title: Application of the SPE reversed phase HPLC/MS technique to determine vitamin B12 bio-active forms in beef.
abstract: Vitamin B12 is an animal origin nutrient of a substantial importance in human diet. Its concentration in foodstuffs is low and its chemical forms are diverse, which significantly hampers its precise determination. The determination method of choice is HPLC (high performance liquid chromatography) coupled with inductively-coupled-plasma mass spectrometry (ICP-MS). The main disadvantage of this method is high instrumentation cost and complexity of handling. The aim of this work was to develop a novel approach for determination of vitamin B12 bio-active forms in beef and beef liver. The proposed method comprises the following steps: (i) vitamin B12 is cleaved off from peptides using thermal denaturation in a weakly acidic environment; (ii) sample is cleaned-up using liquid-liquid extraction and reversed phase solid phase extraction; and finally (iii) vitamin B12 is determined using HPLC and single-quadrupole mass spectrometer with ESI source. Vitamin B12 concentrations in various beef meats were in the 2.84-3.95 μg 100g(-1) range. Average B12 concentration in beef liver was 153,60 μg 100g(-1) (n = 15). Major forms of B12 present in beef meat include adenosine cobalamin (AdoCbl) and in smaller quantities hydroxycobalamin (OHCbl). Major forms of vitamin B12 present in beef liver include OHCbl (48.2%), AdoCbl (33.8%), methylocobalamin (MeCbl, 16.3%), and cyanocobalamin (CNCbl, 1.7%). Thermal treatment noticeably decreases B12 the content in meat. Depending on conditions of treatment, B12 concentrations in the 1.04-2.20 μg 100g(-1) range were found in processed meats.
authors.affiliation: Department of Functional Food and Commodities Faculty of Human Nutrition and Consumer Sciences, Warsaw University of Life Sciences, 159 c Nowoursynowska, 02-0776 Warsaw, Poland. szterkarkadiusz@gmail.com
authors.forename: Arkadiusz
authors.initials: A
authors.lastname: Szterk
authors.forename: Marek
authors.initials: M
authors.lastname: Roszko
authors.forename: Krystian
authors.initials: K
authors.lastname: Małek
authors.forename: Małgorzata
authors.initials: M
authors.lastname: Czerwonka
authors.forename: Bożena
authors.initials: B
authors.lastname: Waszkiewicz-Robak
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008099:Liver / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008460:Meat / Q000032:analysis*; D052616:Solid Phase Extraction / Q000379:methods; D014805:Vitamin B 12 / Q000032:analysis*
chemical_list: D014805:Vitamin B 12