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.
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.
Lebiedzińska
Anna
A
Department of Food Sciences, Medical University of Gdańsk, Al. Gen. J. Hallera 107, Gdańsk 80-416, Poland. aleb@amg.gda.pl
Marszałł
Marcin Leszek
ML
Kuta
Jadwiga
J
Szefer
Piotr
P
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*
D011730:Pyridoxal; D011733:Pyridoxamine; D025101:Vitamin B 6; D011736:Pyridoxine; D014805:Vitamin B 12; D013831:Thiamine