Abstract
By the use of Sep-Pak C18 cartridge, a simple and rapid cleanup method was established for the quantitative determination of pheophorbide a (PB-a) and pyropheophorbide a (PyPB-a) in chlorella tablets. PB-a and PyPB-a were extracted with methanol from chlorella tablets, then the extract was adjusted to methanol-1% acetic acid (7: 3, v/v) and centrifuged. The supernatant was cleaned up by the use of Sep-Pak C18 cartridge, eluted with methanol-0.025M ammonium acetate (88: 12, v/v) and applied to high performance liquid chromatography. They were separated on Radial PAK NOVA-PAK C18 column with the same solvent as a mobile phase. The average recoveries of PB-a and PyPB-a added to chlorella tablets were 88.9% and 96.3%, respectively.In order to estimate chlorophyllase activity in chlorella tablets, the proposed method was used to determine the amount of PB-a increased by incubation for 4h. As PB-a was separated from other chlorophyll degradative products derived during incubation, the proposed method was more specific and accurate than the official one (spectrophotometric determination).
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