Ubbink J B, Serfontein W J, de Villiers L S
J Chromatogr. 1985 Aug 9;342(2):277-84. doi: 10.1016/s0378-4347(00)84518-1.
The determination of pyridoxal-5-phosphate (PLP) and pyridoxal (PL) in plasma requires the availability of dark room facilities, due to the photosensitivity of these vitamin B6 vitamers. The fact that the semicarbazone forms of PL and PLP are more strongly fluorescent than the underivatized B6 vitamers has been exploited in plasma analyses, but it was not previously realised that these semicarbazone forms are also very stable even under conditions that lead to rapid decomposition of free PL and PLP. The stabilisation of PLP and PL obtained in this manner is sufficient and fully adequate to meet the practical requirements of clinical field studies. We report a high-performance liquid chromatographic method for plasma PLP and PL determinations based on precolumn semicarbazone formation and fluorescence detection. The method is sensitive enough for quantitative plasma PLP determinations even in B6-deficient patients.
由于这些维生素B6的光敏感性，血浆中磷酸吡哆醛（PLP）和吡哆醛（PL）的测定需要暗室设备。在血浆分析中，人们利用了PL和PLP的半卡巴腙形式比未衍生化的B6维生素具有更强荧光这一事实，但此前并未意识到这些半卡巴腙形式即使在导致游离PL和PLP快速分解的条件下也非常稳定。以这种方式获得的PLP和PL的稳定性足以满足临床现场研究的实际需求。我们报告了一种基于柱前半卡巴腙形成和荧光检测的高效液相色谱法用于血浆PLP和PL的测定。该方法灵敏度足够高，即使在维生素B6缺乏的患者中也能进行血浆PLP的定量测定。
