反相液相色谱法(RPLC)测定食品中维生素B_6(呲哆胺、呲哆醛、吡哆醇)

APPLICATION OF RPLC TO THE SEPARATION AND DETERMINATION OF VITAMIN B_6-PYRIDOXAMINE, PYRIDOXAL, PYRIDOXINE

应用HPLC反相色谱法对维生素B_6(VB_6)的三种不同同系物(吡哆醇、吡哆胺、吡哆醛)进行分离测定是一种新的VB_6测定法,经充分粉碎的样品,用0.1M硫酸溶液,在120℃下水解30分钟,冷却后经离心、过滤,用ODS-C_(18)反相柱、pH2.0～2.5的硫酸溶液作流动相,在1.0ml/min的流速下,能将吡哆胺、呲哆醛、吡哆醇完全分离,并在激发波长293nm,发射波长395nm时,应用外标法定量,其回收率在99～103％,变异系数在5.0％～5.3％。实验结果表明,HPLC反相色谱法测定VB_6(吡哆胺、吡哆醛、吡哆醇)是一种快速,简便、灵敏度高,重现性好的方法。

A method for the determination of B6-pyridoxamine (PAM), pyridoxal (PAL) and pyridoxine (POL) by RPLC was proposed. The procedure included the addition of 0.1M H2SO4 to the sample, hydrolysis for 30 min at 120℃, centrifuging, filtration and direct analysis by ODS Cl8 column. PAM, PAL and POL were completely separated, when the flow rate of the mobile phase (pH2.0-2.1) was 1.0-1.5ml/min. Quantitation by a fluorescent detector was performed with Exc: 293 nm and Em: 395 nm. The recovery ranged from 99 to 103% with CV 5.0-5.3%. The method was simple, rapid, sensitive and reproducible.