HPLC による穀類中メラトニンの分析
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A qualitative analysis of melatonin(MEL)in 9 species of cereals using a precolumn derivatization method was developed. The extraction method is described as follows. Ten gram of cereal and dichloromethane 40mL was added into 50mL centrifuge tube and mixed with glass rod for 1min. The mixture was centrifuged(2580G, 5min)and the MEL dissolved in supernatant(5mL)was adsorbed to Sep-Pak Silica cartridge. The adsorbed MEL was eluted by dichloromethane/Methanol( 1 : 1 ), and the eluant was evaporated to dryness. For the derivatization of melatonin, 150μL of aqueous 2mol/L Na2CO3, 100μL 100mmol/L H2O2, 50μL 1mmol/L CuSO4 and 700μL water were added to the residue, and the mixture was heated in boiled water for 6min. The synthesized melatonin derivative was extracted with ethyl acetate, evaporated to dryness, dissolved in aqueous 15%(v/v)CH3CN, and subjected to HPLC. The resultant fluorogen was excited at 245nm and the emission wavelength was 380nm, and the melatonin derivative was separated by reversed-phase HPLC in about 18min. Thus obtained extracts from 8 species cereals, brown rice, shiratamako, corn grits, buck wheat flour,foxtail millet, malt, pressed barley and wheat flour were detected MEL derivative, but the extract from corn flour was not detected. Fluorescence excitation and emission spectra of derivative from standard MEL and from 8 species cereals extract seemed to show similar patterns. From these results, it has been clarified that these 8 species cereals are contained MEL.
- 2012-07-01
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