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Highly Selective Fluorometric Determination of Dicarboxylic Acids Based on Intramolecular Excimer-Formfng Derivatization with a Pyrene-Labeling Reagent.

Research Project

Project/Area Number 12672101
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionFukuoka University

Principal Investigator

YAMAGUCHI Masatoshi  Faculty of Pharm. Sci., Fukuoka University Professor, 薬学部, 教授 (50117280)

Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥2,400,000 (Direct Cost: ¥2,400,000)
Keywordsblood / urine / dicarboxylic acids / excimer fluorescence / derivatization / HPLC / グルタル酸血症 / 診断
Research Abstract

A highly selective and sensitive method was developed to determine dicarboxylic acids, based on an intramolecular excimer-forming fluorescence derivatization with a pyrene reagent, 4-(l-pyrene)butanoic acid hydrazide (PBH) in the presence of l-ethyl-3-(3-dimrthylaminopropyDcarbodiimide, followed by reversed phase high performance liquid chromatography (HPLC). Dicarboxylic acids, having two carboxylic acid moieties in a molecule, were converted to the corresponding dipyrene-labeled derivatives by reaction (40ーC, 60 min) with PBH. The derivatives afforded intramolecular excimer fluorescence (450-520 nm) which can clearly be discriminated from the monomer (normal) fluorescence (360-420 nm) emitted from PBH and monopyrene-labeled derivatives of monocarboxylic acid. The structures of the derivatives were confirmed by HPLC with mass spectrometry, and the emission of excimer fluorescence could be proved by spectrofluorometry and time-resolved fluorometry. The PBH derivatives of four dicarboxylic acids [glutaric acid (GA), adipic acid (AA), suberic acid (SubA) and sebacic acid (SebA)] could be separated by reversed phased HPLC on C8 column with linear gradient elution. The detection limits (signal-to-noise ratio of 3) for the dicarboxylic acids were 22 (GA), 25 (AA), 31 (SubA) and 54 (SebA) fmol on column. Furthermore, the present method was so selective that biogenic monocarboxylic acids gave no peak in the chromatogram.

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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