Zur trennung und identifikation der freien porphyrine und deren isomere

• Published in: Journal of Chromatography A 1968, Vol.38, p.106-119
• Main Author: Mundschenk, H
• Format: Article
• Language: ger
• ISSN: 0021-9673
• DOI: 10.1016/0021-9673(68)85013-7

Complex porphyrin mixtures can be analyzed by means of extraction chromatography combined with thin-layer chromatographic and absorption spectrophotometric measurements. The fractions obtained in the partition system (tri- n-butyl phosphate/hydrochloric acid) may be identified as regards the number of carboxylic groups n by determining the peak position of the Soret band (λ p using the correlation between n and λ p. The isometric structure of the porphyrins can be established by thin-layer chromatography with 2,6-lutidine/water of the corresponding coproporphyrin derivatives obtained by decarboxylation. Using the methods described, nearly all porphyrin compounds in the reaction chain from uroporphyrin to coproporphyrin of the isometric series I and III could be identified in a porphyrin sample isolated from the urine of a patient with a hyporegenerative anaemia. Furthermore, several fraction were observed after the extraction and thin-layer chromatographic separations which could not be related to the porphyrins of the isomeric types I and III normally found in porphyrin samples isolated from urine. This was proved by the fact that the λ p values of the single fractions did not agree with the expected values. Therefore, it was assumed that these compounds, belonging to the isomeric series I and III, differ from the normal porphyrins by their substituents. However, further investigations are still necessary to elucidate the structure and determine the assumed substituents of the isolated porphyrins. There is no evidence that porphyrins of the isomeric types II and IV occur in the porphyrin mixture studied. Zur Analyse komplexer Porphyringemische wird eine Kombination verschiedener Untersuchungsmethoden empfohlen, die eine Identifikation der einzelnen Hämpräkursoren durchzuführen gestattet. Die bei der extraktionschromatographischen Auftrennung erhaltenen Fraktionen können bezüglich des Decarboxylierungsgrades über eine Bestimmung der Peaklage der Soretbande λ p , die bereits mit Mengen von min. 0.2 μg durchgeführt werden kann, eingeordnet werden. Zur Ermittlung des Isomerentyps werden die Porphyrine in die entsprechenden Coproporphyrinisomere durch Decarboxylierung überführt, die dünnschichtchromatographisch im 2,6-Lutidin/Wasser-System identifiziert werden. Mit den beschriebenen Verfahren (Extraktionschromatographische Auftrennung der Porphyrine im Verteilungssystem Tri- n-butylphosphat/1 N Salzsäure; Dünnschichtchromatographische Trennung der Porphyrine i