Lim C K, Li F, Peters T J
Division of Clinical Cell Biology, MRC Clinical Research Centre, Harrow, Middx, U.K.
Biochem J. 1987 Oct 1;247(1):229-32. doi: 10.1042/bj2470229.
A reversed-phase h.p.l.c. system is described for the separation of the four type-III heptacarboxylic porphyrinogen isomers. The effects of buffer concentration, pH and type and proportion of organic modifier in the mobile phase on retention and resolution of isomers were studied. Optimum separation on an ODS-Hypersil column was by elution with a ternary mobile phase of acetonitrile, methanol and 1 M-ammonium acetate, pH 5.16 (7:3:90, by vol.). Isomer identification was based on a comparison of their retention times with those of authentic standards, and was further confirmed by h.p.l.c. analysis of the characteristic mixture of three pentacarboxylic porphyrins formed after partial decarboxylation of individual isomers in 0.3 M-HCl at 160 degrees C.
描述了一种反相高效液相色谱系统，用于分离四种III型七羧酸卟啉原异构体。研究了流动相中缓冲液浓度、pH值、有机改性剂的类型和比例对异构体保留和分离度的影响。在ODS-Hypersil柱上的最佳分离是通过用乙腈、甲醇和1M醋酸铵（pH 5.16，体积比7:3:90）的三元流动相洗脱来实现的。异构体的鉴定基于其保留时间与标准品的比较，并通过对各异构体在160℃下于0.3M盐酸中部分脱羧后形成的三种五羧酸卟啉特征混合物进行高效液相色谱分析进一步确认。
