Jacob K, Egeler E, Neumeier D, Knedel M
Institut für Klinische Chemie am Klinikum Grosshadern der Universität München, F.R.G.
J Chromatogr. 1989 May 12;468:329-38. doi: 10.1016/s0021-9673(00)96327-1.
Urinary porphyrins of porphyric patients were isolated as their methyl esters by using a simple, modified thin-layer chromatographic system. Existing methods for the isocratic ion-pair high-performance liquid chromatographic separation of uroporphyrin and coproporphyrin isomers were decisively improved by elevating the column temperatures, changing the types of columns used and modifying the eluent compositions. These techniques were applied to the determination of the isomeric distribution of uroporphyrins and coproporphyrins isolated from urines of patients in the acute or latent phase of acute intermittent porphyria. In these urines relatively high contents of the atypical uroporphyrins II (2-5%) and IV (13-19%) were found. The coproporphyrin fractions contained significantly smaller amounts of the atypical isomers II (1-2%) and IV (2-5%), the presence of which was demonstrated for the first time in such urines. Several mechanisms for the formation of the atypical coproporphyrin isomers are discussed. The isocratic ion-pair separation method served also to control the isomeric purity of uroporphyrin specimens of both natural and synthetic origin.
采用一种简单的改良薄层色谱系统，将卟啉症患者的尿卟啉分离为其甲酯形式。通过提高柱温、改变所用色谱柱类型以及调整洗脱液组成，对现有的尿卟啉和粪卟啉异构体等度离子对高效液相色谱分离方法进行了显著改进。这些技术被应用于测定从急性间歇性卟啉症急性期或潜伏期患者尿液中分离出的尿卟啉和粪卟啉的异构体分布。在这些尿液中，发现非典型尿卟啉II（2 - 5%）和IV（13 - 19%）的含量相对较高。粪卟啉组分中，非典型异构体II（1 - 2%）和IV（2 - 5%）的含量明显较少，首次在这类尿液中证实了它们的存在。讨论了非典型粪卟啉异构体形成的几种机制。等度离子对分离方法还用于控制天然和合成来源的尿卟啉样品的异构体纯度。
