title: Purification, identification, and characterization of methylcobalamin from Spirulina platensis.
abstract: 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.
authors.affiliation: Plant Cell Biotechnology Department, Central Food Technological Research Institute, Mysore, India.
authors.forename: Anantharajappa
authors.initials: A
authors.lastname: Kumudha
authors.forename: Sagaya Selva
authors.initials: SS
authors.lastname: Kumar
authors.forename: Munna Singh
authors.initials: MS
authors.lastname: Thakur
authors.forename: Gokare Aswathanarayana
authors.initials: GA
authors.lastname: Ravishankar
authors.forename: Ravi
authors.initials: R
authors.lastname: Sarada
mesh_terms: D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism; D048668:Lactobacillus delbrueckii / Q000254:growth & development / Q000378:metabolism; D053545:Spirulina / Q000737:chemistry*; D014805:Vitamin B 12 / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D014803:Vitamin B Complex / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D014803:Vitamin B Complex; C019476:mecobalamin; D014805:Vitamin B 12