Hachey D L, Coburn S P, Brown L T, Erbelding W F, DeMark B, Klein P D
Anal Biochem. 1985 Nov 15;151(1):159-68. doi: 10.1016/0003-2697(85)90066-1.
Methods have been developed for the simultaneous quantitative analysis of vitamin B6 forms in biological samples by isotope dilution mass spectrometry using deuterated forms of pyridoxine, pyridoxal, pyridoxamine, and pyridoxic acid. The biological fluid or tissue sample was homogenized and then treated with a cocktail containing appropriate amounts of each deuterated vitamer, as well as the deuterated, phosphorylated vitamer forms. The individual vitamers were isolated from the homogenate by a complex high-performance liquid chromatographic procedure that provided separate fractions for each of the six vitamers found in biological samples. Aldehydic B6 vitamers were reduced to the alcohol form prior to acetylation and analysis by gas chromatography/mass spectrometry (GC/MS). The three resulting vitamers were analyzed by electron ionization GC/MS using a silicone capillary column. The methods have been applied to analysis of vitamin B6 in liver, milk, urine, and feces at levels as low as 0.02 nmol/ml.
已开发出通过同位素稀释质谱法同时定量分析生物样品中维生素B6各形式的方法，该方法使用氘代形式的吡哆醇、吡哆醛、吡哆胺和吡哆酸。将生物体液或组织样品匀浆，然后用含有适量各氘代维生素原以及氘代、磷酸化维生素原形式的混合液处理。通过复杂的高效液相色谱程序从匀浆中分离出各个维生素原，该程序为生物样品中发现的六种维生素原中的每一种提供单独的馏分。醛基B6维生素原在乙酰化和气相色谱/质谱（GC/MS）分析之前还原为醇形式。使用硅胶毛细管柱通过电子电离GC/MS分析得到的三种维生素原。这些方法已应用于分析肝脏、牛奶、尿液和粪便中低至0.02 nmol/ml水平的维生素B6。
