Retinol is an unsaturated fat-soluble isoprenoid alcohol. It is essential for the normal functioning of cells, human growth, and development, and it is found in animal sources in the form of palmitate ester. Chicken liver is an excellent source of retinol and is widely used in the diet of people in developing countries to combat Vitamin A deficiency. Several methods for the quantification of retinol in chicken liver have been reported in the literature. Most of them use more than one solvent for the determination and quantification of this micronutrient. A new method was developed based on a reverse phase system and using isocratic elution in a C18 column (5 mm, 250 × 4.6 mm) coupled to a SPD-20A UV-VIS detector at a wavelength of 325 nm with a single mobile phase, methanol. The method is simple and inexpensive, allowing the rapid extraction, determination, and quantification of retinol in chicken liver samples, with an average retention time of 5.2 min at 23°C, yielding good linearity results (R2 = 0. 9999), standard stock and freeze–thaw process stabilities of 93.2% and 97.5%, respectively, precision with coefficients of variation below 15%, and recovery coefficients ranging between 93% and 101.2%.
Cite this paper
Rüegg, R. A. B. and Dimenstein, R. (2020). Validation of an Analytical Method Based on High-Performance Liquid Chromatography for the Determination of Retinol in Chicken Liver. Open Access Library Journal, 7, e6181. doi: http://dx.doi.org/10.4236/oalib.1106181.
Brasil; Ministério Da Saúde (2009) Pesquisa Nacional de Demografia e Saúde da Criança e da Mulher. PNDS 2006: Dimensões do Processo Reprodutivo e da Saúde da Criança. Centro Brasileiro de Análise e Planejamento. Série G. Estatística e Informação em Saúde.
http://bvsms.saude.gov.br/bvs/publicacoes/pnds_crianca_mulher.pdf
Gallagher, M.L. (2010) Os Nutrientes e Seu Metabolismo. In: Gallagher, M., Ed., Krause: Alimentos, nutrição e dietoterapia, Elsevier, Rio de Janeiro, 68.
Institute of Medicine (IOM) (2001) Dietary Reference Intakes for Vitamin A, Vitamin K, Arsenic, Boron, Chromium, Copper, Iodine, Iron, Manganese, Molybdenum, Nickel, Silicon, Vanadium, and Zinc: A Report of the Panel on Micronutrients. Nutrition Reviews, 55, 319-326.
Howells, L.C. and Livesey, C.T. (1998) A Survey of Vitamin A Concentrations in the Liver of Food-Producing Animals. Food Additives and Contaminants, 15, 10-18.
https://doi.org/10.1080/02652039809374595
Jaskiewicz, T., Sagan, A. and Puzio, I. (2014) Effect of the Camelina sativa Oil on the Performance, Essential Fatty Acid Level in Tissues and Fat-Soluble Vitamins Content in the Livers of Broiler Chickens. Livestock Science, 165, 74-79.
https://doi.org/10.1016/j.livsci.2014.04.003
Kang, K.R., Cherian, G. and Sim, J.S. (2006) Tocopherols, Retinol and Carotenes in Chicken Egg and Tissues as Influenced by Dietary Palm Oil. Journal of Food Science, 63, 592-596. https://doi.org/10.1111/j.1365-2621.1998.tb15792.x
Majchrzak, D., Fabian, E. and Elmadfa, I. (2006) Vitamin A Content (Retinol and Retinyl Esters) in Livers of Different Animals. Food Chemistry, 98, 704-710.
https://doi.org/10.1016/j.foodchem.2005.06.035
Schindler, R., Scholz, M. and Feldheim, W. (1987) Quantitative Determination of Vitamin A in Liver and Liverwurst Using High Pressure Liquid Chromatography (HPLC). Zeitschrift für Lebensmittel-Untersuchung und-Forschung, 185, 208-212.
https://doi.org/10.1007/BF01042048
Surai, P.F. and Kuklenko, T.V. (2000) Effects of Vitamin A on the Antioxidant Systems of the Growing Chicken. Asian-Australasian Journal of Animal Sciences, 13, 1290-1295. https://doi.org/10.5713/ajas.2000.1290
Furr, H.C., Cooper, D.A. and Olson, J.A. (1986) Separation of Retinyl Esters by Non-Aqueous Reversed-Phase High-Performance Liquid Chromatography. Journal of Chromatography, 378, 45-53. https://doi.org/10.1016/S0378-4347(00)80698-2
Kane, M.A., Folias, A.E. and Napoli, J.L. (2008) HPLC/UV Quantitation of Retinal, Retinol, and Retinyl Esters in Serum and Tissues. Analytical Biochemistry, 378, 71-79. https://doi.org/10.1016/j.ab.2008.03.038
Karadas, F., et al. (2005) Effects of Maternal Dietary Supplementation with Three Sources of Carotenoids on the Retinyl Esters of Egg Yolk and Developing Quail Liver. Comparative Biochemistry and Physiology—A Molecular and Integrative Physiology, 140, 430-435. https://doi.org/10.1016/j.cbpb.2005.02.005
Kim, Y.-K. and Quadro, L. (2010) Reverse-Phase High-Performance Liquid Chromatography (HPLC) Analysis of Retinol and Retinyl Esters in Mouse Serum and Tissues. Methods in Molecular Biology, 652, 263-275.
https://doi.org/10.1007/978-1-60327-325-1_15
Satomura, Y., Kimura, M. and Itokawa, Y. (1992) Short Communication Simultaneous Determination of Allantoin and Oxypurines in Biological Fluids by High- Performance Liquid Chromatography. Journal of Chromatograpy, 625, 372-376.
https://doi.org/10.1016/0021-9673(92)85224-H
Shintaku, T., et al. (1998) Detection of Retinyl Palmitate and Retinol in the Liver of Mice Injected with Excessive Amounts of Retinyl Acetate. The Journal of Veterinary Medical Science/The Japanese Society of Veterinary Science, 60, 471-477.
https://doi.org/10.1292/jvms.60.471
Tatum, V. and Chow, C.K. (2005) Rapid Measurement of Retinol, Retinal, 13-Cis-Retinoic Acid and All-Trans-Retinoic Acid by High Performance Liquid Chromatography. Journal of Food and Drug Analysis, 13, 256-259.
Milne, D.B. and Botnen, J. (1986) Retinol, Alpha-Tocopherol, Lycopene, and Alpha- and Beta-Carotene Simultaneously Determined in Plasma by Isocratic Liquid Chromatography. Clinical Chemistry, 32, 874-876.
https://doi.org/10.1093/clinchem/32.5.874
Solomons, N.W. (2012) Vitamin A. In: Present Knowledge in Nutrition, 10th Edition, International Life Sciences Institute (ILSI), Washington DC, 149-185.
https://doi.org/10.1002/9781119946045.ch11
Hosotani, K. and Kitagawa, M. (2003) Improved Simultaneous Determination Method of Beta-Carotene and Retinol with Saponification in Human Serum and Rat Liver. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences, 791, 305-313. https://doi.org/10.1016/S1570-0232(03)00233-2
FDA Food and Drug Administration (2013) Guidance for Industry: Bioanalytical Method Validation. U.S. Department of Health and Human Services, Washington DC, 22.
Brito, N.M., et al. (2003) Validação De Métodos Analíticos: Estratégia E Discussão. Revista de Ecotoxicologia e Meio Ambiente, 13, 129-146.
https://doi.org/10.5380/pes.v13i0.3173
Ribani, M., et al. (2004) Validação em métodos cromatográficos e eletroforéticos. Quimica Nova, 27, 771-780. https://doi.org/10.1590/S0100-40422004000500017
Chen, S. and Kord, A. (2009) Theoretical and Experimental Comparison of Mobile Phase Consumption between Ultra-High-Performance Liquid Chromatography and High Performance Liquid Chromatography. Journal of Chromatography A, 1216, 6204-6209. https://doi.org/10.1016/j.chroma.2009.06.084
Rüegg, R.A.B. and Dimenstein, R. (2018) Effects of Cooking and Defrost Methods on Retinol Concentration of Chicken Liver. Journal of Food and Nutrition Research, 6, 146-151.
Yokota, S. and Oshio, S. (2018) A Simple and Robust Quantitative Analysis of Retinol and Retinyl Palmitate Using a Liquid Chromatographic Isocratic Method. Journal of Food and Drug Analysis, 26, 504-511.
https://doi.org/10.1016/j.jfda.2017.07.002
De Aquino, R.C.P., et al. (2006) Analysis of Retinol Concentrations in Bovine Liver and Its Habitual Consumption by Pregnant Women. Annals of Nutrition and Metabolism, 50, 325-329. https://doi.org/10.1159/000094295