Antioxidant effect of curry leaf (Murraya koenigii) powder on quality of ground and cooked goat meat

Abstract

The antioxidant effect of curry leaf powder (CLP) was determined by assessing the formation of lipid peroxides, free fatty acids (FFA) and thiobarbituric acid substances (TBARS) in raw ground and cooked goat meat patties during refrigerated storage. pH, water holding capacity and cooking loss per cent were not affected by curry leaf powder when added in raw ground goat meat. Fresh goat meat with CLP had acceptable odour up to 5 days whereas in control sample it was up to 3 days. Raw goat meat with CLP had significantly lower free fatty acids content than control during 9 days refrigerated storage. CLP significantly inhibited the rate of lipid peroxides and TBARS formation in raw meat than control. CLP in cooked goat meat patties showed significant antioxidant effect as indicated by TBARS values measured by distillation as well as extraction method. CLP did not affect microbial population in raw and cooked goat meat during entire storage period. These results show that CLP at concentrations as low as 0.2% is a very effective inhibitor of primary and secondary oxidation products in raw ground and cooked goat meat patties and has potential as a natural antioxidant in raw and cooked meat systems.

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International Food Research Journal 18: 559-565 (2011)
Das, A.K., Rajkumar, V. and Dwivedi, D.K.
Goat Products Technology Laboratory, Central Institute for Research on Goats,
Makhdoom, Farah-281122, Mathura, India
Antioxidant effect of curry leaf (Murraya koenigii) powder on quality
of ground and cooked goat meat
Abstract: The antioxidant effect of curry leaf powder (CLP) was determined by assessing the formation of lipid
peroxides, free fatty acids (FFA) and thiobarbituric acid substances (TBARS) in raw ground and cooked goat
meat patties during refrigerated storage. pH, water holding capacity and cooking loss per cent were not affected
by curry leaf powder when added in raw ground goat meat. Fresh goat meat with CLP had acceptable odour up
to 5 days whereas in control sample it was up to 3 days. Raw goat meat with CLP had signicantly lower free
fatty acids content than control during 9 days refrigerated storage. CLP signicantly inhibited the rate of lipid
peroxides and TBARS formation in raw meat than control. CLP in cooked goat meat patties showed signicant
antioxidant effect as indicated by TBARS values measured by distillation as well as extraction method. CLP
did not affect microbial population in raw and cooked goat meat during entire storage period. These results
show that CLP at concentrations as low as 0.2% is a very effective inhibitor of primary and secondary oxidation
products in raw ground and cooked goat meat patties and has potential as a natural antioxidant in raw and
cooked meat systems.
Keywords: Curry leaf powder, antioxidant activity, free fatty acids, goat meat, quality
Introduction
Lipid oxidation is a major cause of muscle food
deterioration as it decreases nutritional properties of
foods since it involves the loss of essentially fatty
acids and vitamins and the generation of potentially
toxic reaction products such as the manoladehyde
(MDA) and cholesterol oxidation products (COPs)
(Morrissey et al., 1998; Tang et al., 2001). In
addition, lipid oxidation affects essential sensory
traits of meat product, causing avour, colour and
texture deterioration (Estevez et al., 2005). The
development of so called warmed over avour (WOF)
in pre-cooked meat products was assumed due to
high storage temperature and/or reheating before
serving. Many substances have been investigated as
potential antioxidant to prevent such lipid oxidation.
However, the image of some synthetic antioxidants
such as butylated hydroxyanisole (BHA), butylated
hydroxytoluene (BHT), tertiary butyl hydroquinone
(TBHQ), and propyl, octyl, and dodecyl gallates
has been worsened by the ndings linking the use
of these compounds to health risk (Tokusoglu and
Basmacioglu, 2004; Das et al., 2006a). Development
of various comminuted meat products offers a
protable utilization of tough meat and ground
meat tends to become more rancid and brown
more rapidly, due to pigment and lipid oxidation.
Moreover, mechanisms for control of lipid oxidation
has become increasingly important with the rise in
popularity of pre-cooked and convenience foods
(Das et al., 2006a). Consequently, substances derived
from the plant kingdom such as dried herbs, grape
seed extract and essential oils have been successfully
used to reduce lipid oxidation in meat products
(Estevez et al., 2005, Brannon, 2008). Many natural
plant extracts contain primarily phenolic compounds,
which are potent antioxidants.
Curry leaf (Murraya koenijii; Rutaceae) is a
leafy spice characterizing authentic Asian-Indian
cuisine and it is used in small quantities for its
distinct aroma as well as for preservation purposes.
Recently some studies have reported that carbazole
alkaloids have several biological activities such
as anticarcinogenic effects in dimethylhydrazine
(DMH) treated rats (Farhath-Khanum et al., 2000),
antiplatelet activity and vasorelaxing effects (Wu et
al., 1998). Chevalier (1996) also reported curry leaf
has medicinal value as traditionally used in eastern
Asia. Interest in greater use of curry leaf has been
stimulated since its high antioxidant potency was
reported and this antioxidant activity is attributed
due to mahanimbine, murrayanol and mahanine from
M .koenigii (Tachibana et al., 2003; Ningappa et al.,
2008). Chowdhury et al. (2001) reported that these
alkaloids shown to have antimicrobial activity against
gram positive and negative bacteria, and fungi. Use
of curry leaf powder as antioxidant in ground and
pre-cooked muscle foods on oxidative stability has
been poorly reported. In this experiment, we used
curry leaf powder instead of different extracts due to
its major use in culinary purpose in the form of green

60 Das, A.K., Rajkumar, V. and Dwivedi, D.K.
International Food Research Journal 18: 559-565
leaf or dry powder. Therefore, this work was design
to investigate the efcacy of CLP as antioxidant and
antimicrobial in ground goat meat and precooked
goat meat patties during refrigerated storage and
to evaluate the TBARS values determined by two
different methods (distillation and extraction).
Materials and Methods
Source of meat samples
The samples, comprising mostly Longissimus
dorsi, Semimembranosus and Semitendinosus muscles
of male (of about 12-16 months age) goat carcasses,
slaughtered as per the approved guidelines of Animal
Ethics Committee in the Institute slaughterhouse,
were collected within 5 hrs of slaughter, packed in
low density polyethylene (LDPE) bags. The samples
were placed in a refrigerator at 4±1°C for about 22
hrs.
Preparation of control and treated meat samples
The meat pieces, after conditioning, were trimmed
off separable fat and connective tissue, cut into small
cubes and minced using automatic meat mincer by
passing through 5 mm plates (Talleres Raman Model
P-22, Barcelona) to obtain ground goat meat (GGM).
The products were collected in sterile mixing bowls.
Ground goat meat was divided into 2 groups. The
curry leaf powder (CLP) was used as nal percentage
of total meat weight: 1) no CLP (control), 2) CLP at
0.2%. CLP was mixed uniformly with ground goat
meat and meat was reground through 5 mm plates
and packaged in LDPE bags, sealed and stored in a
refrigerator 4±1°C up to 9 days. Two packages for
each treatment were selected on sampling days 0, 3,
5, 7 and 9 to determine various physico-chemical and
microbiological quality parameters.
Preparations of spice mix and curry leaf powder
Dried spices namely aniseed, black pepper,
capsicum, caraway seed, cardamom, cinnamon,
clove, coriander, cumin, ginger and turmeric were
procured from local market and oven dried at 50°C
for 2 hrs before grinding and mixing. Spice mix was
stored in polyethyleneterephthalet (PET) bottle for
experimental use. Fresh matured leaves of M. koenigii
plant from Institute campus were collected and sun-
dried after removal of extraneous matter. The leaves
were kept in oven at 50°C for 2 hrs and then ground
mechanically and sieved through a ne mesh (U.S.S.
30 # mesh screen). The curry leaf powder was stored
in a bottle for further use.
Preparation of goat meat patties
The separated lean obtained from the carcasses
was packed in high-density polyethylene bags and
stored at -15±1°C till used for patties preparation.
Before use, meat was thawed overnight at 4±1°C.
Minced goat meat was transferred into a bowl
chopper (Seydelmann, Model type K20, Germany)
for preparation of meat emulsion in batches of 3 kg
using standard recipe (Das et al., 2006b). After the
preparation of emulsion, it was divided into 4 groups:
1). No text ingredients (control), 2) 1.5% spice mix
only, 3) 0.2% CLP only and 4) mixture of spice mix
and CLP. These emulsions were mixed separately in
Hobart paddle type mixer (Model No.50, USA) for
three min.
About 70 g of emulsion was moulded in a
Petridish (75 mm diameter and 15 mm height) to
form patties and was cooked in a pre-heated oven at
180±5°C for 15 minutes after which they were turned
and allowed to get cooked for 10 more minutes till
internal temperature reached about 75 ± 2°C recorded
by a probe thermometer (Labware Scientic, Inc,
USA). The patties were packed in low-density
polyethylene pouches and stored at refrigerated
temperature (4±1ºC). TBARS values of the product
were evaluated using both distillation and extraction
method at an interval of 5 days. The experiment was
replicated thrice.
pH determination
pH of raw ground and cooked meat patties was
determined by blending 10 g of sample with 50 ml of
distilled water for 1 min using a tissue homogenizer
(Model PT-MR-2100, Kinematica AG, Switzerland)
at 8000 rpm for 1 min. The pH of the suspension
was recorded by dipping combined glass electrode
of digital pH meter (Systronics, µ pH system 361,
Delhi, India).
Water holding capacity and cooking loss
Water holding capacity (WHC) of ground goat
meat was estimated by a centrifugation method
(Wardlaw et al., 1973). Cooking loss (CL) percent of
the samples was estimated by heating approximately
25 g of ground meat sample individually placed
inside polyethylene bags at 80°C for 20 min using
a thermostatically controlled water bath. Samples
were cooled for 15 min under running tap water.
They were taken from the bags, dried with lter
paper and weighed. Cooking loss was expressed as
the percentage loss related to the initial weight (Das
et al., 2006a).

Antioxidant effect of curry leaf powder 61
International Food Research Journal 18: 559-565
Colour and odour score
Color score was determined by using a 5-point
scale, where 1=pale pink, 2=pink, 3=pinkish red,
4=bright red and 5= reddish brown. Whereas sensory
score for the meat odour was obtained using a 5-point
scale, where 1=very unpleasant, 2=moderately
unpleasant, 3=moderately pleasant, 4=pleasant and
5=very pleasant (Das et al., 2006a).
Lipid oxidation
Lipid oxidation was monitored by measuring
thiobarbituric acid reactive substances (TBARS),
lipid peroxides and free fatty acids. The distillation
as well as extraction method described by Tarladgis
et al. (1960) and Witte et al. (1970) respectively,
were followed to measure 2-thiobarbituric acid-
reacting substances (TBARS) values of the raw
ground and cooked meat. The values were expressed
as mg malonaldehyde/kg of sample. The methods
described by Koniecko (1979) were followed for the
determination of peroxide value and free fatty acids
(FFA).
Microbiological examination
For microbiological examination, a representative
of 10 g ground meat sample was withdrawn and
homogenized (Model PT-MR-2100, Kinematica AG,
Switzerland) aseptically using 90 ml 0.1% peptone
water (APHA, 1984) and serial dilutions were made
using 0.1% sterile peptone water. Total aerobic
bacteria and psychrotrophs counts were enumerated
on duplicate pour plates of Plate Count Agar (Hi-
Media, Mumbai, India), which were incubated at 37°C
for 48 h and 5°C±1 for 10 to 12 days respectively.
After appropriate incubation, plates having 25 to
250 colony-forming units (CFU) were counted and
multiplied by the dilution factor to determine CFU/g
of meat.
Statistical analysis
Three trials were conducted for each experiment.
The statistical design of the rst experiment was 2
(treatment) × 3 (replication) × 5 (storage periods)
combination with randomized block design and data
were analyzed by analysis of variance (ANOVA)
using Linear Model Procedure of SPSS software 10.
Duncan’s multiple range test (DMRT) and critical
difference (CD) were determined at 5% and 1%
signicant level. The statistical design for the second
experiment (cooked meat patties) was same, except
four treatments were conducted.
Results and Discussion
Curry leaf powder on pH, Water holding capacity
(WHC) and cooking loss
Effect of curry leaf powder on pH, water
holding capacity and cooking loss of ground goat
meat during refrigerated storage is presented in Table
1. Addition of curry leaf powder in ground goat meat
(GGM) did not change the pH. Biswas et al. (2006)
observed that pH was not affected due to curry leaf
powder added in ground poultry meat. pH values of
control and treated samples increased signicantly
with increasing storage period. The increase in pH
during the storage period may be due to growth
of Gram-negative bacteria such as Pseudomonas,
Moraxella, Acinetobacter, etc (McDowell et al.,
1986). Jay (1996) also reported increase in pH during
storage and this could be due to accumulation of
metabolites by bacterial action on protein and amino
acids. The pH recorded in the present study were in
range reported by Verma and Sahoo (2000) in ground
chevon during refrigerated storage.
Water holding capacity (WHC) is known to be
one of the major quality characteristics of fresh meat,
as it affects some major characteristics of cooked
meat such as potential drip loss, technological
quality, appearance and sensory properties. WHC
was not inuenced by CLP. There was a signicant
linear decrease in WHC of the meat samples during
storage period. Similar result was reported in ground
buffalo meat during refrigerated storage (Das et al.,
2006a). Cooking loss (CL) percentage of ground goat
meat was not signicantly different from the control
sample. CL of goat meat increased linearly during
storage period for 9 days. It increased from 43.92 on
day 1 to 51.58 on day 9 of the storage period. This
trend of CL may be due to a decrease in the solubility
of protein during storage and post mortem enzymatic
Table 1. Effect of curry leaf powder on pH, water holding capacity and cooking loss of ground
goat meat during storage
Treatments Storage period (days) Treatment effect
13579SEM
pH NS
Control 5.70c5.84b5.88b6.05a6.09a0.03
CLP 5.69d5.75cd 5.92bc 5.99ab 6.07a0.03
Water holding capacity (%) NS
Control 15.43a12.87b10.99c5.66d3.13e0.85
CLP 15.38a12.88b10.82c5.50d2.79e0.86
Cooking loss (%) NS
Control 43.37e45.76d48.09c50.13b51.67a0.56
CLP 43.92e45.03d47.89c49.67b51.58a0.51
Means bearing different superscripts in a row differ signicantly (P<0.05),
NS= Non-signicant, SEM- Standard Error of the Mean.

62 Das, A.K., Rajkumar, V. and Dwivedi, D.K.
International Food Research Journal 18: 559-565
hydrolysis of ATP (Hamm, 1970).
Curry leaf powder on colour and odour score
Colour and odour of fresh meat are also
important criterion for its acceptability. There was no
signicant difference in colour and odour of control
and CLP treated ground goat meat (Table 2). During
refrigerated storage period, the visual colour declined
signicantly as the storage days progressed. The
colour score of treated sample was 4.62 on day 1,
which decreased to 2.41 whereas in control sample,
it was 4.59 on day 1 and 2.06 on day 9. Das et al.
(2006a) also reported similar results in ground buffalo
meat in refrigerated storage. In the present study, the
visual colour crossed the borderline (3.00) in control
sample after day 5 while the sample containing CLP
had colour appealing till the end of day 7. These
ndings were in agreement with the observation of
Verma and Sahoo (2000) in tocopherol preblended
ground chevon in chilling storage.
CLP though did not signicantly inuence odour
score but improved odour score than control sample.
As indicated in Table 2, the CLP treated sample
was acceptable upto 5 days, odour score being 3.35.
On the contrary, the control sample was below the
acceptable level (3.00) on day 5 that means it was
acceptable up to three days. The odour score started
declining linearly as the storage period increased. Das
et al. (2006a) reported similar trend of odour score
in ground buffalo meat during refrigerated storage.
In general, minced goat meat sample maintained
the desirable dour up to 5 days and lost the odour
sometime between day 5 and 7 (Verma and Sahoo,
2000).
Curry leaf powder on free fatty acids and peroxide
values
Curry leaf powder treated sample had signicantly
lower free fatty acids content (0.31 to 0.71) as
compared to control sample (0.37 to 0.93) (Table 2).
It is clear that addition of 0.2% CLP to ground goat
meat is sufcient to lower the free fatty acids content
during refrigerated storage. Pearson (1968) reported
that minced beef had FFA content in the range of
0.38 to1.74% and a maximum acceptability limit
of 1.8% FFA in view of their progressive increase
during storage. In the present study, FFA content of
minced goat meat was well within this limit. Again
Lea (1962) also reported that avour deterioration
in beef or mutton was noticeable when the acidity
of extracted fat reached 2-3% as oleic acid. FFA
content of meat sample signicantly increased during
refrigerated storage. Previous workers also reported
the increasing trend of FFA content of buffalo meat
(Rao and Kowale, 1988) and goat meat (Verma and
Sahoo, 2000) during 9 days refrigerated storage.
Recently, Das et al. (2008) also reported increase of
FFA during refrigerated storage of goat meat patties.
The peroxide value generally serves as a useful
indicator of the extent of oxidation of lipids, fats and
oils. An advantage of peroxide value determination
is that it directly measures the lipid peroxides,
which are primary lipid oxidation products (Shantha
and Decker, 2001). Similarly, addition of CLP
signicantly reduced the peroxide value of ground
goat meat during storage. There was a signicant
linear increase in peroxide value of the meat samples
during refrigerated storage. In this study, peroxide
value in control sample increased from 1.74 ppm
day 1 to 3.56 ppm on day 9 whereas incase of treated
sample, it increased from 1.52 to 2.82 ppm (Table 2).
Verma and Sahoo (2000) found a positive correlation
between peroxide value and thiobarbituric acid
reactive substances (TBARS) number of chevon
during refrigerated storage.
Table 2. Effect of curry leaf power on colour, odour, free fatty acids and peroxide values of
ground goat meat during storage
Treatments Storage period (days) Treatment effect
1 3 5 7 9 SEM
Colour Score$NS
Control 4.59a4.28b3.57c2.83d2.07e0.18
CLP 4.62a4.38b3.34c3.22d2.41e0.16
Odour Score#NS
Control 4.48a3.72b2.99c2.42d1.82e0.19
CLP 4.32a3.96b3.25c2.86d2.00e0.17
Free fatty acids (%) **
Control 0.37d0.58c0.65c0.78b0.93a0.04
CLP 0.34e0.46d0.55c0.57bc 0.71a0.03
Peroxide values (meq/kg)
Control 1.74a2.21b2.52c3.01d3.56e0.03 **
CLP 1.52a2.04b2.17b2.61c2.82d0.05
Means bearing different superscripts in a row differ signicantly (P<0.05),
**P<0.01; NS= Non-signicant, SEM- Standard Error of the Mean
#Odour score based on 5 point scale, where 1=pale pink, 2=pink, 3=pinkish red, 4=bright red and 5= reddish brown.
$Colour score based on 5 point scale, where 1=very unpleasant, 2= moderately unpleasant, 3=moderately pleasant, 4= pleasant and 5=very pleasant.

Antioxidant effect of curry leaf powder 63
International Food Research Journal 18: 559-565
Curry leaf powder on thiobarbituric acid values of
raw ground goat meat
Figure 1 shows the effect of curry leaf powder on
TBARS values measured by distillation and extraction
method of raw goat meat during refrigerated storage.
Addition of curry leaf powder, into minced goat meat
at a concentration of 0.2% resulted in a signicant
(P<0.05) reduction of TBARS values compared to the
control. There was a linear increase of TBA value in
both control and treated goat meat during refrigerated
storage. TBARS value increased from 0.65 on day
1 to 1.12 on day 9, whereas in control sample it
was 0.74 to 1.43 during storage. Extraction method
showed signicantly lower mean TBARS value than
distillation method. The control meat sample was
not acceptable after day 3 while CLP treated sample
could be acceptable up to day 7 as revealed by TBA
value of 0.99. Control ground goat meat exhibited
signicantly higher rate of TBRAS formation than
CLP containing raw goat meat during storage. The
acceptability limit of TBARS value in this study
was 1.0. Earlier workers reported that meat sample
containing TBARS value less than 1 possesses no off
odour (Tarladgis et al., 1960).
Spice mix (SM) and CLP on TBARS values of
cooked goat meat patties
CLP in cooked goat meat patties showed
signicant antioxidant effect as indicated by TBARS
values estimated by distillation and extraction method
(Fig 2 and Fig 3). In distillation method (DM),
the control sample had signicantly higher lipid
oxidation throughout the storage period than SM,
CLP + SM and CLP treated patties. CLP along with
SM revealed enhanced antioxidant effect than CLP
alone in goat meat patties during 20 days refrigerated
storage. Control patties like raw ground meat showed
signicantly higher rate of TBARS formation than
patties containing CLP during storage. TBARS
value increased from 0.36 on day 1 to 0.89 on day
20 whereas in control sample it was 0.49 to 1.11 mg
malonaldehyde/kg sample during 20 days storage.
Tachibana et al. (2003) reported that carbazole
alkaloids of CLP have potent antioxidant activity.
Enhanced antioxidant activity of CLP extract was
due to higher phenolic content (Ningappa et al.,
2008). Again Lee et al. (2002) noted that enrichment
of phenolic compounds within the plant extract is
correlated with their enhanced antioxidant activity.
In extraction method (EM), similar trend of
antioxidant activity of CLP was observed in goat
meat patties during 20 days storage period. Patties
with CLP and SM had signicantly lower TBARS
value than others. In DM, signicantly higher TBARS
value had been observed in all samples during storage
period than EM. Such difference in TBA value
between these methods was due to estimation of
malonaldehyde at different wavelength (Tarladgis et
al., 1960). Biswas et al. (2006) also report that higher
TBARS values in DM than in EM in cooked chicken
patties during storage. Besides this, heating in DM
increases aldehyde quantities from lipid precursors.
Ulu (2004) reported that heating of samples during
distillation promotes further oxidation leading to
additional malonaldehyde and other TBA reacting
substances. This study reported above clearly
indicates that CLP is heat stable and withstands meat
processing temperatures
Curry leaf powder on microbial quality of raw and
cooked goat meat
Curry leaf powder did not show any antimicrobial
effect when mixed with ground goat meat during
chilling storage. Microbial count increased
signicantly with increase in storage period in both
control and treated sample. The total plate count
ranged from 4.57 to 7.01 log10 cfu/g and 3.92 to
0.30
0.40
0.50
0.60
0.70
0.80
0.90
1.00
1.10
1.20
0 5 10 15 20
Storage period(days)
TBARS value (mg/malonaldehye/kg)
cont rol SM SM + C LP CLP
Figure 2. Effect of spice mix and CLP on TBARS values (distillation
method) of cooked goat meat patties during refrigerated storage
0.20
0.30
0.40
0.50
0.60
0.70
0.80
0.90
1.00
1.10
0 5 10 15 20
Storage period (days)
TBARS value (mg malonaldehyde/kg)
cont rol SM SM + C LP CLP
Figure 3. Effect of spice mix and CLP on TBARS values (extraction
method) of cooked goat meat patties during storage
0.40
0.50
0.60
0.70
0.80
0.90
1.00
1.10
1.20
1.30
1.40
1.50
13579
Storage period
TBARS Number(mg malonaldehyde/kg)
Control C LP Contr ol-ex CLP -ex
Figure 1. Effect of CLP on TBARS value (distillation and extraction
method) of raw ground goat meat during refrigerated storage (ex-
extraction method)

64 Das, A.K., Rajkumar, V. and Dwivedi, D.K.
International Food Research Journal 18: 559-565
6.39 log10 cfu/g in case of psychrotrophs during
9 days storage period. Contrasting to this nding,
Chowdhury et al. (2001) reported the antimicrobial
activity of carbazoles from M. koenigii. Similarly,
in case of cooked goat meat patties, there was no
signicant difference in microbial count between
control and treated patties. Patties with dry spice
mix had slightly lower microbial load but that was
not signicant. Total plate count and psychrotrophs
count observed in this study were well with in the
acceptable limit in cooked meat products. Biswas et
al. (2006) did not nd any antimicrobial effect of
CLP both in fresh and cooked chicken meat.
Conclusion
Use of curry leaf powder inhibits the formation of
free fatty acids, lipid peroxide and thiobarbituric acid
reactive substances in ground goat meat that does not
affect pH, water holding capacity and cooking loss
during storage. Shelf life of the ground goat meat
could be extended up to 5 days as compared to 3 days
in control sample during refrigerated storage. This
study also shows that CLP has effective antioxidant
activity in cooked goat meat patties during storage.
Curry leaf powder seems to have no anti-microbial
effect on fresh as well as cooked goat meat during
storage. However, further studies especially in regard
to use of different CLP extracts as antioxidant and
pro-oxidative effect of NaCl are needed in order to
reinforce this conclusion.
Acknowledgement
The authors would like to thank the Director
of the Institute for providing necessary facilities to
carryout this study and to Mr Radhey Shyam and
Mohd. Sarfaraj for their technical assistance.
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