Abstract
Studies were made on separation and detection of vitamin B_<12> by thin layer chromatography. It has been found that in the case using Aluminium oxide G as the adsorbent and glacial acetic acid・water・methanol・chloroform・n-butanol (1 : 4.5 : 5 : 10 : 25) as the solvent system for development, cyanocobalamin was completely separated, but not hydroxocobalamin. On the other hand using Silica gel G (Keiselgel G) as the adsorbent and gracial acetic acid・water・methanol・chloroform・n-butanol (9 : 11 : 5 : 10 : 25) as the solvent system, vitamin derivatives could be successfully separated, i.e. hydroxocobalamin as Rf 0.05,cyanocobalamin as Rf 0.23,unknown substance as Rf 0.33 and thymdine as Rf 0.7. Detection of spots was made by bioautography on the plate using vitamin B_<12> assay agar medium and Lactobacillus leichmannii ATCC 7830 as a test organism. The detectable minimal amount, using Silica gel G and Aluminium oxide G was found to be 0.005 and 0.025 mμg cyanocobalamin respectively, a far higher sensitivity than chemical procedure (200mμg).
