WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Viraj et al. World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 5.210

Volume 4, Issue 08, 1347-1356 Research Article ISSN 2278 – 4357

ANTIOXIDANT POTENTIAL OF ORGANIC AND CONVENTIONAL

CLOVE (SYZYGIUMAROMATICUM) AND CINNAMON
(CINNAMOMUM ZEYLANICUM)

Viraj Roghelia* and V. H. Patel

Laboratory of Foods and Nutrition, P.G. Department of Home Science, Sardar Patel
University, Vallabh Vidyanagar-388120, Gujarat, India.

ABSTRACT
Article Received on
11 June 2015, Spices are widely used for various culinary preparations in India and
Revised on 01 July 2015, worldwide. Nowadays, spices are also popular for their antioxidants
Accepted on 21 July 2015
and other medicinal properties such as antimicrobial, antidiabetic,
antiallergic, antimutagenic, antiulcerogenic, antipyretic,
*Correspondence for antianaesthetic, antispasmodic and carminative activities. Antioxidants
Author
prevent a specific action of initiation, propagation and oxidizing chain
Viraj Roghelia
reaction. In the present study, organic and conventional cinnamon and
Laboratory of Foods and
Nutrition, P.G. clove were studied for total phenol, flavonoids and antioxidant
Department of Home capacity using standard methods. Results indicated that organic
Science, Sardar Patel cinnamon contained higher content of total phenol, flavonoid,
University, Vallabh
DPPHRSA and FRAP as compared to conventional cinnamon. Similar
Vidyanagar-388120,
results were observed for clove except FRAP as FRAP content of
Gujarat.
conventional clove was found to be higher. Regression analysis
revealed that antioxidant capacity of organic spices could be due to their flavonoid content.
Hence, the study concludes that organic spices could be a better source of dietary
antioxidants.

KEYWORDS: Antioxidant, organic, conventional, clove and cinnamon.

INTRODUCTION
Antioxidants are well known for prevention of various chronic diseases by delaying,
inhibiting and oxidation of lipid and other molecules. They neutralize the free radicals and
prevent many non-communicable diseases. The free radicals are considered as etiological

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factors in the development of non-communicable chronic diseases such as cancer, diabetes,

cardiovascular diseases, auto immune disorders, neurodegenerative disorders, ageing and so
on.[1,2] Plant polyphenols are aromatic hydroxylase compounds, commonly found in
vegetables, fruits and many food sources that form a significant portion of our diet.[3] Many
herbs and spices, which are traditionally used for the food preparation are rich source of
antioxidants.

Clove (Syzygium aromaticum) and Cinnamon (Cinnamomum zeylanicum) are two most
commonly consumed spices in India and worldwide. These two spices are abundant source of
antioxidants that neutralize free radicals or their actions. The essential oil of clove buds is
widely used for its medicinal properties such as antispetic, analgesic, antibacterial, antifungal
activities and it is also important for dental care.[4] Cinnamon is recognised as a functional
food source of antioxidants which help decrease oxidative stress by inhibiting the enzyme 5-
lipooxygenase improving insulin sensitivity.[5] Some studies have mentioned cinnamon
possesses anti-carcinogenic, anti-microbical and anti-inflammaroty effects.[6,7]

These days, consumers are concerned about nutrition, health and food safety. The awareness
about some special aspects such as genetically modified organisms, irradiation of foods and
use of synthetic pesticide residue in foods.[8] Due to this, the popularity of natural foods and
organic foods continues to grow dramatically. Nowadays, many consumers prefer organic to
nonorganic food because of their interest in both a healthier-safer diet and a better
environment.[9,10] The production of organic products is also increasing day by day. India is
the second leading country in among the ten countries with the largest increase of organic
farmland in 2011 as well as India is the leading country with the highest number of organic
producers (547,591 producers) in the world.[11] Organic foods are produced using methods
that do not involve modern synthetic inputs such as synthetic pesticides and chemical
fertilizers, do not contain genetically modified organisms, and are not processed using
irradiation, industrial solvents, or chemical food additives. The use of genetic engineering,
sewage sludge, and irradiation also are prohibited in organic production and processing.[12,13]
The agricultural practices followed in organic and conventional farming varied a lot. Organic
foods are reported with higher bioactive compounds and antioxidant capacity.[14,15,16] The
present study was planned with objectives to analyse non-nutritional bioactive compounds
and antioxidant activity of organic and conventional cinnamon and clove.

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MATERIALS AND METHODS

Samples
Two most commonly consumed spices namely Clove (Syzygium aromaticum) and Cinammon
(Cinnamomum zeylanicum) were selected for the study. Organic samples (having organic
certification) were collected from certified organic store while conventional samples were
procured from local grocery store of Anand (Gujarat, India). Samples were purchased twice
to analyse all the compounds from two different extractions.

Chemicals used
Trolox, Gallic acid, Rutin, Ascorbic acid, Folin cio-calteu, 1,1-Diphenyl-2-picrylhydrazyl
(DPPH) and 2,4,6- Tris (2-pyridyl)-s-triazine (TPTZ) were purchased from Sigma-Aldrich
Ltd.( India).

Preparation of the sample for total phenol, flavonoid and total antioxidant capacity
The samples were powdered and 1 gm of homogenized powdered sample was weighed and
extracted using 80% acidified methanol (pH-2.0) in shaker (Genie Ltd.) for 30 minutes at
room temperature (37°C). The extracts were centrifuged at 7000 rpm for 10 minutes and
supernatant were collected in separate flask. The residues were further extracted by adding
the same solvent and the extraction procedure was repeated for three times. Volume made to
a round number of collected extracts and sample were stored at -20°C and used for total
phenol, flavonoid and antioxidant activity analysis.

Total Phenol content assay

Total phenol content was measured by the spectrophotometric method.[17] Aliquots (0.1ml
and 0.2 ml) sample extracts of the sample were mixed with remaining volume of Distilled
water to made the total volume 1 ml which were further mixed with 1ml of folin cio-calteu
(50% v/v with D/W) and 1 ml of 35 % sodium carbonate reagents. All the tubes were
vortexed (Gilson Ltd.) and kept for 1 hour incubation at 37°C. 2 ml of distilled water was
added to all the tubes after incubation and the absorbance was measured at 620 nm against
D/W as a blank. Gallic acid was used as a standard. Total Phenol content was expressed as
Gallic acid Equivalent.

Flavonoid assay
The concentrated extracts of the samples were used to analyse flavonoid content by
spectrophotometric method. [18] Appropriate aliquots of the sample were taken and made 1 ml

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with 95% methanol. Tubes were further added with 0.1 ml of 10 % aluminium chloride, 0.1
ml of 1M Potassium acetate and 2.8 ml of D/W. After incubation of 30 minutes at 37°C,
absorbance was measured at 415 nm against 95% methanol as a blank. 10 mg% Rutin was
used as a standard. Flavonoid content was expressed as rutin equivalents (mgRE/100g).

Ferric Reducing Antioxidant Power

Total antioxidant capacity of extracts was determined by using the method.[19] Suitable
aliquot of extracted sample was taken in clean and dry test tube and volume was made 300 µl
with distilled water and 1.8 ml of freshly prepared FRAP reagent ( Acetate buffer (pH- 3.6) +
TPTZ solution+ 20 mM Ferric chloride) was added and vortexed. After incubation of 10
minutes at 37°C, the absorbance was read at 593nm. Different aliquots of Trolox were treated
as standard and results were expressed as mg TE/100g.

DPPH radical scavenging activity

1,1- diphenyl, 2-picrylhydrazyl (DPPH) scavenging activity was measured by the
spectrophotometric method.[20] A solution of DPPH in methanol was prepared freshly. About
3 ml aliquot of this solution was mixed with 0.1 ml of the samples. The solutions in the test
tubes were shaken well and incubated in the dark for 15 min at room temperature. The
absorbance was measured at 517 nm against methanol as blank. Control tube containing 1 ml
of methanol and 3 ml of DPPH reagent was also noticed for absorbance. Gallic acid was used
as a standard and Total antioxidant capacity was expressed as Gallic acid Equivalent
(mgGAEq/100g).

Statistical analysis
Mean, standard error of mean, t –test and regression analysis was done to determine the difference
between the means and significance was noted at P < 0.01 level using SPSS (version-16).

RESULT AND DISCUSSION

Polyphenols compounds are a class of free radical terminator. The products of the metal
oxide reduction have a blue colour that exhibits a broad light absorption with a maximum at
765 nm. The intensity of light absorption at that wavelength is proportional to the
concentration of phenols. Total phenol and total flavonoid content of organic and
conventional spices are shown in Table-1. Total phenol content of organic clove and
cinnamon were found to be significant (p<0.01) higher as compared to conventional ones.
Similar result was also noted for flavonoid content of clove. The flavonoid content was found

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higher in organic cinnamon, however no significant difference was noted while comparing
the type of growing system.

Table-1: Total phenol and flavonoid content of organic and conventional spices
Cultivation Total Phenol Flavonoid
Practice ( mgGAE/100g) ( mgRE/100g)
Organic 5039.09**±164.16 2289.58**±332.39
Clove
Conventional 2957.04±119.43 1516.66±165.71
Organic 1642.30**±21.98 1144.79NS±74.36
Cinnamon
Conventional 1063.46±60.45 1057.29±44.94
Values are Mean ± Standard Error of Mean (N=4),
NS-No significant difference,** indicates significant difference between organic and
conventional at p<0.01 level.

The antioxidant activity of phenolic compounds is mainly due to redox properties, which
allow them to act as reducing agents, hydrogen donors, singlet oxygen quenchers, heavy
metal chelators, and hydroxyl radical quenchers.[21] In the present study, antioxidant activity
measured as DPPH radical scavenging activity (DPPHRSA) and Ferric Reducing Antioxidant
Power (FRAP). The results obtained for DPPHRSA and FRAP are presented in table-2. The
DPPHRSA of organic clove and cinnamon was 19.54% and 28.88% higher (p<0.01)
respectively as compared to their counterparts. Ferric reducing antioxidant power of organic
cinnamon was also found significantly (p<0.01) higher by 34.07% than conventional one.
However, no significant difference was observed for FRAP in clove pertaining to cultivation
practices.

Lv et al.(2012)[22] have reported that gallic acid, catechin, EGCG, vanillic acid, syringic acid,
ferulic acid, p-coumaric acid were found higher in organic cinnamon as compared to
conventional cinnamon but significant (p<0.05) difference was noticed for ferulic acid and
syringic acid. It was also noticed that insoluble bound phenolic compounds were found
higher in organic cinnamon and pepper mint. Liyana and shahidi (2012)[23] noted the
importance of bound insoluble phenolic compounds to antioxidant activities. Roghelia and
Patel, (2013)[24] have reported higher antioxidant activities in organic turmeric as compared
to conventional turmeric.

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Table-2: Antioxidant activity of organic and conventional spices

Cultivation DPPHRSA FRAP
Practice ( mgGAE/100g) (mgTE/100g)
Organic 2177.55**±24.37 5656.25NS±54.13
Clove
Conventional 1821.60±35.90 5823.44±64.37
Organic 1514.86**±16.48 2879.71**±141.41
Cinnamon
Conventional 1175.40±32.27 2147.85±178.35
Values are Mean ± Standard Error of Mean (N=4),
NS-No significant difference, ** indicates significant difference between organic and
conventional at p<0.01 level, DPPHRSA: DPPH radical scavenging activity, FRAP: Ferric
Reducing Power Assay

The higher antioxidants in organic spices could be due to variation in agricultural practices
followed in organic and conventional farming. In organic farming plants are unprotected by
pesticide and fungicide. Hence, the plants under organic farming suffer from higher biotic
and abiotic stress. This results into activation of the shikimate pathway especially
modulation in the activity of phenylalanine ammonia lyase (PAL). Consequently, this defense
mechanism leads to increases the antioxidants such as plant polyphenolics.[25,26] Another
reason for higher antioxidant potential in organic crops could be the type of fertilizers used.
The use of synthetic fertilizers in conventional farming makes nitrogen more available for the
plants and this could accelerate plant growth and development. This supportive mechanism
makes plant resources for growth purposes, resulting in a decrease in the production of plant
secondary metabolites such as organic acids, polyphenolics, chlorophyll, and amino acids.[27]

Regression analysis was done to study the correlationship of total phenol and flavonoids with
the antioxidant activity. All the studied correlationships were found positive and significant
for both organic and conventional spices. The higher R2 value was observed for flavonoid-
antioxidant activities in organic spices [figure-1(a)] as compared to conventional spices
[figure-1(b)]. While, the higher R2 value was observed for total phenol- antioxidant activities
in conventional spices as compared to organic ones [figure-2(a) and 2 (b)]. This reveals that
the antioxidant capacity of organic spices could be due to their flavonoid content.

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Figure-1(a): Correlation ship of flavonoid with DPPHRSA and FRAP in organic spices

Figure-1(b): Correlation ship of flavonoid with DPPHRSA and FRAP in conventional

spices

Figure-2(a): Correlation ship of total phenol with DPPHRSA and FRAP in organic
spices

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Figure-2(b): Correlation ship of total phenol with DPPHRSA and FRAP in conventional
spices

CONCLUSION
Antioxidant potential, flavonoid and total phenol were found to be higher in organic spices as
compared to conventional spices. Hence it may be concluded that organic cinnamon and
cloves can be considered as food source for antioxidant activity with reduced threat of
adverse effects of pesticides.

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