Abstract
High-performance thin-layer chromatography (HPTLC) is a separation technique commonly used to identify and quantify compounds in chemical mixtures. Preliminary experiments indicated that Lichrospher silica gel 60 F254s HPTLC sheets were the most suitable for analyzing vitamin B12 compounds. This study revealed the advantages of miniaturized Lichrospher HPTLC for analyzing authentic vitamin B12 compounds as short migration distances (5 cm) and short development times (<45 min) in combination with high separation efficiency and sensitivity (>25 pmol at 254 nm). The practicability of using miniaturized HPTLC was demonstrated by the separation and identification of vitamin B12 compounds purified from foods using an immunoaffinity column.
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References
Scheider Z, Stroiñski A (1987) Biosynthesis of vitamin B12. In: Schneider Z, Stroiñski A (eds) Comprehensive B12. Walter de Gruyter, Berlin, pp 93–110
Watanabe F (2007) Vitamin B12 sources and bioavailability. Exp Biol Med 232:1266–1274
Chen Z, Crippen K, Gulati S, Banerjee R (1994) Purification and kinetic mechanism of a mammalian methionine synthase from pig liver. J Biol Chem 269:27193–27197
Fenton WA, Hack AM, Willard HF, Gertler A, Rosenberg LE (1982) Purification and properties of methylmalonyl coenzyme A mutase from human liver. Arch Biochem 228:323–329
Watanabe F, Katsura H, Takenaka S, Fujita T, Abe K, Tamura Y, Nakatsuka T, Nakano Y (1999) Pseudovitamin B12 is the predominant cobamide of an algal health food, Spirulina tablets. J Agric Food Chem 47:4736–4741
Miyamoto E, Tanioka Y, Nakao T, Barla F, Inui H, Fujita T, Watanabe F, Nakano Y (2006) Purification and characterization of a corrinoid-compound in an edible cyanobacterium Aphanizomenon flos-aquae as a nutritional supplementary food. J Agric Food Chem 54:9604–9607
Allen RH, Stabler SP (2008) Identification and quantitation of cobalamin and cobalamin analogues in human feces. Am J Clin Nutr 87:1324–1335
Watanabe F, Miyamoto E (2003) Hydrophilic vitamins. In: Sherma J, Fried B (eds) Handbook of thin-layer chromatography, 3rd edn. Marcel Dekker, Inc. New York, pp 589–605 (revised and expanded)
Watanabe F, Yabuta Y (2011) Microbiological detection of vitamin B12 and other vitamins. In: Rychlik M (ed) Fortified foods with vitamins—analytical concepts to assure better and safer products. Wiley & Sons Inc, Hoboken, pp 165–171
Tanioka Y, Yabuta Y, Miyamoto E, Inui H, Watanabe F (2008) Analysis of vitamin B12 in food by silica gel 60 TLC and bioautography with vitamin B12-dependent Escherichia coli 215. J Liq Chrom Rel Technol 31:1977–1985
Tanioka Y, Miyamoto E, Yabuta Y, Onishi K, Fujita T, Yamaji R, Misono H, Shigeoka S, Nakano Y, Inui H, Watanabe F (2010) Methyladeninylcobamide functions as the cofactor of methionine synthase in a cyanobacterium, Spirulina plantensis NIES-39. FEBS Lett 584:3223–3226
Watanabe F, Takenaka S, Katsura H, Miyamoto E, Abe K, Tamura Y, Nakatsuka T, Nakano Y (2000) Characterization of a vitamin B12 compound in the edible purple laver, Porphyra yezoensis. Biosi Biotechnol Biochem 64:2712–2715
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Published in the topical collection Miniaturized and New Featured Planar Chromatography and Related Techniques with guest editor Pawel K. Zarzycki.
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Bito, T., Yabuta, Y. & Watanabe, F. Miniaturized HPTLC of Vitamin B12 Compounds in Foods. Chromatographia 76, 1333–1337 (2013). https://doi.org/10.1007/s10337-013-2386-x
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DOI: https://doi.org/10.1007/s10337-013-2386-x