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Development of a new highly sensitive D,L-discriminating amino acid analyzer

Research Project

Project/Area Number 08556016
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section展開研究
Research Field Bioproduction chemistry/Bioorganic chemistry
Research InstitutionTohoku University

Principal Investigator

OHRUI Hiroshi (1997)  Tohoku University, Faculty of Agriculture, Professor, 農学部, 教授 (20100050)

目黒 煕 (1996)  東北大学, 農学部, 教授 (30005590)

Co-Investigator(Kenkyū-buntansha) AKASAKA Kazuaki  Tohoku University, Faculty of Agriculture, Assistant, 農学部, 助手 (10201881)
大類 洋  東北大学, 農学部, 助教授 (20100050)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥6,900,000 (Direct Cost: ¥6,900,000)
Fiscal Year 1997: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1996: ¥5,900,000 (Direct Cost: ¥5,900,000)
Keywords(S) -MNB-carboxylic acid / (S) -TBMB-carboxylic acid / D,L-analysis / sequential analysis / on-line HPLC-exciton CD analysis / 蛍光分析 / アミノ酸 / エナンチオマー / キラル分析 / HPLC / ベンゾジオキソ-ルカルボン酸
Research Abstract

We have focused on the development of a highly sensitive method for the sequential determination of D,L-amino acid from the C-terminus of peptides. We have developed a method which involves the selective hydrolysis of C-terminal psptide bond as the key step. The method is as follows, (1) ethyl esterification of the C-terminal carboxylic acid, (2) NaBH_4 reduction of the ester, (3) selective hydrolysis of the terminal amide bond under acidic conditions via N*O acyl migration, (4) separation of the resulting amino alcohol from peptide and mono-N-MNB carbonylation, (5) HPLC analysis of the mono-N-MNB carbonylated derivatives.
The so-called "new generation reagent" (^+)-MNB carboxylic acid was superior in the D,L-analysis of amino alcohols than the "old" (^+)-TBMB carboxylic acid. However, the mono-N-MNB-carbonylation method needed the authentic samples for unequivocal identification. Therefore, we next studied the exciton-CD monitoring method by N,O-di-O- (S)-TBMB-carbonylated amino alcohols were clearly separated by normal phase column chromatography but their stereochemistry could not be determined by CD monitoring because they did not show the characteristic exciton CD spectra for the specific stereoisomer. Finally, we have found that the amino alcohols can be determined by the on-line HPLC-exciton CD analysis without reference asmples by N-phthaloyl-O-(S)-TBMB-carbonylation. Thus, we have developed a new sensitive and selective method for the determination of D,L of amino acids.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report

Research Products

(16 results)

All Other

All Publications (16 results)

  • [Publications] H.Meguro, H.Ohrui: "“Strategy and design of novel reagents for the fluorometric analysis of biomolecules"" Bisci.Biotech.Biochem.60. 1919-1924 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] E.Itoh, Nishida, T.Togashi, Hiroshi Ohuri, Hiroshi Meguro: "“Simultaneous analysis of enantiomeric amino acids based on derivatization with(S)-(+)-2-t-butyl-2-methyl-1,3-benzodioxole-4-carboxylic acid and high-performance liquid chromatography"" Analytical Sciences. 12. 551-556 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Ohrui, E.Itoh, Y.Nishida, H.Horie, H.Meguro: "“Potential D,L-amino scid sequence analysis of peptides from the C-teminus"" Biosci.Biotech.Biochem.61. 392-395 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Nishida, J-H.Kim, H.Ohrui, H.Meguro: "“On-line HPLC-exciton CD analysis using a chiral benzoyl agent,(S)-TBMB carboxylic acid;A promising approach toward selective of enantimeric diols and diamines"" J.Am.Chem.Soc.119. 1484-1485 (1197)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] K.Shimizu, J.-H.Kim, K.Akasaka, H.Ohrui: "“Application of(S)-TBMB carboxylic acid-based on line HPLC-exciton CD analysis to acyclic vicinal alcohols and anino alcohols"" Chirality. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hiroshi Meguro and Hiroshi Ohrui: ""Strategy and design of novel reagents for the fluorometric analysis of biomolecules"" Biosci.Biotech.Biochem.60 (12). 1919-1924 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Eigo Itoh, Yoshihiro Nishida, Yukie Togashi, Hiroshi Ohrui and Hiroshi Meguro: ""Simultaneous analysis of enantiomeric amino acids based on derivatization with (S) - (+) -2-t-butyl-2-methyl-1,3-benzodioxole-4-carboxylic acid and high-performance liquid chromatography"" Analytical Sciences. 12 (8). 551-556 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Hiroshi Ohrui, Eigo Itoh, Yoshihiro Nishida, Hiroko Horie and Hiroshi Meguro: ""Potential D,L-amino acid sequence analysis of peptides from the C-teminus"" Biosci.Biotech.Biochem.61 (2). 392-395 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Yoshihiro Nishida, Jeong-Hwan Kim, Hiroshi Ohrui and Hiroshi Meguro: ""On-line HPLC-exction CD analysis using a chiral benzoyl agent, (S) -TBMB carboxylic acid : A promising approach toward selective identification of enantiomeric diols and diamines"" J.Am.Chem.Soc.199. 1484-1485 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kumi Shimizu, Jeong-Hwan Kim, Kazuaki Akasaka and Hiroshi Ohrui: ""Application of (S) -TBMB carboxylic acid-based on-line HPLC-exciton CD analysis to acyclic vicinal alcohols and amino alcohols"" Chirality. (in press). (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Meguro, H.Ohrui: ""Strategy and design of novel reagents for the fluorometric analysis of biomolecules"" Biosci.Biotech.Biochem. 60・12. 1919-1924 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] E.Itoh, Y.Nishida, Y.Togashi, Hiroshi Ohrui, Hiroshi Meguro: ""Simultaneous analysis of enantiomeric amino acids based on derivatization with(S)-(+)-2-t-butyl-2-methyl-1,3-benzodioxole-4-carboxylic acid and high-performance liquid chromatography"" Analytical Sciences. 12・8. 551-556 (1996)

    • Related Report
      1997 Annual Research Report
  • [Publications] H.Ohrui, E.Itoh, Y.Nishida, H.Horie, H Meguro: ""Potential D,L-amino acid sequence analysis of peptides from the C-teminus"" Biosci.Biotech.Biochem.61・2. 392-395 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Y.Nishida, J.-H.Kim, H.Ohrui, H.Meguro: ""On-line HPLC-exciton CD analysis using a chiral benzoyl agent,(S)-TBMB carboxylic acid:A promising approach toward selective identification of enantiomeric diols and diamines"" J.Am.Chem.Soc.119. 1484-1485 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] K.Shimizu, J.-H.Kim, K.Akasaka, H.Ohrui: ""Application of (S)-TBMB carboxylic acid-based on-line HPLC-exciton CD analysis to acyclic vicinal alcohols and amino alcohols"" Chirality. (in press).

    • Related Report
      1997 Annual Research Report
  • [Publications] E. Itoh, Y. Nishida, Y. Togasi, H. Ohrui, H. Meguro: "Simultaneous Analysis of Enantiomeric Amino Acids Based on Derivatization with (S)- (+) -2-t-Butyl-2-methyl-1, 3-benzodioxole-4-carboxylic Acid and High-Performance Liquid Chromatography" Analytical Sciences. 12 (8). 551-556 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-03-31   Modified: 2016-04-21  

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