| MS Description
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Tissue samples were cut into small pieces … Tissue samples were cut into small pieces and extracted with ten volumes (w/v) of 2% acetic acid in water at 100°C for 10 min. The extracts were centrifuged at 15,000 g for 10 min, and filtered through a Cosmonice Filter S (pore size 0.5 mm, filter diameter 13 mm, Nacalai Tesque, Kyoto). The resulting filtrates were subjected to analyses. For the quantification of Trp, the filtrate (10 ml) was analyzed with an HPLC system (Hitachi L7000 series) coupled with a UV detector (Hitachi L7400). Chromatography was performed with a COSMOSIL 5C18AR-II column (1504.6 mm; particle size, five mm, Nacalai Tesque) and a solvent mixture of methanol and 0.1% phosphoric acid in water at a flow rate of 0.8 ml min1 at 35°C. The ratio of methanol to 0.1% phosphate in water was programmed as: 10:90, v/v, for 5 min, and then from 10:90 to 70:30 over 30 min. The detection wavelength was set at UV280 nm. The levels of anthranilic acid were determined by a previously reported method (Dubouzet et al. 2007). ly reported method (Dubouzet et al. 2007).
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