title: Reversed-phase high-performance liquid chromatography method with coulometric electrochemical and ultraviolet detection for the quantification of vitamins B(1) (thiamine), B(6) (pyridoxamine, pyridoxal and pyridoxine) and B(12) in animal and plant foods.
abstract: A rapid and sensitive high-performance liquid chromatography (HPLC) method with coulometric electrochemical and ultraviolet detections for analysis of vitamin B (B(1), B(6) and B(12)) in animal and plant foods has been developed. A combination of acid digestion and enzymatic extraction to release protein bound and phosphorylated vitamins followed by HPLC analysis was applied. The analyses were carried out on a LC18 column 5 microm (25 cm x 4.6 mm), using the mobile phase consisting of methanol-phosphate buffer (10:90) and 0.018 M trimethylamine, pH 3.55, following at 1.0 ml min(-1). The method offers excellent linearity with regression coefficient r>0.998, good repeatability and reproducibility and relatively short analysis time (17 min). The relative standard deviation (RSD) of the intermediate precision was satisfactory for all the vitamins studied and amounted to <7.3%. The repeatability of the method was <4.7%. The limits of quantification (LOQ) for pyridoxamine, pyridoxal, pyridoxine, vitamins B(1) and B(12) in seafood products were as follows: 2.1, 2.01, 0.99, 62.0 and 0.11 ng ml(-1), respectively. The mean recovery values from a food products spiked with all five vitamins ranged from 92.3 to 101.3%, with a relative standard deviation less than 3.4%. The proposed separation and detection procedures were successfully applied for the simultaneous determination of vitamins B(1), B(6) and B(12) in fruit juices and vitamins B(6) and B(12) in seafood.
authors.affiliation: Department of Food Sciences, Medical University of Gdańsk, Al. Gen. J. Hallera 107, Gdańsk 80-416, Poland. aleb@amg.gda.pl
authors.forename: Anna
authors.initials: A
authors.lastname: Lebiedzińska
authors.forename: Marcin Leszek
authors.initials: ML
authors.lastname: Marszałł
authors.forename: Jadwiga
authors.initials: J
authors.lastname: Kuta
authors.forename: Piotr
authors.initials: P
authors.lastname: Szefer
mesh_terms: D000818:Animals; D001628:Beverages / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005638:Fruit / Q000737:chemistry; D011730:Pyridoxal / Q000032:analysis; D011733:Pyridoxamine / Q000032:analysis; D011736:Pyridoxine / Q000032:analysis; D015203:Reproducibility of Results; D017747:Seafood / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013831:Thiamine / Q000032:analysis*; D014805:Vitamin B 12 / Q000032:analysis*; D025101:Vitamin B 6 / Q000032:analysis*
chemical_list: D011730:Pyridoxal; D011733:Pyridoxamine; D025101:Vitamin B 6; D011736:Pyridoxine; D014805:Vitamin B 12; D013831:Thiamine