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Development of a sensitive enzymatic assay for free fatty acid esters of 5'-guanosine monophosphate in food and environmental samples.
We have developed a sensitive enzymatic assay for the assay of 5'-guanosine monophosphate (5'-GMP) esterified with short-chain fatty acids. The developed assay is based on the specific hydrolysis of 5'-GMP esters by a 5'-guanosine monophosphatase. The hydrolysis is followed by a colorimetric measurement of an enzymatically generated product: 4-methylumbelliferone. Experiments with complex food matrices containing free fatty acids yielded an initial rate of 0.25 pmol/min/microl 5'-GMP esterification activity. The hydrolysis reaction is linear for the assay over a range from 0 to 24 pmol of 5'-GMP. The developed method allows for accurate measurement of fatty acid esters of 5'-GMP in food and environmental samples including hydrolyzed fish muscle and eggs and lake sediments. The developed method was linear for determination of 5'-GMP esters containing up to 12 carbons.This invention relates to a steering apparatus, particularly a steering apparatus for use in a vehicle, such as an automobile.
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