Separation of protoporphyrins and related compounds by reversed-phase liquid chromatography.
We describe the separation of protoporphyrin and related porphyrins by reversed-phase "high-performance" liquid chromatography, with fluorometric detection. We used the method to demonstrate that acid hydrolysis of the dimethyl ester of protoporphyrin IX is complete in 2 to 3 h and is followed by the acid-catalyzed conversion of protoporphyrin IX to a chemical species that chromatographic evidence indicates to be hematoporphyrin IX. In addition, the method was used to evaluate the purity of a commercial preparation of protoporphyrin IX and was also demonstrated to have the sensitivity and specificity needed for measuring the protoporphyrin IX content of whole-blood.
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D011166:Porphyrins / Q000097:blood*; D011524:Protoporphyrins / Q000097:blood* / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
D011166:Porphyrins; D011524:Protoporphyrins