Abstract
A derivative ratio spectrophotometric method was used for the simultaneous determination of β-carotene and astaxanthin produced from Phaffia rhodozyma. Absorbencies of a series of the standard carotenoids in the range of 441 nm to 490 nm demonstrated that their absorptive spectra accorded with Beer’s law and that the additivity when the concentrations of β-carotene and astaxanthin and their mixture were within the range of 0 to 5 μg/ml, 0 to 6 μg/ml, and 0 to 6 μg/ml, respectively. When the wavelength interval (Δλ) at 2 nm was selected to calculate the first derivative ratio spectra values, the first derivative amplitudes at 461 nm and 466 nm were suitable for quantitatively determining β-carotene and astaxanthin, respectively. Effect of divisor on derivative ratio spectra could be neglected; any concentration used as divisor in range of 1.0 to 4.0 μg/ml is ideal for calculating the derivative ratio spectra values of the two carotenoids. Calibration graphs were established for β-carotene within 0–6.0 μg/ml and for astaxanthin within 0–5.0 μg/ml with their corresponding regressive equations in: y=−0.0082x–0.0002 and y=0.0146x–0.0006, respectively. R-square values in excess of 0.999 indicated the good linearity of the calibration graphs. Sample recovery rates were found satisfactory (>99%) with relative standard deviations (RSD) of less than 5%. This method was successfully applied to simultaneous determination of β-carotene and astaxanthin in the laboratory-prepared mixtures and the extract from the Phaffia rhodozyma culture
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References
Berzas-Nevado, J.J., Guiberteau, C.C., Salinas, F., 1992. Spectrophotometric resolution of ternary mixtures of salicylaldehyde, 3-hydroxy benzaldehyde and 4-hydroxy benzaldehyde by the derivative ratio spectrum zero crossing method.Talanta,39:547–553.
Britton, G., 1995. Structure and properties of carotenoids in relation to function.FASEB J., 9:1551–1558.
Calo, P., Velazquez, J.B., Sieiro, C., Blanco, P., Longo, E., Villa, T.G., 1995. Analysis of astaxanthin and other carotenoids from severalPhaffia rhodozyma Mutants.J. Agric. Food Chem.,43(5):1396–1399.
Cruz, J.M., Parjao, J.C., 1998. Improved astaxanthin production byXanthophyllomyces dendrohous growing on enzymatic wood hydrolysated containing glucose and cellobiose.Food Chemistry,63(4):479–484.
Garcia, J.M., Hermandez, O., Jimenez, A.I., Jimenez, F., Arias, J.J., 1995. A contribution to the derivative ratio spectrum method.Analytica Chimica Acta,317:83–93.
Johnson, E.A., 1991. Astaxathin from microbiol sources.Critical Reveiws in Biotechnology,11(4):297–326.
Johnson, E.A., 2003.Phaffia rhodozyma: Colorful odyssey.Int Microbiol.,6:169–174.
Liang, Y.Z., 1996. White, Gray and Black Multicomponents Systems and Their Chemometric Algorithms. China Hunan Publishing House of Science and Technology, Hunan, China (in Chinese).
Ong, A.S.H., Tee, E.S., 1992. Natural sources of carotenoids from plants and oils.Meth. Enzymol. 213:142–167.
Parajo, J.C., Santos, V., Vazquez, M., 1997. Co-production of carotenoids and xylitol byXanthophyllomyces dendrorhous (Phaffia rhodozyma).Biotechnology Letters,19(2):139–141.
Parajo, J.C., Santos, V., Vazquez, M., 1998. Production of carotenoids byPhaffia rhodozyma growing on media made from Hemicellulosic Hydrolysates of Eucalyptus globules wood.Biotechnology and Bioengineering,59(4):501–506.
Paul, D.F., Yutaka, M., Misawa, N., 1997. In vitro characterization of astaxanthin biosynthetic enzymes.Journal of Bioengineering and Chemistry,272(10):6128–6135.
Ramirez, J., Gutierrez, H., Gschaedler, A., 2001. Optimization of astaxanthin production byPhaffia rhodozymathrough factorial design and response surface methodology.Journal of Biotechnology,88:259–268.
Salinas, F., Berzas-Nevado, J.J., Espinosa, M.A., 1990. A new spectrophotometric method for quantitative multicomponent analysis resolution of mixtures of salicylic and salicyluric acids.Talanta,37:347–351.
Sedmak, J.J., Weerasinghe, D.K., Jolly, S.O., 1990. Extraction and quantification of astaxanthin fromPhaffia rhodozyma.Biotechnol. Tech.,4:107–112.
Tangeras, A., Slinde, E., 1994. Coloring of Salmonids in Aquaculture: The YeastPhaffia Rhodozyma as a Source of Astaxanthin.In: Martin, A.M. (Ed.), Fisheries Processing: Biotechnological Application. Chapman & Hall, London, p. 391–431.
Verdoes, J.C., Misawa, N., Van Ooyen, A.J.J., 1999. Clonig and characterization of the astaxanthin biosynthetic gene encoding phytoene desaturase ofXanthphyllomyces dendrorhous.Biotechnol. Bioeng.,63:750–755.
Wang, Q.Y., Li, Q.S., 1997. Nature Carotenoids: Advancements of Studies, Production and Applications. China Press of Pharmaceutical Science and Technology, Beijing, China (in Chinese).
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Project (No. 20276064) supported by the National Natural Science Foundation of China
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Ni, H., He, Gq., Ruan, H. et al. Application of derivative ratio spectrophotometry for determination of β-carotene and astaxanthin from Phaffia rhodozyma extract. J Zheijang Univ Sci B 6, 514–522 (2005). https://doi.org/10.1631/jzus.2005.B0514
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DOI: https://doi.org/10.1631/jzus.2005.B0514