Purification, Identification, and Characterization of Methylcobalamin from Spirulina platensis.

The present study reports methylcobalamin in Spirulina platensis using high-performance liquid
chromatography (HPLC), thin-layer chromatography (TLC), microbiological assay, chemiluminescence
assay, liquid chromatography-mass spectrometry (LC-MS), and tandem mass spectrometry (MS/MS).
Extraction of vitamin B12 from S. platensis was carried out without using cyanide. Partial purification was
achieved using Amberlite XAD-2 followed by elution with 80% (v/v) methanol. Activated charcoal
facilitated removal of impurities in S. platensis extract and in further purification of vitamin B12. The
purified fraction was identified to contain methylcobalamin as analyzed by HPLC and TLC. Authenticity
of methylcobalamin was further confirmed by LC-MS and MS/MS. Quantitation of methylcobalamin in a
test sample of S. platensis biomass was performed using microbiological assay and chemiluminescence
assay and was found to be 38.5 ( 2 and 35.7 ( 2 μg/100 g of dry biomass, respectively.