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KMID : 0614019850010010049
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Journal of Pharmaceutical Sciences (C.N.U.)
1985 Volume.1 No. 1 p.49 ~ p.56
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Assay for Catechol-O-Methyltransferase in Erythrocytes Using a New Fluorogenic Substrate, 2-(3, 4-Dihydroxy phenyl)-Naphthol [1,2-d] Thiazole
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Hitoshi Nohta
Shunya Noma/Yosuke Ohkura and 유병태
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Abstract
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A highly sensitive method for the assay of catechol-O-methyltransferase in erythrocytes is described, which employs high-performance liquid chromatography with fluorescence detection. A newly synthesized catechol compound, 2-(3,4-dihydroxyphenyl)naptho[1,2-d]-thiazole is used as a highly fluorogenic substrate for catechol-O-methyltransferase; the m-and p-methylated products formed enzymatically from the substrate under the optimum conditions, after extraction with n-hexane-chloroform, are separated by normal-phase chromatography on LiChrosorb Si 100. The limits of detection for m-and p-methylated products are 3 pmol per assay tube (60 fmol per injection volume of 20 ㎕) in each case. The ratio of m-and p-methylated products was 0.54. This method requires as little as 50㎕ of human erythrocytes.
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