title: Technical note: development and validation of a method using ultra performance liquid chromatography coupled with tandem mass spectrometry for determination of vitamin B12 concentrations in milk and dairy products.
abstract: A method using ultra performance liquid chromatography coupled with tandem mass spectrometry was developed to measure cobalamins in naturally enriched raw milk and to evaluate their fate during thermal treatments and along the process of cheese making. After addition of methotrexate as internal standard, samples were submitted to heat treatment in the presence of cyanide, which converts all the less-stable cobalamins into cyanocobalamin; then, purification was performed by a solid-phase extraction step. Reverse-phase ultra performance liquid chromatography separation coupled with tandem mass spectrometry provided a fast and reliable determination. Mass spectrometric analysis was carried out in multiple reaction monitoring mode. The monitored transitions were m/z 678.36 → 147.10 and 678.36 → 359.30 for vitamin B12 and m/z 455.22 → 175.13 and 455.22 → 308.22 for methotrexate (internal standard). The limit of quantification was 2 ng/g. The method showed good linearity from 2 to 20 ng/g (R(2) ≥ 0.98) and intra- and interday precisions were always less than 19%.
authors.affiliation: Laboratory of Analytical Bio-Agroalimentary Chemistry (CABA-Lab), Department of Veterinary Medical Sciences, Alma Mater Studiorum - University of Bologna, Via Tolara di Sopra 50, 40064 Ozzano Emilia (BO), Italy.
authors.forename: E
authors.initials: E
authors.lastname: Zironi
authors.forename: T
authors.initials: T
authors.lastname: Gazzotti
authors.forename: A
authors.initials: A
authors.lastname: Barbarossa
authors.forename: C
authors.initials: C
authors.lastname: Devicienti
authors.forename: M
authors.initials: M
authors.lastname: Scardilli
authors.forename: G
authors.initials: G
authors.lastname: Pagliuca
mesh_terms: D000818:Animals; D002611:Cheese / Q000032:analysis; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D003611:Dairy Products / Q000032:analysis*; D005524:Food Technology; D006358:Hot Temperature; D008892:Milk / Q000737:chemistry*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014805:Vitamin B 12 / Q000032:analysis*
chemical_list: D014805:Vitamin B 12