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Research Article
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Antioxidative Effects of Pomposia Extract, on Lipid Oxidation and Quality of Ground Beef during Refrigerated Storage |
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Rehab F.M. Ali
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ABSTRACT
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The aim of the current investigation was to investigate the effect of pomposia juice and extracts compared to BHT as synthetic antioxidant on lipid oxidation and quality of ground beef during refrigerated storage at 0± 0.5°C for up to 10 days. Two varieties of pomposia were evaluated the first one is Rajamun, the second one is known as Kaatha. In the present investigation based on the antioxidant and free radical scavenging activities results, the highest antioxidant and free radical scavenging activities was shown by the crude juice of Kaatha, therefore 0.5, 0.75 and 1% of either crude juice of Kaatha and BHT were added to minced meat to evaluate it's effects on the lipid peroxidation of ground beef during storage process. TBA test as quality assurance test was conducted at the beginning of the experiment and after 2, 4, 6, 8 and 10 days of storage experiment. The results of this study showed that the crude juice and ethanolic extract of Kaatha variety had significantly (p<0.05) the highest levels of total polyphenols and antioxidant activity. The most effective radical scavenging activity was showed by crude juice of Kaatha was 85.22% while the least effective was the aqueous extract of Rajamun. Control samples had significantly (p = 0.05) the highest TBA value was 1.98 mg malondialdehyde (MDA) kg-1, while beef samples mixed with 1% of pomposia juice had significantly (p = 0.05) the lowest TBA values was 0.79 mg (MDA) kg-1 at the end of the storage period. All samples showed good overall acceptability; in all cases the values were higher than 6. However samples mixed with 0.75 and 1.00 of crude juice of Kaatha had significantly (p = 0.05) the highest scores of over all acceptability were 8.59 and 8.48, respectively. The obtained results indicated that polyphenolic compounds present in pomposia juice and ethanolic extract of pomposia (Kaatha) had high antioxidative effect in reducing the formation of hydroperoxides during refrigerated storage, in the same time the results of sensory evaluation suggests that the crude juice of pomposia (Kaatha) can be successfully used as natural antioxidant and colourant for meat products.
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Received: March 05, 2010;
Accepted: June 02, 2010;
Published: July 05, 2010
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INTRODUCTION
Fresh meat products are commonly marketed at refrigerated temperatures (2-5°C).
However, many undesirable changes of the products can occur during refrigeration
due to microbial growth and lipid oxidation, which give rise to quality reduction,
meat spoilage and economic loss. Lipid oxidation is a major cause of quality
deterioration in meat and its cooked products. Ground meat tends to become rancid
and brown more rapidly, due to pigment and lipid oxidation. An oxidative reaction
in muscle foods leads to degradation of lipid and proteins, resulting in deterioration
of flavour, texture and nutritive value and is considered as one of the major
problems in the development of new convenient meat products and processes (Gray
and Pearson, 1987). Lipid oxidation is often responsible for quality loss
via formation of rancid flavour (Asghar et al., 1988)
and is affected by the duration and temperature of storage of meat (Sun
et al., 2002; Smet et al., 2005; Tan
and Chen, 2005).
Moreover, mechanisms for the control of lipid oxidation in meats have become
increasingly important with the rise in popularity of pre-cooked and convenience
foods. Many substances have been investigated as potential antioxidants to prevent
such lipid oxidation. These include chemical substances such as Butylated Hydroxyanisole
(BHA) and Butylated Hydroxytoluene (BHT). However, the use of synthetic antioxidants
in food has been decreased due to their suspected action as promoters of carcinogenesis,
as well for the general consumer rejection of synthetic food additives (Namiki,
1990; Djenane et al., 2004; Yanai
et al., 2004). Several studies indicated that the use of synthetic
antioxidants has begun to be restricted because of their health risks and toxicity
(Williams et al., 1990; Duve
and White, 1991; Buxiang and Fukuhara, 1997; Hirose
et al., 1998; Farag et al., 2006). Therefore,
the importance of replacing synthetic antioxidants with natural antioxidants
from oilseeds, spices and other plant materials has greatly increased. In the
past few years, various plant materials containing phenolic compounds have been
demonstrated to be effective antioxidants in model systems. Flavonoids, the
most potent antioxidative compounds of plant phenolics occur in vegetables,
fruits, berries, herbs and tea leaves (Shahidi and Wanasundara,
1992; Kandaswami and Middleton, 1997; Skrede
and Wrolstad, 2002; Das et al., 2006; Shaker,
2006; Farag et al., 2006, 2007;
El-Anany, 2007). Non-meat ingredients that have antioxidative
and/or reducing activity can stabilize meat color thus extending shelf life
of meat and meat products (Greene et al., 1971;
Mitsumoto et al., 1991a,b).
Jamun (Syzygium cuminii L.) is a very common, large evergreen beautiful
tree of Indo-Pak subcontinent. It belongs to Myrtaceae plant family. Its fruit
is generally round in shape, deep purple or bluish in colour, having juicy,
sweet pulp and a small stone. The keeping quality of fruit is very poor so the
fruit is to be utilized within 24 h after picking (Shukla
et al., 1991). Jamun fruit is one of those fruits that contain variety
of important nutritional and medicinal elements. The fruit syrup is very useful
for curing diarrhea. It is carminative and diuretic, apart from having cooling
and digestive properties. It also contains Anthocyanins, flavenoids and other
useful ingredients.
The aims of the current investigation were to evaluate antioxidant efficiency of crud juice and crude extracts of two varieties of pomposia by using β-carotene bleaching and free radical scavenging (DPPH) methods in comparison with BHT as synthetic antioxidant, evaluating the effect of adding various levels of the highest antioxidant efficiency of pomposia extracts on the quality of ground beef by determination the changes in TBA value during refrigerated storage compared to BHT and to evaluate the effect of adding different levels of pomposia extract on sensory properties of cooked ground beef at the end of storage period.
MATERIALS AND METHODS
Materials: Two varieties of ripened and freshly harvested pomposia, (Syzyygium
cumini) season, July 2008, the first one Rajamun is big oblong, deep purple
in colour having pink to grey juice, sweet flesh with small seeds. The second
one is known as Kaatha which has small fruits with comparatively big seed and
flesh acidic in taste were obtained from the farm of Faculty of Agriculture,
Cairo University, Giza, Egypt.
The fresh beef (Flank) was purchased from a local market, Giza, Egypt. The fat content of the beef was 15.5%. Chemicals: 2-Thiobarbituric Acid (TBA), 1,1-Diphenyl-2-picrylhydrazyl (DPPH), polyoxy-ethyene-sorbitanmonolaurate (Tween 20), β-carotene β-carotene and linoleic acid were purchased from Sigma Chemical Co. (St. Louis, MO, USA). All chemical used were of analytical reagent grade. Preparation of crud juice and extracts of pomposia (Syzygium cumini): Pomposia fruit Rajamun and Kaatha varieties were cleaned, cut into pieces and then pressed by means of the hydraulic laboratory press model C S/N 37000-156 Freds from Carver (WI, USA). The resultant crude juices were centrifuged at 4000 rpm for 30 min, the supernatants were lyophilized using a freeze-dryer (Labconco Corporation Kansas city, Missouri 64132 USA). The concentrated juice was kept in a brown bottle at-18°C until further use.
Ethanolic and aqueous extracts of Pomposia fruit were prepared according to
the method described by Sanbongi et al. (1998).
The cleaned pieces of Pomposia fruits were diluted with 70% ethanol (1: 7 v/v).
The mixture was stirred with magnetic stirrer for 2 h. at ambient temperature.
The resultant mixture was left overnight and filtered through a Whatman No.
4 filter paper (Whatman International Ltd., Maidstone, UK), the remaining residue
was re-extracted as described above. The filtrate was concentrated using a rotary
evaporator (Buchi, Switzerland) at 40°C under pressure to remove ethanol
residues. In addition, distilled water was used instead of ethanol to prepare
aqueous extracts of Pomposia fruit. The ethanolic and aqueous extracts were
lyophilized using a freeze-dryer (Labconco corporation Kansas city, Missouri
64132 USA). The concentrated juice was kept in a brown bottle at-18°Cuntil
further use.
Preparation of ground beef: Beef meat (5 kg) was cut into small pieces and homogenized in stainless steel blender (Univest Co., 6th of October city, Industrial zone No. 3, Giza, Egypt). Ground beef were mixed by latex gloved hands with 0.5, 0.75 and 1% of either the highest antioxidant efficiency of pomposia extracts and BHT. Minced beef without additives was run as control. The abovementioned samples were packed in polyethylene bags, each bag contain 250 g and stored at 0± 0.5°C in refrigerator for 10 days.
Analytical methods
Determination of moisture content: A known weight of ground beef
(5 g) was dried in an oven at 105°C until a constant weight was reached
(AOAC, 2000).
Determination of ash content: The ash content of ground beef was determined
by muffle furnace at 550°C until a constant weight was obtained (AOAC,
2000).
Determination of crude protein: The total nitrogen of ground beef was
determined by the usual Kjeldahl method (AOAC, 2000).
The crude protein was then calculated by multiplying the total nitrogen by a
factor of 6.25.
Determination of total crude lipids: The crude lipids of ground beef
was determined according method of Bligh and Dyer (1959)
using a mixture of chloroform-methanol (2: 1, v/v).
Nitrogen Free Extract (NFE): The Nitrogen Free Extract (NFE) was calculated by difference.
Determination of total polyphenols: Total polyphenols were determined
according to the method of Jayaprakasha et al. (2003).
An aliquot from pomposia juice and extracts (0.1 mL) were dissolved in a 10
mL mixture of acetone and water (6:4 v/v). Sample (0.2 mL) was mixed with 1.0
mL of ten-fold diluted Folin-Ciocalteu reagent and 0.8 mL of 75 g L-1
sodium carbonate solution. After standing for 30 min at room temperature, the
absorbance was measured at 725 nm. Phenolic contents were calculated on the
basis of the standard curve for Gallic Acid (GAL).The results were expressed
as mg of gallic acid equivalent per g of dry extract.
Determination of free radical scavenging activity (DPPH): The antioxidant
activity of crude juice and extracts of pomposia were evaluated by using the
2, 2'diphenylpicrylhydrazyl (DPPH) assay (Cuendet et
al., 1997; Burits and Bucar, 2000). Fifty microliters
of the extracts were added to 5 ml of a 0.004% (w/v) of DPPH in methanol (100%
v/v). After, a 30 min incubation period at room temperature the absorbance at
517 nm was compared to DPPH in methanol without an extract sample (blank). The
percent inhibition of free radical formation (I%) was calculated as:
Determination of antioxidant activity: The antioxidant activity of crude
juice and extracts of pomposia were by using β-carotene bleaching method,
according to the method described by (Jayaprakasha et
al., 2001). One milliliter of β-carotene solution (0.2 mg β-carotene
mL-1 of chloroform) was transferred into a round-bottom flask containing
0.02 mL of linoleic acid and 0.2 mL of Tween 20. The Chloroform was evaporated
at 40°C using rotary evaporator. The resultant mixture was immediately diluted
with 100 mL of deionized water and mixed for 2 min to form emulsion. A mixture
prepared similarly without β-carotene was used as a blank. A control, containing
0.2 mL of 70% ethanol instead of extract was also prepared. Five milliliter
of the emulsion was transferred into test tube containing 0.2 mL of extract.
The tubes were placed at 50°C in water bath for 2 h. The absorbance was
recorded at 470 nm using UV-Visible spectrophotometer ((Shimadzu, Kyoto, Japan).
The percentage antioxidant activity was calculated based on the following equation:
where, A0 and A°0 are the absorbance values measured at zero time of incubation for extracts and control, respectively. At and A°t are the absorbance values for extracts and control at t = 120 min.
Thiobarbituric Acid-reactive Substances (TBARS): The TBARS values were
determined in triplicate samples by the extraction method of Mielnik
et al. (2006). For extraction, 10 g meat was homogenized with 30
mL of a 7.5% aqueous solution of trichloroacetic acid (TCA). After filtration
(vacuum filtered through a Whatman No. 41 filter paper (Whatman International
Ltd., Maidstone, UK), 5.0 mL of extract was mixed with 5.0 mL of 0.02 mol L-1
aqueous Thiobarbituric acid (TBA) in a stoppered test tube. The samples were
incubated at 100°C for 35 min in a water-bath and subsequently cooled for
10 min in cold water. Absorbance was measured at 532 nm by using UV-Visible
spectrophotometer (Shimadzu, Kyoto, Japan). against a blank containing 5 mL
distilled water and 5 mL TBA reagent. Results expressed as milligrams malondialdehyde
kg-1 meat.
Cooking and sensory evaluation: Minced beef samples were mixed with
1% salt, pressed into a mould (8 cm diameter, 0.5 height) and cooked for 30
min in water bath at 75°C until an internal temperature of 75°C was
reached. Cooked beef samples were left to be warm then served for sensory evaluation.
Sensory evaluation method was conducted according to the method described by
Mansour and Khalil (2000), cooked beef samples were
served warm to 10-membered trained panel (Staff of Food Technology Research
Institute, Agriculture Research Center, Giza, Egypt) and to 40 of nonspecialist
panelists randomly recruited within the student and lecturers of Biochemistry
Department, Faculty of Agriculture, Cairo University, without care of age or
sex. The panelists were subjected to sensory evaluation using a 9-point hedonic
scale for taste, color, odor and overall-acceptability. A numerical basis as
a sort of evaluation from 1 to 9 was used where (1 = dislike extremely, 2 =
dislike very much, 3 = dislike moderately, 4 = dislike slightly, 5 = neither
like nor dislike, 6 = like slightly, 7 = like moderately, 8 = like very much,
9 = like extremely).
Statistical analysis: The data of the present study were subjected to analysis of variance and the least significant difference test, in order to compare the mean values of the investigated parameters. RESULTS AND DISCUSSION Pomposia polyphenols: Jamun fruit is one of those fruits that contain variety of important nutritional and medicinal elements. It contains Anthocyanins, flavenoids and other useful ingredients. Crude juice and ethanolic extract of Kaatha variety had significantly (p<0.01) the highest levels of total polyphenols were 16.32 and 15.45 mg g-1, respectively, whereas aqueous extract had significantly (p<0.05) the lowest values of total polyphenols were 5.42 and 6.66 for the two varieties of pomposia Rajamun and Kaatha, respectively (Table 1).
Antioxidant activity and free radical scavenging activity (DPPH): Antioxidant
tests could be based on the evaluation of lipid peroxidation or on the measurement
of free radical scavenging potency (hydrogen-donating ability). The radical
scavengers donate hydrogen to free radicals, leading to non toxic species and
therefore to inhibition of the propagation phase of lipid oxidation. The use
of DPPH radical provides an easy, rapid and convenient method to evaluate the
antioxidants and radical scavengers (Soler-Rivas et al.,
2000; Kansci et al., 2003; Argolo
et al., 2004; Roginski and Lissi, 2005).
Therefore, in the present investigation, pomposia juice and extracts were evaluated
for their antioxidant activity and radical scavenging activity by β-carotene
bleaching and DPPH techniques. Crude juice and ethanolic extract of Kaatha variety
had significantly (p<0.05) the highest antioxidant activity were 90.00 and
89.00%, respectively, however, the lowest antioxidant activities was shown by
aqueous extract of Rajamun was 62.00% (Table 1). One could
report that there is high relationship between the content of total polyphenols
of extract and its antioxidant activity, hence the content of total polyphenol
of extract is increased as antioxidant activity of extract increased. The most
effective radical scavenging activity was shown by crude juice of Kaatha was
85.22% while the least effective was the aqueous extract of Rajamun. Anyway,
crude juice and ethanolic extracts of pomposia showed relatively high value
of radical scavenging activity ranged from 67.67 to 85.22% (Table
1). This high antioxidant capacity may be due to the high concentration
of phenolics in pomposia extracts. These compounds are an important group of
natural antioxidants with possible beneficial effects on human health (Meyers
et al., 2003). They can participate in protection against the harmful
action of reactive oxygen species, mainly oxygen free radicals. These compounds
are known also to possess antioxidant activity due to its ability to reduce
free radical stability via electron or hydrogen-donating mechanism. (Lee
et al., 1999; Ruberto and Baratta, 2000).
Table 1: |
Polyphenolic content, antioxidant and free radical scavenging
activities of pomposia juice and extracts |
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Values are expressed as the mean of three determinations.
Values followed by different letter are significantly different at (p<0.05).
LSD refers to least significant difference test |
The antioxidants activity of polyphenols is principally based on the redox
properties of their hydroxyl groups and the structural relationships between
different parts of their chemical structure (Rice-Evans et
al., 1996).
Chemical composition of ground beef: The moisture content of ground
beef was 65.39%. The protein content, crude fat and ash content of ground beef
were 51.91, 37.71 and 3.06%, respectively. The Nitrogen Free Extract (NFE) was
7.33%. The obtained results indicate that beef contain relatively high levels
of fat which consider a major cause of quality deterioration in meat and its
cooked products (Table 2). These results are in good agreement
with those obtained by Ismail and Yee (2006) and Abd
El-Hamied et al. (2009).
Changes of TBA value of ground beef during refrigerated storage: Ground
meat tends to become rancid and brown more rapidly, due to pigment and lipid
oxidation. An oxidative reaction in muscle foods leads to degradation of lipid
and proteins, resulting in deterioration of flavour, texture and nutritive value
and is considered as one of the major problems in the development of new convenient
meat products and processes (Gray and Pearson, 1987).
In the present investigation based on the antioxidant and free radical scavenging
activities results, the highest antioxidant and free radical scavenging activities
was shown by the crude juice of Kaatha, therefore 0.5, 0.75 and 1% of either
crude juice of Kaatha and BHT. were added to minced meat to evaluate it's effects
on the lipid peroxidation of ground beef during storage process.
Table 2: |
Chemical composition of minced beef (based on dry weight
basis) |
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Values are expressed as the mean of three determinations.
Values followed by different letter are significantly different at (p<0.05).
LSD refers to least significant difference test |
Generally, TBA values increased gradually and significantly (p<0.05) during
storage period. The phospholipids in muscle membrane provide an ideal substrate
for lipid peroxidation. Iron bound to negatively charged phospholipids promotes
lipid peroxidation, resulting in generation of warmed-over flavor (Empson
et al., 1991). Both nonheme and heme iron in beef muscle can catalyze
lipid peroxidation (Kanner et al., 1988; Love,
1983; Monahan et al., 1993). However, mixing
minced beef with various levels of crude juice of Kaatha caused a significant
(p = 0.05) reduction of TBA values compared to control sample, These inhibitory
effects of crude juice of Kaatha were dose-dependent. Control samples had significantly
(p = 0.05) the highest TBA value was 1.98 mg malondialdehyde (MDA) kg -1
at the end of the storage period, the highest TBARS values of the control sample
at the end of storage days might be due to an interaction between the natural
substances (for example, polyunsaturated fatty acids) and catalysts (for example,
iron ion) from the meat tissue during storage (Decker and
Hutlin, 1990; Kim et al., 2000), while beef
samples mixed with 1% of pomposia juice had significantly (p = 0.05) the lowest
TBA values was 0.79 mg malondialdehyde (MDA) kg -1 at the end of
the storage period. No significant (p>0.05) differences were observed in
TBA values of ground beef mixed with 0.75% crude juice of Kaatha and that sample
treated with 1.00% of synthetic antioxidant BHT. TBARS numbers in treated samples
were much lower than the control sample. The inhibitory effect of pomposia juice
on lipid oxidation might be due to scavenging of free radicals and chelating
of transition metals (Chan et al., 1994; Lee
and Hendricks, 1997; Niki, 1991). The amounts of
TBARS as mg MDA kg-1 meat formed at the end of storage period for
control and treated samples were far below the critical value of 3 mg kg -1
at which rancidity is detected (Wong et al., 1995),
while Greene and Cumuze (1982) considered a TBARS range
0.6-2.0 mg kg -1 to be the minimum detectable level for oxidized
flavour in ground beef. The obtained results indicate high antioxidative effect
of polyphenolic compounds present in pomposia extract in reducing the formation
of hydroperoxides during storage process (Table 3) These results
are in good agreement with those obtained by Ismail and
Yee (2006) who studied the effects of extracts of cocoa shell, Roselle seeds
and a combination of them on the lipid oxidation of cooked beef and they reported
that the tested extracts were more effective than BHT and tocopherols in inhibiting
lipid oxidation of cooked-refrigerated beef and Azizah et
al. (1999), who found that phenolic compounds of cocoa shells exhibited
a strong antioxidant activity compared to synthetic antioxidant.
In this respect the results of Abd El-Hamied et al.
(2009) observed that the addition of rosemary, sage and their combination
showed high antioxidative effects during refrigerated and frozen storage of
minced meat also Ali (2010) showed that mixing oil with
various levels of pomposia extracts caused significant decrease of the formation
of secondary products during frying process.
Sensory evaluation: Sensory characteristics of cooked minced beef as affected by adding different levels of pomposia juice and BHT are presented in Table 4. Treated sample had relatively high value of taste, odor, colour and overall acceptability compared to control sample.. No significant (p>0.05) differences were observed in taste between samples mixed with 0.5 and 0.75% of crude juice of Kaatha and those mixed with 0.05% of BHT The addition of pomposia juice affected significantly (p<0.05) colour parameter of the final product, minced beef mixed with 1.00% of pomposia juice had significantly (p = 0.05) the highest value of colour was 9.00 whereas control samples had significantly (p = 0.05) the lowest value was 5.00.
Odor also is one of the most important quality factors of acceptance for cooked
meat products. The results presented in Table 4 show that
samples mixed with 0.5 of crude juice of Kaatha or BHT had significantly (p
= 0.05) the highest value of odor was 9.00 and 8.94, respectively whereas control
samples and those samples mixed with 1.00% of BHT had significantly (p = 0.05)
the lowest odor values of were 7.50 and 7.44, respectively. All samples showed
good overall acceptability; in all cases the values were higher than 6. However
samples mixed with 0.75 and 1.00 of crude juice of Kaatha had significantly
(p = 0.05) the highest scores of over all acceptability were 8.59 and 8.48,
respectively. The results of sensory evaluation suggests that the crude juice
of pomposia (Kaatha) can be successfully used as natural antioxidant and colourant.
Table 3: |
Effect of crude juice of Kaatha and BHT on TBA No. (mg MDA
kg-1 meat) of minced beef during refrigerated storage at 0°C |
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Values are expressed as the mean of three determinations.
Values followed by different letter are significantly different at (p<0.05).
LSD refers to least significant difference test |
Table 4: |
Sensory evaluation of cooked minced beef mixed with different
levels of crude juice of pomposia and BHT |
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Values are expressed as the mean of three determinations.
Values followed by different letter are significantly different at (p<0.05).
LSD refers to least significant difference test |
CONCLUSIONS
The obtained results showed that the most effective radical scavenging activity was shown by crude juice of Kaatha was 85.22% while the least effective was the aqueous extract of Rajamun. Anyway, crude juice and ethanolic extracts of pomposia showed relatively high value of radical scavenging activity ranged from 67.67 to 85.22%. polyphenolic compounds present in pomposia juice and ethanolic extract of pomposia (Kaatha) had high antioxidative effect in reducing the formation of hydroperoxides during refrigerated storage, in the same time the results of sensory evaluation suggests that the crude juice of pomposia (Kaatha) can be successfully used as natural antioxidant and colourant for meat products.
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