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Int. J. Agrl. Sc. & Vet. Med. 2013 Venkatasai Kumar T et al., 2013

EFFECT OF METHANOLIC EXTRACTSOF CAESALPINIA SAPPAN ON PRESERVATION

OF CHICKEN MEAT

*Corresponding Author: Venkatasai Kumar T, [email protected]

Research Paper

ISSN 2320-3730 www.ijasvm.comVol. 1, No. 3, August 2013

© 2013 Meghana Publications. All Rights Reserved

Int. J. Agrl.Sc & Vet.Med. 2013

1 Department of Veterinary Biochemistry, College of Veterinary Science, Tirupati.2 Department of Veterinary Public Health and Epidemiology, College of Veterinary Science, Proddatur.

INTRODUCTIONOxidation in fresh meat can lead to discoloration

due to formation of metmyoglobin from either

myoglobin (Potter, 1986) or oxymyoglobin

(O’Grady et al., 2001). Oxidative processes lead

to the degradation of lipids and proteins and are

one of the primary mechanisms of quality

deterioration and limiting the shelf life in meat and

meat products (Lui et al., 2010). Oxidative

processes also responsible for the reactive

oxygen species (ROS) that cause oxidative

changes in carbohydrate, DNA, lipids and protein

(Koşar et al., 2008). Free radicals are responsible

for causing various diseases, such as heart

diseases, stroke and cancer, as well as for aging

process (Mothana et al., 2008).

An antioxidant can quench reactive free

radicals thus preventing the oxidation of other

molecules and may therefore have health

promoting effects. Antioxidants can delay or inhibit

the oxidation propagation of oxidizing chain

reactions in the oxidation process (Zheng and

Wang, 2001) and considered as important

nutraceuticals because of many health benefits

Venkatasai Kumar T1*, Eswara Prasad P1, Padmaja K1 and Jagadeesh Babu A2

The present study was conducted to evaluate the antimicrobial and antioxidant activity ofmethanolic extracts of Caesalpinia sappan. Chicken meat samples obtained from local retailmarket were treated with methanolic extracts of C.sappan alone and in combination with antibiotic(Chlortetracycline), acetic acid and chlorinated water to study the antioxidant effect and reductionin the surface microflora. Various parameters like pH, lipid peroxidation (TBARS), Peroxide value,Free Fatty Acids (FFA), Standard Plate count, Yeast and mould count, Psychrophilic counts,Coliform count, MRSA and Salmonella counts were evaluated. It was observed that methanolicextract of C. sappan and methanolic extract of C. sappan with antibiotic significantly exhibitedbeneficial effects on preservation of meat as compared to other conventional treatments.

Keywords: Heart wood, Meat preservation, Rancidity, Oxidation

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(Valko et al., 2007). Meat products containing

natural antioxidants, as opposed to synthetic

derivatives, are more desirable from a consumer

viewpoint (Mitsumoto et al., 2005). Moreover,

natural antioxidants are reported to be more

powerful than the synthetic antioxidants,

especially, rosemary, sage, and green tea

extracts. Therefore, natural antioxidants are very

important for human health (Bozkurt, 2006). The

search for natural antioxidants and other

preparations of plant origin to achieve this

objective has been intensified and many plants

are reported to posses free radical scavenging

and antimicrobial activity.

Caesalpinia sappan (heart wood) exhibited

strong antioxidant activity when its ethylacetate

and methanolic extracts were studied (Badami

et al., 2003). Hence an attempt is made to study

the free radical scavenging and antimicrobial

activity of C. sappan and its effect on meat

preservation.

MATERIALS AND METHODSa. Preparation of Methanolic andAqueous Extracts of CaesalpiniaSappan Stem Bark

The stem bark of the C. sappan was made into

coarse powder. 100 g of stem bark powder was

soaked in 500 ml of methanol (95% v/v) in a round

bottomed flask and stirred occasionally for 72 h.

The contents were filtered using muslin cloth

followed by Whatmann No. 1 filter paper. The

methanolic filterate was evaporated using rotar

evaporator at 64°C. The yield obtained was 10%

w/w.

b. Treatment of Raw ChickenSamples

The raw chicken samples were obtained from

local retail meat market. Each sample was

treated with methanolic extract of C. sappan plus

antibiotic (T1) , methanolic extract of C. sappan

(T2) , acetic acid (T

3) and chlorinated water (T

4)

and effect of the treatments were evaluated using

different parameters.

b. pH

The pH of the samples was determined by

following the procedure of Jay (1964).

c. 2-Thiobarbituric Acid ReactiveSubstance Value (TBARS)

TBARS value was determined based on

procedure of Witte et al. (1970).

d. Free Fatty Acids (FFA)

The methodology adopted by Pearson (1973) was

followed to determine the free fatty acids present

in the refrigerated and frozen samples.

e. Peroxide Value

The procedure outlined by AOAC (2005), was

followed to determine the peroxide value in the

refrigerated and frozen samples.

f. Microbial Counts

The mesophilic, the psychrophilic and the yeast

and mould counts per gram of chicken meat

samples at refrigerated (4±1°C) temperature were

estimated as per the techniques recommended

by Chestnut et al. (1977).

h. Statistical Analysis

Statistical analysis of the data was carried out as

per the SPSS (Version 10.0) software.

RESULTS AND DISCUSSIONThe raw chicken meat along with different

treatments were stored at refrigeration (5±1°C)

and the quality of meat was analyzed at 2 days

interval.

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The pH was determined to assess the storage

stability of meat. It was observed that the pH

increased significantly during refrigerated storage

for 8 days which might be due to the accumulation

of metabolites by bacterial action on meat in

addition to protein and amino acid degradation

resulting in the formation of ammonia and

consequent increase in pH. TBARS assay

measures the quantity of malonaldehyde which

is an oxidative breakdown product formed mainly

from peroxidised polyunsaturated fatty acids. In

the present study, the overall mean TBA values

revealed that treatments were significantly

different from each other and methanolic extract

of C. sappan showed significantly lower values

than control and other treatments during

refrigeration storage periods. This might be due

to the presence of large amount of phenolics,

flavonoids in heart wood of C. sappan (Saraya et

al., 2009) which act as reductants by donating

electrons that react with free radicals to convert

them to more stable products and terminate free

radical chain reactions. The results are in

accordance with Naveena et al. (2007).

Free fatty acid content can be considered as

an indicator of lipid hydrolysis and flavor of the

product. The overall mean free fatty acid values

(percent oleic acid) increased gradually with

increasing storage periods. This increase might

be due to progressive hydrolysis of lipids during

storage. The results are in agreement with Kanatt

et al. (1998). Methanolic extract of C. sappan

exhibited significantly lower values than other

treatments, the results were in agreement with

Saraya et al. (2009) who reported that free radical

scavenging activity of sappan wood extracts

determined by 1,1-diphenyl 2-picryl hydrazyl

(DPPH) radical assay showed good antioxidant

activity. An advantage of peroxide value

determination is that it directly measures the lipid

peroxides which are primary lipid oxidation

products. Methanolic extract of C. sappan treated

samples showed significantly lower values than

others. The results were in agreement with

Saraya et al. (2009) who reported that free radical

scavenging activity of sappan wood extracts

determined by 1,1-diphenyl 2-picryl hydrazyl

(DPPH) radical assay showed good antioxidant

activity.

The mean values of total plate count of minced

chicken meat were significantly influenced by

treatments and between days of refrigerated

storage. A significant difference in total plate

counts was observed between treatments and

between storage periods. The overall mean

bacterial count (log10

cfu/g) increased significantly

with increase in storage period during refrigeration.

This might be due to the permissive temperature

and relative availability of moisture and nutrients

for the growth of mesophilic bacteria. These

results are in accordance with Nag et al. (1998).

Among the treatments methanolic extact of C.

sappan plus antibiotic showed significantly lower

count than others. The results were in agreement

with Saraya et al. (2009) who reported that chilli

paste treated with C. sappan showed significant

reduction in total plate count.

The overall mean psychrophillic count (log10

cfu/g) increased significantly with increase in

storage period during refrigeration. All treatments

differed significantly from control. Among

treatments methanolic extract of C. sappan plus

antibiotic showed significantly lower count. The

results were in agreement with Siriporn et al.

(2000) who reported similar f indings with

ethylalcohol extract of C. sappan.

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The overall mean Yeast and Mould count (log10

cfu/g) increased significantly with increase in

storage period during refrigeration. Methanolic

extract of C. sappan showed significantly lower

count than other treatments. The results were in

agreement with Niranjan Reddy et al. (2003) who

reported that protosappanin A , isolated from C.

sappan showed antifungal activity.

The overall mean coliform count (log10

cfu/g)

increased significantly (P<0.05) with increase in

storage period during refrigeration. All treatments

differed significantly from control. Among

treatments methanolic extract of C. sappan plus

antibiotic showed significantly lower count. The

results were in agreement with Saraya et al.

(2009) who reported that MPN coliform and E.

coli were less than 3 MPN/g. Darwish and Aburjai

(2010) reported that methanolic extracts of plant

material significantly enhanced the inhibitory

effects of chloramphenicol , doxycycline against

both standard and resistant strains of E. coli.

Among treatments methanolic extract of C.

sappan plus antibiotic showed significantly lower

count. The results were in agreement with Saraya

et al. (2009) who reported that there was no

growth of S. aureus during the period of

preservation. Adwan and Among treatments

methanolic extract of C. sappan plus antibiotic

showed significantly lower count. The results

were in agreement with Saraya et al. (2009) who

reported that there was no growth of S.

typhimurium through the period of preservation.

CONCLUSIONBased on the results obtained in the present study

it can be concluded that methanolic extract of C.

sappan plus antibiotic (chlorteracyclines) and

methanolic extract of C. sappan showed better

antioxidant and antimicrobial activities than

conventional methods. It is also evident through

this study that the treatments like C. sappan plus

antibiotic (T1) and C. sappan methanolic extract

(T2) did not differ from each other in their action

to reduce the microbial load and improve

antioxidant activity on chicken meat samples. In

future, more basic research is needed to

elucidate the mechanism of actions and isolation

of its active ingredients.

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