Abstract
We aim to establish a simple comprehensive analytical method for sugar phosphates to know the phosphate metabolism in plants. For this purpose, we applied anion exchange separation and electrochemical detection (HPAE-PAD) method for Arabidopsis. It is necessary to inactivate enzymes immediately after sampling for applying the chromatography. We tried to examine the extraction by hydrochloric acid, TCA, microwave, ultra filtration or heating to inactivate enzymes. The extraction by hydrochloric acid and TCA caused anion overloading in anion exchange column, and it was difficult to determine sugar phosphates. Changes in metabolites levels were observed during the sample homogenization using ultra filtration or heating. Heating or microwave application after the lyophilization of frozen sample made better detection of sugar phosphates. Sugar phosphates in Arabidopsis which was grown in different Pi concentration media, were determined by this extraction method.
