Analysis of Metalloporphyrins Using Cyclodextrin Stationary Phases With Photodiode Array UV Detection

Abstract

Abstract A method for the high-performance liquid chromaographic separation of metalloporphyrins using cyclodextrin column with photodiode array UV detection is described. The isocratic separation of hemin, protoporphyrin IX, Mn protoporphyrin IX, cobalt protoporphyrin IX, Sn protoporphyrin IX, Zn protoporphyrin IX was achieved in less than 10 min using β-cyclodextrin stationary phases and a mobile phase consisted of a mixture of 28 ml of 5 mM ammonia solution and 90 ml of acetone (v/v, 28:90). The apparent pH of the mobile phase was 7.9. The chromatographic behavior of tinprotoporphyrin IX is markedly different from the transition metals-protoporphyrin IX in cyclodextrin column. The effects of ammonia concentration and the composition of the mobile phase were studied to optimize the separation of the compounds. The concentration of ammonia solution in the mobile phase affects the solvent strength and selectivity of the mobile phase.