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Annals of Agricultural Science (2014) 59(2), 213220

H O S T E D BY
Faculty of Agriculture, Ain Shams University

Annals of Agricultural Science


www.elsevier.com/locate/aoas

Oxidative stability of ghee as aected by natural


antioxidants extracted from food processing wastes
Gehan A. El-Shourbagy *, Kahled M. El-Zahar

Food Science Department, Faculty of Agriculture, Zagazig University, 44519 Zagazig, Egypt

Received 14 August 2014; revised 25 August 2014; accepted 1 September 2014


Available online 8 December 2014

KEYWORDS Abstract In this research, bioactive compounds found in peanut skins (PS), pomegranate peels
Ghee; (PP) and olive pomace (OP) were extracted using ethanol (80%), ethyl acetate and n-hexane. Eth-
Food processing wastes; anol extract showed slightly better antioxidant characteristics compared with ethyl acetate and hex-
Natural antioxidants; ane extracts. Extracts showed varying degrees of antioxidant potential in different test systems in a
Stability indices; dose-dependent manner, whereas, antioxidant capacity of extracts was found to be in parallel with
BHA their higher phenolic contents. Total phenolic compounds (as gallic acid equivalent) ranged between
0.89 and 16.6, 1.83 and 261, and 1.56 and 124 mg gallic acid/g extract for OP, PS and PP, respec-
tively. Ethanol extracts of different by-products were added to ghee at concentrations of 200, 400
and 600 ppm, respectively. BHA was also added to ghee at a concentration of 200 ppm for compar-
ison. All samples were incubated at 63 C for 21 days. Ethanol extracts of PS, OP and PP gave good
antioxidant activity during accelerated oxidative incubation of ghee. The results revealed that
ethanolic extracts under study, at a concentration of 200 ppm, could be used to retard fat
auto-oxidation.
2014 Production and hosting by Elsevier B.V. on behalf of Faculty of Agriculture, Ain Shams
University. Open access under CC BY-NC-ND license.

Introduction has been reassessed because of possible toxic or carcinogenic


components formed during their degradation (Jo et al., 2006;
Antioxidants (natural and synthetic) play a signicant role in Pitchaon et al., 2007). Consequently, the search for endoge-
retarding lipid oxidation reactions in food products. The detri- nous protective ingredients in foods has been intensied
mental effects of excessive lipid oxidation such as formation of wherein their utilization requires only manipulation of food
off-avors and undesirable oxidized chemical compounds formulations. A number of natural antioxidants have been
(aldehydes, ketones and organic acids) are well known (Saad added during food processing and have elongated the shelf life
et al., 2007). Synthetic antioxidants (e.g., TBHQ, BHA and and oxidative stability of stored products (Chenn et al., 2008;
BHT) are widely used as food additives, but their application Ebrahimabadi et al., 2010; Jang et al., 2012; Xiaowei et al.,
2011).
A huge amount of plant biomass wastes are produced
* Corresponding author. Mobile: +20 01005469481.
yearly as by-products from the agro-food industries. These
E-mail address: gehan53@yahoo.com (G.A. El-Shourbagy).
wastes are attractive sources of natural antioxidants. The high
Peer review under responsibility of Faculty of Agriculture, Ain-Shams
concentration of phenolic compounds present in peels, skins
University.
http://dx.doi.org/10.1016/j.aoas.2014.11.008
0570-1783 2014 Production and hosting by Elsevier B.V. on behalf of Faculty of Agriculture, Ain Shams University.
Open access under CC BY-NC-ND license.
214 G.A. El-Shourbagy, K.M. El-Zahar

and seeds supports the utilization of these residues as a source (Olea europaea L.) pomace (OP) was obtained from Food
of natural antioxidants. Phenolic compounds exhibit a wide Technology Research Institute (Agricultural Research Center,
range of physiological properties such as anti-allergenic, anti- Giza, Egypt). Peanut (Arachis hypogaea L.) skins (PS) were
atherogenic, anti-inammatory, anti-microbial, antioxidant, obtained from the 10th of Ramadan City. Pomegranate
anti-thermobiotic, cardio protective and vasodilatory effects (Punica granatum L.) fruits (PP), were obtained from local
(Balasundran et al., 2006). Phenolics could be extracted by market (Zagazig, Egypt), washed with distilled water and man-
water or solvents and the extraction conditions need to be opti- ually peeled. Pomegranate peels (PP) were collected then rinsed
mized with respect to solvent polarity and physical conditions with distilled water and considered as a by-product. The start-
(Nepote et al., 2005). In addition, research has indicated that ing materials were dried in an air draft drying oven (40 C)
natural phenolic compounds can be extracted from raw mate- until the moisture content became 12% or less. By-products
rials or waste products of food industry (Peschel et al., 2006). were ground and sieved through 60 mesh sieve and nally
Studies were conducted to investigate antioxidant proper- cooled or kept at 4 C until the extractions were carried out.
ties of peanut, peanut kernels, peanut hulls and peanut-based Butylated hydroxy anisole (BHA), 1,1-diphenyl-2-pic-
products (Yu et al., 2005; Wang et al., 2007). Peanut skins were rylhydrazyl (DPPH), gallic acid and quercin were purchased
demonstrated to be rich in phenolics and other health promot- from Sigma (St. Louis, MO, USA). All other chemicals and
ing compounds (Yu et al., 2005; Wang et al., 2007; Monagas reagents were of the highest purity available.
et al., 2009). The olive oil industry generates large quantities
of a deleterious by-product known as olive pomace. Olive Preparation of extracts
pomace has broad spectrum toxicity against some microorgan-
isms, plants, insects, animals and human cells (Obied et al., Dried materials were extracted with different solvents, namely
2007; Aldini et al., 2006). Nevertheless, olive pomace consid- ethanol (80%), ethyl acetate and n-hexane at a ratio of 10:1 (v/
ered as a potential source for natural antioxidants w, 10 mL solvent: 1 g raw material) in closed vessels by stirring
(Niaounakis and Halvadakis, 2004; Aldini et al., 2006). A wide at room temperature (25 C) for 4 h followed by ltration
range of phenolic compounds have been identied in virgin oil through Whatmann #1 lter paper. The residues were re-
(Suarez et al., 2010) wherein only ca. 2% of the total phenols extracted again under the same conditions. All vessels were
found in olive fruits are transferred to the extracted olive oil. wrapped with aluminum foil to prevent light degradation dur-
The rest of phenolic compounds (98%) are retained in the olive ing extraction (Yu et al., 2005). N-hexane and ethyl acetate
waste cake. Pomegranate has been used extensively in the folk extracts were evaporated in a rotary evaporator (Buchi-water
medicine of many cultures and its consumption has grown tre- bath-B-480, Switzerland) below 40 C, while ethanol 80%
mendously especially in the last decades (Li et al., 2006; Cam extracts were freeze-dried (Thermo Electron Corporation-
et al., 2009). The peels of some fruits have higher antioxidant Heto Power Dry LL 300 Freeze Dryer, Czechoslovak). The
activity than pulps (Guo et al., 2003; Fuhrman et al., 2005). dried extracts after evaporation of solvents were weighed to
Pomegranate is a good example for this type of fruits wherein determine the yield and stored at 20 C until used.
their peels constitute approximately 40% of the whole fruit
and are rich in ellagic acid derivatives (Cerda et al., 2003; Determination of total phenolic compounds (TPC)
Seeram et al., 2005).
From an environmental and economical perspective, it is
The concentration of TPC in different extracts was measured
very important that plant by-products produced by agro-food
using UV spectrophotometer (Jenway-UVVIS Spectropho-
industry be used in food industry. However, ghee, the most
tometer), based on a colorimetric oxidation/reduction reac-
famous traditional dairy product in Egypt and many countries
tion, as described by Skerget et al. (2005) using Folin
in Middle East, undergoes oxidative degradation during stor-
Ciocalteu reagent. Specically, 0.5 mL of diluted extract
age, resulting in alteration of major quality parameters affect-
(10 mg in 10 mL solvent) was mixed with 2.5 mL of FolinCio-
ing its suitability for consumption. Development of rancidity
calteu reagent (diluted 10 times with distilled water) and 2 mL
reduces the shelf life of the product, which ultimately affects
of Na2CO3 (75 g/1 L). The sample was incubated for 5 min at
consumer acceptability (Mehta, 2006; Mariod et al., 2010;
50 C then cooled. For a control sample, 0.5 mL of distilled
Pawar et al., 2012). Therefore, the objectives of this study
water was used. The absorbance was measured at 760 nm.
were; (1) to evaluate different extracts from peanut skins
Total phenolic content expressed as gallic acid equivalent
(PS), pomegranate peels (PP) and olive pomace (OP) as a
(GAE) was calculated, and the results were expressed as a
source of natural antioxidants, (2) to characterize the compo-
mg GAE g1 extract.
sition and content of phenolics in different extracts and (3) to
evaluate the efciency of using agro food wastes ethanolic
Identication of phenolic acids using HPLC
extracts in improving the quality, overall acceptance and oxi-
dative stability of ghee during storage under thermal oxidative
conditions. Phenolic acids of the dried extracts were identied according to
the method described by Mattila et al. (2000). HPLC (Hewllet
Packard series 1050, USA) equipped with autosampling, injec-
Materials and methods tor, solvent degasser, UV detector set at 330 nm and quarter
HP pump (series 1050) was used. Column (C18 hypersil
Materials BDS) with particle size 5 lm was used. The separation was car-
ried out with methanol and acetonitrile as a mobile phase at
Plant biomass wastes, as a by-product of food industries, com- ow rate of 1 mL/min. The column temperature was per-
monly found in Egypt, were used in this investigation. Olive formed at room temperature (25 C) throughout the
Oxidative stability of ghee as affected by natural antioxidants 215

experiment. Identication and quantication were carried out oxidation reaction passed through deionized water, in which
based on calibrations of the standards prepared from phenolic conductivity values were detected. Heating block was held con-
acids dissolved in a mobile phase. Retention time and peak stant at 130 C. A rate of air ow through liquid butter oil
area were used for calculation of phenolic acid compounds (ghee) was 10 l/h. Prior to the testing, frozen samples of ghee
by the data analysis of Hewllet Packared Software. were thawed at 4050 C, and a 3 0.0020 g of ghee sample
was taken for the analysis according to AOCS (1997).
Radical scavenging activity (RSA) of extracts
Statistical analysis
The electron donation ability of the obtained extracts was
measured by bleaching of the purple colored solution of DPPH Statistical analysis for the obtained data was carried out using
according to the method of Hanato et al. (1988). One hundred SPSS version 20 computer program (Dominick and Derrick,
lL of each extracts (10 mg extract/10 mL solvent) was added 2001). All data were expressed by means and standard devia-
to 3 mL of 0.1 mM DPPH dissolved in ethyl acetate, ethanol tions of three replicates and were compared using one-way
and hexane according to the solvent used for extraction. After ANOVAs and least signicant difference (LSD) values with
incubation period of 30, 60 and 120 min at room temperature, different letters within the same column differ signicantly at
the absorbance was determined against a control at 517 nm P < 0.010.05.
(Gulcin et al., 2004). Percentage of antioxidant activity of
DPPH was calculated as follows: Results and discussion
Antioxidant activity Inhibition%
Yield of different food processing waste extracts
Acontrol  Asample =Acontrol   100
where Acontrol is the absorbance of the control reaction and The yield of antioxidant extracts with different solvents varied
Asample is the absorbance in the presence of extract. BHA from 0.1 to 42.5 g extract/100 g wastes (Fig. 1). PS, PP and OP
was used as a positive control. Samples were analyzed in had the highest yield when extracted with ethanol 80% fol-
triplicate. lowed by hexane and ethyl acetate, respectively. Variation in
the extraction yields of different extracts might be attributed
Stability of ghee enriched with food processing wastes ethanolic to differences in polarity of compounds found in plants such
extracts differences have been reported by Prakasha et al. (2001).

Total phenolic compounds (TPC)


The butter, used for preparing ghee in the present study, was
made from pasteurized and un-ripened buffaloes cream. The
butter was converted into ghee by boiling according to the TPC of different food processing waste extracts were deter-
method described by Fahmi (1961). Ghee samples were divided mined on the basis that the FolinCiocalteu method measures
into eleven portions and treated as follows: the reduction of the reagent by phenolic compounds via the
formation of a blue complex that can be measured at 760 nm
Portion (1) was kept without additives and was considered against GAE as a standard. The amount of TPC varied in
to be as a negative control (C).
Portion (2) was treated with 200 ppm BHA and was consid- hexane extract ethylacetate extract ethanol 80% extract
ered to be as positive control (C1). 50
Extraction yield (g/100g)

45
Portions (3, 4 and 5) were treated with 200, 400 and
40
600 ppm of PS ethanolic extract respectively, (T1, T2 35
and T3). 30
25
Portions (6, 7 and 8) were treated with 200, 400 and
20
600 ppm of PP ethanolic extract respectively, (T4, T5 15
and T6). 10
5
Portions (9, 10 and 11) were treated with 200, 400 and
0
600 ppm of OP ethanolic extract respectively, (T7, T8 Olive pomace Pomegranate peels Peanut skin
and T9).
Fig. 1 Yield of antioxidant extracts (g/100 g) for different food
All samples were incubated in an oven at 63 1 C to processing wastes.
accelerate the oxidation for 21 days. Samples were analyzed
every three days for peroxide value (PV), acid value (AV),
and 2-thiobarbituric acid (TBA) value. AV and PV were deter- Table 1 Total phenolic compounds (mg gallic acid/g extract)
mined according to AOAC (2005), while TBA was assessed in different extracts.
according to Fernandez-Lopez et al. (2005).
By-product Concentration (mg gallic acid/g extract)
Hexane Ethyl acetate Ethanol 80%
Oxidation stability test of ghee using Rancimat equipment
Olive pomace (OP) 16.63 0.89 12.23
Pomegranate peels (PP) 1.56 12.49 124.23
Determination of an oxidative stability of ghee by Rancimat Peanut skins (PS) 1.83 5.69 261.69
equipment (USA, model 617), was based on volatile acids from
216 G.A. El-Shourbagy, K.M. El-Zahar

Zero time after 30 min after 1 hour after 2 hours


Table 2 Concentration of identied phenolic compounds
120
(mg/g dry matter) in food processing waste extracts as (A)

Antioxidant activity %
100
determined by HPLC.
80
Peanut skin Olive pomace Pomegranate peels
60
Gallic 0.07 0.01 0.0
Pyrogallol 0.90 0.27 12.64 40
Chlorogenic 2.58 0.06 0.84 20
Protocatechuic 5.07 1.0 1.86
Catechol 1.86 0.0 0.0 0
Olive pomace Pomegranate Peanut skins BHA
Caeic 0.54 0.0 0.0 peels
Vanillic 0.19 0.04 0.34 Hexane extracts
Catachin 10.64 0.0 0.0
Caeine 0.69 0.04 0.21 120
(B)

Antioxidant activity %
Ellagic 1.40 0.46 5.90 100
P-Coumaric 0.0 0.0 0.05
80
Benzoic 0.0 0.32 0.0
Syringic 0.0 0.01 0.0 60

40

20

the different extracts, ranging from 0.89 to 261.69 mg GAE g1 0


Olive pomace Pomegranate Peanut skins BHA
extract (Table 1). In general, the results stated that ethanol peels
80% and ethyl acetate were better than hexane in extracting Ethylacetate extracts
phenolics from PP and PS owing to their higher polarity and
good solubility (Siddhuraju and Becker, 2003; Kequan and Antioxidant activity % 120
Liangli, 2004). On the other side, hexane extracted the highest 100 (C)
amount of phenolics from OP followed by ethanol 80% as 80
shown in Table 1.
60

Identied phenolic compounds of food processing wastes 40

20

Table 2 shows the percentage of identied phenolic com- 0


Olive pomace Pomegranate Peanut skins BHA
pounds in PS, PP and OP. There was a great variation among peels
the components identied in each waste by-product. Phenolic Ethanol 80% extracts
compounds identied in PS were pyrogallol, protocatechuic,
catachin and ellagic acid with amounts ranging from 0.07 to Fig. 2 Radical scavenging activity (RSA) of hexane (A), ethyl
10.64 mg/g. The main phenolics identied in OP were pyrogal- acetate (B) and ethanol 80% (C) waste extracts in DPPH radical
lol, gallic acid, vanillic acid, protocatechuic, ellagic acid with scavenging activity system compared with BHA.

Table 3 Effect of ethanol extracts at different concentration on the PV (meq.O2/kg fat) of ghee during storage.*
Treatments Storage period (days)
0 3 6 9 12 15 18 21
a a a a a a a
C 0.87 0.02 1.95 0.2 2.22 0.04 3.65 0.06 4.42 0.05 5.33 0.03 7.61 0.02 10.21a 0.02
C1 0.85a 0.01 0.93d 0.01 1.13de 0.03 1.26de 0.02 1.82e 0.03 2.43d 0.03 3.16de 0.01 5.32d 0.03
T1 0.85a 0.01 0.92d 0.03 1.11ef 0.02 1.24de 0.05 1.82e 0.06 2.43d 0.4 3.14de 0.04 5.32d 0.05
T2 0.85a 0.02 0.91d 0.02 1.08f 0.02 1.23de 0.03 1.80e 0.03 2.41d 0.04 3.15de 0.05 5.28d 0.04
T3 0.86a 0.02 0.91d 0.01 1.08f 0.02 1.22e 0.05 1.80e 0.03 2.40d 0.03 3.12e 0.06 5.27d 0.03
T4 0.85a 0.04 0.91d 0.02 1.12def 0.04 1.25de 0.05 1.80e 0.06 2.41d 0.05 3.15de 0.03 5.32d 0.04
T5 0.86a 0.02 0.92d 0.03 1.09ef 0.03 1.24de 00.1 1.81e 0.08 2.40d 0.03 3.15de 0.04 5.29d 0.02
T6 0.86a 0.02 0.92d 0.03 1.09ef 0.02 1.22e 0.03 1.79e 0.03 2.38d 0.02 3.14de 0.03 5.28d 0.02
T7 0.85a 0.03 1.22b 0.01 2.00b 0.03 3.03b 0.02 3.93b 0.04 4.42b 0.04 5.14b 0.02 8.17b 0.04
T8 0.86a 0.01 1.03c 0.02 1.37c 0.02 2.32c 0.03 2.41c 0.04 3.61c 0.03 4.81c 0.6 6.29c 0.02
T9 0.86a 0.03 0.93d 0.04 1.16d 0.02 1.30d 0.02 1.93d 0.05 2.43d 0.04 3.19d 0.03 5.33d 0.04
LSD 0.037 0.040 0.042 0.061 0.082 0.061 0.063 0.055
C: control without antioxidants.
C1: ghee treated with 200 ppm BHA (positive control).
T1, T2, T3: ghee treated with 200, 400 and 600 ppm PS extracts, respectively.
T4, T5, T6: ghee treated with 200, 400 and 600 ppm PP extracts, respectively.
T7, T8, T9: ghee treated with 200, 400 and 600 ppm OP extracts, respectively.
*
Means with different letters within the same column differ signicantly at P < 0.010.05.
Oxidative stability of ghee as affected by natural antioxidants 217

amount ranging from 0.01 to 1.0 mg/g. The major phenolic


compounds identied in PP were pyrogallol, gallic acid, cou-

0.101a 0.004
0.053a 0.004
0.058a 0.004
0.056a 0.003
0.052a 0.004
0.058a 0.003
0.056a 0.003
0.210a 0.007
0.085a 0.002
0.079a 0.002
0.061a 0.002
maric acid, caffeine with amount ranging from 0.05 to
12.64 mg/g. Balasundran et al. (2006) stated that the antioxi-
dant activity of phenolic compounds depends on the structure,

0.14159
in particular the number and positions of the hydroxyl groups
21

of the nature of substitution on the aromatic rings. Moure


et al. (2001) reported that the antioxidant compounds from
residual sources could be used for increasing the stability of
0.048cd 0.003
0.049cd 0.004
0.049cd 0.003

0.052cd 0.004
0.050cd 0.001
0.048d 0.001

0.048d 0.003
0.071b 0.004
0.068b 0.002
0.086a 0.002

0.053c 0.003
foods by preventing lipid peroxidation and also for protecting
oxidative damage in living systems by scavenging oxygen rad-

0.00496
icals. It is well known that total antioxidant activity of waste
extracts was lineearly proportional to the concentration of
18

total phenolics.
0.062ab 0.004

0.054ab 0.003
0.051ab 0.004

RSA against DPPH


0.041b 0.003
0.041b 0.003
0.042b 0.004
0.040b 0.004
0.043b 0.002
0.043b 0.003
0.040b 0.002

0.177a 0.002

The results of RSA of various extracts are represented in


0.11641

Fig. 2. The results clearly indicated that all extracts exhibited


Effect of ethanol extracts at different concentration on the TBA (mg malonaldehyde/kg fat) of ghee during storage.*

15

antioxidant activity. The extracts that contained high amount


of TPC (Table 1) showed high RSA. In general, ethanolic
extracts followed by hexane then ethyl acetate extracts showed
0.053ab 0.002

0.050ab 0.001
0.042ab 0.003
0.035b 0.002

0.035b 0.002
0.034b 0.004
0.038b 0.004
0.036b 0.002
0.033b 0.002

0.039b 0.002
0.135a 0.002

RSA as strong as that of BHA (Fig. 2AC). It has been proved


that the antioxidant activity of plant extracts is mainly ascrib-
0.08617

able to the concentration of phenolic compounds in the plant


(Heim et al., 2002). The extract RSA with different solvents
12

varied from 91.4% to 5.50% after 120 min incubation. The


highest RSA was observed with PP ethanol 80%, hexane and
0.030 cd 0.003
0.027cd 0..002
0.027cd 0.002
0.030cd 0.005
0.029cd 0.004

0.031cd 0.003

ethyl acetate extracts with respective values of 80%, 38%


0.026d 0.001

0.041b 0.003
0.040b 0.002
0.051a 0.006

0.032c 0.003

and 35%, respectively. Ethanol 80%, hexane and ethyl acetate


extracts of PS had values of 85%, 22% and 19%, respectively.
0.00554

In addition, hexane, ethanol 80% and ethyl acetate extracts of


Means with different letters within the same column differ signicantly at P < 0.010.05.

OP had values of 24%, 20% and 18%, respectively. The results


9

of the DPPH radical scavenging assay suggest that compo-


nents involving the extracts are capable of scavenging free rad-
0.015a 0.002
0.015a 0.002
0.015a 0.002
0.013a 0.003
0.013a 0.002
0.015a 0.002
0.015a 0.001
0.015a 0.001
0.015a 0.001
0.015a 0.001
0.015a 0.002

icals via electron- or hydrogen-donating mechanisms and thus


might be able to prevent the initiation of deleterious free rad-
T7, T8, T9: ghee treated with 200, 400 and 600 ppm OP extracts, respectively.
T4, T5, T6: ghee treated with 200, 400 and 600 ppm PP extracts, respectively.
T1, T2, T3: ghee treated with 200, 400 and 600 ppm PS extracts, respectively.
0.0028

ical mediated chain reactions in susceptible matrices. This fur-


ther shows the capability of the extracts to scavenge different
6

free radicals in different systems, indicating that they may be


useful therapeutic agents for treating radical-related patholog-
0.013a 0.001
0.041a 0.001
0.013a 0.001
0.013a 0.003
0.013a 0.001
0.014a 0.002
0.012a 0.001
0.011a 0.002
0.052a 0.006
0.013a 0.001
0.013a 0.001

ical damage. The effect of antioxidants on DPPH radical-


scavenging is thought to be due to their hydrogen-donating
ability, DPPH is a stable free radical and accepts an electron
0.0431

C1: ghee treated with 200 ppm BHA (positive control).


Storage period (days)
3
0.013a 0.001
0.014a 0.001
0.013a 0.001
0.014a 0.001
0.011a 0.001
0.012a 0.002
0.012a 0.002
0.011a 0.002
0.013a 0.002
0.013a 0.001
0.013a 0.001
0.02602
C: control without antioxidants.
0
Treatments
Table 4

LSD

Fig. 3 Rancimat of ghee treated with different food processing


C1
T1
T2
T3
T4
T5
T6
T7
T8
T9

*
C

waste extracts at 130 C.


218 G.A. El-Shourbagy, K.M. El-Zahar

or hydrogen radical to become a stable molecule (Gulcin et al.,


2004). Free radicals involved in the process of lipid

0.101a 0.004
0.053a 0.004
0.058a 0.004
0.056a 0.003
0.052a 0.004
0.058a 0.003
0.056a 0.003
0.210a 0.007
0.085a 0.002
0.079a 0.002
0.061a 0.002
peroxidation are considered to play a major role in numerous
chronic pathologies such as cancer and cardiovascular diseases
(Dorman et al., 2003).

0.14159
21
Oxidative stability of ghee enriched with food processing wastes
ethanolic extracts peroxide values

0.048cd 0.003
0.049cd 0.004
0.049cd 0.003

0.052cd 0.004
0.050cd 0.001
0.048d 0.001

0.048d 0.003
0.071b 0.004
0.068b 0.002
0.086a 0.002

0.053c 0.003
Data presented in Table 3 show that the PV of control ghee
samples increased during the accelerated incubation up to

0.00496
21 days. The others samples enriched with BHA and natural
antioxidants had the lower PV values than the control sample

18
during storage under accelerated incubation at 63 C for
21 days. The results obtained in this work reected the impact

0.062ab 0.004

0.054ab 0.003
0.051ab 0.004
0.041b 0.003
0.041b 0.003
0.042b 0.004
0.040b 0.004
0.043b 0.002
0.043b 0.003
0.040b 0.002

0.177a 0.002
of these extracts, as natural antioxidants, in retarding of ghee
oxidation. The order of efciency in inhibiting oxidation was

0.11641
in the order PS, PP and nally OP extract. These results are
in agreement with Puravankara et al. (2000) and Pankaj

15
et al. (2013) who concluded that addition of ethanolic extract
of T. arjuna bark at 7% by the weight was highly effective in

0.053ab 0.002

0.050ab 0.001
0.042ab 0.003
retarding the auto-oxidation of both cow and buffalo ghee

0.035b 0.002

0.035b 0.002
0.034b 0.004
0.038b 0.004
0.036b 0.002
0.033b 0.002

0.039b 0.002
0.135a 0.002
during storage and ethanolic extract of Arjuna has signicant
Effect of ethanolic extracts at different concentrations on AV (mg KOH/g fat) of ghee during storage.*
ability to enhance the antioxidant potential of ghee. The onset

0.08617
of rancidity in ghee is mainly due to the oxidation of unsatu-
rated glycerides leading to development of peroxides and/or
due to hydrolysis of glycerides resulting in increased levels of 12
free fatty acids (FFA).
0.027cd 0.002
0.030cd 0.005
0.029cd 0.004

0.030cd 0.003
0.027cd 0.002

0.031cd 0.003
0.026d 0.001

0.041b 0.003
0.040b 0.002
0.051a 0.006

0.032c 0.003
Thiobarbituric acid (TBA) value

0.00554
It is well known that TBA values are taken as an index to eval-

Means with different letters within the same column differ signicantly at P < 0.010.05.
9

uate the advance of oxidation changes occurred in oil and fats.


The addition of extracts as natural antioxidants to ghee
0.015a 0.002
0.015a 0.002
0.015a 0.002
0.013a 0.003
0.013a 0.002
0.015a 0.002
0.015a 0.001
0.015a 0.001
0.015a 0.001
0.015a 0.001
0.015a 0.002
retarded the oxidative changes during accelerated storage
(Table 4). This means that the formation of malonaldehyde,
which affects the formation of pink color intensity from the

T7, T8, T9: ghee treated with 200, 400 and 600 ppm OP extracts, respectively.
T4, T5, T6: ghee treated with 200, 400 and 600 ppm PP extracts, respectively.
T1, T2, T3: ghee treated with 200, 400 and 600 ppm PS extracts, respectively.
reaction of TBA material with malonaldehyde, took place at 0.0028
a relatively lower rate in treated ghee samples. However, the
6

control ghee samples showed higher TBA values throughout


0.013a 0.001
0.041a 0.001
0.013a 0.001
0.013a 0.003
0.013a 0.001
0.014a 0.002
0.012a 0.001
0.011a 0.002
0.052a 0.006
0.013a 0.001
0.013a 0.001

the accelerated incubation period. These structural require-


ments were supported by the powerful antioxidant activity of
the well known BHA. Phenolic compounds act as hydrogen
0.0431

or electron donors to the reaction mixture and therefore the


C1: ghee treated with 200 ppm BHA (positive control).
Storage period (days)

formation of hydro peroxides is decreased. The slow formation


3

of conjugated dienes and consequently the secondary products


0.013a 0.001
0.014a 0.001
0.013a 0.001
0.014a 0.001
0.011a 0.001
0.012a 0.002
0.012a 0.002
0.011a 0.002
0.013a 0.002
0.013a 0.001
0.013a 0.001

by extracts and their major compounds indicated that these


materials may be acted as hydrogen donors to proxy radicals.
Thus, retarding the autoxidation of linoleic acid by chain rad-
0.02602

ical termination has been reported by Farag et al. (1989),


Ozkanli and Kaya (2007) and Mohdali (2010).
C: control without antioxidants.
0

Acid value

Data in Table 5 show that the AV remained without noticeable


changes within the rst six days of storage at 63 C for all
treatments including the control. Slight increases in AV were
Treatments

observed until 15 days of storage period then considerable


Table 5

increase in AV was recorded up to the end of storage period


LSD

(21 days) for all samples including the control. Data presented
C1
T1
T2
T3
T4
T5
T6
T7
T8
T9

*
C

in Table 4 show clearly that the AV of stored ghee was


Oxidative stability of ghee as affected by natural antioxidants 219

noticeably affected by enrichment with by-products ethanolic extracts could be used as preservative ingredients in the food
extracts. PS ethanol 80% extracts (200 ppm) showed the low- and/or pharmaceutical industries.
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