Analysis of tumour-localizing haematoporphyrin derivative by high-performance liquid chromatography and fast-atom bombardment mass spectrometry.
Reversed-phase chromatography using a MOS-Hypersil (C8) column with methanol-1 M ammonium acetate buffer (pH 4.6) (60:40) as mobile phase has been developed for the isolation of tumour-localizing haematoporphyrin derivative (HPD). The system effectively resolved the diastereoisomers of haematoporphyrin and its acetyl derivatives. The chromatography peaks were identified by fast-atom bombardment mass spectrometry and were confirmed by chemical synthesis. The main components of HPD before alkaline hydrolysis were diacetylhaematoporphyrin, 8-(1-acetoxyethyl)haematoporphyrin and 3-(1-acetoxyethyl)haematoporphyrin with small amounts of haematoporphyrin, 8-(1-hydroxyethyl)-3-vinyldeuteroporphyrin, 3-(1-hydroxyethyl)-8-vinyldeuteroporphyrin, 8-(1-acetoxyethyl)-3-vinyldeuteroporphyrin, 3-(1-acetoxyethyl)-8-vinyldeuteroporphyrin and protoporphyrin. After hydrolysis with 0.1 M sodium hydroxide, the main components were haematoporphyrin, hydroxyethylvinyldeuteroporphyrins, and protoporphyrin.
J C
JC
Meijers
C K
CK
Lim
A M
AM
Lawson
T J
TJ
Peters
D000107:Acetylation; D002851:Chromatography, High Pressure Liquid; D006415:Hematoporphyrins / Q000032:analysis*; D006801:Humans; D006868:Hydrolysis; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D009369:Neoplasms / Q000175:diagnosis*; D013237:Stereoisomerism
D006415:Hematoporphyrins; D007202:Indicators and Reagents