Journal of the Saudi Society of Agricultural Sciences (2015) 14, 9199

King Saud University

Journal of the Saudi Society of Agricultural Sciences

www.ksu.edu.sa
www.sciencedirect.com

FULL LENGTH ARTICLE

Antioxidant activities of phenolics, avonoids

and vitamin C in two cultivars of fennel
(Foeniculum vulgare Mill.) in responses to organic
and bio-organic fertilizers
Zeinab A. Salama a,*, Farouk K. El Baz a, Alaa A. Gaafar a,
Mohamed Fathy Zaki b
a
b

Plant Biochemistry Department, National Research Centre, Dokki, Cairo, Egypt

Vegetables Crops Department, National Research Centre, Dokki, Cairo, Egypt

Received 18 June 2013; accepted 30 October 2013

Available online 8 November 2013

KEYWORDS
Sweet fennel;
Antioxidant activity;
Organic fertilizer;
Bio-organic fertilizer;
Phenolics;
Flavonoids;
Vit. C

Abstract Field experiment was conducted to study the effect of organic and bio-organic fertilizers
on dry weight; yield, total phenolics (TPC), total avonoids (TFC), vitamin C and on their antioxidant activities of two sweet fennel cultivars Dolce and Zefa no. Results strongly showed that there
were signicant differences between sweet fennel cultivars. Generally the highest values of all
parameters were obtained when fennel plants were supplemented with 50% NPK + 50% organic
fertilizer and bio fertilizer when compared with control treatment. The highest values of TPC,
TFC and Vit. C were recorded by Zefa no cultivar when received 50% NPK + 50% organic treatment. The antioxidant activities of both cultivars were evaluated and Dolce cultivar showed the
highest DPPH scavenging activity expressed as IC50 compared with Zefa no cultivar. In addition,
Dolce cultivars exhibited the highest value for Fe2+-chelating activities for organic and bio-organic
fertilizers followed by Zefa no when compared to control treatment. Dolce cultivar generally
showed superiority than Zefa no in all measured parameters.
2013 King Saud University. Production and hosting by Elsevier B.V. All rights reserved.

1. Introduction
* Corresponding author. Tel.: +20 0235867416; fax: +20
00237610850.
E-mail addresses: zeinabsalama70@hotmail.com, dr.zeinabsalama70@gmail.com (Z.A. Salama).
Peer review under responsibility of King Saud University.

Production and hosting by Elsevier

Recently, an increasing interest in the cultivation and the

production of un-traditional vegetable crops has been noticed in Egypt in order to cover the increasing demand of
the local consumption as well as export purposes. Among
them sweet fennel (Foeniculum vulgare Mill.) is one of the
most promising new crops in Egypt. Also, Ehsanipoura
et al. (2012) found that nitrogen is one of the most important

1658-077X 2013 King Saud University. Production and hosting by Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.jssas.2013.10.004

92
nutrients needed for plant growth and seed yield, essential
oil contents of seed and foliage and ber, protein and ash
contents of seeds. It is considered as the most important
economic medicinal and aromatic plant grown within the
Mediterranean region (Kandil, 2002). The cultivated area
in Egypt is about 20003000 fed and its export value
amounts to 10 million US $ according to the Ministry of
Agriculture Statistics.
Fennel is a member of the family Apiaceae. It is classied
into two sub species vulgare and piperitum. The most important cultivated fennel cultivars belong to subspecies vulgare.
Sweet fennel is a variety group with inated leaf bases which
form an edible bulb-like structure (swollen base). Due to
medicinal and aromatic compounds content of fennel, it is
widely used in culinary preparation as a avoring agent,
food and beverages and perfumery industries, scenting soaps
and cosmetics industries as well as in confectionery. Also in
phototherapy and pharmaceutical purposes (Blumenthal
et al., 2000). It is also considered as a spice due to terpenic
compounds isolated from its fruit volatile oil (Abdallah
et al., 1978) .Vegetative growth, edible bulb yield and quality of sweet fennel plants were strongly affected by cultivars
as reported by Atta-Aly (2001), Paschalina et al. (2006), Osman (2009), Zaki et al. (2009).
Many investigators proved that phosphorus source even
as mineral or organic is urgently needed to enhance plant
growth, bulb yield and produce acceptable amounts of
essential oils with sufcient quality in sweet fennel plants
(Kandil, 2002; Rai et al., 2002; Kapoor et al., 2004; Osman,
2009 and Zaki et al., 2010). Moreover, Abd El-Salam (1999)
indicated that increasing phosphorus fertilization rates
caused an increase in plant height, number of branches/
plant, dry weight of different plant organs in sweet fennel
plants. Also, Ehsanipoura et al. (2012) reported that nitrogen is the most important nutrient needed for plant growth,
seed yield, essential oil contents of fennel.
Organic fertilization has a stimulatory effect on accumulation of phenolics in fennel. It is well known that, the higher concentrations of phenolics can be explained by the role
of organic fertilizers in the biosynthesis which induces the
acetate shikimate pathway, resulting in higher production
of avonoids and phenolics (Sousa et al., 2008). Also, because of the higher photo-pathogenic stress in organic farming, which in turn may have abiotic stress, and causes an
increase of phenolics and avonoids and their antioxidant
activity grown organically (Young et. al., 2000).
This study was undertaken to study the enhancement of
total phenolics (TPC), total avonoids (TFC), Vit. C and their
antioxidant activities in sweet fennel in response to organic
and bio-organic fertilizers.

2. Materials and methods

2.1. Chemicals
All chemicals and solvents used were HPLC spectral grade,
and obtained from Sigma (St. Louis, USA) from Merck
Shcuchrdt (Munich, Germany).

Z.A. Salama et al.

2.2. Plant materials
Two cultivars of sweet fennel (Foeniculum vulgar, Mill.) which
belong to family Apiaceae (Umbelliferae) were Dulce and Zefa
no which were obtained from Topstar Co., Holland.
2.3. Field experiment
A eld experiment was carried out at El-Saff, Hellwan Governorate, Egypt, on newly reclaimed soil. The aim is to study the
effect of organic and bio-organic fertilizers on phenolic avonoids, glucosinolates and Vit. C contents and their antioxidant
activities of two cultivars of fennel compared with NPK as
control treatment.
2.3.1. Agriculture conditions
Seeds of sweet fennel were sown in foam trays lled with a mixture of peat moss and vermiculite (1:1 volume). Seedlings at
45 days old were transplanted in the open eld on the center
of raw and spacing of 50 cm between plants. Ditches of 20 cm
depth and 40 cm width were prepared in the sites of drip irrigation lines; chicken manure, rocks phosphate, calcium super
phosphate and agricultural sulfer were mixed and were added
and covered with soil before planting. Plot area was 3.5 lengths
and 3.0 widths in three raw which equals to 10.5 m2.
2.3.2. Fertilizer treatments
2.3.2.1. Mineral NPK fertilizer (control). Basic fertilizers were
mixed with the soil in doses of ammonium sulfate (21.5% N) at
the rate of 100 N2 kg/fed; calcium super phosphate (15.5%
P2O5), and rock phosphate (30% P) at the rate of 60 P2O5
kg /fed and potassium sulfate (48% K2O) at the rate of 40
K2O kg/fed. In addition, agricultural sulfur was used at the
rate of 200 kg/fed.
2.3.2.2. Organic fertilizer. Complementary between 50% organic (chicken manure) at the rate of 100 N2 unit/fed
(5.850 m3/fed) and 50% mineral (NPK) was used. NPK is
formed from ammonium sulfate (21.5% N) as a source of nitrogen, at the rate of 100 N2 kg/fed; calcium super phosphate
(15.5% P2O5) and rock phosphate (30% P) were used at the rate
of 60 P2O5 kg/fed; Potassium sulfate (48% K2O) was used at the
rate of 40 K2O kg/fed. In addition, agricultural sulfur was used
at the rate of 200 kg/fed based on the recommended fertilizer
requirements according to the Ministry of Agriculture.
Ammonium sulfate and potassium sulfate were splinted
into three equal doses and applied as dressing (0, 30 and
60 days after transplanting) beside plants.
2.3.2.3. Bio-organic fertilizer. It contained a mixture of N2-xing bacteria Azotobacter Chrococcum and phosphate dissolving
bacteria Bacillus megaterium (1:1 v/v). Bio-organic fertilizer
was applied to the plant once after 2, 5 and 8 weeks of transplanting at the level of 10 ml/plant at concentration of
109 cell/ml. The mixture was injected near the root of the plants.
The bio-organic fertilizer was kindly supported by the Agriculture Microbiology Department, National Research Centre.
In all treatments, Drip irrigation line was spread over the
ditches and soil was irrigated continuously 3 days before
transplanting.

Antioxidant activities of phenolics, avonoids and vitamin C

2.4. Preparation of plant samples for chemical analysis
The bulbs, harvested at fresh harvest maturity stage (uniformly
green, compact) were put in polyethylene bags and transported
immediately to the laboratory, washed, chopped, and the
moisture content in the two cultivars was determined using
oven at 105 C, for 24 h. Moisture was calculated while
expressing the data on fresh weight basis.
2.4.1. Preparation of fresh plant methanolic extract

93
shaken vigorously and allowed to stand at room temperature
for 30 min. The butylated 4-ydroxyl toluene (BHT) was used
as a positive control; and negative control contained the entire
reaction reagent except the extracts. Then the absorbance was
measured at 517 nm against blank (methanol pure). Lower
absorbance of the reaction mixture indicated higher free radical scavenging activity.
The capacity to scavenge the DPPH radical was calculated
using the following equation:
DPPH scavenging effectInhibition%

The edible parts (fennel) were extracted by liquid nitrogen. Ten

grams of frozen tissues were ground in mortar with pestle and
extracted 100 ml of 80% methanol by maceration (48 h).
2.5. Determination of total phenolic content (TPC)s
Total phenolic content (TPC) of fresh sweet fennel extracts was
spectrophotometrically determined by Folin Ciocalteu reagent
assay according to Singleton and Rossi (1965), using gallic acid
as a standard compound for the preparation of calibration
curve (20120 mg/l) Total phenolic content of samples was
measured at 670 nm and expressed as mg gallic acid equivalents
(GAE)/g dry weight. All samples were analyzed in triplicates.
2.6. Determination of total avonoid content (TFC)
Total avonoid content (TFC) of fresh sweet fennel extracts
was spectrophotometrically determined by the aluminum chloride method using quercetin as a standard (Zhishen et al.,
1999). After incubation at room temperature samples were
measured at 512 nm and expressed as mg quercetin equivalents
(QE)/g fresh weight. Samples were analyzed in triplicates.

Ac  AS =Ac  100
where Ac was the absorbance of the control reaction and As the
absorbance in the presence of the plant methanolic extract.
IC50: its concentrations for 50% inhibition
2.9. Ferrous chelating activity
The chelating of ferrous ions of sweet fennel was estimated
according to Decker and Welch (1990). 5 ml of 80% sweet fennel methanolic extract, or EDTA solution as a positive control
at different concentrations (5, 10, 20, 30 lg/ml) were spiked
with 0.1 ml of 2 mM FeCl2 and 0.2 ml of 5 mM ferrozine solution. The reaction mixture was left for 10 min at room temperature then the absorbance was measured at 562 nm. The
percentage of ferrous ion chelating ability was calculated using
the following equation:
Iron chelating activity Inhibition% Ac  AS =Ac  100
where Ac was the absorbance of the control reaction and As the
absorbance in the presence of the plant methanolic extract.
2.10. Statistical analysis

2.7. Determination of vitamin C (ascorbic acid)

Ascorbic acid content in fresh samples was measured colorimetrically according to Bajaj and Kaur (1981).

Data were statistically analyzed using Costat statistical package data according to Snedecore and Cochran (1980).
3. Results and discussion

2.8. Measurements of antioxidant capacity

2.8.1. DPPH Free radical scavenging assay

3.1. Inuence of organic and bio-organic fertilization on dry

weight and yield of two sweet fennel cultivars

The method described by Blois (2002) was used to assess the

(1,1-diphenyl-2-picrylhydrazyl) DPPH radical scavenging
activity of sweet fennel methanolic extract. 1 ml of 0.1 mM
DPPH was added to 3 ml of fennel methanolic extracts at different concentrations (5, 10, 20, 30 lg/ml). The mixture was

From the data presented in Table 1 it is clear that the yield of

bulbs in both cultivars of fennel was highly increased as a result of organic and bioorganic treatments. Organic and bio-organic fertilizer exhibited the highest value of yield (7.40 and

Table 1

Effect of organic and bioorganic fertilizer treatments on weight and yield of sweet fennel cultivars.

Cultivars

Treatments

Bulbs dry weight %

Yield (ton/fed)

Yield increases %

Dolce

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio

8.03a 0.07
8.34ab 0.17
8.51ab 0.36

5.90a 0.15
7.40c 0.12
8.85e 0.09

100
125
150

Zefa no

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio

8.02a 0.39
8.32ab 0.32
8.92b 0.23
0.52

7.07b 0.10
8.08d 0.09
9.16f 0.09
0.19

100
114
130

L.S.D at 0.05

All values are the means of three replicates and are signicantly different at p P 0.05.

94

Z.A. Salama et al.

Table 2 Effect of organic and bio-organic fertilizer treatments on total phenolics (TPC), total avonoid (TFC) and Vit. C of sweet
fennel cultivars.
Cultivars

Treatments

TPC
(mga/g DW)

TPC
increase %

TFC
(mgb/g DW)

TFC
increase %

Vit. C
(mg/100 g FW)

Vit. C
increase %

Dolce

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio

4.94b 0.15
6.96e 0.18
5.71c 0.12

100
141
116

3.13b 0.37
4.89d 0.36
3.87c 0.14

100
156
124

31.49ab 0.623
34.00c 1.35
32.88bc 0.60

100
108
104

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio
L.S.D at 0.05

4.03a 0.09
6.14d 0.18
5.09b 0.11
0.27

100
152
126

2.00a 0.22
4.05c 0.29
3.03b 0.13
0.51

100
202
152

30.45a 0.63
33.69bc 1.19
32.44abc 1.03
1.64

100
111
107

Zefa no

All values are the means of three replicates and are signicantly different at p P 0.05 + Standard deviation.
a
As gallic acid.
b
As quercetin.

Table 3 Effect of organic and bio-organic fertilization on yield of total phenolic, total avonoid and vitamin C contents of bulbs in
sweet fennel cultivars.
Cultivars

Treatments

TPC
yield kg/fed

TPC yield
increases %

TFC
yield kg/fed

TFC yield
increases %

Vit. C
yield kg/fed

Vit. C yield
increases %

Dolce

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio

2.34a 0.08
4.29b 0.22
4.30b 0.24

100
183
184

1.48a 0.16
3.02c 0.33
2.92bc 0.20

100
204
197

1.86a 0.07
2.52c 0.13
2.91e 0.06

100
135
156

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio
L.S.D at 0.05

2.29a 0.12
4.12b 0.18
4.15b 0.14
0.31

100
179
181

1.14a 0.18
2.72bc 0.21
2.48b 0.07
0.38

100
239
218

2.15b 0.04
2.72d 0.08
2.97e 0.09
0.14

100
126
138

Zefa no

TPC, TFC and TGsC (yields kg/fed) = {Concentration (kg/ton) yield of orets as DW (ton/fed)}/10.

8.85 ton/fed) of Dolce cultivar and (8.08 and 9.16 ton/fed) of

Zefa no cultivar, respectively compared to control (5.90 and
7.07 ton/fed) of both cultivars. The yield increased by 25%
and 50% in Dolce cultivar and by 14% and 30% in Zefa no
cultivar by the application of organic and bioorganic fertilizers, respectively.
However, bulbs dry weight % showed no signicant response to all kinds of fertilization (Table 1). It was also
found by Badawi et al. (2005) that the yield of sweet fennel
was widely increased by bio-organic fertilization. On the
other hand the use of excessive amount of inorganic fertilizer may cause a problem for human health and environment (Arisha and Bradisi, 1999). Badawi et al. (2005)
added that the best means of maintaining soil fertility and
productivity is the addition of organic manure either alone
or with the addition of biofertilizers and mineral fertilizers.
Agamy (2004)showed that bio-organic fertilization increased
the vegetative growth, mineral content and dry mater accumulation of sweet fennel plant. Since yield is the function of
the vegetative growth, photosynthesis activity, mineral and
dry matter accumulationin .In this concern Pavla and pokluda (2008), showed that the organic fertilizers have positive
effects due to the improvement of the plant nutritional status. El-Desuki et al. (2001) found that the application of
compost at 12 ton/fed caused signicant increase in vegetative growth parameters, N, P, K content, total bulb yield
and oil content of bulb.

3.2. Inuence of organic and bio-organic fertilization on total

phenolics (TPC) and total avonoids content (TFC)
Results in (Table 2) showed that organic and bio-organic fertilization increased TPC in Dolce cultivar from 4.94 (control)
to 6.96 and 5.71 mg/g DW respectively.
The increase reached more than 41% and 16%, respectively. However, in Zefa no cultivars TPC increased from
4.03 mg/g DW in control to 6.14 and 5.09 mg/g DW in organic
and bio-organic, respectively, (52% and 26% increases, respectively). On the other hand, TFC increased in response to organic and bio-organic treatments by 56% and 24%,
respectively in Dolce cultivar and 102% and 52% in response
to organic and bio-organic treatments respectively in Zefa no
cultivar compared with control treatment (see Table 3).
These results are in agreement with those obtained by Liu
et al. (2008) who found that the total phenolic content of ethanol extract of fennel was 7.54 mg/g. Yoo et al. (2008) reported
that the total phenolics of fennel 70% methanolic extract was
7.01 mg/g DW and the total avonoids content of the same extract was 3.29 mg/g DW.
3.3. 3-Inuence of organic and bio-organic fertilizers on total
vitamin C content
Vitamin C contents of two sweet fennel cultivars treated with
organic and bioorganic fertilizers are illustrated in Table 2.

Antioxidant activities of phenolics, avonoids and vitamin C

95

80
64.57

70

56.11

58.09

IC50 g/ml

60

49.66
45.25

50

42.47

40
30
20

10.07

10
0
Dolce NPK

Dolce
Organic

Dolce
Bioorganic

Zefa fino
NPK

Zefa fino
Organic

Zefa fino
Bioorganic

BHT
Standard

Fertilizer treatments of sweet fennel cultivars

Figure 1 IC50 of DPPHfree radical of 80% methanolic extract of Sweet fennel cultivars in response to organic and bio-organic
fertilizers.

y = -0.1201x + 11.812

y = -0.1174x + 9.6669

R2 = 0.9718

R2 = 0.9527
8

TFC mg/g as DW.

TPC mg/g as DW.

8
6
4
2
0
40

50

60

6
4
2
0
40

70

50

IC 50 g/m
(Sweet fennel)

60

70

IC 50 g/ml
(Sweet fennel)

Vitamin C mg/100 g FW.

y = -0.1577x + 40.801
R2 = 0.9663
35
34
33
32
31
30
40

45

50

55

60

65

70

IC 50 g/ml
(Sweet fennel)

Figure 2

Correlation between IC50 and phenlics (a), avonoids (b) and vitamin C (c).

As shown in Table 2 Dolce cultivar contained 34.00

and 32.88 mg/100 g FW, in response to organic and bioorganic fertilizers, respectively. The increase was 8% and 4%
compared to control (31.5 mg/100 g FW). While, Zefa no
cultivars treated with organic and bio-organic fertilizers
exhibited more increase for Vit. C (11% and 7% increases,
respectively) compared to control treatments. Regarding
the variations in the vitamin C content of the two cultivars of sweet fennel, Badawi et al. (2005) reported that
vitamin C content ranged from 21.83 to 48.04 mg/100 g
FW when organic fertilizers from different sources (chicken
manure, cattle manure, compost, town refuse, half cattle + half mineral and mineral fertilization) were applied.

Koudela and Petrikova (2008) studied the content of vitamin C of three cultivars of sweet fennel and found that
the values of vitamin C content ranged from 87 to
347 mg/kg fresh matter. The cultivar Precoce di bologna recorded the highest values in summer and autumn, followed
by cultivar Zefa no while, the lowest amount of vitamin
C was recorded by cultivar Rudy F1. Barros et al. determined the total content of vitamin C in different parts
of fennel and found that the shoots seem to have the
highest ascorbic acid content (570.89 0.01 mg/g) which
is in agreement with the highest content in phenolis
(65.85 0.74 mg/g) and radical scavenging activity found
in this part.

96

Z.A. Salama et al.

Table 4 Iron chelating activity by different concentrations of 80% plant methanolic extracts of sweet fennel cultivars in response to
organic and bio-organic fertilizers.
Cultivars

Treatments

Inhibition %
5 lg/ml

10 lg/ml
b

20 lg/ml
a

30 lg/ml

Dolce

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio

9.72 0.12
15.28d 0.58
12.24b 0.50

16.25 0.20
21.20e 0.42
19.44c 0.42

28.57 0.20
36.80d 0.42
29.92b 0.35

46.00b 0.12
52.88e 0.50
47.84c 0.29

Zefa no

100% NPK (Control)

50% NPK + 50% Organic
50% NPK + 50% Organic + Bio

9.13a 0.23
13.12c 0.37
11.60b 0.24
19.02e 0.06
0.72

15.42a 0.27
20.40d 0.29
19.04c 0.42
44.35f 0.11
0.59

28.16a 0.16
32.40c 0.29
28.56a 0.42
82.32e 0.17
0.57

43.47a 0.24
49.92d 0.35
46.32b 0.35
96.49f 0.06
0.58

EDTA standard
L.S.D at 0.05

All values are the means of three replicates and are signicantly different at p P 0.05 standard deviation.

4. Effect of organic and bio-organic fertilization on yield of

TPC, TFC and Vit. C
From the results given in Table 3 the yield of TPC in sweet fennel cultivars increased by organic and bio fertilizer application.
In Dolce cultivars (TPC) was increased by more than 83% and
84% followed by Zefa no 79% and 81% compared to control
treatment. Similar trend was found in TFC of sweet fennel
cultivars treated with organic fertilizer,where it revealed high
responses estimated at about 104% and 97% in Dolce cultivar
and 139% and 118% in Zefa no cultivars, compared to control treatment. Similar trend was detected in case of Vit. C,
however biofertilizer application showed the highest values in
case of Vit. C. However bio fertilizer application showed the
highest values. In addition, the inuence of organic and bio-organic fertilizer on hydroxyl radicals generated at or near DNA
may eventually result in strand breakage of DNA (Floyed,
1981) and contribute to signicant biological effects such as
carcinogensis, mutagenesis and cytotoxicity (Swartz, 1984).
The avonoids widely used as therapeutic agents are also
known to act as strong superoxide radical O2 scavengers
(Baumann et al., 1980), Singlit oxygen O2 quenchers (Sorata
et al., 1984). Flavonoids also react with peroxy radicals involving termination of chain reaction during the autooxidation of
polyunsaturated fatty acids (Torel et al., 1986).
4.1. Antioxidant activity
4.1.1. Scavenging activity on DPPH radicals
DPPH has been widely used for the determination of antioxidant activity of different plants, vegetables, and fruit extracts
(Goupy et al., 1999; Yu, 1994). The main values of radical
DPPH scavenging activity expressed as IC50 against DPPH
radical of two sweet fennel cultivars Dolce and Zefa no in response to organic and bio-organic fertilizers compared to BHT
as a synthetic antioxidant agent are illustrated in Fig. 1.
All values are the means of three replicates and are signicantly different at p P 0.05 standard deviation. IC50: Its
concentration for 50% inhibition.
Dolce cultivars treated with organic and bio-organic fertilizers demonstrated the highest IC50 (42.47 and 49.66 lg/ml,
respectively) in comparison with control (58.09 lg/ml). In
addition, the values of IC50 for methanolic extracts of Zefa no
cultivars treated with organic and bio-organic fertilizers were

45.25 and 56.11 lg/ml compared to control NPK (64.57 lg/

ml) while, BHT exhibited the highest scavenging activity
(10.07 lg/ml). The methanolic extracts of both sweet fennel
cultivars were able to scavenge DPPH radical. These results
revealed that Dolce cultivar exhibited the highest activity when
compared with, Zefa no cultivar. Mata et al. (2007) evaluated
the antioxidant activity of ethanol and water extracts of fennel
plant .The ethanolic extract showed the highest radical
scavenging activity value (IC50 = 12 lg/ml) while, butylated
hydroxytoluene (BHT) which was used as standard gave
IC50 = 15.7 lg/ml and water extract gave IC50 = 48.0 lg/ml.
The correlation coefcient between IC50 and total phenolic
content (TPC), total avonoid content (TFC), and vitamin C
of sweet fennel cultivars is shown in Fig. 2. Results given in
Fig. 2 showed that TPC, TFC and vitamin C are highly correlated with DPPH scavenging activity assay (R2 = 0.9718),
(R2 = 0.9527) and (R2 = 0.9663), respectively (Fig. 2ac).
The results revealed that TPC, TFC and Vit. C strongly contributed to the antioxidant scavenging activity of sweet fennel
methanolic extracts. These ndings further supported the positive relationship between TP, TF and Vit. C and antioxidant
activity of different plant species. Ghasemzadeh et al. (2012)
reported the strong positive relationship between TP and antioxidant activity that appears to be the trend in many plant
species.
The importance of phenolic compounds as antioxidant
in the maintenance of health and protection against coronary heart disease and cancer is also of raising interest
among scientists, food manufactures and consumers. This
trend leads the future toward functional food with specic
health effects (Loliger, 1991). Serani et al. (1998) and Carbonneau et al. (1998) and Carbanaro et al. (2002) also suggested that avonoids and phenolics have a preventive role
in the development of cancer and heart disease. Consumption of controlled diets high in the fruits and vegetables increased signicantly the antioxidant capacity of plasma
(Cao et al., 1998). Moreover, epidemiological studies
showed that signicant negative correlation was found between the intake of fruits and vegetables and heart disease
mortality (Knekt et al., 1996). Carbanaro et al., 2002 and
Floridi et al. (2003) showed that the presence of phenolics
in food is particularly important for their oxidative stability
and antimicrobial protection. From clinical point of view,
there is numerous evidence of sweet fennel to alleviate
diseases.

Antioxidant activities of phenolics, avonoids and vitamin C

97

y = 0.3071x - 9.1792

y = 0.2979x - 10.738
R 2 = 0.9522

TFC mg/g as DW.

TPC mg/g as DW.

R 2 = 0.9851

6
4
2
0
40

45

50

55

6
4
2
0
40

Iron chelating at 30 g/ml

(Sweet fennl)

45

50

55

Iron chelating at 30 g/ml

(Sweet fennl)

Vitamin C mg/100 g FW.

y = 0.3848x + 14.121
R2 = 0.8921
35
34
33
32
31
30
40

45

50

55

Iron chelating at 30 g/ml

(Sweet fennl)

Figure 3 Correlation between iron chelating activity at 30 lg/mand phenolics (a), avonoid (b) and vitamin C (c) of sweet fennel
cultivars in response to organic and bio-organic fertilizers.

4.2. Ferrous ion-chelating activity of sweet fennel cultivars

5. Conclusion

The effect of fertilization treatments on ferrous ion chelating

power of the 80% methanolic extracts of sweet fennel cultivars
(Dolce and Zefa no) is shown in Table 4. The Fe2+-chelating
activities of organic and bio-organic fertilizers of both sweet
fennel cultivars (Dolce and Zefa no) and EDTA standard
were increased by increasing the concentration of methanolic
extracts.
Organic treatment at the concentration of 30 lg/ml
showed the highest chelating activity of both cultivars (Dolce
and Zefa no) (52.88% and 49.92%, respectively) followed
by bio-organic treatment (47.84% and 46.32%, respectively)
compared to NPK (control) (46.00% and 43.47%,
respectively).
Dolce cultivars exhibited the highest value for Fe2+-chelating activities for organic and bio-organic fertilizers (52.88 and
47.84 lg/ml, respectively) followed by Zefa no (49.92 and
46.32 lg/ml, respectively) with the same fertilizer treatments
compared to control (46.00 and 43.5 lg/ml, respectively).
The EDTA standard gave the highest chelating activity at all
concentrations used compared with sweet fennel cultivars at
all treatments (Table 4). The correlation between Fe2+-chelating activity at concentration 30 lg/ml and measured parameters in 80% methanolic extract of sweet fennel are presented
in Fig. 3.
Correlation between Fe2+-chelating activity at concentration 30 lg/ml and TPC had a very strong correlation
coefcient of R2 = 0.9851 Fig. 3a, R2 = 0.9522 with avonoids content (Fig. 3b) and R2 = 0.8921 with vitamin C
(Fig. 3c).

Based on the above mentioned results, it could be concluded

that Dolce cultivar was generally superior than Zefa no cultivar in all measured parameters.The observed antioxidant of
methanolic extract of sweet fennel cultivars may be due to
the presence of phenolics, avonoids, Vit. C and other natural
active constituents. There have been many studies showing
that different extracts of fennel have diuretic (Wright et al.,
2007), antimicrobial (Ruberto et al., 2000), hepatoprotective
(Ozbek et al., 2004) and anti-inammatory (Choi and Hwang,
2004) activities. Earlier investigation of sweet fennel led to the
isolation of phenolic components with antihypertensive activity (Nyemba et al., 1995 and Ono et al., 1996).
This study supports the potential application of sweet fennel as potent natural sources for antioxidants and/or
nutraceuticals.
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