Influence of Synthetic Antioxidants on Extraction of All-trans-Lutein from Spinach under Air and Nitrogen Atmosphere


Extraction is most commonly used sample preparation technique in quantitative determination of all-trans-lutein in spinach. In our study RP-HPLC was applied to identify the constituents of spinach extract and to quantify all-trans-lutein content with the mobile phase consisted of acetone and water. Due to sensitivity of all-trans-lutein, we compared effects of different extraction conditions; air and nitrogen atmosphere and ethanol, ethanol with added 0.1% TBHQ and ethanol with added 0.1% BHT as solvents. Results for the all-trans-lutein content in the spinach samples and spinach samples with standard addition obtained in air and in nitrogen confirmed the induced degradation of all-trans-lutein in oxygen from the air. Ethanol containing synthetic antioxidant TBHQ under nitrogen atmosphere gave the highest yield of extraction and recovery of all-trans-lutein from spinach.

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K. Cernelic, M. Prosek, A. Golc-Wondra, Z. Rodic, B. Simonovska and M. Puklavec, "Influence of Synthetic Antioxidants on Extraction of All-trans-Lutein from Spinach under Air and Nitrogen Atmosphere," Food and Nutrition Sciences, Vol. 4 No. 2, 2013, pp. 195-200. doi: 10.4236/fns.2013.42027.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] G. Sandmann, “Carotenoid Biosynthesis and Biotechnological Application,” Archives of Biochemistry and Biophysics, Vol. 385, No. 1, 2001, pp. 4-12. doi:10.1006/abbi.2000.2170
[2] D. M. Deming and J. W. Erdman Jr., “Mammalian Carotenoid Absorption and Metabolism,” Pure and Applied Chemistry, Vol. 71, No. 12, 1999, pp. 2213-2223. doi:10.1351/pac199971122213
[3] F. Khachik, J. S. Bertram, M.-T. Huang, J. W. Fahey and P. Talalay, “Dietary Carotenoids and Their Metabolites as Potentially Useful Chemoprotective Agents against Cancer,” In: L. Packer, M. Hiramatsu and T. Yoshikawa, Eds., Antioksidant Food Supplements in Human Health, Academic Press, San Diego, 1999, pp. 203-229. doi:10.1016/B978-012543590-1/50015-9
[4] F. Khachik, G. R. Beecher and J. C. Smith Jr., “Lutein, Lycopene, and Their Oxidative Metabolites in Chemoprevention of Cancer,” Journal of Cellular Biochemistry, Vol. 59, No. S22, 1995; pp. 236-246. doi:10.1002/jcb.240590830
[5] D. Ren and S. Zhang, “Separation and Identification of the Yellow Carotenoids in Potamogeton crispus L.,” Food Chemistry, Vol. 106, No. 1, 2008, pp. 410-414. doi:10.1016/j.foodchem.2007.05.074
[6] A. Alves-Rodrigues and A. Shao, “The Science behind Lutein,” Toxicology Letters, Vol. 150, No. 1, 2004, pp. 57-83. doi:10.1016/j.toxlet.2003.10.031
[7] W. Stahl and H. Sies, “Bioactivity and Protective Effects of Natural Carotenoids,” Biochimica et Biophysica Acta, Vol. 1740, No. 2, 2005, pp. 101-107. doi:10.1016/j.bbadis.2004.12.006
[8] R. L. Roberts, J. Green and B. Lewis, “Lutein and Zeaxanthin in Eye and Skin Health,” Clinics in Dermatology, Vol. 27, No. 2, 2009, pp. 195-201. doi:10.1016/j.clindermatol.2008.01.011
[9] A. Perry, H. Rasmussen and E. J. Johnson, “Xanthophyll (Lutein, Zeaxanthin) Content in Fruits, Vegetables and Corn and Egg Products,” Journal of Food Composition and Analysis, Vol. 22, No. 1, 2009, pp. 9-15. doi:10.1016/j.jfca.2008.07.006
[10] O. F. O’Connell, L. Ryan and N. M. O’Brien, “Xanthophyll Carotenoids Are More Bioaccessible from Fruits than Dark Green Vegetables,” Nutrition Research, Vol. 27, No. 5, 2007, pp. 258-264. doi:10.1016/j.nutres.2007.04.002
[11] R. Aman, J. Biehl, R. Carle, J. Conrad, U. Beifuss and A. Schieber, “Application of HPLC Coupled with DAD, APcI-MS and NMR to the Analysis of Lutein and Zeaxanthin Stereoisomers in Thermally Processed Vegetables,” Food Chemistry, Vol. 92, No. 4, 2005, pp. 753-763. doi:10.1016/j.foodchem.2004.10.031
[12] B. Stephen Inbaraj, J. T. Chien and B. H. Chen, “Improved High Performance Liquid Chromatographic Method for Determination of Carotenoids in the Microalga Chlorella pyrenoidosa,” Journal of Chromatography A, Vol. 1102, No. 1-2, 2006, pp. 193-199. doi:10.1016/j.chroma.2005.10.055
[13] B. Gandul-Rojas, M. R. Cepero and M. I. Mínguez-Mosquera, “Chlorophyll and Carotenoid Patterns in Olive Fruits, Olea europaea Cv. Arbequina,” Journal of Agricultural and Food Chemistry, Vol. 47, No. 6, 1999, pp. 2207-2212. doi:10.1021/jf981158u
[14] J. Oliver and A. Palou, “Chromatographic Determination of Carotenoids in Foods,” Journal of Chromatography A, Vol. 881, No. 1-2, 2000, pp. 543-555. doi:10.1016/S0021-9673(00)00329-0
[15] S. Orset, G. C. Leach, R. Morais and A. J. Young, “SprayDrying of the Microalga Dunaliella salina: Effects on Beta-Carotene Content and Isomer Composition,” Journal of Agricultural and Food Chemistry, Vol. 47, No. 11, 1999, pp. 4782-4790. doi:10.1021/jf990571e
[16] A. M. Nuutila, K. Kammiovirta and K. M. Oksman-Caldentey, “Comparison of Methods for the Hydrolysis of Flavonoids and Phenolic Acids from Onion and Spinach for HPLC Analysis,” Food Chemistry, Vol. 76, No. 4, 2002, pp. 519-525. doi:10.1016/S0308-8146(01)00305-3
[17] P.-P. Hao, J.-R. Ni, W.-L. Sun and W. Huang, “Determination of Tertiary Butylhydroquinone in Edible Vegetable Oil by Liquid Chromatography/Ion Trap Mass Spectrometry,” Food Chemistry, Vol. 105, No. 4, 2007, pp. 1732-1737. doi:10.1016/j.foodchem.2007.04.058
[18] K. Cernelic, B. Simonovska and I. Vovk, “The Influence of Adding Antioxidants TBHQ and BHT to Ethanol at Extraction of Lutein from Spinach,” Abstract Book and Final Programme of 10th International Nutrition & Diagnostics Conference, Prague, 4-7 September 2010, p. 101.
[19] N. E. Craft and J. H. Soares Jr., “Relative Solubility, Stability, and Absorptivity of Lutein and Beta-Carotene in Organic Solvents,” Journal of Agricultural and Food Chemistry, Vol. 40, No. 3, 1992, pp. 431-434. doi:10.1021/jf00015a013
[20] M. Dachtler, T. Glaser, K. Kohler and K. Albert, “Combined HPLC-MS and HPLC-NMR On-Line Coupling for the Separation and Determination of Lutein and Zeaxanthin Stereoisomers in Spinach and in Retina,” Analytical Chemistry, Vol. 73, No. 3, 2001, pp. 667-674. doi:10.1021/ac000635g
[21] A. Bunea, M. Andjelkovic, C. Socaciu, O. Bobis, M. Neacsu, R. Verhe and J. Van Camp, “Total and Individual Carotenoids and Phenolic Acids Content in Fresh, Refrigerated and Processed Spinach (Spinacia oleracea L.), Food Chemistry, Vol. 108, No. 2, 2008, pp. 649-666. doi:10.1016/j.foodchem.2007.11.056
[22] U. Kidmose, P. Knuthsen, M. Edelenbos, U. Justesen and E. Hegelund, “Carotenoids and Flavonoids in Organically Grown Spinach (Spinacia oleracea L) Genotypes after Deep Frozen Storage,” Journal of the Science of Food and Agriculture, Vol. 81, No. 9, 2001, pp. 918-923. doi:10.1002/jsfa.902
[23] R. Tsao and R. Yang, “Lutein in Selected Canadian Crops and Agri-Food Processing by-Products,” Journal of Chromatography A, Vol. 1112, No. 1-2, 2006, pp. 202-208. doi:10.1016/j.chroma.2005.09.088
[24] D. A. Kopsell, M. G. Lefsrud, D. E. Kopsell, A. J. Wenzel, C. Gerweck and J. Curran-Celentano, “Spinach Cultigen Variation for Tissue Carotenoid Concentrations Influences Human Serum Carotenoid Levels and Macular Pigment Optical Density Following a 12-Week Dietary Intervention,” Journal of Agriculture and Food Chemistry, Vol. 54, No. 21, 2006, pp. 7998-8005. doi:10.1021/jf0614802
[25] F. B?hm, R. Edge and T. G. Truscott, “Interactions of Dietary Carotenoids with Singlet Oxygen (1O2) and Free Radicals: Potential Effects for Human Health,” Acta Viochimica Polonica, Vol. 59, No. 1, 2012, pp. 27-33.
[26] Y. Liu, C. O. Perera and V. Suresh, “Comparison of Three Chosen Vegetables with Others from South East Asia for Their Lutein and Zeaxanthin Content,” Food Chemistry, Vol. 101, No. 4, 2007, pp. 1533-1539. doi:10.1016/j.foodchem.2006.04.005

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