Argoudelis C J
University of Illinois, Department of Food Science, Urbana 61801.
J Chromatogr. 1988 Feb 26;424(2):315-23. doi: 10.1016/s0378-4347(00)81108-1.
A simple, sensitive and fast isocratic high-performance liquid chromatographic method has been developed for the separation of all six biologically active forms (vitamers) of vitamin B6. The separation is accomplished using a strong cation-exchange column and a mobile phase of 0.1 M ammonium dihydrogenphosphate adjusted to pH 4.0. All six vitamers are separated within 20 min at a flowrate of 1 ml/min. The concentration of the vitamers is determined with a fluorescence detector (excitation 290 nm; emission 389 nm). The within-run precision of the method expressed as the coefficient of variation is below 5% at the 25 pmol level. Pyridoxal 5'-phosphate can be determined using either pre- or post-column derivatization with sodium bisulfite. Application of the method to cell-free yeast culture media is presented.
已开发出一种简单、灵敏且快速的等度高效液相色谱法，用于分离维生素B6的所有六种生物活性形式（维生素异构体）。使用强阳离子交换柱和pH值调至4.0的0.1 M磷酸二氢铵流动相进行分离。在流速为1 ml/min时，所有六种维生素异构体在20分钟内分离完毕。维生素异构体的浓度用荧光检测器（激发波长290 nm；发射波长389 nm）测定。该方法在25 pmol水平下的批内精密度以变异系数表示低于5%。可以使用亚硫酸氢钠进行柱前或柱后衍生化来测定5'-磷酸吡哆醛。本文介绍了该方法在无细胞酵母培养基中的应用。
