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Effects of green tea-and amla extracts on quality and melanosis of Indian

white prawn (Fenneropenaeus indicus, Milne Edwards, 1837) during chilled
storage

Article  in  Aquaculture and Fisheries · October 2020

DOI: 10.1016/j.aaf.2020.09.003

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Effects of green tea- and amla extracts on quality and melanosis of Indian
white prawn (Fenneropenaeus indicus, Milne Edwards, 1837) during
chilled storage
Aimen Firdous a, Einar Ringø b, Preetham Elumalai a, *
a
Department of Fish Processing Technology (Biochemistry), Kerala University of Fisheries and Ocean Studies, Panangad, Kochi, 682 506, Kerala, India
b
Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT, The Arctic University of Norway, Tromsø, Norway

A R T I C L E I N F O A B S T R A C T

Keywords: Effect of ethanol extracts of green tea (Camellia sinensis L.) and amla (Phyllanthus emblica Linn) were investigated
Indian white prawn on quality and melanosis of chilled stored Indian white prawn (Fenneropenaeus indicus) during 28 days. Extracts
Amla extract were subjected to antioxidant assays viz.1,1-diphenyl-2-picryl hydrazyl radical reducing power methods (DPPH),
Green tea extract
total antioxidant capacity (TAC), total phenolic content (TPC) and ferric reducing antioxidant power(FRAP) to
Lipid oxidation
Melanosis
evaluate antioxidant potentiality and fourier-transform infrared spectroscopy (FT-IR) to identify organic con­
Quality stituents. Polyphenol oxidase (PPO) inhibition was assessed to check the efficacy of the extracts as anti-
melanogenic agents. Biochemical (total volatile nitrogen, free fatty acid and peroxide values), bacteriological
(aerobic counts), melanosis inhibition and sensory quality of chilled stored shrimp were addressed to investigate
the efficacy of extracts as preservative and anti-melanogenic remedy. Free reducing power of green tea - and
amla extracts were in a range of 28.72–65.67%and 17.38–66.95%, respectively. Phenolic content level was
almost same for green tea and amla extract (2.46 ± 0.002 and 2.51 ± 0.036 mg GAE/gram). Total antioxidant
capacity of green tea (210.33 ± 4.63 mg EqAsc/g) was slightly higher than that of amla extracts (145.56 ± 1.98
mg EqAsc/g). FRAP value revealed that green tea(477.49 ± 3.25 mgE Fe (II)/g) had more ferric reducing power
than amla (324.39 ± 5.85 mgE Fe (II)/g).FT-IR analysis revealed the presence of essential organic bioactive
compounds, which play an important role in reducing lipid oxidation and quality loss, and both extracts possess
an encouraging PPO inhibition ability. Treatment by green tea - and amla extracts on chilled stored shrimp
showed promising effects on biochemical and microbiological parameters followed by melanosis inhibition and
enhanced sensory attributes. Treated Indian white prawn with green tea – and amla extract revealed significantly
(P < 0.05) lower value of biochemical indices and microbial load during chilled storage compared to untreated
sample.

1. Introduction polyunsaturated fatty acids e.g. n-3 (204 mg/100g shrimp meat) and n-6
(106 mg/100g shrimp meat), essential amino acids, vitamin B12, sele­
One of the most preferred seafood item is shrimp, with an estimated nium, zinc and other vital micro -and macro minerals, and a high protein
production of 4 million tonnes in 2018-19 (FAO, 2019), and accounts for content (19g/100 g shrimp meat) (Dayal et al., 2013). However,
41.1% in quantity and 68.46% of total income in Indian seafood export compared to poultry products, shrimp has very short storage life due to
trade (MPEDA, 2017-18). Indian white prawn (Fenneropenaeus indicus, its biochemical composition (lipid and amino acids), muscle type and
Milne Edwards, 1837) is important in coastal fisheries, as well as it has microbiological degradation due to higher muscle-pH than meat, and
an important position in the export seafood market in the tropics and the microbiota. Rapid autolysis and lipid oxidation via enzymatic and
subtropics due to its high price (MPEDA 2015). Furthermore, Indian non-enzymatic pathways and black spot (melanosis) formation are other
white prawn is considered as an important species in the rice field major concerns for their limited shelf life. Melanosis is the oxidation of
shrimp farming (pokkali) of the Kerala coast of southwest India (Ranjith, phenol to quinine in the presence of polyphenoloxidase (PPO) which
Karunakaran, & Sekhar, 2018). It contains a high amount of undergoes polymerisation to form black spots on head, exoskeleton and

* Corresponding author. Department of Processing Technology, Kerala University of Fisheries and Ocean Studies (KUFOS), Panangad, Kochi, 682506, Kerala, India.
E-mail address: [email protected] (P. Elumalai).

https://doi.org/10.1016/j.aaf.2020.09.003
Received 13 May 2019; Received in revised form 10 June 2020; Accepted 2 September 2020
2468-550X/© 2020 Shanghai Ocean University. Published by Elsevier B.V. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/).

Please cite this article as: Aimen Firdous, Aquaculture and Fisheries, https://doi.org/10.1016/j.aaf.2020.09.003
A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

uropod, as it causes loss in seafood market (Martinez-Alverez, Montero, leaves, fresh amla fruit were purchased from the local market of Kochi,
& del Carmen Gómez-Guillén, 2005) due to unappealing appearance and Indian white prawn packed with ice in a ratio of 2:1 was brought
from the consumers point of view. from the fish-landing centre at Kalamukku, Kerala.
To combat these problems, the industry has been dependent on
synthetic chemicals, but they can cause hazard health effects. The 2.2. Extraction of green tea and amla
growing health concern among consumers, has forced the processing
industry to switch onto natural remedies to prevent the quality loss of Green tea leaves was extracted as described by Nirmal & Benjakul
the seafood product, instead of using chemical additives, which can (2011a, 2011b), and properly grinded tea leaves were sieved through
cause many chronic health problems and can be carcinogenic above 80 mm mesh. Twenty five gram of the uniformly grinded powder was
certain levels (McEvily, Iyengar, & Otwell, 1991). Recently, Xu et al. mixed with 1L 80% ethanol (tea powder: ethanol = 1:20, w/v) and kept
(2017) conducted an experiment by using plant extract as a potential in magnetic stirrer followed by water bath for 2 h (Rotex Plus, W.ven­
source of antioxidant in food. The authors noticed that extracts from gola, Kerala, India) at 40 ◦ C. After filtering, the filtrate was subjected to
different medicinal plants could be an emerging source to ameliorate the rotary evaporator (IKA HB10) for 60 min at 40 ◦ C to get concentrated
damage caused by oxidative stress by inhibiting the initiation or prop­ filtrate in two successive batches by refluxing in ethanol for
agation of oxidative chain reaction; acts as free radical scavengers and re-extraction followed by drying in a hot air oven at 40 ◦ C for 12 h, and
they comprehended on green tea extraction technologies for natural stored in the dark at 4 ◦ C in an airtight glass container.
antioxidants and their effects at cellular based level. Phenolic compound Extraction of amla was carried out as described by Ahmad, Meh­
e.g. catechin, ferulic acid, lead seed (Leucaena leucocephala), grape seed mood, and Mohammad (1998) with a slight modification. Briefly, amla
(Vitis vinifera), rosemary extract (Rosmarinus officinalis), pomegranate fruit was made seedless and chopped and kept in drier at 40 ◦ C until
peel (Punica granatum) extract are potent anti-melanogenic agents properly dried and to be crushed, and 10g crushed amla powder was
which can lower the PPO formation and enhance the storage time of then mixed with 50 mL of 95% ethanol (amla powder: ethanol = 1:5,
shrimp (Nirmal & Benjakul, 2009a, 2009b, 2011a, 2011b; Gokoglu & w/v) and stirred continuously to obtain the maximum solvent dissolved
Yerlikaya, 2008). constituents. Filtrate was thereafter concentrated by using rotary
According to Vinson (2000) and Cheng (2004), green tea (Camel­ evaporator for 20 min at 40 ◦ C under reduced pressure followed by two
liasinensisL.) is one of the healthiest beverages in the world, as fresh times re-extraction. Concentrated amla extract, was further subjected to
green tea leaves contains 36% polyphenols, of which catechins is the a hot air oven for proper drying at 45 ◦ C for 12 h, and thereafter stored at
major component (Tadesse et al., 2015). In addition, 4 ◦ C in the dark in an airtight glass container, until further use.
epigallocatechin-3-gallate (EGCG), epigallocatechin (EGC),and epi­
catechin (EC) are well known catechins constituents present in green 2.3. Phytochemical screening of the extracts
tea, while other catechins viz. gallocatechin (GC), gallocatechingallate
(GCG) and catechingallate (CG) are present in lesser amount and other Extracts were subjected to Phytochemical screening to determine the
components in green tea include; different sterols, vitamins, triterpe­ presence of tannins and saponins. Ferric chloride test and Frothing Test
noids and other aroma chemicals (Vishnoi, Bodla, & Kant, 2018).The were performed to check out the presence of tannin and saponin
most significant property of tea catechins are their antioxidant potential respectively by the method described by Auwal et al. (2014). For tannin
that can sequester metal ions and scavenge free radicals. Some studies test, 2 mL of the ethanolic extracts were added to a few drops of 10%
have revealed that green tea prevents Parkinson’s disease as well as to Ferric chloride solution (light yellow). The appearance of blackish blue
combat against colon cancer (Koo & Cho, 2004) and improve kidney colour confirmed the presence of Gallic tannins and a green-blackish
functions (Mowafy, Salem, Al-Gayyar, El-Mesery, & El-Azab, 2011). colour indicated presence of catechol tannins. On the other hand, for
Amla (Phyllanthusemblica Linn) is a well-known fruit due to its ethno Frothing Test, 3 mL of the ethanolic extracts were mixed with 10 mL of
medical value(Gaire & Subedi, 2014), and contains nutrients and distilled water in a clean dry test tube and closed it with a stopper and
phenolic constituents such as tannins, phyllembilic acid, rutin and cur­ shaken vigorously for about 5 min, it was kept for 30 min at room
cuminoids, vitamin C and several minerals (Fujii et al., 2013). Several temperature and checked for a presence of honey - comb froth, which
studies reported that amla is a potent candidate against oxidative was the indication of the presence of saponins.
stresses (Kim et al., 2005; Reddy, Padmavathi, Paramahamsa, & Vara­ To check the presence of flavonoid presence, both the extracts were
dacharyulu, 2010; Sharma, Sharma, & Kumar, 2009; Yokozawa et al., subjected to Ferric chloride test and Sodium hydroxide test. For the
2007), and enables antioxidant defence mechanism, and increases the former one, about 0.5 of each portion was boiled with distilled water
levels of antioxidant enzymes like catalase, superoxide dismutase, GSH and then filtered. To 2 ml of the filtrate, few drops of 10% ferric chloride
reductase, GSH peroxidase, and GSH S-transferase (Shukla, Vashistha, & solution were then added. A green-blue or violet colouration indicated
Singh, 2009). the presence of a phenolic hydroxyl group (Trease & Evans, 2002, pp.
To our knowledge, the use of amla to prevent melanosis as well as 95–96). For the alter one, few quantity of the each portion was dissolved
other quality loss of Indian white prawn is not investigated, in addition in water and filtered; to this 2 ml of the 10% aqueous sodium hydroxide
to the effect of ethanol extract of green tea on overall quality of chilled was later added to produce a yellow colouration. A change in colour
stored Indian white prawn. Hence, the current study address to inves­ from yellow to colourless on addition of dilute hydrochloric acid was an
tigate the efficacy of green tea and amla extracts on improvement of indication for the presence of flavonoids (Trease & Evans, 2002, pp.
chilled stored shrimp quality with respect to biochemical, bacteriolog­ 95–96).
ical, melanosis inhibition and sensory quality.
2.4. Antioxidant activity evaluation
2. Materials and methods
2.4.1. DPPH scavenging assay
2.1. Chemicals and raw materials Free radical scavenging ability of both the extracts was assessed by
the DPPH assay as described previously (Ohnishi et al., 1994). Eight
2,2 diphenyl 1 picryl hydroxyl (DPPH), gallic acid, ammonium concentrations (10 μg/ml, 20 μg/ml, 30 μg/ml, 40 μg/ml, 50 μg/ml, 80
molybdate, 2,4,6, tripyridyl 1,3,5 Follin ciocalteu (FC) reagent, TPTZ (2, μg/ml, 100 μg/ml, 120 μg/ml, 150 μg/ml) of pure concentrated extracts
4, 6-tri (2-pyridyl)-s-triazine), L-DOPA, ascorbic acid, nutrient agar were were mixed with 3 mL DPPH (0.1 mM/mL in ethanol) solution followed
obtained from HiMedia (Associated Scientific Company, Kochi, Kerala). by 30 min incubation in dark condition. Absorbance of the mixture was
Other analytical reagents used were procured by Merck. Dry green tea recorded at 517 nm using UV-Spectrophotometer (UV–1800

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A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

SHIMADZU). Ascorbic acid was used as standard and radical scavenging for 15 days. 50 g of grinded shell was mixed with 150 mL extracting
percentage was determined as described by Ohnishi et al. (1994) using buffer (0.05 M Sodium phosphate buffer having 1 M NaCl and 0.2%
the following formula, Brij-35, pH 7.2). The mixture was stirred for 30 min at 4 ◦ C by using a
magnetic stirrer followed by centrifugation at 8.000 rpm at 4 ◦ C for 30
% ​ of ​ DPPH ​ scavenging = (Acontrol–A ​ sample)/(Acontrol) × 100
min. Supernatant was collected, added ammonium sulphate until it
reached 40% saturation. Thereafter, it was stored for 30 min at 4 ◦ C,and
2.4.2. Total phenolic content (TPC) determination centrifuged at 12.500 rpm at 4 ◦ C for 30 min. Pellet was collected and
Estimation of total phenolic content present in the extracts were added with minimum quantity of 0.05 M sodium phosphate to get dis­
estimated spectrophotometrically by using Follin ciocalteu (FC) reagent solved and thereafter centrifuged once more at 3.000 rpm at 4 ◦ C for 30
following the standard method described by McDonald, Prenzler, min. Supernatant was collected and used as “Crude PPO extract”.
Antolovich, and Robards (2001) with slight modifications. In the present
study, gallic acid was used to obtain standard calibration curve. One mL 2.4.6.2. Inhibitory activity of green tea extract and amla extract on PPO.
of aliquots were added 5 mL FC reagent and sodium carbonate (4 mL, Inhibitory activity was carried out as described by Nirmal & Benjakul
0.7M) followed by 30 min absorbance period. Absorbance values were (2009a, 2009b). Hundred μL of five different concentrations of GTE and
recorded at 765 nm using spectrophotometer and the standard curve AE (0.2%, 0.4%, 1%, 2% and 4% w/v) was mixed with 100 μL of pre­
was plotted according to McDonald et al. (2001) pared crude PPO extract to get the final concentrations of 0.1%, 0.2%,
Phenolic content (T) of both the extracts was expressed in mg GAE/g. 0.5%, 1% and 2% w/v respectively and incubated for 30 min at room
The following formula was used to calculate the TPC, temperature. 400 μL of phosphate buffer was then added which was
followed by 600 μL of pre incubation (45 ◦ C) with 15 mM L-DOPA. Re­
T = CV/M
action mixture was left for 3 min at 45 ◦ C and absorbance measured at
475 nm. Control used was without extract. Relative activity, expressed
Where C = Gallic acid concentration (mg/ml) from plotted standard
as percentage was calculated using the following formula:
curve; V = volume of extract taken (ml); M = weight of sample (g).
PPo activity in the presence of extracts × 100
Realativeactivity (%) =
2.4.3. Total antioxidant activity (TAC) evaluation PPO activity of control
Determination of total antioxidant capacity was performed by the
phosphomolybdenum method as reported previously (Prieto, Pineda, & 2.5. Shrimp sample preparation
Aguilar, 1999). 0.3 mL of aliquot from each extract (1 mg/mL) was
added 2.7 mL of phosphomolybdenum reagent, which was prepared Whole Indian white prawns were divided into three slots; green tea
with 28 mM sodium phosphate and 4 mM ammonium molybdate in 0.6 treated -, amla treated -, and control slot (without any treatments). For
M sulphuric acid followed by an incubation of 90 min at 95 ◦ C in a water the green tea - and amla treated slots, 50 g/L extracts were used, and
bath. Thereafter, it was cooled to room temperature, and absorbance soaked for 15 min. Treated shrimps were drained for 3 min at 4 ◦ C and
recorded at optical density (OD), OD695. A blank sample (without adding thereafter all the six slots were packed separately in 12μ polyester
any test sample) was analysed simultaneously along with green tea and laminated with polyethylene bags of 20 × 15 cm dimension and 420 mm
amla test samples. TAC results were expressed as equivalent of standard thickness and stored at 4 ◦ C throughout the experiment.
ascorbic acid (mg Asc/g of dry sample) using the ascorbic acid standard
calibration curve. 2.6. Proximate composition analysis of Indian white prawn

2.4.4. Evaluation of ferric reducing antioxidant power (FRAP) assay All proximate analysis was performed by standard methods of AOAC
Ferric reducing antioxidant power of green tea and amla extracts (2005); moisture content analysis (AOAC – 925.10), total fat content by
were analysed spectrophotometrically by the procedure described by soxhlet extraction (AOAC – 2003.05), ash content by combustion pro­
Benzie and Strain (1996). The FRAP reagent was prepared by mixing cedure (AOAC – 923.03) and protein by the micro Kjeldahl method.
acetate buffer (300 mmol/L, pH 3.6), 10 mmol/L TPTZ solution in 40
mmol/L HCl and 20 mmol/L FeCl3 solution in proportions of 10:1:1
2.7. Chemical analyses
(v/v), respectively. Freshly prepared FRAP reagent was warmed to 37 ◦ C
in a water bath before use. The samples were added to the FRAP reagent.
2.7.1. Total volatile base nitrogen content
The absorbance of the reaction mixture was than recorded at OD593 after
TVB-N determination was done using Conway micro-diffusion
4 min FeSO4was used to prepare the standard curve. The results were
method (Conway, 1933) and value obtained in the experiment was
expressed as mgEq Fe (II)/g dry weight of extracts.
expressed as mg TVB-N/100 g muscle.

2.4.5. FT-IR analysis of green tea and amla ethanol extract

2.7.2. Free fatty acid (FFA) content
Green tea and amla ethanol extracts were scanned in the range of
Free fatty acid analysis of the samples was carried out by the stan­
4000-400/cm with a resolution of 4/cm. The FT-IR spectra of these two
dard protocol of AOAC (1975) and the results expressed as percentage of
samples were recorded in FT-IR spectrometer (Thermo Nicolet, Avatar
oleic acid.
370) using DTGS detector and the KBr beam splitter with ZnSe crystal at
45◦ angle (Thermo Nicolet). Two mg of each sample was used with
2.7.3. Peroxide value (PV) evaluation
respective extraction solvents, and terminated after 20 s. An average of
Peroxide value was assessed by standard AOCS (1989) protocol and
three scans was used to minimise the error.
value was expressed as milliEquivalents/kg of fat.

2.4.6. Assessment of green tea and amla extract efficacy on polyphenols

2.8. Bacteriological analyses
oxidase (PPO) activity

2.8.1. Aerobic bacterial count

2.4.6.1. Crude PPO extract from shell of shrimp. PPO isolation was car­
Twenty-five gram of aseptically minced shrimp samples were
ried out as described by Simpson, Marshall, and Otwell (1987) with
transferred to stomacher bags, containing 225 mL of 0.85% physiolog­
slight modification. Briefly, shell of 30 shrimps was separated, cleaned
ical sterile saline, and mixed for 2 min in a stomacher blender until
with potable water and grinded in cooled condition, and kept at − 20 ◦ C
uniformity. Hundred μL of 10− 1, 10− 2, 10− 3, 10− 4 and 10− 5 dilutions

3
A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

were plated on nutrient agar plates with 0.5% salt, and incubated at 35◦ on addition of dilute hydrochloric acid was an indication for the pres­
± 2 ◦ C for 48±2h for measuring aerobic plate count (mesophilic bac­ ence of flavonoid.
teria)as described by Songsaeng, Sophanodora, Kaewsrithong, and
Ohshima (2010), and the values expressed as CFU/g muscle. 3.2. Antioxidant capacity of green tea and amla extract

2.9. Melanosis assessment 3.2.1. DPPH scavenging assay

The DPPH scavenging assay is an effective antioxidant assay as it
Melanosis was assessed at every 4th day interval basis by the mela­ confirms the ability of the extracted components to scavenge free radi­
nosis scale reported by Montero, Lopez-Caballero, and Pérez-Mateos cals (Ohnishi et al., 1994). This is based on hydrogen donation as donor
(2001), and all samples and the control were given a specific code and of hydrogen molecule acts as an antioxidant agent, and compounds can
each package contained ten numbers of shrimp. Score 0 represented be measured as free radical scavengers and DPPH free radical has the
absence of blackening; 2 represented blackening up to 20% of shrimp ability of transferring electron and producing violet colour solution in
exoskeleton. Likewise, score 4 was assigned for moderate blackening or methanol (Poli et al., 2003). The degree of radical-scavenging potential
20–40% of the shrimp body, 6 for 40–60% and 8 for severe (60–80% of the samples was denoted by the degree of discolouration of the so­
shrimp body) blackening and 10 was assigned for overall (80–100%) lution according to Blois (1958). DPPH scavenging percentage ability
blackening. The same personnel assigned for melanosis evaluation also and IC 50(effective concentration) of green tea and amla ethanol ex­
evaluated sensory quality evaluation. tracts are shown in Tables 2 and 3, respectively. The different concen­
trations of green tea extract (10–150 μg/mL) showed a range of
2.10. Sensory evaluation percentage inhibition between 28.7% and 65.7%, whereas amla at
similar concentrations revealed percentage of radical scavenging ability
Sensory evaluation was carried out, every 4th day interval basis between 17.4% and 66.9%. The result of the present study shows a
according to the 9-point hadonic scale (Meilgaard, Civille, & Carr, parallel correlation of scavenging ability with the highest extract con­
1999). All samples and the control were given a specific code and each centrations, and is in accordance with Kumari and Khatkar (2016).
package contained ten shrimp. Score 9 represented the superior quality Theyreported IC 50 values for antioxidant activity of amla fresh fruit
or like extremely and 1 represented the poorest quality or dislike within a range of 46.7–359.7 μg/mL. Nanjo et al. (1996) reported the
extremely. Likewise, 7 point for moderate likeness, 5 for neutral likeness highest antioxidant capacity for ethanol extract followed by aqueous
and 3 signified moderately disliking. Sensory quality was assessed by a 5 extracts for green tea. Manjulatha, Manjari, Sarita, Imam, and Setty
member expert panel. (2014) reported DPPH radicals of amla with a percentage inhibition
range of 75.59 ± 2.98 to 72.25 ± 4.63% at the highest concentration of
2.11. Statistical analyses 200 μg/mL and IC50 values range of 41.05 to 32.66 μg/mL, which
support the results of the present study. The present study revealed that
Each test was carried out in triplicate, and performed in a completely both extracts have appreciable level of free radical scavenging ability.
randomised design (CRD). The T-test used Graph pad prism 6 and SPSS Hence, it shows the potentiality to be natural antioxidant to reduce
package, and P < 0.05 was considered as statistically significant. Mean oxidative damage during chilled storage.
and Standard error (SE) at 5% was level of significance.
3.2.2. Total phenolic content (TPC)
3. Results and discussion Phenolic compound is the largest and ubiquitous group among all
plant metabolites, which have aromatic ring possessing hydroxyl group
3.1. Phytochemical screening of the extracts (Singh et al., 2007). However, a previous study revealed that, natural
antioxidant comprises of different types of polyphenols mainly flavo­
Phytochemical screening of the ethanolic extracts of green tea and noid groups along with phenolic acid and tocopherol in plant source (Ali
amla revealed the presence of various bioactive components as sum­ et al., 2008). These phenolic groups play the pivotal role in scavenging
marised in Table 1. It indicates the presence of tannin, saponin, flavo­ reactive oxygen constituents (Pourmorad et al., 2006).
noid in the ethanolic extract of Green tea leaves and amla fruit. The The gallic acid standard solution of different concentration (25–200
occurrence of blackish blue colour upon adding 10% Ferric chloride μg/mL) was subjected at 765 nm. A regression co-efficient (R2) = 0.998
solution (light yellow) showed the presence of tannin in green tea and and a slope (m) = 0.003 were obtained with an intercept = 0.187. The
amla. Similarly honey-comb frothing of GTE and Amla extract while equation of standard curve is y = 0.003x + 0.187 (Fig. 1). The total
shaking vigorously with 10 ml distilled water for about 5 min and phenolic content of green tea and amla both ethanol extract were found
allowed to stand for 30 min indicated the presence of saponin. For to be promising with a value of 2.4699 ± 0.002 mg Eq GAE/g dry weight
flavonoid qualitative test, Ferric chloride test and Sodium hydroxide test of green tea extract and 2.5151 ± 0.036 mg Eq GAE/g dry weight of
both were performed and they showed positive result as green-blue amla extract (p < 0.05). Total phenolic content of green tea and amla are
colouration appeared for both extract upon adding few drops of 10% summarised in Table 3. The present study revealed both green tea and
ferric chloride solution and a change in colour from yellow to colourless amla have almost similar quantity of phenolic content (~2.5 mg Eq

Table 1 Table 2
Phytochemical screening of Green tea and amla ethanolic extract. Percentage (%age) of DPPH radical scavenging ability of green tea and amla
ethanol extracts at different concentration.
Phytochemical Type of test Appearance Green tea Amla
constituent extract extract Concentration (μg/mL) Green tea extract (% RS) Amla extract (% RS)

Tannin Ferric chloride Blackish blue þ þ 10 28.72 17.38

test 20 44.83 28.91
Saponin Frothing test Honey comb þ þ 30 48.68 37.77
like froth 50 51.48 51.29
Flavonoid Ferric chloride Green blue þ þ 80 54.64 55.07
test (10%) 100 59.54 64.67
Sodium Yellow to þ þ 120 62.69 65.06
hydroxide test colourless 150 65.67 66.95

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A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

Table 3 of several compounds like polyphenols, ellagic acid, gallic acid and
IC50, TPC, TAC and FRAP of green tea and amla ethanol extract. tannins in green tea. Another reason could be high antioxidant capacity,
Extracts IC 50 TPC (mg Eq TAC (mg FRAP (mgE Fe due to the presence of hydroxyl (OH) groups on the β-ring as in EGCG,
Quercetin/100g) EqAsc/g) (II)/g) GCG, EGC and GC in green tea, which are suggested to have more ability
Green tea 60.33 2.46 ± .002 210.33 ± 477.49 ± 3.25 to scavenge free radicals(Almajano, Carbo, Jiménez, & Gordon, 2008).
extract 4.63 Haji et al. (2008) observed almost similar TAC value as that revealed in
Amla 74.89 2.51 ± .036 145.56 ± 324.396 ± 5.85 the present study, as the authors displayed that 1 g of green tea is
extract 1.98 equivalent to 50–275 mg of pure ascorbic acid (vitamin C), or 156–813
Values are mean ± S.D, n = 10. mg vitamin E, which implicate that green tea is an efficient source of
antioxidant. It was previously displayed that green tea is able to inhibit
lipid peroxidation by their free radical scavenging activity (Soong &
Barlow, 2004).
Manjulatha et al. (2014) revealed TAC in amla in a range of 249.81
± 0.63 to 733.02 ± 2.1 mg ascorbic acid equivalent/g, and is in
agreement with the results of the present study. Amla comprise a higher
amount of abundant antioxidant (Antony, Merina, Sheeba, & Mukka­
dan, 2006), and it is believed that amla can be a potent alternative
source of synthetic additives to prevent lipid oxidation in food pro­
cessing sector (Liu, Qiu, Ding, & Yao, 2008). Reason behind this high
amount of total antioxidant may be due to the presence of various
polyphenols i.e. kaempferol, ellagic acid and gallic acid (Hab­
ib-ur-Rehman et al.,2007).

3.2.4. Ferric reducing antioxidant power (FRAP) assay

FRAP is a widely used antioxidant assay which works based on the
Fig. 1. Total phenolic content standard curve of Gallic acid Concentration principal of redox-linked colorimetric reaction, where ferric ion (Fe3+)
(μg/ml). converts to ferrous (Fe2+) ion at a lower pH, and forms coloured ferrous-
probe complex from a colourless ferric-probe complex (Firuzi et al.,
GAE/g− 1). 2005). The antioxidant activity is parallel to the polyphenols content of
The TPC of green tea revealed in the present study is similar to the the solvent extracts. Polyphenols are more efficient reducing agents for
TPC value of 2.53 mg GAE/g for green tea reported by Ramírez-Ar­ ferric iron (Wong, Leong, & Koh, 2006), and possess a highly positive
istizabal, Ortíz, Restrepo-Aristizabal, and Salinas-Villada (2017). Haji relationship between total phenols and antioxidant activity (Oktay,
et al. (2008) observed similar value of green tea total phenolic content Gülçin, & Küfrevioğlu, 2003). The reducing power of the ethanol extract
(0.196 ± 0.012 mg GAE/gram) due to the high antioxidant activity of of green tea and amla are revealed in Table 3. Amla ethanol extract
green tea. Phenolic contents from green tea extracts correlate with showed FRAP values of 324.39 ± 5.85 mgE Fe (II)/g,while green tea
radical scavenging activity (Li et al., 2009), and this explains their ethanol extract revealed 477.49 ± 3.25 mgE Fe (II)/g, and the values
reducing capability by hydrogen donation as well as singlet oxygen were significantly (P < 0.05) different. Amla sought seems to have more
quenching. Tsai, Tsai, Yu, and Ho (2007) reported that phenolic com­ reducing power compare to green tea extract, and this is probably due to
pounds in green tea have strong antioxidant capacity, as it possesses the high concentration of polyphenols present in amla compare to green
flavan-3-ols and can prevent oxidative stress by chelating available free tea (Habib-ur-Rehman et al., 2007).The present study observed a good
ferrous ions. correlation between DPPH scavenging ability, and ferrous ion reducing
The TPC value of 2.51 ± 0.036 of amla revealed in the present study ability that implies the efficacy of both the extract to be a potent
is similar as that reported by Mukhinder (2017). Agarwal, Kumar, reducing agent to fight against oxidation. This trend is in agreement
Gupta, and Upadhyaya (2012) reported that amla possesses a high with the finding of Chen, Lin, and Hsieh (2007).
amount of total phenolic content, and it increases with higher concen­
tration of extraction solvent. Polyphenols content varies with solvent, 3.3. FT-IR analysis
degree of polarity of the solvents and compatibility of compound with
the solvents (Teh et al., 2014; Zhang et al., 2007). Hossain, Barry-Ryan, In the present study, green tea - and amla ethanol extracts were
Martin-Diana, and Brunton (2010) stated that fresh sample revealed subjected to FT-IR analysis. Fig. 2 a and b show the FT-IR spectral profile
lower value of TPC compared to processed fruits, due to that the outer for green tea and amla respectively. Main peaks range from 3271.88
layers contains higher phenolic content than seed or pulp (Chism & cm− 1 to 432.30 cm− 1 for green tea and 3278.53 cm− 1 to 766.71 cm− 1 for
Haard, 1996). Green tea and amla extracts comprise of good amount of amla are showed with extended marking. It was revealed that 3271.88,
phenolic compounds which enriches their antioxidant activity. 1694.89, 1606.84, 1552.85,1518.0, 1448.80, 1338.63, 1235.77,
1144.14, 1033.96, 821.78, 763.34 and 432.30 cm− 1 main peaks for
3.2.3. Total antioxidant capacity (TAC) green tea and 3278.53, 1719.59, 1613.68, 1544.60, 1343.10, 1213.26,
Total antioxidant capacity by phosphomolybdenum method is based 1039.34, 918.96, 871.32, 812.71 and 766.71 cm− 1 main peaks for amla
on the principle of green phosphate complex MO (V) formation from MO were clearly recorded in fingerprint region of FT-IR. In case of green tea,
(VI) in the presence of sample analysed (Subhashini, Thangathirupathi, 3271.88 cm− 1, the value could be due to vibration of –OH stretching
& Lavanya, 2011). The phosphomolybdenum method is quantitative bands (Nakashini, 1969; Silverstein et al., 1998). 1694.89 cm− 1 is due to
since TAC is expressed as the numbers of equivalent of ascorbic acid. It is C–– O stretching of protein and 1606.84 cm− 1 is due to CH2 symmetric
otherwise termed as PM (phosphomolybdenum) assay, and is widely stretching (Toyran, Zorlu, & Severcan, 2005). Range of 1500–1600
used as a routine laboratory method to quantify the total antioxidant cm− 1 indicates the presence of flavonoids, polyphenols and catechins
capacity of certain plant extract (Prieto et al., 1999). TAC values of green (Lam, Proctor, Howard, & Cho, 2005). 1448.80, 1338.63, 1235.77
tea and amla ethanol extracts are revealed in Table 3. Green tea showed cm− 1is mainly due to –CH in plane bending vibration of allylic group,
more total antioxidant capacity (210.33 mgE Asc/g) compared to amla –CH stretching vibration of gem-dimethyl group and C–O–C stretching
(145.56 mgE Asc/g) (p < 0.05). The reason for this could be the presence vibration band of ether group, respectively. 1144.14 cm− 1 indicates the

5
A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

Fig. 2. a) FT-IR spectra of green tea ethanol extract sample and b) amla ethanol extract sample. (For interpretation of the references to colour in this figure legend,
the reader is referred to the Web version of this article.)

presence of alcohols, esters, carboxylic acids groups in sample. Peaks of

Table 4
1033.96, 821.78 and 763.34 cm− 1 could be due to the presence of –CH
Chemical groups analysed in the extracts as per the main peaks obtained by
in plane bending vibration of trans or E-alkene, and cis or Z-alkene
FTIR.
presence as suggested previously (Nakashini, 1969; Silverstein et al.,
Chemical group Green tea extract Amla extract
1998). In case of amla, 3278.53 cm− 1 is probably due to O–H stretch,
H–bonded, which signifies the presence of phenols and alcohols (Orčić, Flavonoids + +
Mimica-Dukić, Francišković, Petrović, & Jovin, 2011). The spectral data Poly phenols + +
Catechins
of 1719.59, 1613.68 and 1544.60 cm− 1 may be due to the presence of
+ +
Allylic + –
–CH symmetric/asymmetric aliphatic bond, ketones/carbonyl groups, Dimethyl + –
phenyl ring and aromatic ring C–C stretching. Similarly, visual intensity Ethers + –
estimates for the spectral band of 1343.10, 1213.26 and 1039.34 cm− 1 is Esters + +
Carboxylic acids
mainly due to C–O (stretching) – ester, CO stretching and aromatic ring + –
Alkenes + –
stretching. The occurrence of spectral wavelength of 918.96, 871.32, Phenols – +
812.71 and 766.71 cm− 1 is possible due to HC– – CH aromatic amides as Ketones – +
previously suggested by Miyazawa, Shimanouchi, and Mizushima Carbonyls – +
(1956).The main chemical groups present in the extracts analysed by Aromatic Amides – +

FTIR are summarised in Table 4. +: Present, -: absent.

6
A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

3.4. Polyphenols oxidase (PPO) activity unacceptable. TVB-N of all groups showed a parallel increment with
storage time (P < 0.05). At start, the initial TVB-N value was 2.38 ±
Polyphenol oxidase is an endogenous enzyme present in shellfish 0.009 mgN/100 g, but at day 28th, the control slot showed an increased
(McEvily et al., 1991) and the major cause of melanosis reaction causing TVB-N value, 34.39 ±.04 mgN/100 g. However, on day 28th of chilled
blacking of shrimp head and cephalothorax. The effect of green tea and storage at 4 ◦ C, treatment of green tea revealed aTVB-N value of 27.84 ±
amla extract solution on inhibition of PPO from Indian white prawn is 0.009 mgN/100 g, and amla treated sample 28.67 ± 0.012 mgN/100 g
showed in Fig. 3.The present study revealed its efficacy in dose depen­ (Fig. 4), which indicates that the control is unacceptable, in contrast to
dent manner (P < 0.05). At similar concentration level, amla revealed treated shrimps.
better relative activity percentage compared to green tea (P < 0.05). In the present study, a good correlation between TVB content and the
Although, at 0.2% concentration both the extract solutions showed bacterial load was revealed (Table 5) and this finding correspond well
similar results (P > 0.05). Nevertheless, present study revealed an with that revealed by Ozogul et al. (2010) as they displayed that un­
encouraging increase in PPO inhibition of amla extract at 2% concen­ treated fish sample was declared as rejected on 13th day of chilled
tration compared to the green tea extract (P < 0.05). A previous study storage. However, chilled storage samples treated with rosemary extract
revealed that, interaction of enzyme with phenolic compound could was acceptable until day 17. Furthermore, the findings in the present
prevent PPO activity (Janovitz-Klapp, Richard, Goupy, & Nicolas, study are in accordance with Nirmal & Benjakul (2011a, 2011b) in their
1990). Nirmal & Benjakul (2009a, 2009b) reported that catechins study on Pacific white shrimp (Litopenaeus vannamei) treated with lead
showed PPO inhibitory activity in a dose-dependent manner, and it was seed extract. Viji et al. (2015) reported a similar trend in a study with
assumed that catechins might act as a competitive inhibitor for PPO due chill stored Indian mackerel and fish treated with mint (Mentha arvensis)
to similarity to L-DOPA structure, which is a substrate for PPO. The leaf and citrus (Citrusaurantium) peel extracts.
present study suggest that green tea and amla extract are effective to The findings of the present study is also in agreement with previous
control PPO effects on chilled stored shrimp, and can be used as an reports observing an beneficial effect by using plant extracts to improve
alternative natural anti-melanogenic substance. TVB-N values of chilled stored fish products (Gao et al., 2014; Pezeshk,
Rezaei, & Hosseini, 2011).
3.5. Proximate analysis
3.6.2. Fee fatty acid content (FFA)
FFA is considered as an indicative tool for quality assessment of
In the present study, the proximate composition of Indian white
chilled stored fish and shellfish. In the current study, significant (P <
prawn was: 71.60 ± 2.54% moisture, 10.32 ± 0.86% crude protein,
0.05) differences (P < 0.05) in FFA contents were observed among the
8.24 ± 0.11% crude fat, 1.96 ± 1.21% ash. Variations in chemical
treatment groups throughout the storage period (Fig. 5). In control
composition are not unusual as Karuppasamy et al. (2013) reported
sample, increased the FFA value from 0.017 ± 0.001% (expressed as
variations in chemical composition of penaeid shrimp, mainly in protein
percentage of oleic acid) to 0.137 ± 0.002%, whereas values of 0.017 ±
and moisture.
0.001% to 0.112 ± 0.001%were revealed for green tea treated sample,
and 0.024 ± 0.001 %to 0.115 ± 0.001% for amla treated samples
3.6. Evaluation of green tea extract and amla extract effect on the quality (Fig. 5). This trend is in accordance with the previous finding of Goko­
of chilled stored Indian white prawn glu, Yerlikaya, Topuz, and Buyukbenli (2012), where FFA value of fish
croquette was 2.75 ± 0.12% in the control, while a value of 1.83 ±
3.6.1. Total volatile base nitrogen (TVB-N) 0.01% was observed in croquettes treated with tomato extract after a
TVB-N is the most reliable parameter for evaluation of freshness storage period of four months.
indices of fish and shellfish and indicates sign of spoilage with produc­ Free fatty acids are derived by enzymatic or non-enzymatic hydro­
tion of trimethylamine (TMA). A level of 35–40 mg TVB-N/100g of fish lysis of lipids, particularly phospholipids and triglyceride, and are
muscle is an acceptable limit (Lakshmanan & Fung, 2000). High TVB-N located primarily in the cell membrane (Serdaroğlu & Felekoğlu, 2005).
values is considered as spoiled and unfit for consumption as trimethy­ Both lipase and phospholipase enzymes have significant activity by
lamineoxide (TMAO) is reduced to TMA, dimethylamine and formal­ producing FFA in various fish and shellfish species stored at − 12 or
dehyde by the action of endogenous enzymes or by spoilage bacteria. − 14 ◦ C (Olley, Pirie, & Watson, 1962). In an early study, interaction
TMA causes off odour (Regenstein, 1982) and high levels are regarded as during frozen storage of various fish species revealed that, accumulation
of FFA increased with prolonged storage time, and at elevated frozen
storage temperature (Dyer & Dingle, 1961). In the present study, it was
observed a lower lipid hydrolysis rate in both treated slots irrespective of
their antioxidant contents (Fig. 5).This finding is in accordance to that
revealed by Sarah, Hadiseh, Gholamhossein, and Bahareh (2010) where

Fig. 3. Effect of green tea - and amla extract at different levels on the inhibition
of polyphenoloxidase from shell of Indian white prawn. Bars represent the
standard error (n = 3). Different capital letters on the bars within the same GTE Fig. 4. Total volatile base content of Indian white prawn treated with green tea
or AE together with the control indicate the significant differences (P < 0.05). – and amla extract, and control during 28 days of storage at 4 ◦ C. Bars represent
(For interpretation of the references to colour in this figure legend, the reader is the standard error (n = 3). (For interpretation of the references to colour in this
referred to the Web version of this article.) figure legend, the reader is referred to the Web version of this article.)

7
A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

Table 5 milliEquivalents/kg from the initial value of 0.66 ± 0.008 and 0.68 ±
Determination of aerobic bacterial counts of Indian white shrimp samples 0.0004 milliEquivalents/kg, respectively. These PV values were signif­
treated with green tea, amla and control during the 28 days storage study. icant (P < 0.05) different between Indian white prawn treated with
1st day 7th day 14th day 21st day 28th day green tea and amla extracts and control (Fig. 6). Haghparast, Kashiri,
Control (CFU/ 3.00 £ 6.00 £ 4.02 £ 9.0 £ 2.1 £
Alipour, and Shabanpour (2011) reported a similar trend of PV values,
mL) 104 105 107 107 108 where green tea treated Persian sturgeon revealed a lesser increase in PV
Green tea (CFU/ 2.03 £ 1.6 × 105 2.00 × 2.0 × 3.4 × vs. control during chilled storage.
mL) 104 105 106 106
Amla (CFU/mL) 2.10 × 3.40 × 4.9 × 105 6.7 × 2.6 ×
3.7. Aerobic bacterial count (ABC)
104 105 106 107

Values are mean ± S.D, n = 10. One important reason why chilled stored shrimp quick spoilage oc­
curs is spoilage bacteria, which causes bad odour, unattractiveness and
makes shrimp not acceptable for consumption, and the ABC indicate the
bacteriological load (Huss, 1988; Viji et al., 2015). ABC of the treated
groups and control group during 28 days storage are showed in Table 5.
On the 1stday, total bacterial count was in a range of log 4.30–4.47 cfu/g
for all slots. During the storage, aerobic bacterial counts increased
significantly (P < 0.05). However, the trend was significantly lower (P
< 0.05) for the amla and green tea treated groups. At the 28th day of
storage, the ABC of the control group raised to log 8.32 cfu/g, whereas,
green tea treated sample showed a load of log 6.53 cfu/g and amla log
6.82 cfu/g (Table 5) which was significantly (P < 0.05) lower vs. the
control group. This log reduction showed a good correlation with sen­
sory analysis score. The ABC support the chemical quality analyses
Fig. 5. Free fatty acid content of Indian white prawn treated with green tea – (TVB-N, FFA and PV) well, and showed almost similar level of quality
and amla extract,and control during 28 days of storage at 4 ◦ C. Bars represent retardation in both treatment groups.
the standard error (n = 3). (For interpretation of the references to colour in this A previous study by Del Nobile et al. (2009) revealed that lemon
figure legend, the reader is referred to the Web version of this article.) extract and thymol in combination with modified atmospheric pack­
aging (MAP) significantly (P < 0.05) reduced the bacterial load vs. the
refrigerated Persian sturgeon (Acipenser persicus) treated with green tea control. ABCs revealed in the present study, indicates that green tea and
and onion juice lower the FFA content compared to the untreated group amla have potential inhibitory effect on spoilage bacterial growth. This
after 8 days of storage. Similar findings were reported in studies by trend is in accordance with Ozogul et al. (2010) using sardine (Sardinella
different authors evaluating the effect of different antioxidants gibbosa) fillet treated with rosemary extract vs. the untreated group.
(Aubourg, Pérez-Alonso, & Gallardo, 2004; Serdaroğlu & Felekoğlu,
2005; Chaijan, Benjakul, Visessanguan, & Faustman, 2006; Lugasi et al., 3.8. Effect of green tea and amla on melanosis formation of Indian white
2007; Sahari, Nazemroaya, & Rezaei, 2009; Taheri, Motalebi, & Fazlara, prawn at chilled storage
2012).
Melanosis score for Indian white prawn treated with green tea and
3.6.3. Peroxide value (PV) amla extract and the control during 28 days of chilled storage study are
Fish lipid is very prone to oxidation as hydroperoxides it produced showed in Fig. 7.On the 1st day, all samples displayed no melanosis, and
due to its rich content of highly unsaturated fatty acid. It oxidises iodide no differences in appearances were revealed. During storage, melanosis
to iodine or iron (II) to iron (III), can be estimated by PV evaluation formation showed an significant (P < 0.05)increasing trend, and is in
(primary oxidation compounds) in terms of milliEquivalent peroxide/kg accordance to the finding of Nirmal & Benjakul (2011a, 2011b) showing
extracted fat from sample (Stine, Harland, Coulter, & Jenness, 1954). that catechins in green tea have a potential effect on melanosis forma­
Detecting spoilage at earlier stage is difficult as this compound is odour tion in Pacific white shrimp.
free, but at later stages, it breaks down and forms auto-oxidative con­ The present study noticed that, the green tea and amla treated groups
stituents, a sign of early phase of auto-oxidation, which leads to severe remained appealing until 18th to 19thday and 16th to 18th day,
oxidative spoilage. Breakdown products i.e. aldehydes, ketones and al­ respectively to the panellists as blackening was moderate (40–60%) and
cohols are volatile in nature and causes off odour, and indicates rancid score obtained was within 4–6 range whereas, control reached a score
stage of product. PV plays an important role in early stage of oxidation as
it increase hydroperoxide formation rate, higher than the decomposition
rate, but once it reach maximum level, it starts decreasing due to lower
availability of substrates and instability of peroxide molecules (De
Abreu, Losada, Maroto, & Cruz, 2011). Alghazeer, Saeed, and Howell
(2008) stated that green tea (250 ppm and 500 ppm) treated Atlantic
mackerel (Scomber scombrus) fillets revealed lower peroxide and hy­
droperoxide rate during 8 weeks of storage at − 10 ◦ C compared to
control. In addition, it is worth to mention that several studies proved
that plant extract could efficiently lower peroxide formation by delaying
secondary reactions of the hydroperoxides (De Abreu et al., 2011;
Pezeshk et al., 2011; Shi, Cui, Yin, Luo, & Zhou, 2014; Viji et al., 2015).
In the present study, PV increased from 0.81 ± 0.004 milli­
Equivalents/kg to 18.46 ± 0.0004 milliEquivalents/kg in the control Fig. 6. Peroxide value (PV) of Indian white prawn treated with green tea – and
group, while green tea and amla treated samples revealed a significantly amla extract, and control during 28 days of storage at 4 ◦ C. Bars represent the
(P < 0.05) lower level of 13.82 ± 0.004 and 15.47 ± 0.009 standard error (n = 3). (For interpretation of the references to colour in this
figure legend, the reader is referred to the Web version of this article.)

8
A. Firdous et al. Aquaculture and Fisheries xxx (xxxx) xxx

Fig. 7. Melanosis score of Indian white prawn treated with green tea – and
amla extract,and control during 28 days of storage at 4 ◦ C. Bars represent the Fig. 8. Sensory score in hedonic scale of Indian white prawn treated with green
standard error (n = 3). (For interpretation of the references to colour in this tea – and amla extract and control during 28 days of storage at 4 ◦ C. Bars
figure legend, the reader is referred to the Web version of this article.) represent the standard error (n = 3). (For interpretation of the references to
colour in this figure legend, the reader is referred to the Web version of
above 8 which implies severe blackening (≥80%)from the 14th day and this article.)
onwards (P < 0.05).Therefore, an extract dose of 50 g/L of green tea and
amla extract significantly(P < 0.05) reduced melanosis of Indian white bacterial counts were significantly lower in treated Indian white prawn
shrimp during chilled storage. This result is in accordance with the compared to the control. Treatment with green tea and amla extract can
finding of Gokoglu and Yerlikaya (2008), where significant (P < 0.05) efficiently retard melanosis development and ameliorate the sensory
inhibitory effect of grape seed treatment on melanosis formation in quality of shrimp in chilled storage as overall acceptance of treated
shrimp was revealed. The authors noticed that the treated group showed sample was encouraging compared to untreated group. Dip treatment in
acceptable condition up to the 2nd day of storage, but the control group ethanol extract of green tea and amla enhance the storage quality and
revealed a heavy blackening by the 3rd day of storage, and was prolong the shelf life of Indian white prawn during chilled storage
considered as unacceptable by the sensory panel. effectively. The present study also revealed that green tea and amla
extracts are alternative natural preservative for fishery products instead
3.9. Effect of green tea and amla on sensory quality of Indian white of the abuse of synthetic additives.
prawn at chilled storage Hence, treatment with green tea and amla extracts are suggested as
an alternate aid to prevent overall quality loss in shrimp during post
The changes in sensory qualities of Indian white prawn treated by mortem handling and subsequent storage.
green tea, amla and the control during chilled storage are showed in
Fig. 8. On the 1stday, all samples had the score near 9 in the hedonic Author’s contributions
scale and the same likeness was observed in all treatments (P > 0.05).
During storage, the spoilage rate was significantly (P < 0.05) faster for Aimen Fidrous was involved in the concept design, data analysis and
the control group. The sensory scores of green tea extract and amla execution of this project. Preetham Elumalai was involved in Supervi­
extract treated lots were significantly repressed, compared to the control sion, Conceptualization and acquisition of data’s. Further improvement
(P < 0.05). of manuscript by critical reviewing and finalization of the manuscript
Generally, shelf life ends when essential sensory parameters such as together with Einar Ringø.
off-odour and flavour become pungent or putrid due to bacterial activity
(Huss, 1988) and even appearance of shrimp becomes unaccepted from Declaration of competing interest
consumer point of view. It is rejected, when it reach a score below 3–4
score in the hedonic scale. The present study revealed that, green tea and The authors have no competing interests. The research did not
amla treatment was acceptable until 18th to 20th day and 17th to 19th receive any specific grant from funding agencies in the public, com­
day, respectively due to texture and odour. Appearance of treated lots mercial, or not-for-profit sectors.
was good to acceptable and score obtained was within 5–6 range
whereas, control received score of below 4 before the treated groups (P Acknowledgements
< 0.05). Thus, from the data indicated that the treated groups retained
their good quality characteristics. The authors extend their sincere appreciation to Kerala University of
A study conducted by Fang et al. (2013) on Pacific white shrimp Fisheries and Ocean Studies, Department of Fish Processing Technology
using pomegranate peel extract revealed that a decrease of sensory score for assistance with the experiments.
was significantly inhibited in pomegranate peel extract treated sample
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