Keywords
spectrophotometry
Artemia
aquaculture nutrition
How to Cite
Abstract
Carotenoids play an essential role in aquaculture by supporting growth, survival, and stress tolerance in cultured species, while also contributing to product coloration, an important indicator of its quality and market value. Reliable monitoring of carotenoid levels in cultured organisms and feeds, as well as the development of effective feed enrichment strategies, requires standardized analytical procedures. This study provides a comparative evaluation of sample preparation methods and calculation approaches used for spectrophotometric determination of total carotenoids, using Artemia as a model organism. The results show that extending acetone extraction from one to three days increases the measured carotenoid concentration in fresh samples, whereas the opposite trend is observed in frozen samples, where prolonged extraction leads to pigment degradation. Additionally, the use of different calculation formulas can produce multi‑fold discrepancies in carotenoid concentrations derived from identical absorbance data. These findings highlight the need for unified protocols for sample preparation, extraction, and data processing to ensure consistency and comparability of carotenoid measurements in aquaculture research and practice.
References
Abdollahi Y, Ahmadifard N, Agh N, Rahmanifarah K, Hejazi MA (2019) β Carotene enriched Artemia as a natural carotenoid improved skin pigmentation and enhanced the mucus immune responses of platyfish Xiphophorus maculatus. Aquaculture International 27:1847–1858. https://doi.org/10.1007/s10499-019-00437-8
Anuevo GM (2014) Evaluation of green tea enriched Artemia sp. as food and antioxidant activity enhancer for freshwater ornamental fishes. Dissertation, University of Tsukuba
Çankırılıgil EC, Berik N, Çakmak E, Özel OT, Erbay EA (2022) Dietary carotenoids influence growth, fillet pigmentation, and quality characteristics of Black Sea trout (Salmo labrax Pallas, 1814). Thalassas 38:793–809. https://doi.org/10.1007/s41208-022-00407-7
Casella P, Iovine A, Mehariya S, Marino T, Musmarra D, Molino A (2020) Smart method for carotenoids characterization in Haematococcus pluvialis red phase and evaluation of astaxanthin thermal stability. Antioxidants 9:422. https://doi.org/10.3390/antiox9050422
Cheban L, Khudyi O, Prusińska M, Duda A, Khuda L, Wiszniewski G, Kushniryk O, Kapusta A (2020) Survival, proximate composition, and proteolytic activity of Artemia salina bioencapsulated with different algal monocultures. Fisheries & Aquatic Life 28:205–215. https://doi.org/10.2478/aopf-2020-0025
De Carvalho CC, Caramujo MJ (2017) Carotenoids in aquatic ecosystems and aquaculture: a colorful business with implications for human health. Frontiers in Marine Science 4:93. https://doi.org/10.3389/fmars.2017.00093
Elbahnaswy S, Elshopakey GE (2024) Recent progress in practical applications of a potential carotenoid astaxanthin in aquaculture industry: a review. Fish Physiology and Biochemistry 50:97–126. https://doi.org/10.1007/s10695-022-01167-0
Elshafey AE, Khalafalla MM, Zaid AAA, Mohamed RA, Abdel-Rahim MM (2023) Source diversity of Artemia enrichment boosts goldfish (Carassius auratus) performance, β carotene content, pigmentation, immune physiological and transcriptomic responses. Scientific Reports 13:21801. https://doi.org/10.1038/s41598-023-48621-4
Feizi P, Jafari SM (2025) Optimization of astaxanthin extraction from red (Gracilaria corticata) and brown (Sargassum polycystum) macroalgae through ultrasonication and microwave processing. Ultrasonics Sonochemistry 121:107556. https://doi.org/10.1016/j.ultsonch.2025.107556
Hata M, Hata M (1969) Carotenoid metabolism in Artemia salina L. Comparative Biochemistry and Physiology 29:985–994. https://doi.org/10.1016/0010-406x(69)91000-7
Hooshmand H, Shabanpour B, Moosavi-Nasab M, Golmakani MT (2017) Optimization of carotenoids extraction from blue crab (Portunus pelagicus) and shrimp (Penaeus semisulcatus) wastes using organic solvents and vegetable oils. Journal of Food Processing and Preservation 41:e13171. https://doi.org/10.1111/jfpp.13171
Hsu WJ, Chichester CO, Davies BH (1970) The metabolism of β carotene and other carotenoids in the brine shrimp, Artemia salina L. (Crustacea: Branchiopoda). Comparative Biochemistry and Physiology 32:69–79. https://doi.org/10.1016/0010-406x(70)90156-8
Huang J, Hui B (2020) Feed induced variation in the carotenoid composition of brine shrimp. eFood 1:247–253. https://doi.org/10.2991/efood.k.200522.001
Johnson EA, An GH (1991) Astaxanthin from microbial sources. Critical Reviews in Biotechnology 11:297–326. https://doi.org/10.3109/07388559109040622
Kanokrung A, Ratananikom K, Watanadilok R (2024) Effect of dietary carotenoids on survival, growth, and carotenoid accumulation in mandarinfish, Synchiropus splendidus (Herre, 1927). Journal of Fisheries and Environment 48:38–47. https://doi.org/10.34044/j.jfe.2024.48.2.03
Kumar PA, Marian MP (2006) Studies on carotenoid in Artemia parthenogenitica. Romanian Biotechnological Letters 11:2733–2737.
Lichtenthaler HK, Wellburn AR (1983) Determinations of total carotenoids and chlorophylls a and b of leaf extracts in different solvents. Biochemical Society Transactions 11:591–592. https://doi.org/10.1042/bst0110591
Manikandan K, Felix N, Prabu E (2020) A review on the application and effect of carotenoids with respect to canthaxanthin in the culture of fishes and crustaceans. International Journal of Fisheries and Aquatic Studies 8:128–133. https://doi.org/10.22271/fish.2020.v8.i5b.2314
Maoka T (2020) Carotenoids as natural functional pigments. Journal of Natural Medicines 74:1–16. https://doi.org/10.1007/s11418-019-01364-x
Maoka T, Kawashima Y, Takaki M (2018) Structures of yellow xanthophylls and metabolism of astaxanthin in the prawn Penaeus japonicus. Journal of Oleo Science 67:1425–1433. https://doi.org/10.5650/jos.ess18103
Millán-Almaraz MI, López-Peraza DJ, Nieves-Soto M, Peraza-Yee MM (2023) Effect of 4 microalgal diets on the proximal composition, chlorophyll concentration, and total carotenoid content in Artemia franciscana. Ciencias Marinas 49:e3381. https://doi.org/10.7773/cm.y2023.3381
Minakova AA, Bazarnova NG, Minakov DV, Mikushina IV, Dubrovina AE (2022) Obtaining astaxanthin from Artemia salina cysts for the development of functional food products. Polzunovskiy Vestnik 1:167–173. https://doi.org/10.25712/astu.2072-8921.2022.04.022
Nelis HJ, Lavens P, Van Steenberge MMZ, Sorgeloos P, Criel GR, de Leenheer A (1988) Qualitative and quantitative changes in the carotenoids during development of the brine shrimp Artemia. Journal of Lipid Research 29:491–499. https://doi.org/10.1016/S0022-2275(20)38522-9
Ownagh E, Agh N, Noori F (2015) Comparison of the growth, survival and nutritional value of Artemia using various agricultural by products and unicellular algae Dunaliella salina. Iranian Journal of Fisheries Sciences 14:358–368.
Parisenti J, Beirão LH, Tramonte VLCG, da Silva FO, da Silveira Brito CC, Moreira CC (2011) Preference ranking of colour in raw and cooked shrimps. International Journal of Food Science and Technology 46:2558–2561. https://doi.org/10.1111/j.1365-2621.2011.02781.x
Rahman MM, Shikdar IK (2025) Assessing growth performance and flesh quality of silver carp (Hypophthalmichthys molitrix) with natural carotenoids in feeds. Sustainable Aquatic Research 4:248–259. https://doi.org/10.5281/zenodo.18005874
Rosenau S, Wolgast T, Altmann B, Risius A (2023) Consumer preference for altered color of rainbow trout (Oncorhynchus mykiss) fillet induced by Spirulina (Arthrospira platensis). Aquaculture 572:739522. https://doi.org/10.1016/j.aquaculture.2023.739522
Safari O, Atash MMS (2015) The effects of dietary supplement of annatto (Bixa orellana) seed meal on blood carotenoid content and fillet color stability in rainbow trout (Oncorhynchus mykiss). Aquaculture 437:275–281. https://doi.org/10.1016/j.aquaculture.2014.12.012
Steine G, Alfnes F, Rora MB (2004) Consumer attitudes towards salmon color: a stated choice experiment with pictures. Marine Resource Economics 1–20.
Svensson PA, Wong BBM (2011) Carotenoid based signals in behavioural ecology: a review. Behaviour 148:131–189. https://doi.org/10.1163/000579510x548673
Tan K, Zhang H, Lim LS, Ma H, Li S, Zheng HT (2020) Roles of carotenoids in invertebrate immunology. Frontiers in Immunology 10:3041. https://doi.org/10.3389/fimmu.2019.03041
Tan K, Zhang H, Zheng H (2024) Carotenoid content and composition: A special focus on commercially important fish and shellfish. Critical Reviews in Food Science and Nutrition 64:544–561. https://doi.org/10.1080/10408398.2022.2106937
Vahdat S, Esmaeili Fereidouni A, Khalesi MK (2018) Long term effects of vermicompost manure leachate (powder) inclusions on growth and survival, biochemical composition, total carotenoids, and broodstock reproductive performance of Artemia franciscana (Kellogg, 1906). Aquaculture International 26:569–588. https://doi.org/10.1007/s10499-017-0232-0
Vikas PA, Chakraborty K, Sajeshkumar NK, Thomas PC, Sanil NK, Vijayan KK (2014) Quality of six Indian populations of Artemia franciscana for larval finfish culture. Journal of Applied Aquaculture 26:271–291. https://doi.org/10.1080/10454438.2014.942124
Wang W, Ma Z, Li W, Xue Y, Moss AS, Wu M (2024) Impact of β carotene enrichment on carotenoid composition and gene expression in Artemia metanauplii. Metabolites 14:676. https://doi.org/10.3390/metabo14120676
Xue Y, Jiang G, Shu H, Wang W, Huang X (2024) Effects of temperature and salinity on the growth, reproduction, and carotenoid accumulation in Artemia sinica and transcriptome analysis. Fishes 9:437. https://doi.org/10.3390/fishes9110437
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