title: Ultra-high performance liquid chromatographic and mass spectrometric analysis of active vitamin B12 in cells of Propionibacterium and fermented cereal matrices.
abstract: A sensitive and selective method is needed to analyse in situ produced vitamin B12 in plant-based materials, potential new dietary sources of vitamin B12. A UHPLC/UV method was developed and validated for the determination of human active vitamin B12 in cell extracts of Propionibacterium freudenreichii subsp. shermanii and after immunoaffinity purification in extracts of cereal matrices fermented by P. freudenreichii. An Acquity HSS T3 C18 column resulted in a baseline separation, a calibration curve of excellent linearity and a low limit of detection (0.075 ng/5 μL injection). As confirmed by UHPLC-MS, the active vitamin B12 could be separated from pseudovitamin B12. The recovery of vitamin B12 from purified spiked cereal matrices was good (>90%; RSD<5%). A nutritionally relevant amount of active vitamin B12 was produced by P. freudenreichii in cereal malt matrices (up to 1.9 μg/100 g) in 24h at 28 °C.
authors: Department of Food and Environmental Sciences, P.O. Box 27, University of Helsinki, FI-00014 University of Helsinki, Finland. Electronic address: bhawani.chamlagain@helsinki.fi.
authors: Bhawani
authors: B
authors: Chamlagain
authors: Department of Food and Environmental Sciences, P.O. Box 27, University of Helsinki, FI-00014 University of Helsinki, Finland.
authors: Minnamari
authors: M
authors: Edelmann
authors: Department of Food and Environmental Sciences, P.O. Box 27, University of Helsinki, FI-00014 University of Helsinki, Finland.
authors: Susanna
authors: S
authors: Kariluoto
authors: Department of Food and Environmental Sciences, P.O. Box 27, University of Helsinki, FI-00014 University of Helsinki, Finland.
authors: Velimatti
authors: V
authors: Ollilainen
authors: Department of Food and Environmental Sciences, P.O. Box 27, University of Helsinki, FI-00014 University of Helsinki, Finland.
authors: Vieno
authors: V
authors: Piironen
mesh_terms: D019149:Bioreactors; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D015169:Colony Count, Microbial; D002523:Edible Grain / Q000737:chemistry*; D005285:Fermentation; D005511:Food Handling; D005516:Food Microbiology; D057230:Limit of Detection; D011424:Propionibacterium / Q000378:metabolism*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry; D014805:Vitamin B 12 / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis
chemical_list: C005602:pseudovitamin B12; D014805:Vitamin B 12
keywords: Fermented cereal matrix; Immunoaffinity purification; Propionibacterium freudenreichii; UHPLC; UPLC; Vitamin B12