International Journal of Horticulture, 2017, Vol.7, No.

4, 26-32
http://ijh.biopublisher.ca

Research Report Open Access

Determination of Phytochemical and Antioxidant Activities in Edible Flowers
M.R. Dhiman , Sandeep Kumar, Chander Parkash, Raj Kumar, Siddharth Moudgil and Sunita Sharma
ICAR-Indian Agricultural Research Institute, Regional Station, Katrain, Kullu-Valley- 175129, HP, India
Corresponding email: mrarjun01@gmail.com
International Journal of Horticulture, 2017, Vol. 7, No. 4 doi: 10.5376/ijh.2017.07.0004
Received: 19 Nov., 2016
Accepted: 25 Jan., 2017
Published: 27 Feb., 2017
Copyright ©2017 Dhiman et al., This is an open access article published under the terms of the Creative Commons Attribution License, which permits
unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited .
Preferred citation for this article:
Dhiman M.R., Kumar S., Parkash C., Kumar R., Moudgil S., and Sharma S., 2017, Determination of phytochemical and antioxidant activities in edible flowers,
International Journal of Horticulture, 7(4): 26-32 (doi: 10.5376/ijh.2017.07.0004)

Abstract In the present study, edible flower samples belonging to 14 different genera were evaluated by employing various in vitro
antioxidant assay such as total antioxidant activity determination by CUPRAC and FRAP, total phenolic content and plant pigments viz.
Lycopene and β-carotene to investigate their antioxidant potential with specific Phytochemicals composition. The results showed that
extracts Vinca Red flower exhibited highest antioxidant capacity viz. CUPRAC 32.33 µmol trolox/g followed by Vinca Pink 30.46 while,
least was observed in Gomphrina 4.983 µmol while its counterpart FRAP was highest (2.07 µmol /g) in poppy as well as in Vinca pink
and least (0.22) in wild salvia. The total phenolic content (TPC) was found to be highest in Pusa Narangi (2282.54 µg GAE/gfw). A
negative correlation was found between phenols and three carotenes. β- Carotene was found to have negative correlation with the three
antioxidant factors (CUPRAC, FRAP and Phenols) while lycopene and total caroteniods showed a positive correlation with CUPRAC
and FRAP. The results revealed the presence of antioxidants, phenols and caroteniods in most of the flowers confirming flowers as a
potential source of these phytonutrient.
Keywords Edible flowers; Phytochemicals; Antioxidant; Phenols; Lycopene; Carotene

1 Introduction
Flower is an important part of plant which contains a great variety of natural antioxidants, such as phenolic acids,
flavanoids, anthocyanins and many other phenolic compounds (Kaur et al., 2006). Due to their appealing and
desirable aesthetic aspects edible flowers are gaining renewed interest as rich source of bio active compounds,
globally. Proliferating awareness among consumers as resulting from global intent has contributed to the
comeback of the early lifestyles, in which edible flowers played an important role (Kopec and Balik, 2008).
Besides their anti-oxidative properties and anti-carcinogenic effect, phenolic acids and flavanoids have long been
recognized to possess anti-allergic, anti-inflammatory and antimicrobial activities as well (Robard et al., 1999). A
high nutritional value, antioxidant capacity and attractive appearance pre determines edible flowers to be a new
and promising foodstuff species for a wider use in human nutrition. Flowers produce a wide array of secondary
metabolites, serving both as food stuffs and medicinal products which are of interest for several Parma and
nutraceutical industries (Bungihan and Matias, 2013).

Phytochemicals are naturally occurring and biologically active plant compounds that have potential disease
inhibiting capabilities due to their antioxidant effects. The high antioxidant capacity of these flowers is correlated
with the level of flavanoids which corresponds to the medicinal properties of these flowers (Mato et al., 2000).
Studies have demonstrated a positive correlation between the qualities of phenolic content to antioxidant
properties. Oxidative stress in human results from imbalance of antioxidant in the body caused by factors as
pollution, diet, chronic infections and so on (Agarwal et al., 2005). Antioxidants play an essential role in the
prevention of disease and have capacity to reduce oxidative stress by chelating trace elements or scavenging free
radicals and protecting antioxidant defenses (Banerjee et al., 2005). Now days, the application of plant based
anti-oxidant or natural antioxidants is replacing synthetic molecules because of toxicities associated with the latter
(Cruz et al., 2001).

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As described in ancient literature, the use of edible flowers for dish preparation has been an atavistic custom.
Besides their use for fresh consumption and garnishing, edible flowers can also be consumed dried, in cocktails,
canned in sugars and preserved in distillated etc. (Neugtebauerova and Vabkova, 2009). Thus a drift towards
natural sources for mitigating these problems has gradually come into vogue. The usage of plants in the system of
medicine as antimicrobial, antiseptic and as cure for infection and sores (Bungihan and Matias, 2013), is widely
accepted by nearly 80 percent of the people throughout the world. Hence, research of indentifying specific plants
for their bioactivity and bio efficiency, is being conducted widely. However, the toxicity of the flower extracts
with high antioxidant activity needs to be tested, so as to determine the daily intake limits (Kaisoon et al., 2011).

Generally, flowers are nearly neglected or rarely studied for their biological properties. Thus analyses for the
presence of these considerable secondary metabolites, antioxidants and anti-bacterial profiles of ornaments plants
is worthwhile for the establishment of their potentials as nutraceutical, cosmeceutical and pharmaceutical
ingredient / additive (Bungihan and Matias, 2013). Keeping in view the importance of Phytochemicals, the present
study was planned to examine and extract various Phytochemicals viz. lycopene, phenols, caroteniods etc. present
in flowers crops grown under Indian condition and their antioxidant potential with specific Phytochemicals
composition. In this study, a total of 14 flowering species, grown locally, were evaluated for their antioxidant
potency and phytoconstituents.

2 Material and Methods

2.1 Basic experimental material and laboratory analysis
Flower samples belonging to 14 different genera were collected fresh from the common plant present and grown
in and around Katrain region of H.P. (India). The collected samples were carries to the lab within a maximum of 6
hours for experimentation. Their names are present in Table 1. True to type representative samples from, each
replication were collected at appropriate stages of harvesting. These were chopped, homogenized and a fresh
sample of 5 g of each was stored immediately under refrigerated conditions (-20°C) until assay. The 5 g sample
was further homogenized in 15 ml absolute ethanol to prepare the ethanol extract, which was further centrifuged
at 10,000 rpm for 15 mins at 4 degree to obtain the supernatant, which is then stored at -20°C.

Table 1 Mean performance of different edible flower crops for different quality traits
Sr. No. Crop CUPRAC FRAP Phenols Lycopene Total carotenoids β-Carotene
1. Calendula yellow 12.02 1.06 1205.91 21.85 37.83 6.98
2. Candula orange 12.26 0.88 973.59 41.94 66.64 10.79
3. Poppy 8.75 2.07 1726.33 28.06 54.22 7.80
4. Vinca red 32.33 1.39 1559.29 28.29 30.36 6.56
5. Vinca pink 30.46 2.07 2041.95 20.70 16.85 4.40
6. Coriopsis 27.48 1.10 2208.30 14.93 25.63 10.97
7. Morning glory 15.53 1.28 1404.59 3.32 3.24 4.09
8. Shoe flwoer 19.21 1.26 1300.82 4.09 4.19 5.38
9. Mirabilis 23.07 0.78 2079.72 2.78 4.43 5.64
10. Coriopsis 411 8.56 0.96 1742.85 7.64 18.11 7.83
11. Ghomprina 4.98 0.37 1590.21 0.55 5.45 6.99
12. Salvia blue 7.35 0.42 1581.28 0.69 7.46 9.66
13. Wild salvia 5.93 0.22 1804.05 0.65 5.83 7.50
14. Pusa Narangi 10.80 0.71 2282.54 16.32 13.45 5.83
C.D.(0.05) 0.42 0.05 0.48 0.08 0.10 0.11
±SE(d) 0.20 0.02 0.23 0.04 0.05 0.06
C.V. (%) 1.58 2.85 0.02 0.35 0.30 0.94

2.2 Antioxidant capacities and total phenolic assay

Measuring the antioxidant activity / capacity level of plants is carried out for the meaningful comparison of the
antioxidant content of several plants. The parent CUPRAC (Cupric Reducing Antioxidant Capacity) method of

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antioxidant measurement, is based on the absorbance measurement of the Cu (I) - neucoproine (Nc) chelate
formed as a result of the redox reaction of chain breaking antioxidants with the CUPRAC reagent, Cu (II) Nc
where absorbance is recorded at the maximal light absorption wavelength of 450 nm; thus this is an electron
transfer (ET) based method. The method described by Apak et al. 2006 was followed with minor modifications.
For CUPRAC analysis, 100 µl samples were mixed with 4ml of CUPRAC reagent (1 ml of neucoproine; 1 ml
ammonium acetate; 1 ml copper chloride and 1 ml distilled water; pH 7.4). Then the absorbance was recorded at
450 nm in spectrophotometer.

Similarly, FRAP (Ferric Reducing Ability of Plasma) was performed based on the procedure described by Benzie
and Strain (1999) with slight modifications. For this, 100 µl of the distilled sample was added to 3 ml of the FRAP
reagent and the reaction was monitored after 4 mins at 593 nm. The results were expressed as µmol Fe (II)/g fresh
weight of the sample.

Total phenolic contents were determined with Folin–Cicalteau method (Singleton and Rossi 1965). Modifications
were done accordingly for the amount of sample present. Briefly, 0.50 ml extract was mixed with 2.5 ml of 1:10
diluted Folin–Cicalteau reagent. After 4 min, 2 ml of saturated sodium carbonate solution was added. The mixture
was incubated in dark for 2 h at room temperature. The resulting complex was measured at 760 nm at the
spectrophotometer for absorbance. Gallic acid was used as a standard for the calibration, and the results were
expressed as mg of Gallic acid equivalents (mg GAE) per 100 g fresh weight (FW) of sample.

2.3 Plant pigment content

Lycopene: it is an important phytonutrient and consumers are becoming increasingly aware of the health benefits
of its consumption. For determination of Lycopene content 5 g of sample was taken and crushed with Acetone
until the residue becomes colourless. Filtrate was transferred in the separating funnel containing 20 ml of
petroleum ether. 2-3 drops of Sodium sulphate was added in separating funnel. Then 20 ml of petroleum ether was
added to make 2 separate phases. Lower phase was re extracted with additional petroleum ether till it became
colourless. Final volume was made up to 50 ml and absorbance was taken at 503 nm and 452 nm using petroleum
ether as blank.
Absorbance (1 unit) = 3.1206 µg Lycopene/ml
3.1206  OD at 503 nm  Volume made  dilution  100
Lycopene (mg in 100 g sample) 
Weight of Sample  1000

OD at 452 nm  13.9  10 4  100

β - Carotene (μg /100 g) 
Weight of Sample  1000

Statistical Analysis:
Statistical analyses were conducted using OP Stat software and SPSS. The variability estimates were worked out
through Analysis of Variance (ANOVA) in a completely randomized Design, while correlation coefficients were
determined by co variance and variance between the traits.

3 Results and Discussion

Phenolic compounds are widely distributed in fruits, vegetables and cereals. Plants vary widely both in their
phenolic composition and content which are controlled both genetically and environmentally (Awika and Rooney,
2004). These components have received considerable attention due to their antioxidant activities and free radical
scavenging capacity, which potentially have beneficial implications in human health (Imeh and Khokar, 2002).
The Analysis of Variance revealed highly significant difference amongst the flower species for all the
Phytochemicals under study, almost all the species contained good amount of antioxidants, Phenol content
ranging CUPRAC 4.98→32.33 µM trolox/ g and FRAP 0.22→2.07 µM trolox/ g, while TPC varied from 973.59
→ 2282.54 µg Gallic acid/ gfw. Vinca Red flower exhibited highest antioxidant capacity viz. CUPRAC 32.33
µmol trolox/g followed by Vinca Pink 30.46 while, least was observed in Gomphrina.4.983 µmol while its
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counterpart FRAP was highest (2.07 µmol /g) in poppy as well as in Vinca pink and least (0.22) in wild salvia.
The total phenolic content (TPC) was found to be highest in African Marigold cv.Pusa Narangi (2282.54 µg
GAE/gfw). Kaisoon et al. (2011) also reported high antioxidant activities from the edible flowers from Thailand.
Similarly Rop et al. (2012) consider edible flower to be a new and promising food stuff species for a wider user in
human nutrition, due to high antioxidant capacity.

From Table 2, a positive significant correlation was found among the phenol content and the antioxidant capacity
viz. CUPRAC (0.262= r) and FRRAP (0.027= r). Similar results were reported by Zeng et al. (2014) who reported
a statistically significant relationship between polyphenolic content and the antioxidant capacity of 19 Chinese
edible flowers. Also previous reports that phenolic compound were major antioxidant constituents in medicinal
herbs, vegetables, fruits and spices gave similar results (Cai et al., 2004; Huang et al., 2010). Caroteniods, which
have a polyisoprenoid structure, are generally found in plants, algae, photosynthetic bacteria, yeast and moulds.
They plays an important role in human nutrition, by acting as a precursor of vitamin A. lycopene and β- carotene
are among the caroteniods, popular to consumers. β- Carotene belongs to their carotene class, which is one of the
most abundantly found in diet and is used as food colourant.

Table 2 Pearson correlation coefficients among different quality traits measured in 14 edible flower crops
Traits CUPRAC FRAP Phenols Lycopene Total carotenoids β-Carotene
*
CUPRAC 1.000 0.538 0.262 0.271 0.003 -0.230
FRAP 1.000 0.027 0.509 0.391 -0.309
Phenols 1.000 -0.269 -0.381 -0.130
Lycopene 1.000 0.921** 0.304
Carotenoids 1.000 0.519
β-Carotene 1.000
Note: *Significant at p<0.05; **Significant at p<0.01

The proposed method was applied for the analysis of total caroteniods, Lycopene and β- carotene content from
different edible flowers. The amount of total caroteniods varied from 3.24 µg/100g in Morning Glory to 66.64
µg/100g in Calendula Orange. Similarly the Lycopene content was found as low as 0.55 mg/100g in Gomphrina
up to 41.94 mg/100g in Calendula Orange whereas Coreopsis had highest amount of β- carotene content (10.97
µg/100g) while Vinca Pink contained least(4.09 µg/100g).

In the correlation study among the caroteniods, a highly statistically significant correlation was found between
Lycopene and total caroteniods. A similar correlation was found amongβ- carotene and total caroteniods. A
negative correlation was found between phenols and three carotenes. β- Carotene was found to have negative
correlation with the three antioxidant factors (CUPRAC, FRAP and Phenols) while lycopene and total caroteniods
showed a positive correlation with CUPRAC and FRAP. Prakash et al. (2015) in their study also found out
Lycopene, β- carotene and Total caroteniods having Positive Association with each other. Kaulmann et al. (2014)
also reported positive correlation between FRAP and phenols

Grouping of Genotypes into various clusters

A hierarchical cluster analysis has been shown in the dendrogram constructed in Figure 1. The variations observed
in the cluster means also points out to the degree of variability and divided the 14 species into two major groups A
and B. Group A was further divided into two subgroups viz. A1 and A2. The subgroup A1 comprised of three
species i.e. 1, 8 and 2, while subgroup A2 accommodated seven species i.e. 3, 10, 13,4,11, 12 and 7. In the
meanwhile, group B was subdivided into subgroups B1 and B2 which comprised of 2 species each viz. 5 and 9 &
6 and 4 respectively.

In the present study two species of group A2 viz. 11 and 12 were found to be genetically most similar in regards
to the six quality traits considered. These are the closest substitute present to each other in the nutrient
composition present in them.

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Result of PCA indicated that first two components having Eigen value greater than one retained in the analysis
because of the substantial amount of variation amongst them. The two components had a variance of 42.07 and
30.62 percent aggregating to total of 72.69 percent of the total variation explained (Table 3).

Figure 1 Dendrogram showing clustering pattern of 14 edible flower crops based on six quality traits constructed using complete
linkage Euclidean distance method

Table 3 Eigen vectors for first three principle components of quantitatively measured traits in eight SI lines of cabbage
Principle Component*
Traits
PC1# PC2
CUPRAC 0.244 0.798
FRAP 0.564 0.707
Phenols -0.390 0.456
Lycopene 0.959 0.061
Carotenoids 0.958 -0.218
β-Carotene 0.399 -0.664
Eigen Value 2.52 1.83
Percentage of variance 42.07 30.62
Cumulative % of variance 42.07 72.69
Note: #PC: Principal component; *Extracted through principle component analysis; **
Bold value indicates the highest Eigen vector
for the corresponding trait amongst the three principal components

The first factor (PC1) had the highest positive values for lycopene (0.959), caroteniods (0.958) and β- carotene
(0.399) content; while the second factor was found superior for CUPRAC (0.798), FRAP (0.707) and Phenols
(0.456). The positive values of different traits in three components indicated its importance in divergence among
the 14 different species of edible flowers whereas, negative value showed the least contribution to divergence.

Further loading of different traits based on first two principle components indicated that CUPRAC, FRAP and
Lycopene are the main components of divergence among the 14 species of edible flowers, whereas, Phenols,
caroteniods and carotenes had lesser contributions, comparatively in the divergence. Banerjee et al. (2013) had
also applied the unweighted pair group clustering through dendrogram showing inter-relationships between food
flowers species and grouped them into two high level clusters based on activity.

4 Conclusions
The current study on the antioxidants ability and phytoconstituents of 14 species of edible flowers provided the
results supporting their usage as nutrient rich raw material. The results revealed the presence of antioxidants,
phenols and caroteniods in most of the flowers confirming flowers as a potential source of these phytonutrient.
However, there stand needs to assess their toxicological and pharmacological effects and establish their safety
limits. Furthermore, there are still lots of flowers which could be given consideration. The obtained results should

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contribute to the popularization of edible flowers as a new and prospective source for food, pharma and
nutraceutical industry; as well as a promising object of human nutrition.
Refrences
Agarwal A., Gupta S., and Sharma R.K., 2005, Role of oxidative stress in female reproduction, Reproductive Biology and Endocrinology, 3: 28
https://doi.org/10.1186/1477-7827-3-28
PMid:16018814 PMCid:PMC1215514
Apak R., GüçlüK., Özyürek M., Esin Karademir S., and Erçağ E., 2006, The cupric ion reducing antioxidant capacity and polyphenolic content of some herbal
teas, International Journal of Food Sciences and Nutrition, 57(5-6), 292-304
https://doi.org/10.1080/09637480600798132
PMid:17135020
Awika J.M. and Rooney L.W., 2004, Sorghum phytochemicals and their potential impact on human health, Phytochemistry, 65: 1199-1221
https://doi.org/10.1016/j.phytochem.2004.04.001
PMid:15184005
Banerjee A., Dasgupta N., and De B., 2005, In vitro study of antioxidant activity of Syzygium cumini fruit, Food chemistry, 90(4), 727-733
https://doi.org/10.1016/j.foodchem.2004.04.033
Banerjee A., and De B., 2013, Comparative study of antioxidant activity of the food flowers of West Bengal, India, International Journal of Food Properties,
16(1), 193-204
https://doi.org/10.1080/10942912.2010.535188
Benzie I.F., and Strain J.J., 1999, Ferric reducing/antioxidant power assay: Direct measure of total antioxidant activity of biological fluids and modified version
for simultaneous measurement of total antioxidant power and ascorbic acid concentration, Methods Enzymol, 299: 15-27
https://doi.org/10.1016/S0076-6879(99)99005-5
Bungihan M.E., and Matias C.A., 2013, Determination of antioxidant, phytochemical and antibacterial profiles of flowers from selected ornamental plants in
Nueva Vizcaya , Philippines, Journal of Agriculture Science and Technology, 3: 833-841
Cai Y., Luo Q., Sun M., and Corke H., 2004, Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with
anticancer, Life sciences, 74(17), 2157-2184
https://doi.org/10.1016/j.lfs.2003.09.047
PMid:14969719
Moure A., Cruz J.M., Franco ., om ngue . ., ineiro ., om ngue ., ... and Parajó J.C., 2001, Natural antioxidants from residual sources, Food
chemistry, 72(2), 145-171
https://doi.org/10.1016/S0308-8146(00)00223-5
Huang W.Y., Cai Y.Z., Corke H., and Sun M., 2010, Survey of antioxidant capacity and nutritional quality of selected edible and medicinal fruit plants in Hong
Kong, Journal of Food Composition and Analysis, 23(6), 510-517
https://doi.org/10.1016/j.jfca.2009.12.006
Imeh U. and Khokar S., 2002, Distribution of conjugated and free phenols in fruits: antioxidant activity and cultivar variation, Journal of Agricultural and Food
Chemistry, 50(22), 6301-6306
https://doi.org/10.1021/jf020342j
PMid:12381107
Kaisoon O., Siriamornpun S., Weerapreeyakul N., and Meeso N., 2011, Phenolic compounds and antioxidant activities of edible flowers from Thailand, Journal
of functional foods, 3(2), 88-99
https://doi.org/10.1016/j.jff.2011.03.002
Kaulmann A., Jonville M.C., Schneider Y.J., Hoffmann L., and Bohn T., 2014, Carotenoids, polyphenols and micronutrient profiles of Brassica oleraceae and
plum varieties and their contribution to measures of total antioxidant capacity, Food chemistry, 155, 240-250
https://doi.org/10.1016/j.foodchem.2014.01.070
PMid:24594181
Kaur G., Alam M.S., Jabbar Z., Javed K., and Athar M., 2006, Evaluation of antioxidant activity of Cassia siamea flowers, Journal of ethnopharmacology,
108(3), 340-348
https://doi.org/10.1016/j.jep.2006.05.021
PMid:16846707
Kopec K. and Balik J., 2008, Kvalitologie zahradnickych produktu, Brno. MZLU., 34-40
Mato M., Onozaki T., Ozeki Y., Higeta D., Itoh Y., Yoshimoto Y., ... and Shibata M., 2000, Flavonoid biosynthesis in white-flowered Sim carnations (Dianthus
caryophyllus), Scientia Horticulturae, 84(3), 333-347
https://doi.org/10.1016/S0304-4238(99)00140-5
Neugebauerova J. and Vabkova J., 2009, Jedle kvety soucast food styling, Zahradnictvi, 83: 22-24
Parkash C., Dey S.S., Kumar R., Dhiman M.R., Dhiman M., Kumar S., and Kumar V., 2015, Comparative analysis for antioxidant capacities, important plant
pigments and total phenolic contents in different Brassica vegetables, Molecular Plant Breeding, 6
https://doi.org/10.5376/mpb.2015.06.0010
31
International Journal of Horticulture, 2017, Vol.7, No. 4, 26-32
http://ijh.biopublisher.ca

Robards K., Prenzler P.D., Tucker G., Swatsitang P., and Glover W., 1999, Phenolic compounds and their role in oxidative processes in fruits, Food chemistry,
66(4), 401-436
https://doi.org/10.1016/S0308-8146(99)00093-X
Rop O., Jurikova T., Mlcek J., Kramarova D., and Sengee Z., 2009, Antioxidant activity and selected nutritional values of plums (Prunus domestica L.) typical
of the White Carpathian Mountains, Scientia Horticulturae, 122(4), 545-549
https://doi.org/10.1016/j.scienta.2009.06.036
Singleton V.L., and Rossi J.A., 1965, Colorimetry of total phenolics with phosphomolybdic-phosphotungstic acid reagents, American journal of Enology and
Viticulture, 16(3), 144-158
Zeng Y., Deng M., Lv Z., and Peng Y., 2014, Evaluation of antioxidant activities of extracts from 19 Chinese edible flowers, SpringerPlus, 3(1), 315
https://doi.org/10.1186/2193-1801-3-315
PMid:25013750 PMCid:PMC4082252

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