The rapid determination of erythrocyte porphyrins using reversed-phase high performance liquid chromatography.
The extraction, separation and quantification of free acid erythrocyte porphyrins are described. The porphyrins are extracted from whole blood using 3:1 ethyl acetate/acetic acid (v/v) and separated using the reversed-phase mode of high performance liquid chromatography (RPLC). The compounds are detected on-line spectrofluorometrically using an excitation wavelength of 404 nm and an emission cut-off filter of 550 nm. Excellent resolution of the porphyrin free acids is achieved in less than 6 min. The analytical recoveries for protoporphyrin IX and zinc protoporphyrin IX were greater than 90% with relative standard deviations for day-to-day analysis under 6%.
D000085:Acetates; D019342:Acetic Acid; D002851:Chromatography, High Pressure Liquid; D003306:Coproporphyrins / Q000097:blood; D004912:Erythrocytes / Q000032:analysis*; D006801:Humans; D008652:Mesoporphyrins / Q000097:blood; D011166:Porphyrins / Q000097:blood* / Q000302:isolation & purification; D011524:Protoporphyrins / Q000097:blood
D000085:Acetates; D003306:Coproporphyrins; D008652:Mesoporphyrins; D011166:Porphyrins; D011524:Protoporphyrins; C031468:mesoporphyrin IX; C031467:coproporphyrin I; C007650:ethyl acetate; C028025:protoporphyrin IX; D019342:Acetic Acid