PURPOSE: To enable microdetermination of a pyrimidine compd. which has a pyrimidine ring in the chemical structure and in which at least one of the nitrogen raw material in the ring is not substd. by subjecting the product obtd. by treating a sample with 4- bromomethyl-7-methoxy cumarine in the presence of crown ethers to fluorescent high- speed liquid chromatography.
CONSTITUTION: The compd. to be measured is a pyrimidine compd. which has a pyrimidine ring in the chemical structure and in which at least one of the nitrogen raw material in the ring has no substituent. Such compd. is exemplified by the base, nucleotide and nucleocide of a pyrimidine nucleic acid and the embodiment thereof is enumerated by uracil, thymine, cytosine, etc. The quantitative determination thereof is accomplished by treating the sample with 4-bromomethyl-7-methoxy cumarine in the presence of crown ethers. The pyrimidine compd. in the sample to be measured reacts with the 4- bromomethyl-7-methoxy cumarine as a result of such treatment and forms a fluorescence-emitting compd. The fluorescence-emitting compd. is the compd. in which the methyl group is bound to the nitrogen atom site having no substituent in the pyrimidine ring. Such fluorescence-emitting compd. is subjected to fluorescent high-speed liquid chromatography and is determined quantitatively by using a fluorescence detector.
YOSHIDA SHIGERU
IWAMOTO MAKI
CHIYOU TOSHIO
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