African Journal of Basic & Applied Sciences 8 (6): 321-323, 2016
ISSN 2079-2034
© IDOSI Publications, 2016
DOI: 10.5829/idosi.ajbas.2016.321.323
The Comparison Survey Antioxidant Power and Content of Extracted Astaxanthin from
Shrimp Waste with Acid, Alkaline and Enzymatic Methods
Ahmadi Asghar, Salehi Ali, Ghorbani Mozhgan, Sadighara Parisa and Asadi Sharifi Tayebehe
Department of Environmental Health Engineering, Division in Food Safety and Hygiene,
Tehran University of Medical Sciences, Tehran, Iran
Abstract: Shrimp waste is one of the important sources of natural carotenoids. In This study describes
pigments and carotenoid extraction from shrimp wastes under different methods. And, evaluated protective
effects of carotenoids on lipid peroxidation of sunflower oil. Level of total carotenoids by alkaline extraction
method was significantly (P < 0.05) higher than other methods. Also, the highest inhibition of lipid peroxidation
was obtained in alkaline extraction method. These results demonstrated that good yields and antioxidant
activity can be obtained alkaline treatment.
Key words: Astaxanthin Shrimp Waste Extraction Methods
INTRODUCTION acceptability of foods by causing undesirable changes
in flavour, texture, appearance and nutritional quality
Global production of fish and shrimp increased [6].
significantly in the recent years. Much of the increase in Since the synthetic antioxidants have possible health
later years is attributable to shrimp farming. Asia plays a effects and toxicity, searching natural preservatives for
leading role in shrimp farming, accounting for almost replacing has been attracted by food industries [7].
80% of world shrimp production [1-3]. According to the Carotenoids are one of major component of the
Iranian Fisheries Organization, the shrimp farming was crustaceans waste [8]. One of the important
about 13,000 tons in the year 2013-14 in Iran. characteristics of carotenoids is their ability to act as
About 48-56% of shrimp total body weight is waste antioxidants. Astaxanthin (3, 30-dihydroxy-b, b-carotene-
[4]. Shrimp waste is the main byproduct in fishery 4, 40-dione) is one of the most important pigments that
industries [3]. Use of shrimp wastes has been of interest its antioxidant properties have been reported in many
to researchers for two reasons: The first these wastes are studies. Owing to its outstanding antioxidant activity
environmental contaminants. Second these are the and attractive pink colour can be used as a colourant and
important sources of natural carotenoids [5]. Therefore antioxidant in food and medicine[9, 10]
reuse of this waste will perform the prevention of Carotenoid has been extracted various method. The
environmental pollution and can be used to produce recovery and extraction of these valuable components
useful compounds. from the shrimp waste would improve the economics of
The major carotenoids in the wastes of shrimps are the shrimp farming process [4]. As well as the use of
astaxanthin, Zeaxanthin, lutein and beta-carotene [4]. carotenoids as antioxidant in the food industry is
Astaxanthin, the main carotenoid found in shrimp, it is affordable.
responsible for the color and antioxidant activity [1]. The present study was undertaken to find out the
Its antioxidant activity was reported to be higher than antioxidant activity in the composition of carotenoid
that of carotene and lutein and is higher than tocopherol pigments during Acidic, alkaline and enzymatic
against certain reactive oxygen species [3]. techniques extraction from shrimp wastes.
Lipid oxidation are major deterioration causes of the Incidentally, carotenoid ability to prevent the
quality of seafood and can decreases consumer oxidation of sunflower oil was studied.
Corresponding Author: Asadi Sharifi Tayebehe, Department of Environmental Health Engineering, Division in Food Safety and
Hygiene, Tehran University of Medical Sciences, Tehran, Iran. E-mail: t-asadish@razi.tums.ac.ir.
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African J. Basic & Appl. Sci., 8 (6): 321-323, 2016
MATERIALS AND METHODS samples were heated at 90°C for 15 min. The absorbance
of the supernatants was measured at 532 nm.All of
In this study, first the carotenoid extraction during experiments were performed in triplicate [13].
enzymatic, alkaline and acidic methods, then its
antioxidant effects on sunflower oil was investigated by Statistical Analysis: The results were evaluated by
spectrophotometry. the analysis of variance and the differences between
the means by Tukey’s test using the SPSS Statistics
Collection of Samples: The shrimp wastes were obtained ver23.
from the processing plants. Then, the wastes were air
dried in the shade and turned into powder and stored at RESULTS AND DISCUSSION
-20°C until use.
Table 1 shows the amount of carotenoids extracted
Fractionation of Carotenoids: At first, the samples from shrimp wastes by Acidic, alkaline and enzymatic
dissolved in 0.1 N HCL for 24h at room temperature for methods. The concentration of carotenoid pigment in the
demineralization treatment. In order to extract extracts was calculated using the standard curve obtained
carotenoids,three methods were used. by commercial carotene. Y= 6.3994x + 0.0096, R2= 0.9998
Amount of total carotenoid by alkaline extraction
Alcalase Deproteinization: Approximately 0.5 g of method was significantly (P < 0.05) higher than other
samples were solved in 20 mL of the sodium hydroxide methods. Accordingly, it can be introduced as an
(1 N) at 90°C for 2h. Then the pH being maintained at 8.5 applicable method for carotenoid extraction. Alkaline
by the addition of 1 N NaOH. The insoluble fraction was extraction of shrimp waste could be a potential source for
centrifuged and the supernatant was used for experiments pigments. The recovery of carotenoids extraction from the
[11, 12]. waste would improve the economics of the shrimp
processing plant. The use of organic solvents are not safe
Enzymatic Deproteinization: In this method 10% of extraction methods Sachindra et al. [4]. Our study showed
trypsin was added to samples. pH was adjusted to 8 for that alkaline extraction was more effective as compared
enzyme activity and heated at 37 °C for 4 h. In this step, with other methods and the extraction of pigment from the
the carotenoid-protein compounds are removed from shrimp waste could be efficiently and economically
wastes. Then, the solution was centrifuged and the achieved by this method Jeddi et al. [8].
supernatant was used for determination of total The protective effects of extracted carotenoids from
carotenoids [1, 3]. shrimp wastes on lipid peroxidation of sunflower oil is
displayed in Table 2. The highest inhibition of lipid
Acidic Deproteinization: In this method the wastes were peroxidation was obtained when carotenoids were
solved in 20% Trichloroacetic acid (TCA (for 24h. extracted using alkaline treatment. The high inhibition of
lipid peroxidation in alkaline treatment method could be
Determination of Total Carotenoids: The total amount of due to carotenoid content and its alkaline conditions [14].
carotenoids were determined by b carotene standard Therefore, the carotenoids separated from the protein
curve and by spectrophotometric method at 470 nm. complex. Results similar to our data have also been
The total carotenoid content of the samples was reported. Studies on Extraction of shrimp waste pigments
calculated on the basis of the standard curve of by enzymatic and alkaline treatment demonstrated that the
carotene [13]. carotenoids extracted from wastes inhibited lipid
peroxidation through enzymatic hydrolysis [5, 12].
Inhibition of Lipid Peroxidation: Inhibition of lipid Therefore, in alkaline condition enzymatic hydrolysis
peroxidation was evaluated by oxidation inhibition of done much better.
sunflower oil. At first, 15 tubes were selected and in 5 Among the carotenoids, astaxanthin as an antioxidant
rows were named (A-B-C-D-E). Then 1 ml of sunflower oil, is more effective than -carotene and other carotenoids to
1 ml lecithin and 0.2 CuSo4 added to all tubes. And were destroy free radicals.Astaxanthin extraction techniques
shaken for 24 hours. The solutions mixed with 20 % from crustacean wastes have include fermentation,
trichloroacetic acid. The samples were centrifuged. enzymes, organic solvents (Such as acetone, methanol,
Thiobarbituric acid was added to the supernatant and the alcohol), hydrochloric acid and edible oils [15].
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African J. Basic & Appl. Sci., 8 (6): 321-323, 2016
Table 1: Level of total carotenoid 5. Babu, C.M., R. Chakrabarti, and K.R.S. Sambasivarao,
Extraction Method Carotenoid Content (ppm) 2008. Enzymatic isolation of carotenoid-protein
Carotenoid extracted by alkaline treatment 0.07±0.001
complex from shrimp head waste and its use as a
Carotenoid extracted by enzyme 0.028±0.001
source of carotenoids. LWT-Food Science and
Carotenoid extracted by acid 0.005±0.002
Values are presented as Mean ± SD
Technology, 41(2): 227-235.
6. German, J.B., S.E. Chen, and J.E. Kinsella, 1985. Lipid
Table 2: Level of lipid peroxidation oxidation in fish tissue. Enzymic initiation via
Level of lipid peroxidation lipoxygenase. Journal of Agricultural and Food
Samples (µM MDA/g sample) Chemistry, 33(4): 680-683.
Control (with no carotenoids 8.46±0.6a,b 7. Maruthiah, T. and A. Palavesam, 2017.
extracted and synthetic carotenoids)
Characterization of haloalkalophilic organic solvent
Synthetic carotenoids + Sunflower oil 3.9±0.4a
Acidic carotenoid + Sunflower oil 6.9±0.2
tolerant protease for chitin extraction from shrimp
Alkaline carotenoid + Sunflower oil 5.06±0.3b shell waste. International Journal of Biological
Enzymatic carotenoid + Sunflower oil 5.51±0.19 Macromolecules.
Values are presented as Mean ± SD 8. Sachindra, N., et al., 2007. Recovery of carotenoids
a,b
Means within a column with differ significantly (P<0.05) from ensilaged shrimp waste. Bioresource
technology, 98(8): 1642-1646.
In our study, alkaline extraction method was more 9. Jeddi, M.Z., G.J. Khaniki, and P. Sadighara, 2013.
effective than other methods to achieve carotenoids from Optimization of extraction of carotenoids from shrimp
shrimp waste. waste. Global Veterinaria, 10(6): 636-637.
10. Quan, C. and C. Turner, 2009. Extraction of
CONCLUSION astaxanthin from shrimp waste using pressurized hot
ethanol. Chromatographia, 70(1-2): 247-251.
Shrimp wastes are an important source of 11. De Holanda, H.D. and F.M. Netto, 2006. Recovery of
carotenoids, particularly that of astaxanthin and its components from shrimp (Xiphopenaeus kroyeri)
esters. The crude carotenoids extract obtained by processing waste by enzymatic hydrolysis. Journal of
enzymatic, acidic and alkaline extraction of shrimp Food Science, 71(5): C298-C303.
processing discards, and its fractions were evaluated for 12. Jafari, A.M., et al., 2012. Extraction of shrimp waste
their antioxidant activity. The results revealed the strong pigments by enzymatic and alkaline treatment:
antioxidant activity of the carotenoids obtained from evaluation by inhibition of lipid peroxidation.
alkaline extraction method. The antioxidant activity of Journal of Material Cycles and Waste Management,
shrimp carotenoid extract could be used for natural 14(4): 411-413.
antioxidant in food and biomedical applications. 13. Gómez-Estaca, J., et al., 2017. Characterization and
storage stability of astaxanthin esters, fatty acid
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