Methodology for porphyrin isolation by high-performance countercurrent chromatography.

Countercurrent chromatography is a versatile technique for the isolation of a wide variety of plant substances. However, little attention has been devoted to the application of this technique for the isolation of porphyrins. This class of compounds are of great importance in the medical area and in photocatalysis due to their heterocyclic structure, composed of four modified pyrrol subunits interconnected on their a carbon atoms by methinic bridges. The methanol extract of Gallesia integrifolia was partitioned using different solvents; the dichloromethane fraction was then submitted to countercurrent chromatography. The solvent system composed of n-hexane, ethyl acetate, methanol, and water (1:2.5:2.5:1) was chosen to perform the chromatographic analysis due to the enhanced solubility and the best distribution coefficients of the target compounds. Two porphyrins were isolated by this method and identified as 13(2)-hydroxypheophorbide a methyl ester and pheophorbide a, methyl ester. The solvent system proposed provided good distribution coefficients for both substances (1.27 and 1.87, respectively), and a high resolution factor.

A new isocoumarin from Cajanus cajan (Fabaceae).

A new isocoumarin, 3-phenyl-8-hydroxy-6-methoxy-5-gamma,gamma-dimethylallyl-isocoumarin, named cajavilmina (1) and eight known compounds: a-amirenone (2), beta-amirenone (3), lupenone (4), 5-hydroxy-7-methoxydihydroflavone (5), longistilin C (6), 3-hydroxy-5-methoxystilbene (7), beta-sitosterol (8) and stigmasterol (9) were identified in a dichloromethane fraction from Cajanus cajan leaves. Structures were elucidated by analysis of spectral data, mainly those afforded by 1H, NOEDIFF and 13C NMR (1D and 2D NMR HMQC, HMBC and COSY) and mass spectra.