functional groups of molecules bound specifically along
the surface of TiO2 NPs. Figure 5 shows the FTIR spectra
of green synthesized TiO2 NPs and leaf powder of E. var-
iegata. The spectrum was recorded in the range 4000–
400 cm–1. The peaks at 3418 and 1628 cm–1 are due to O–H
Figure 4. Tauc plot of TiO2 NPs. stretching and bending vibration of surface-adsorbed water
respectively30. This hydroxyl group bound to the surface
of TiO2 was assumed to trigger the photocatalytic activity
(Shimadzu UV-2450 PC) in the range 200–900 nm. Figure 3 of NPs. FTIR spectrum of green-synthesized NPs also
shows the absorption spectrum of aqueous extract of E. showed characteristic bands at 2836 cm–1, indicating the
variegata leaves and the synthesized TiO2 NPs. The ab- secondary amines and 2932 cm–1 confirming the hydrogen-
sorption maximum at 317.6 nm supports the formation of bonded alcohols. The strong peak noticed at 1589 cm–1 rep-
TiO2 NPs (Figure 2). resented aliphatic of the nitro compound with stretching of
N–O and the peak at 1071 cm–1 represented ethers. The
absorption band with stretching mode of Ti–O in the finger-
Tauc plot band-gap energy calculation
print region 755 cm–1, which corresponded to the anatase
phase of TiO2 (ref. 31). Thus, the FTIR results success-
The band-gap energy can be determined using the Tauc
fully confirmed the formation of anatase TiO2, which is
relation. The dependence of the optical band gap (Eg) on
formed due to the reduction of TTIP precursor by phyto-
the absorption coefficient and the incident photon energy
chemical components present in the extract of E. variegata
(hν) is given by the following relation
leaves. These components have several organic functional
groups which attach to the surface of TTIP and cause a
αhν = A(hν – Eg)n,
reduction of the same. Thus, they act as a good reducing
as well as capping agents32.
where A is the optical constant, α the absorption coefficient,
h the Planck’s constant, ν the frequency of the incident
photon and n is the number characterizing the nature of X-ray diffraction studies
the transition process (here n = 2 for direct transition and
n = 1/2 for indirect transition). The value of Eg was obtai- Figure 6 shows the XRD pattern of synthesized TiO2 NPs.
ned by plotting hν versus (αhν)2 and then extrapolating The peaks appearing at 2θ values of 25.28°, 37.82°,
the linear region on the energy axis (Figure 4). 48.01°, 53.89°, 55.03° and 62.64° correspond to the Bragg

62 CURRENT SCIENCE, VOL. 123, NO. 1, 10 JULY 2022

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Table 1. Structural and geometrical parameters of titanium dioxide (TiO2)
Height Relative
Angle 2θ (cts) hkl FWHM d value (Å) intensity (%)
25.289 22.97581 101 0.76653 3.51891 100.0
37.829 4.96466 004 1.42267 2.37907 26.6
48.005 7.78938 200 0.94946 1.89436 34.0
53.897 5.42721 105 0.87328 1.69856 24.3
55.030 5.20181 211 1.03566 1.67297 20.8
62.641 3.8286 204 1.24493 1.36642 7.1

Figure 5. Fourier transform infrared spectra of (a) Erythrina variegata leaf extract and (b) TiO2 NPs.

The maximum peak appeared at 2θ value of 25.28°, which

corresponds to an average size of TiO2 of 7.91 nm. Table 1
lists the structural and geometrical parameters of TiO2 NPs.

Scanning electron microscopy

The morphological features of the synthesized TiO2 NPs

were studied by SEM analysis. SEM images confirmed
that the synthesized TiO2 NPs were tetragonal in shape.
Figure 7 shows the physical morphology, particle size and
aspect ratio of the synthesized NPs in the lowest and
highest magnification of 20.00 and 50.00 kX respectively34.

Figure 6. X-ray diffraction pattern of TiO2 NPs. Energy-dispersive X-ray spectroscopy

The EDX spectrum of TiO2 showed peaks for titanium (Ti),

reflection of the (101), (004), (200), (105), (211) and (204) oxygen (O) and carbon (C) elements as well as the atomic
planes respectively. The above diffraction peaks indicate and weight percentages. The observed peak of C was due
the tetragonal body-centred structure of TiO2 NPs33. Sharp to the presence of minerals in the E. variegata extract.
and broad diffraction peaks were observed, which level This study confirms the active participation of E. varie-
the crystallinity and the nanosized crystallite. The data gata elements in the synthesis of TiO2 NPs. It also sup-
are in good agreement with JCPDS card no. 84-1285. The ports the high purity of the synthesized sample35. Figure 8
average size of TiO2 NPs was calculated using the Debye– shows the EDAX results.
Scherrer equation.
Photocatalytic activity of TiO2 on methylene blue
D = Kλ/β cos θ, dye solution

where D is the crystalline size under UV radiation, K the In this study, methylene blue dye solution was degraded
shape factor (0.9), λ the wavelength of X-rays (1.5406 Å), using green-synthesized TiO2 NPs and UV irradiation (multi-
β the full width at half maximum and θ is the Bragg angle. lamp photo reactor at 365 nm). UV–Vis double-beam
CURRENT SCIENCE, VOL. 123, NO. 1, 10 JULY 2022 63
RESEARCH ARTICLES

Figure 7. Scanning electron microscopic images of TiO2 NPs.

tected by gradual change in the colour of the dye solution

from deep blue to almost colourless. The characteristic ab-
sorption peak for methylene blue was noticed at 661 nm.
The control exhibited no change in colour during expo-
sure in the multi-lamp photo reactor (UV light source @
365 nm). Degradation of the dye in the presence of TiO2
NPs was verified by the decrease in peak intensity (at
661 nm) and the appearance of a new peak at 645 nm dur-
ing 210 min of exposure. Figure 9 shows the optical den-
sity graph. The optimum absorbance peak of methylene
blue dye observed at a wavelength of 661 nm decreased
from 1.193 to 0.39. This in turn shows the tendency of the
green-synthesized TiO2 NPs to adsorb and degrade the
pollutant. The formation of a new peak at 645 nm indica-
Figure 8. Energy-dispersive X-ray spectrum of TiO2 NPs.
tes that the chemical configuration of methylene blue dye
starts to break up by UV–light. After 210 min under UV
exposure the dye is found to degrade completely and ef-
fectively. When compared to the nano TiO2 prepared from
other plants extract36.

Antibacterial activity of TiO2 NPs

The antibacterial action of green-synthesized TiO2 NPs

was examined for four different pathogenic strains (Strepto-
coccus, Staphylococcus, E. coli and P. aeruginosa). Using
the disk diffusion method, zones of inhibition for Gram-
positive and Gram-negative bacteria were found to be 7,
2, 8 and 14 mm in Figure 10. The results reveal that TiO2
NPs synthesized using E. variegata leaves showed maxi-
mum antibacterial activity against P. aeruginosa com-
pared to the other bacteria37,38 in Table 2.

Conclusion
Figure 9. a, UV–Vis spectra of methylene blue dye degradation as a
function of time. b, Variation of methylene blue dye degradation with and The present study focuses on the green-synthesis of a tran-
without TiO2 NPs at different time intervals. c, Decolourization image. sition metal of TiO2 NPs using E. variegata leaf extract.
The formation of NPs was confirmed by a colour change
of the precipitate from pale yellow to sandal colour. It exhi-
spectrophotometer was used to measure absorbance of the bited the maximum absorbance at 317.6 nm in UV spec-
dye solution before and after degradation at regular time tral data. E. variegata leaf extract plays a prominent role
intervals of exposure. Dye degradation was visually de- in reducing TTIP to TiO2 NPs. The energy-band gap of
64 CURRENT SCIENCE, VOL. 123, NO. 1, 10 JULY 2022
 RESEARCH ARTICLES

Figure 10. Well diffusion assay to assess bactericidal potential of biosynthesized TiO2 NPs prepared from
Erythrina variegata leaf extract against various bacterial strains, viz. Streptococcus, Staphylococcus, Staphy-
lococus aureus, Escherichia coli and Pseudomonas aeruginosa. Bar graph represents antibacterial activity.

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