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Viraj et al.
World Journal of Pharmacy and Pharmaceutical Sciences
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ANTIOXIDANT POTENTIAL OF ORGANIC AND CONVENTIONAL CLOVE (SYZYGIUMAROMATICUM) AND CINNAMON (CINNAMOMUM ZEYLANICUM)
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.

Article Received on 11 June 2015, Revised on 01 July 2015, Accepted on 21 July
Article Received on
11 June 2015,
Revised on 01 July 2015,
Accepted on 21 July 2015
*Correspondence for Author Viraj Roghelia Laboratory of Foods and Nutrition, P.G. Department of Home Science,
*Correspondence for
Author
Viraj Roghelia
Laboratory of Foods and
Nutrition, P.G.
Department of Home
Science, Sardar Patel
University, Vallabh
Vidyanagar-388120,
Gujarat.

ABSTRACT Spices are widely used for various culinary preparations in India and worldwide. Nowadays, spices are also popular for their antioxidants and other medicinal properties such as antimicrobial, antidiabetic, antiallergic, antimutagenic, antiulcerogenic, antipyretic, antianaesthetic, antispasmodic and carminative activities. Antioxidants prevent a specific action of initiation, propagation and oxidizing chain reaction. In the present study, organic and conventional cinnamon and clove were studied for total phenol, flavonoids and antioxidant capacity using standard methods. Results indicated that organic cinnamon contained higher content of total phenol, flavonoid, DPPHRSA and FRAP as compared to conventional cinnamon. Similar

results were observed for clove except FRAP as FRAP content of 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)

Clove

Organic

5039.09**±164.16

2289.58**±332.39

Conventional

2957.04±119.43

1516.66±165.71

Cinnamon

Organic

1642.30**±21.98

1144.79

NS ±74.36

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)

Clove

Organic

2177.55**±24.37

5656.25

NS ±54.13

Conventional

1821.60±35.90

5823.44±64.37

Cinnamon

Organic

1514.86**±16.48

2879.71**±141.41

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 R 2 value was observed for flavonoid-

antioxidant activities in organic spices [figure-1(a)] as compared to conventional spices

[figure-1(b)]. While, the higher R 2 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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al. World Journal of Pharmacy and Pharmaceutical Sciences Figure-1(a): Correlation ship of flavonoid with DPPHRSA and

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

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

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

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

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

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al. World Journal of Pharmacy and Pharmaceutical Sciences Figure-2(b): Correlation ship of total phenol with DPPHRSA

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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