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Determination of 2-Aminoethylphosphonic Acid and Its Y-iM ethyl Derivative in Animal Tissues by Gas Chromatography with Flame Photometric Detection
Authors:Hiroyuki Kataoka  Norihisa Sakiyama  Masami Makita
Institution:Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama 700, Japan
Abstract:A sensitive and selective method was developed for the determination of 2-aminoethylphosphonic acid (AEP) and W-methyl AEP in animal tissues by gas chromatography (GC). These compounds were converted into their A-isobutoxycarbonyl methyl ester derivatives and measured by GC with flame photometric detection (FPd-GC), using 0.5 % FFAP on Uniport HP as the GC column packing. The calibration curves for AEP and A-methyl AEP in the range of 0.02 ~ 2 μg were linear, and the detection limit was about 20 pg as an injection amount. AEP and A^-methyl AEP in animal tissues were found in the free form and bound form with lipid and other biological macromolecules, and they could be measured without any influence from coexistent substances by FPd-GC. The recoveries of AEP and A'-methyl AEP added to the tissue samples were 92 —105 %, and their reproducibility was found to be satisfactory. The distribution of these compounds in various animals was also studied by using this new method.
Keywords:diheme cytochrome c 4  physicochemical property  recombinant protein  Vibrio parahaemolyticus
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