Phytochemical and biological screening is necessary for the isolation of functional groups which lead to the discovery of new active compounds. In the present work, the crude aqueous extract of Daucus carota were tested for phytochemical screening and HPLC analysis which indicated the presence of various class of bioactive secondary metabolite such as terpenoids, flavonoids, saponins, steroids and glycosides. HPLC analysis was performed to quantify the β-carotene content in pulp and aerial extract of Daucus carota.

1. IJSRD - International Journal for Scientific Research & Development| Vol. 2, Issue 07, 2014 | ISSN (online): 2321-0613
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Comparative Study on Phytochemical Screening and HPLC Analysis of
Daucus Carota Pulp and Aerial Parts
V. Kavitha1
N. Gunavathy2
1
M.Phil Scholar 2
Assistant Professor
1,2
Department of Chemistry
1,2
Nirmala College for College, Coimbatore, Tamilnadu
Abstract— Phytochemical and biological screening is
necessary for the isolation of functional groups which lead
to the discovery of new active compounds. In the present
work, the crude aqueous extract of Daucus carota were
tested for phytochemical screening and HPLC analysis
which indicated the presence of various class of bioactive
secondary metabolite such as terpenoids, flavonoids,
saponins, steroids and glycosides. HPLC analysis was
performed to quantify the β-carotene content in pulp and
aerial extract of Daucus carota.
Keywords: active compounds, Daucus carota , pulp, aerial,
phytochemical screening, HPLC
I. INTRODUCTION
Plants are richest source for phytochemicals and
nutraceuticals. It plays a role as an alternative medicine for
drugs. Phytochemicals are primary and secondary
compounds found in plants. These active substances in
plants have been used for medicinal purposes from the
prehistoric time, as home remedies, an over-the-counter
drug products, and dietary supplement.
Chlorophyll, proteins and common sugars are the
primary constituents and secondary compounds have
terpenoid, alkaloids and phenolic compounds [1]. The active
phytoconstituents of plant have become keen interest in
recent time due to its resourceful applications. Secondary
metabolites present in smaller quantities in higher plants,
include the alkaloids, steroids, flavonoids, terpenoids,
tannins, and many others [2]. According to (Li et al) [3],
phenolic compounds is one of the most numerous groups of
substances in plant kingdom ranging from simple molecules,
such as phenolic acids, to complex compounds, such as
tannins.
Flavonoids constitute a group of natural
compounds that occur in fruits, vegetables, wine, tea,
chocolate and other cocoa products. Daily dietary intake of
flavonoids and similar polyphenols exceeds that of
antioxidative vitamins and provitamins [4]. Phytochemicals
accumulate in different parts of the plants, such as in the
roots, stems, leaves, flowers, fruits or seeds.
Daucus carota Linn. has been used for treatment of anti-
diarrhea, anti-infection, anti-high blood cholesterol, anti-
inflammation, anti-seizure, anti-fungal anti-bacteria and
anti-cancer [5-11]. Previous studies revealed the presence of
phenolic, flavonoid, carotenoid and anthocyanin in
ethylacetate and methanolic extract of Daucus carota roots
during a phytochemical screening [12-17]. Survey of
literature provided information on Daucus carota pulp.
Until now, we have no information about Daucus carota
aerial extract.
The objective of study involved the phytochemical
evaluation and HPLC analysis of Daucus carota plant.
Extraction methods are used for separation of active
constituents of plant tissues using solvents through standard
procedures. Different isolation methods are available to
identify active compounds present in crude extract like thin
layer chromatography (TLC), gas chromatography (GC),
high performance liquid chromatography (HPLC) and more.
But the easiest and simplest method is phytochemical
screening. The phytochemical screening method employed
in this study was to find out qualitatively and quantitatively
the phytochemical constituents in the pulp and aerial extract
of Daucus carota plant (Fig. 1). The techniques played may
have significant role in the search for additional resources of
raw material for pharmaceutical industry.
Plant: Daucus carota
Aerial part - DCA
Pulp - DCP
Fig. 1: hotograph of Daucus carota parts used as corrosion
inhibitor
II. MATERIALS AND METHODS
A. Collection of plant materials
The study was carried out on the Daucus carota aerial part
(DCA) and pulp (DCP). The samples were obtained from
the commercial market, Coimbatore, India. The aerial part
were separated from the fruit pulp, cleaned and made into
small pieces, washed and then was air dried under shade
(Fig. 2. and 3). The dried samples were ground into powder
using an electronic blender and the fine powder was stored
in air tight container and was used for phytochemical
analysis.
Fig. 2: Dried pulp materials of Daucus carota (DCP)

2. Comparative Study on Phytochemical Screening and HPLC Analysis of Daucus Carota Pulp and Aerial Parts
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Fig. 3: tried aerial parts of Daucus carota (DCA)
B. Phytochemical Screening
Dried powder of Daucus carota pulp and aerial parts were
subjected to preliminary phytochemical screening. The
methods of analysis were carried out using standard
qualitative methods as described by various researchers [18,
19]. The samples were screened for the presence of
compounds at T Stanes Phytopharma Lab, Coimbatore,
India.
C. Qualitative Phytochemical Analysis
1) Detection of reducing sugar
2 ml of the extract was added to Fehling’s solution and
boiled for 5 min. Formation of a brick red precipitate
indicated the presence of reducing sugar.
2) Detection of Alkaloids
Extracts were treated with Mayer’s reagent and formation of
cream coloured precipitate indicated the presence of
alkaloids.
3) Detection of Saponins
Extracts were diluted with 20 ml of distilled water and
shaken in a graduated cylinder for 15 min. Formation of 1
cm layer of foam indicated the presence of saponins.
4) Detection of Tannins
To 2 ml of the extract, few drops of 1% lead acetate were
added and the formation of yellowish precipitate indicated
the presence of tannins.
5) Detection of Flavonoids
To a small quantity of the extract dilute sulphuric acid was
added. The appearance of orange colour indicated the
presence of flavonoids.
6) Detection of Total phenol
Extract with 3% ferric chloride resulted in deep blue colour.
This indicated the presence of phenol.
7) Detection of Steroids
2 ml of the extract with 2 ml of chloroform, acetic acid and
1 ml of concentrated sulphuric acid resulted in violet to blue
green formation, indicating the presence of steroids.
8) HPLC analysis
HPLC analysis was performed for DCP and DCA extract
using HPLC Waters system, equipped with 515 HPLC pump
and waters 2998 photodiode array detector at T Stanes
Phytopharma Lab, Coimbatore. The compounds were
identified by comparing retention time and reference
compounds.
III. RESULTS AND DISCUSSIONS
A. Qualitative Phytochemical Analysis
Much plant analysis is devoted to the isolation and
identification of secondary constituents in a particular
species or group of species or species with the expectation
that some of the constituents may be novel or of an unusual
structure. Many compounds that occur in plant tissues are
quite labile and almost inevitably may undergo change
during extraction. The polarity of the solvent for extraction
may influence the group of bioactive compounds obtained
from the Daucus carota material. The phytochemicals
present in pulp and aerial extract of Daucus carota in
aqueous medium are summarized in Table.1.
S. No. Phytochemical DCP DCA
Reducing sugar +++ +
Alkaloids ++ +
Protein ++ ++
Phenols ++ ++
Flavonoids + +
Amino acids ++ ++
Tannin + ++
Steroids - -
Glycosides + +
Saponins - -
Table 1 Phytochemical constituents present in the extract of
Daucus carota
Key: “+++” active compound copiously present,
“++” active compound moderately present, “+”active
compound present, “-“ active compound absent
The Table 1 indicated that protein, phenol and
amino acids were moderately present in the DCP and DCA
extract. DCP and DCA extracts showed the active presence
of flavonoids and glycosides. DCA showed active presence
of reducing sugar and alkaloids. The presence of phenols,
flavonoids, alkaloids, glycosides, and tannins was
confirmed. The exceptional factor was reducing sugar
content seems to be high in DCP than DCA extract of
Daucus carota. Tannin composition was moderately present
in DCA where as in DCP; it showed an active presence of
tannin.
Phenolic compounds are the largest group of
phytochemicals and account for most of antioxidant activity
of in plants [20]. Phenolic compounds are a large and
complex group of chemical constituents found in plants
[21]. Phenolics have been shown to contain higher levels of
antioxidant activities [22].
Flavonoids are the largest group of plant phenols
and the most studied [23]. Flavonoids have been stated to
possess many useful properties, containing anti-
inflammatory activity, enzyme inhibition, antimicrobial
activity, oestrogenic activity, anti-allergic activity,
antioxidant activity, vascular activity and cytotoxic
antitumor activity [24]. Flavonoids constitute a wide range
of substances that play important role in protecting
biological systems against the harmful effects of oxidative
processes on macromolecules, such as carbohydrates,
proteins, lipids and DNA [25].
Tannins are active detoxifying agents by
precipitating the protein component and hence inhibiting
their growth. This effect agrees with the mode of action of
aminoglycoside antibiotic (e.g streptomycin in which they
first latch onto surface receptor on the bacteria before
exerting their action). Other uses of tannins are to stop
haemorrhage, to treat diarrhea, as well as local burns

3. Comparative Study on Phytochemical Screening and HPLC Analysis of Daucus Carota Pulp and Aerial Parts
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treatment where they precipitate proteins in the burned area,
thus forming a protective layer [26].
Glycosides are non-volatile and serve as defense
mechanisms of plants against predation by many
microorganisms and herbivores [27]. At lower
concentrations, tannins inhibit the growth of
microorganisms and act as anti-fungal agents; at higher
concentrations they act by coagulating the protoplasm of the
microorganism [28]. Alkaloids are significant for the
protecting and survival of plant because they ensure their
survival against micro-organisms (antibacterial and
antifungal activities), insects and herbivores (feeding
deterrens) and also against other plants by means of
allelopathically active chemicals [29]. The tannin-containing
plant extracts are used as astringents, against diarrhoea, as
diuretics, against stomach and duodenal tumours [30], and
as anti-inflammatory, antiseptic, antioxidant and
haemostatic pharmaceuticals [31].
It is clearly evident from the tabulation that the
other phytoconstituents like steroids and saponins were
absent in parts of Daucus carota. The presence of
phytocompound in turn helps in development of new bio
products as corrosion inhibitor for future.
B. HPLC analysis
1) Analysis of Daucus carota Pulp Extract
β-carotene is the predominant compound in Daucus carota
pulp and leaf [32]. The analysis was performed to quantify
and identify the β-carotene in DCP. The phenolic
compounds were eluted with 89% acetonitrile and 11%
dichloromethane as mobile phase at flow rate of 1 mL/min
with injection volume 10 µL/min using detection
wavelength at 254 nm. The extract was quantified and
chromatogram of the pulp extract was done to determine the
β-carotene composition in the pulp. The HPLC fingerprint
chromatogram of DCP produced 17 peaks in phenolic
compounds (Fig. 4).
Peaks of phenolic compounds were identified by
comparison of retention times with those of authentic
reference compounds. β-carotene appeared at 3.653 min in
the chromatogram. Phenolic compounds also contribute for
the inhibition efficiency of DCP extract. The composition of
β-carotene was quantified in DCP extract and found to be
2.47 mg/100 g.
Fig. 4: HPLC chromatogram of DCP extract
2) Analysis of Daucus carota Aerial Extract
The phenolic compounds were eluted with 89% acetonitrile
and 11% dichloromethane as mobile phase at flow rate of 1
mL/min with injection volume 10 µL/min using detection
wavelength at 254 nm. The HPLC fingerprint chromatogram
of DCA produced 13 peaks in phenolic compounds (Fig. 5).
Peaks of Phenolic compounds were identified by
comparison of retention times with those of authentic
reference compounds. β-carotene appeared at 3.687 min in
the chromatogram. The plant extract propose inhibition on
mild steel due to presence of effective organic molecules as
functional groups. Phenolic compounds also contribute for
the inhibition efficiency of DCA extract. The composition of
β-carotene was quantified in DCA extract and found to be
4.26 mg/100 g.
Fig. 5: HPLC chromatogram of DCA extract
β-carotene is a precursor (inactive form) to vitamin
A via the action of beta-carotene 15,15'-monooxygenase
[33]. β-carotene is used to decrease asthma symptoms
caused by exercise; to prevent certain cancers, heart disease,
cataracts, and age related macular degeneration (AMD); and
to treat AIDS, alcoholism, Alzheimer’s disease, depression,
epilepsy, headache, heartburn, high blood pressure,
infertility, Parkinson’s disease, rheumatoid arthritis,
schizophrenia, and skin disorders including psoriasis and
vitiligo. Β-carotene is an effective treatment for EPP [34]. β
-carotene is also in used in malnourished (underfed) women
to reduce the chance of death and night blindness during
pregnancy, as well as diarrhea and fever after giving birth.
Some people who sunburn easily, including those with an
inherited disease called erythropoietin protoporphyria (EPP)
[35], use beta-carotene to reduce the risk of sunburn. Blood
concentrations of carotenoids including are more strongly
associated with reduced breast cancer risk than are
carotenoids assessed by dietary questionnaires [36].
IV. CONCLUSION
The phytochemical screening on Daucus carota pulp and
aerial peel in aqueous extract showed the presence of active
metabolites of viable importance which act as the precursors
for the production of secondary metabolites. HPLC analysis
showed that β-carotene content was more in aerial extract
compared to pulp extract which suggest Daucus carota
aerial part has an economic importance and cannot be
neglected. These in turn help in the development of new
compounds which could provide an alternative therapeutic
importance.
ACKNOWLEDGEMENT
Authors are highly thankful to Management and Principal of
Nirmala College for Women, (Autonomous), Coimbatore,
for their kind support and encouragement.
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