Original Research Papers
An Endangered Medicinal Plant, Plumbago zeylanica L.
and Its Substitute, Dyerophytum indicum (Gibbs ex Wight)
Kuntze: A Comparative HPTLC, HPLC Study, and Screening
of Antioxidant Activity
Richa Tyagi*, Ekta Menghani, and Gaurav Sharma
Key Words:
Plumbago zeylanica
Dyerophytum indicum
High-performance thin-layer chromatography
High-performance liquid chromatography
Endangered species
Antioxidant activity
Summary
Plumbago zeylanica L. (PZ) is a significant medicinal plant in
Ayurveda, so is used for the treatment of various disorders. The
main active part of this plant is the root, and due to its inherent uses,
it has been exploited hence becoming an endangered species. Dyero-
phytum indicum (Gibbs ex Wight) Kuntze (DI), mainly considered
as a substitute of PZ, is also an important folklore medicine, used
in many health problems. Both plants are much similar in their
physical as well as chemical properties. However, an effective val-
idation is required before declaration of substitution. In the pres-
ent study, quantitative and qualitative estimations were performed
on both plants with the help of modern analytical techniques. Si-
multaneous high-performance liquid chromatography (HPLC)
and high-performance thin-layer chromatography (HPTLC) and
quantitative determinations of β-sitosterol have been performed
for comparing both plants, i.e., PZ and DI. HPTLC fingerprinting
analysis was also performed comparatively in different plant parts
of PZ and DI. Successive extracts from different plant parts were
evaluated for TLC separation profile of secondary metabolites.
A comparative polyphenolic content- and antioxidant screening
was evaluated to check the free radical scavenging effect of both
plants (leaf, stem, and root) in comparison with the standards gal-
lic and ascorbic acid.
1 Introduction
Besides knowing the facts about the significance of medicinal
plants, more than 90% of the medicinal plants are picked up from
wild sources and it is reported that, sometimes, people tried to
reseed them [1, 2]. In more than 75% of the reported cases, a
destructive harvesting was noticed as people need different parts
of the plants like root, leaves, stem, and bark and, sometimes, the
R. Tyagi and G. Sharma, Department of Life Sciences, Suresh Gyan Vihar Uni-
versity, Mahal Jagatpura, Jaipur 302017, India; and E. Menghani, Department of
Biotechnology, JECRC University, Sitapura Industrial Area Extn, Ramchandra-
pura, Vidhani Village, Jaipur 303905, India.
E-mail: richatyagi31@gmail.com
Journal of Planar Chromatography 30 (2017) 1, 17–24
Journal of Planar Chromatography 29 (2016) 3
0933-4173/$ 20.00 © Akadémiai Kiadó, Budapest
whole plant, too. Furthermore, people also disturb the natural
environment by factors like suburbanization, development, and
habitat destruction that lead to the destruction of wild plant life.
According to a survey conducted by the International Union for
Conservation of Nature (IUCN), there are 33,799 plant species
out of which 381 are extinct, 372 are in the danger zone, 6523
are in the list of endangered plants, and the remaining plants
are rare species [3]. As reported, there are approximately 200
medicinal plants in the Southern and Northern regions of India
[4]. The evergreen rainy forest of South India is rich in therapeu-
tic plants, and Plumbago zeylanica L. and Plumbago rosea L.
are also recorded in this region, but both curative plants are on
the edge of extinction; the prime reasons are deforestations and
the adversarial conditions of the environment [5].
Deforestation has became a major problem because herbal med-
icines have touched a decline phase because of the low density
of plants in a number of forests, which drops the availability
of fresh plant material for medicines. Up to the present day,
approximately 50% of the tropical forests and animal diversity
have been destructed because of various reasons. Unfortu-
nately, some of these medicinal plants are on the edge of extinc-
tion and some plants have been lost due to the lack of awareness
and deforestation. This is the reason that valuable herbal medic-
inal plants are becoming rare and precious. In India, deforest-
ation is performed at an annual rate of 1.5 million hectares per
year. Now, only 8% are left as against a requirement of 33% of
the geographical zone. Unfortunately, some ancient knowledge
and treasured plants are under the threshold of extinction at a
shocking rate. As reported in The Red Data Book, in India,
around 427 plant species have been recorded, out of which 28
species are extinct, 100 are exceptional species, 34 species are
not sufficiently known, 81 are susceptible, and 124 are consid-
ered as endangered species [6]. Thus, to preserve the knowl-
edge of herbal medicines, the conservation of the species the
mankind has received from past generations is important. Some
plants are facing the threat of extinction and endangerment
issues. Thus, there is a need for the conservation of these plants
[7]. Further, it should be compulsory to accept common univer-
sal methods to preserve the plants and to promote new ideas for
protecting plants on an ecological basis in the future [8].
DOI: 10.1556/1006.2017.30.1.2
17
A Comparative HPTLC, HPLC Study and Screening of Antioxidant Activity
P. zeylanica (PZ), family Plumbaginaceae (known as “lead
wort-white flowered” and “Ceylon leadwort”), is inherent in
the Southern regions of Asia [9]; it is spread in humid and
subtropical countries of the world and found in huge quan-
tities in the mid region of India to the Western part of Ben-
gal, Maharashtra, and Uttar Pradesh, and to specific regions
of Southern India [10]. It is commonly known as Chitrak [11,
12], an ancient herbal plant that created the base for Ayurvedic
medicine for numerous ailments over a long period of time.
It is widely nurtured in our country and also cultivated at a
commercial level.
PZ is documented for its pharmaceutical activities with impe-
rious biochemical complexes [13‒16]. It is reported to contain
phenolic composites, triterpenoids, steroids, coumarins, gly-
cosides, starches, saponins, tannins, alkaloids, naphthoqui-
nones, flavonoids, fatty acids, amino acids [17‒20], seselin [21],
5-methoxyseselin [22], and suberosin [23] as well as xanthyle-
tin and xanthoxyletin which are present in its roots [24]. Among
all, “plumbagin” is an active chemical of the plant that mainly
accumulates in the root, where l-dopa [25], plumbagin (naph-
thoquinone), droseron, chitranone, triterpenoid, and anthraqui-
none are present. The HPTLC quantification and validation of
the plumbagin content were also performed in polyherbal oil
formulations containing PZ as an ingredient [26].
Since the ancient times, PZ has been utilized for skin diseases
(leprosy, scabies, dermatitis, pimples, lesions, and pustules),
contaminations, and abdominal worm, and many scientists
believed that malaria, rheumatism, abdominal worm, anemia,
shock, toxic inflammation, and furunculous scabies can be
cured with the plant [27–31]. Ayurveda mentioned Chitrak as
a rejuvenator, and it is acclaimed for its bitter taste. Chitrak is
also used in combination with vata, sunthi, and kutaja, and it
is a very beneficial medicine for diarrhea, associated with
abdominal pain. The decoction of the plant is helpful to recover
abdominal sicknesses like splenomegaly, hepatomegaly, and
ascites. It increases digestion and appetite [32].
Dyerophytum indicum (DI) can be found in dense forest areas,
near bushes, on dry rocks, and mostly on hilly places. It is pres-
ent in waste lands of arid regions and, sometimes, it is found
on marine shores including salty planes, particularly in tropical
Asia and at some places of western Asia. The flowers are red in
color and bloom all year round. The individual flowers are up
to ½ inches.
DI is a small plant or herb, classified also as a shrub sometimes.
It attains a height of 1-2 m and it is a flowering plant. The plants
and their extracts have been used to cure many diseases for cen-
turies. People use DI to cure asthma and chest infections. Pastes
from DI root are applied to cure skin contaminations, scabies,
and moles. The root pastes are applied on the forehead to cure
headache as well. As reported, the early shoots and stems are
mostly utilized for therapeutic uses; these parts are used as
dried powders for different breathing alignments. The early
shoots and branches of this plant are highly salty and they are
used in cooking in rural areas at times of salt shortage.
The present study deals with the ethnomedicinal background
of DI. Since the Charak era, DI has been used as a wonderful
substitute for PZ and as a potential alternative to cure various
ailments. Therefore, it is high time to validate DI which has
never been experimentally demonstrated as a potential thera-
peutic agent in lieu of PZ.
18
2 Experimental
2.1 Materials and Methods
For collecting test plants, the arid zone of Rajasthan, India, was
selected; DI (whole plant) and PZ (whole plant) were collected
from the district Mt. Abu, India, in February, 2011. As reported,
the test plants were used by the tribal community of Mt. Abu
for different medicinal needs. Both plants were submitted
to the Herbarium (Department of Botany) at the University
of Rajasthan. The identification numbers for the plants were
given as D. indicum (Gibbs ex Wight) Kuntze – leaf, root, and
stem (RUBL211590) and P. zeylanica L. – leaf, root, and stem
(RUBL211589). All the reference standards were procured from
Sigma-Aldrich (St. Louis, MO, USA), and the other chemicals
from Merck (Darmstadt, Germany).
2.2 Extraction of Test Plants
The fresh plant material was thoroughly washed with water to
remove all debris and it was shade dried; the material was then
powdered using electric grinder at 20 mesh size. The test sam-
ples were Soxhlet-extracted in petroleum ether, benzene, chlo-
roform, ethyl acetate, methanol, and distilled water for up to
6 h. Then, the extracts were filtered, dried, and finally weighed
separately for further investigations [33].
2.3 Physiochemical Investigations
Basic physiochemical tests were performed in relation to
extractive values, loss on drying, and ash values [34, 35].
2.4 Total Polyphenolic Content
The total phenolic content (TPC) was evaluated by a colorimetric
Folin‒Ciocalteu method. Briefly, 1 mL of deionized water and
0.5 mL of different extracts of known dilution (1 mg mL−1) were
added to reaction vials. The Folin‒Ciocalteu reagent (0.5  mL)
was added to the solutions and it was allowed to react for 6 min.
Then, 2.5 mL of 7% sodium carbonate solution was added to each
reaction vial, and the mixture was diluted to 6 mL with deionized
water. The color was allowed to develop at room temperature for
20 min, and the final absorbance was read at 760 nm using a
Thermo UV1 spectrophotometer after 90 min. A  calibration
curve was prepared using different concentrations of standard
gallic acid solutions, by running an analysis each time. The total
phenolic contents in the samples were calculated from the stand-
ard calibration curves, and the results were expressed in % w/w
gallic acid equivalents of the dry extract. Each sample was meas-
ured in triplicate, and the mean was taken [36].
2.5 Antioxidant Activity
2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging
activities of the extracts were investigated. Briefly, to a meth-
anolic solution of DPPH (100 mM, 2 mL), an amount of 2.0
mL of test sample dissolved in methanol was added at different
concentrations (0.05–0.2 mg  mL−1 for DI and 0.5–2 mg  mL−1
for PZ). Equal amount of methanol was added to the control.
Absorbance was recorded at 517 nm at 15 min. The scavenging
activity was calculated using the formula of scavenging activity
(%) = [(A517 of control − A517 of sample) / A517 of control] ×
100. Ascorbic acid was used as a positive control [36, 37].
Journal of Planar Chromatography 30 (2017) 1
 A Comparative HPTLC, HPLC Study and Screening of Antioxidant Activity

2.6 HPTLC Investigations Simultaneous and successive extraction was performed on the
High-performance thin-layer chromatographic study was per- leaf, root, and stem of both plants (DI and PZ). The fractions
formed on different successive extracts of PZ and DI. The basic were extracted in different solvents followed by their increas-
comparison of their secondary metabolites and TLC patterns ing polarities, starting from benzene, chloroform, ethyl acetate,
in different extracts was developed. The spots were visualized methanol, and distilled water. Successive extract values are pre-
in visible light by chemical derivatization or staining of plates sented in Figure 2.
using 0.5% anisaldehyde‒sulfuric acid reagent and dried at Quantitative determinations of total polyphenols performed on
105°C for 15 min. methanolic extract of both plants (leaf, stem, and root) showed
Thin-layer chromatography was performed on Merck silica gel significant amounts of polyphenolic content (equivalent to
60G F254 (20 cm × 20 cm) plates (catalog No. 100390). Succes- gallic acid) which reflects the possible presence of secondary
sive extracted samples and standard compounds β-sitosterol of
known concentrations were applied to the plate as 6-mm-wide
bands positioned 10 mm from the bottom and 15 mm from the
side of the plate, using a CAMAG (Muttenz, Switzerland) Lino-
mat V automated TLC applicator with nitrogen flow providing a
delivery speed of 150 nL s−1 from the applicator syringe. These
conditions were kept constant throughout the analysis of sam-
ples. Following sample application, the layers were developed in
a CAMAG twin-trough glass chamber which was presaturated
for 20 min with the solvent system: acetone‒hexane (5:15) till the
proper separation of the bands up to 8 cm height. After develop-
ment, the TLC plates were dried with an air dryer. Standard and
test spots were visualized in visible light by chemical derivati-
Figure 1
zation with dipping in 0.5% anisaldehyde‒sulfuric acid reagent
and dried at 105°C for 15 min. β-Sitosterol was quantified using Total ash value, acid insoluble ash, water, and alcohol soluble ex-
a CAMAG TLC Scanner equipped with CAMAG winCATS tractive, loss on drying in leaf, root, and stem of PZ and DI. All re-
sults are in percentage (%).
software. The following scan conditions were applied: slit width,
6 mm × 0.45 mm; wavelength, 600 nm; and absorption–reflection
mode. In order to prepare calibration curves, a stock solution of
β-sitosterol in methanol (0.1 mg mL−1) was prepared, and various
volumes of the solution were analyzed by HPTLC; the calibration
curves of peak area vs. concentration were also plotted [38, 39].

2.7 HPLC Investigations

High-performance liquid chromatographic analysis was per-
formed on a liquid chromatography system of Thermo Scientific
(Waltham, MA, USA) on an Ultimate 3000 system equipped
with quaternary gradient pump system (pressure, 620 bars), an
online degasser, injection port with loop of 20 µL, ultraviolet–
Figure 2
visible (UV–vis) detector, and Chromeleon 6.8 software. Each
analysis was repeated three times, and the respective retention Successive extractive values in different solvents of leaf, root, and
stem of PZ and DI. All results are in percentage (%).
time and the area under the curve have been averaged. HPLC
conditions were as follows: Column Dionex Acclaim 120 A,
C-18 (5 mm, 4.6 mm × 250 mm); solvent system, isocratic sys-
tem methanol–acetonitrile (15:85); flow rate, 1 mL min−1; col-
umn temperature, ambient; injection volume, 20 µL; standard
concentration, 0.1 mg mL−1; sample concentration, 25 mg mL−1;
and UV–vis detection, 254 nm [40].

2.8 Statistical Analysis

The results are expressed as mean ± SEM of three determi-
nants. The comparison among the groups was tested by one-
way ANOVA. P < 0.05 values were considered significant.

3 Results and Discussion

The physiochemical parameters of different parts of both DI Conc. (mg mL –1)
and PZ, ash value, and extractive values were determined. The
observed values are shown in Figure 1. The results were very Figure 3
much comparable between PZ and DI. Total polyphenol content; linear curve of gallic acid.

Journal of Planar Chromatography 30 (2017) 1 19

 A Comparative HPTLC, HPLC Study and Screening of Antioxidant Activity

Table 1
Total polyphenolic content and antioxidant activity of different parts
of PZ and DI, where n = 3 and *P < 0.05, **P < 0.01, and ***P < 0.001.
Total polyphenolic content
(equivalent to gallic acid)

(IC 50 in mg mL−1 ± SD)

P value for comparison

P value for comparison

Methanolic extracts

scavenging activity
between PZ and DI

between PZ and DI
DPPH free radical
(% mean ± SD)

PZ leaf 1.74 ± 0.13 3.75 ± 0.032 Conc. (mg mL –1)

** ***
Figure 4
DI leaf 2.65 ± 0.21 3.33  ± 0.045
The antioxidant activity of ascorbic acid.
PZ stem 2.43 ± 0.18 1.2  ± 0.025
* ***
DI stem 2. 58 ± 0.23 0.94  ± 0.0078

PZ root 2.85 ± 0.09 1.11 ± 0.021

* ***
DI root 3.05 ± 0.19 0.108  ± 0.006

Ascorbic acid Not applicable – 0.00317 ± 0.0001 –

metabolites as antioxidant compounds in both PZ and DI. The

standard curve of gallic acid deduced at the straight-line equa-
tion of y = 122.5x + 0.016 with a correlation coefficient (R2) of
0.996, shown in Figure 3. The quantitative results in PZ and DI
samples are given in Table 1. Conc. (mg mL –1)
The DPPH free radical scavenging activity was performed on Figure 5
the methanolic extracts of leaf, stem, and roots of both plants,
The antioxidant activity of different samples of PZ and DI. Graph
PZ and DI. DPPH radicals reacted with different reducing
of % inhibition versus concentration of sample in mg mL–1. R, root;
agents losing color with a number of electrons consumed which
L, leaf; S, stem.
was measured in the visible region spectrophotometrically at

Table 2
Straight-line equation and correlation coefficient found in the DPPH
free radical scavenging activity of different samples of PZ and DI.

Sample Straight-line equation Correlation coefficient (R2)

PZ root y = 34.35x + 11.78 0.968

PZ leaf y = 14.22x + 3.37 0.999

PZ stem y = 33.74x + 9.29 0.996

DI root y = 355.8x + 11.58 0.944

DI leaf y = 14.23x + 2.55 0.994

Figure 6
DI stem y = 31.40x + 20.30 0.983 TLC profile at white light of derivatized PZ successive extracted
plant parts. PZSPE, Plumbago zeylanica stem pet. ether; LPE, leaf
Ascorbic acid y = 13.79x − 1.272 0.996 pet. ether; BE, benzene; CH, chloroform; EA, ethyl acetate; ME,
methanol; Beta Sito, β-sitosterol.

20 Journal of Planar Chromatography 30 (2017) 1

 A Comparative HPTLC, HPLC Study and Screening of Antioxidant Activity

517 nm. The comparison of IC 50 values between PZ and DI

Figure 7
TLC profile at 366 nm of derivatized PZ successive extracted plant
parts. PZSPE, Plumbago zeylanica stem pet. ether; LPE, leaf pet.
ether; BE, benzene; CH, chloroform; EA, ethyl acetate; ME, metha-
nol; Beta Sito, β-sitosterol.
in Table 1 clearly reflects that DI is more active to scavenge the
DPPH free radical than PZ. The free radical scavenging activity
was found to be concentration dependent (Figure 3); the related
straight line equation and correlation coefficient (R2) are shown
in Table 2. The comparison with standard ascorbic acid (stand-
ard curve in Figure 4) and other methanolic extracts follows
the increasing % inhibition with increased concentration and,
thus, follows a straight-line equation. The comparative profile
of antioxidant activity in graphical representation is shown in
Figure 5. The root extract of DI showed the best antioxidant
activity followed by PZ root, DI and PZ leaf, and stem extract.
Moreover, the different parts of both plants have the same corre-
lation in antioxidant activity, but DI is more potent in scaveng-
ing DPPH free radical.
The HPTLC analysis of different successive extracts is shown
in Figures 6‒9. The related TLC profile differentiated on the
basis of different plant parts in both PZ and DI. The comparative
evaluation of the secondary metabolite extraction on the basis
of the separation profile of different extracts in their Rf values is
shown in Tables 3 and 4. Most of the Rf values and spot colors
were found common in both plants, which clearly designates a
very similar separation profile on TLC and strongly supports
the presence of the same secondary metabolites.
The quantitative determination of biomarkers was also regarded
as an important tool for the standardization and compara-
tive evaluation between two different plants. β-Sitosterol was
taken as standard reference marker for its quantitative deter-
mination in PZ and DI. HPTLC and HPLC quantification were

Table 3
Comparison of R f values of different successive extracts of PZ and
Figure 8 DI. TLC plate derivatized with anisaldehyde–sulfuric acid and visu-
alized in visible day light.
TLC profile at white light of DI successive extracted plant parts.
SPE, Dyerophytum indicum stem pet. ether; LPE, leaf pet. ether;
BE, benzene; CH, chloroform; EA, ethyl acetate; ME, methanol; Rf values on TLC plate
B-SITO, β-sitosterol.
Plant extract
Plumbago zeylanica Dyerophytum indicum
(solvent)

Stem Leaf Stem Leaf

Petroleum ether 0.42, 0.56, 0.08, 0.2, 0.35, 0.56, 0.35, 0.56,
0.61 0.38, 0.42, 0.57, 0.60 0.64
0.45, 0.54,
0.56, 0.59,
0.61, 0.63

Benzene 0.14, 0.18, 0.42, 0.45, 0.14, 0.18, 0.14, 0.35,

0.29, 0.34, 0.56 0.35, 0.56, 0.44, 0.54,
0.42, 0.45, 0.51, 0.64 0.56, 0.54,
0.56, 0.61 0.56, 0.67

Chloroform 0.05, 0.18, 0.24, 0.4, 0.14, 0.35, 0.14, 0.35,

0.56 0.42, 0.56 0.57, 0.94 0.57, 0.94

Ethyl acetate No spot 0.24, 0.56 0.14, 0.35, 0.34, 0.57,

Figure 9
observed 0.57, 0.94 0.60
TLC profile at 366 nm of derivatized DI successive extracted plant
parts. SPE, Dyerophytum indica stem pet. ether; LPE, leaf pet. Methanol 0.14, 0.56 No spot 0.14, 0.35 No spot
ether; BE, benzene; CH, chloroform; EA, ethyl acetate; ME, metha- observed observed
nol; B-SITO, β-sitosterol.

Journal of Planar Chromatography 30 (2017) 1 21

A Comparative HPTLC, HPLC Study and Screening of Antioxidant Activity

Table 4
Comparison of R f values of different successive extracts of PZ and
DI. TLC plate derivatized with anisaldehyde–sulfuric acid and visu-
alized in UV 366 nm.

Rf values on TLC plate

Plant extract
Plumbago zeylanica Dyerophytum indicum
(solvent)

Stem Leaf Stem Leaf

Petroleum ether 0.42, 0.56, 0.18, 0.2, 0.35, 0.55, 0.35, 0.56,
0.61, 0.64, 0.24, 0.42, 0.56, 0.64 0.64
0.78 0.45, 0.50,
0.56, 0.59, Figure 10
0.61, 0.63
HPTLC profile of reference standard β-sitosterol; TLC profile of dif-
ferent concentrations of β-sitosterol; five-point calibration curve.
Benzene 0.14, 0.18, 0.24, 0.50, 0.14, 0.18, 0.14, 0.35,
0.29, 0.34, 0.61 0.35, 0.55, 0.44, 0.54,
0.42, 0.45, 0.57, 0.64 0.57, 0.67
0.50, 0.54,
0.56

Chloroform 0.24, 0.49, 0.24, 0.4, 0.14, 0.54, 0.14, 0.54,

0.54 0.42, 0.56 0.57, 0.94 0.57, 0.64,
0.73

Ethyl acetate 0.54 0.24, 0.49, 0.14, 0.35, 0.54, 0.64,

0.56, 0.76 0.54, 0.57, 0.73, 0.94
0.64, 0.73,
0.94

Methanol 0.56, 0.76 0.76 No spot No spot

Figure 11
observed observed
Spectral analysis of β-sitosterol with all tracks.

Figure 12
HPLC chromatograms in overlaid position for the quantitative determination of β-sitosterol in different extracts. 1, Comparison of standard
with PZLPE (Plumbago leaf petroleum ether extract) and DPLPE (Dyerophytum leaf petroleum ether extract); 2, comparison of standard with
PZSBE (Plumbago stem benzene extract) and DPSBE (Dyerophytum stem benzene extract); 3, comparison of standard with PZSEA (Plumba-
go stem ethyl acetate extract) and DPSEA (Dyerophytum stem ethyl acetate extract).

22 Journal of Planar Chromatography 30 (2017) 1


Table 5
Quantitative determination and comparison of β-sitosterol by
HPTLC and HPLC profiles. Values are in mean ± SD.
Extracts/samples HPTLC results (% w/w) HPLC results (% w/w)
PZLPE 0.285 ± 0.038 0.277 ± 0.023
DPLPE 0.161 ± 0.031 0.154 ± 0.021
PZSBE 0.279 ± 0.018 0.288 ± 0.019
DPSBE 0.160 ± 0.030 0.163 ± 0.025
PZSEA 0.182 ± 0.019 0.176 ± 0.028
DPSEA 0.143 ± 0.033 0.150 ± 0.024
Values are in mean ± SD; PZ: Plumbago zeylanica; DP: Dyerophytum
indicum; LPE: leaf petroleum ether extract; SBE: stem benzene extract;
SEA: stem ethyl acetate extract
performed on both plants. The results of standard curve, TLC
profile, and HPTLC spectral analysis are shown in Figures 10
and 11. HPLC chromatograms and the separation profile of dif-
ferent extracts containing β-sitosterol have been compared and
evaluated in Figure 12. The results of HPLC and HPTLC quan-
titative determinations in both plants are compared in Table 5.
4 Conclusion
The above study is concluded with some important research
outcomes for confirming the scientific validations for the com-
parison between an original and a substitute plant species.
Medicinal plants are getting depleted and becoming endan-
gered with time, and the only way to preserve and regenerate
them is through plant genetic engineering. However, some-
times these tools are more costly and typical to be executed.
In contrast, the substitution of an original plant species with its
same species and genus plant is much easy to correlate, with
the same medicinal benefits. The present study is an evalua-
tion of analytical findings in both plants, PZ and DI. Moreover,
the results were very much similar and correlated when com-
parison of each parameter took place. The HPTLC and HPLC
quantification studies of β-sitosterol have shown comparative
results between both, that is, instruments and plant extracts.
Other parameters, HPTLC fingerprint and antioxidant activity
showed that DI has probably the same chemical separation pro-
file and is much more effective in free radical scavenging activ-
ity than PZ. Thus, the above scientific outcome supports our
study to correlate D. indicum as a substitute of P. zeylanica.
Acknowledgments
The authors are thankful to Mr. Sunil Sharma (Chancellor) and
Dr. Sudhanshu Sharma (Chief Mentor) of Suresh Gyan Vihar
University, Jaipur, India, for providing stage for this research.
Journal of Planar Chromatography 30 (2017) 1
A Comparative HPTLC, HPLC Study and Screening of Antioxidant Activity
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Ms received: June 22, 2016
Accepted: November 27, 2016
Journal of Planar Chromatography 30 (2017) 1