Extraction and Determination of Astaxanthin Pigment From Haematococcus pluvialis Microalgae

Document Type : Original Article

Authors

Academic Center for Education, Culture, and Research (ACECR)-Mashhad Branch, Mashhad, Iran

Abstract

Astaxanthin and β-Carotene are well-known carotenoids globally, covering more than half of the market demand for carotenoids. Haematococcus pluvialis microalgae are one of the most important sources of natural astaxanthin, consisting of up to 4% of its dry weight. The most critical challenge for this microalgae is the breakdown of the wall and the extraction of the pigment. In this study, chemical methods, including acid, acetone, and ionic solution, and physical processes such as ultrasound waves and magnetic stirrer, were used to break down the cell wall and measure total astaxanthin in H. pluvialis, respectively. Due to the rapid oxidation of the pigment, in the next step, to extract and store astaxanthin from damaged cells, use olive oil. A spectrophotometer examined astaxanthin, monoester, and diester derivatives, and their amount was determined by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC). The results showed that using acid treatment, ultrasound waves, and extraction by acetone is the best method to measure the amount of astaxanthin in the algae. The HPLC results also showed that the amount of astaxanthin monoester (88.44%) was higher than the free forms (3.76%) and diester (7.82%) in the total content of extracted astaxanthin. In addition, the amount of total astaxanthin in the H. pluvialis was about 1.6% of the dry weight of the algae

Keywords


Basiony M, Ouyang L, Wang D, Yu J, Zhou L, Zhu M, Wang X, Feng J, Dai J, Shen Y. (2022). Optimization of microbial cell factories for astaxanthin production: Biosynthesis and regulations, engineering strategies and fermentation optimization strategies. Synthetic and Systems Biotechnology. 7: 689-704.
Borowitzka MA. (2013). High-value products from microalgae—their development and commercialization. Journal of Applied Phycology. 25: 743-756.
Cheng YS, Labavitch J, Vandergheynst J. (2015). Elevated CO2 concentration impacts the cell wall polysaccharide composition of green microalgae of the genus Chlorella. Letters in Applied Microbiology. 60: 1-7.
Choi SA, Oh YK, Lee J, Sim SJ, Hong ME, Park JY, Kim MS, Kim SW, Lee JS. (2019). High-efficiency cell disruption and astaxanthin recovery from Haematococcus pluvialis cyst cells using room-temperature imidazolium-based ionic liquid/water mixtures. Bioresource Technology. 274: 120-126.
Desai RK, Streefland M, Wijffels RH, EppinK MH. (2016). Novel astaxanthin extraction from Haematococcus pluvialis using cell permeabilizing ionic liquids. Green Chemistry. 18: 1261-1267.
Elumalai S, Santhose BI, Kanna GR. (2014). Extraction of carotenoid and thin layer chromatography (TLC), GC-MS, FT-IR, and HPLC analysis of pharmaceutically important pigment astaxanthin from a new strain of Haematococcus pluvialis. Weekly Science Research Journal. 2: 2321-7871.
Kang CD and Sim SJ. (2008). Direct extraction of astaxanthin from Haematococcus culture using vegetable oils. Biotechnology Letters. 30: 441-444.
Keykha akhar F, Fakhrfeshani M, Alipour H, Ameri M. (2021). Micro Algal pigments: An introduction to their biosynthesis, applications and genetic engineering. Journal of Plant Molecular Breeding.
Kim DY, Vijayan D, Praveenkumar R, Han JI, Lee K, Park JY, Chang WS, Lee JS, Oh YK. (2016). Cell-wall disruption and lipid/astaxanthin extraction from microalgae: Chlorella and Haematococcus. Bioresource Technology, 199, 300-310.
Koller M, Muhr A, Braunegg G. (2014). Microalgae as versatile cellular factories for valued products. Algal research. 6, 52-63.
Li Y, Miao F, Geng Y, Lu D, Zhang C, Zeng M. (2012). Accurate quantification of astaxanthin from Haematococcus crude extract spectrophotometrically. Chinese Journal of Oceanology and Limnology. 30: 627-637.
Liu ZW, Zeng XA, Cheng JH, Liu DB, Aadil RM. (2018). The efficiency and comparison of novel techniques for cell wall disruption in astaxanthin extraction from Haematococcus pluvialis. International Journal of Food Science and Technology. 53: 2212-2219.
Machmudah S, Shotipruk A, Goto M, Sasaki M. Hirose T. (2006). Extraction of astaxanthin from Haematococcus pluvialis using supercritical CO2 and ethanol as entrainer. Industrial and Engineering Chemistry Research. 45: 3652-3657.
Molino A, Rimauro J, Casella P, Cerbone A, Larocca V, Chianese S, Karatza D, Mehariya S, Ferraro A, Hristoforou E. (2018). Extraction of astaxanthin from the microalga Haematococcus pluvialis in red phase by using generally recognized as safe solvents and accelerated extraction. Journal of biotechnology. 283: 51-61.
Noroozi M, Omar H, Napis S, Hejazi MA, Tan SG. (2012). Comparative biodiversity and effect of different media on growth and astaxanthin content of nine geographical strains of Haematococcus pluvialis. African Journal of Biotechnology. 11: 15049-15059.
Orona-Navar A, Aguilar-Hernández I, Cerdán-Pasarán A, López-Luke T, Rodríguez-Delgado M, Cárdenas-Chávez D, Cepeda-Pérez E, Ornelas-Soto N. (2017). Astaxanthin from Haematococcus pluvialis as a natural photosensitizer for dye-sensitized solar cells. Algal Research. 26: 15-24.
Panis G and Carreon JR. (2016). Commercial astaxanthin production derived by green alga Haematococcus pluvialis: A microalgae process model and a techno-economic assessment all through the production line. Algal Research, 18, 175-190.
Panis, G. & Carreon, J. R. 2016. Commercial astaxanthin production derived by green alga Haematococcus pluvialis: A microalgae process model and a techno-economic assessment all through the production line. Algal Research. 18: 175-190.
Reyes FA, Mendiola JA, Ibanez E, Del Valle JM. (2014). Astaxanthin extraction from Haematococcus pluvialis using CO2-expanded ethanol. The Journal of Supercritical Fluids. 92: 75-83.
Saini RK and Keum YS. (2018). Carotenoid extraction methods: A review of recent developments. Food chemistry. 240: 90-103.
Shah M, Mahfuzur R, Liang Y, Cheng JJ, Daroch M. (2016). Astaxanthin-producing green microalga Haematococcus pluvialis: from a single cell to high-value commercial products. Frontiers in Plant Science. 7: 531.
Suseela M and Toppo K. (2006). Haematococcus pluvialis–A green alga, richest natural source of astaxanthin. Current Science. 90: 1602-1603.