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2000 Fiscal Year Final Research Report Summary

Establishment and practical use of reference method for glyco-haemoglobin by mass spectrometry

Research Project

Project/Area Number 09557220
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section展開研究
Research Field Laboratory medicine
Research InstitutionOsaka Medical College

Principal Investigator

SHIMIZU Akira  Faculty of Medicine, Professor, 医学部, 教授 (00028581)

Co-Investigator(Kenkyū-buntansha) YAGI Masayuki  Arkray Co., Ltd., Director, 技術研究部, 係長
KISHIKAWA Masahiko  Faculty of Medicine, Osaka Medical Research Assistant, 医学部, 助手 (40268199)
NAKANISHI Toyofumi  Faculty of Medicine, Osaka Medical College Associate Professor, 医学部, 講師 (10247843)
Project Period (FY) 1997 – 2000
KeywordsHbAlc / electrospray ionization / mass spectrometry / abnormal Hb / HPLC / immunoassays / Glu-C / ESI / MS
Research Abstract

We established a reference method for glyco-haemoglobin (HbAlc), which is a most popular index for control of diabetes mellitus. The N-terminal residues of the Hb β-chains, which are released by enzymatic cleavage of the intact Hb molecule with endoproteinase Glu-C, were measured by electrospray ionization mass spectrometry(ESI/MS). The procedure was originally described by Kobolt et al., and we modified several points. We calculated the peak ratio of glycated to non-glycated hexapeptide by monitoring both singly and doubly protonated ions ('weighed sum method'). Secondly, We prepared a calibrator by mixing synthetic peptides, instead of mixing purified HbAlc and HbA_0. The coefficient of variation (CV) of intra- and inter-assay by our method was less than 2.0%. We studied the structure of various abnormal Hbs also using ESI/MS Most cases were requested to elucidate the cause of the unexpected values of HbAlc measured by HPLC.We measured HbAlc in samples with abnormal Hbs using 3 commercial systems and the ESI/MS method, and we clarified the error of HbAlc values both by HPLC and immunoassays for 25 kinds, 49 cases with abnormal Hbs.

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Toyofumi Nakanishi: "Effect of hemoglobin variants on routine glycohemoglobin measurements assessed by a mass spectrometric method."Clinical Chemistry. 46. 1689-1692 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masahiko Kishikawa: "Detection by mass spectrometry of highly increased amount of S-sulfonated transthyretin in serum from a patient with molybdenum cofactor deficiency."Pediatric Reasearch. 47. 492-494 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toyofumi Nakanishi: "Determination of ionization efficiency of glycated and non-glycated peptides from the N-terminal of hemoglobin β-chain by ESI mass spectrometry"J Chromatography B. 46. 83-89 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toyofumi Nakanishi: "Ratio of the ion peak intensity of glycated to non-glycated hexapetides from the N-terminal of hemoglobin β-chain measured by LC-ESI MS."J Mass Spectrom .Soc.Jpn. 47. 389-391 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ayako Miyazaki: "Compound heterozygosity for β^+-thalassemia -31 (A→G) and a variant with low oxygen affinity, Hb Sagami [β139 (H17) Asn→Thr]."Hemoglobin. 23. 257-271 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Saraswathi Mandapati: "Relative quantification of glycated Cu-Zn superoxide dismutase in erythrocytes by electrospray ionization mass spectrometry"Biochim Biophys Acta. 1426. 483-490 (1999)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akira Shimizu: "Determination of an accurate ratio of a peptide and its adduct by ESIMS : calculation of the weighted sum of singly and doubly charged ion intensity."John Wiley & Sons,Ltd.,Chichester,UK (In press). 2

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Akira Shimizu: "Detection and characterization of aberrant blood proteins by LC/ESI/MS."Elsevier Science Publishers B.V.,Amsterdam. 14 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toyofumi Nakanishi, Ayako Miyzazki, Masahiko Kishikawa, Mamoru Yasuda, Yoshiyuki Tokuchi, Yasutaka Kanada and Akira Shimizu: "Quatification of glycated hemoglobin by electrospray ionization mass spectrometry"J Mass Spectrom. 32. 773-778 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toyofumi Nakanishi, Ayako Miyazaki, Msahiko Kishikawa, Akira Shimizu, O.Kishida, S.Sumi, T.Tsubakio and Kiyohiro Imai: "A new hemoglobin variant found during Hb Alc measurement : Hb Hokusetsu [β52 (D3) Asp→Gly]"Hemoglobin. 22(4). 355-371 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akira Shimizu, Toyofumi Nakanishi, Masahiko Kishikawa and Ayako Miyazaki: "Detection and characterization of aberrant blood proteins by LC/ESI/MS"Advances in Mass Spectrometry Vol.14, Editors : E.J.Karjalainen, et al.. Elsevier Science B.V.. 405-418 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ayako Miyazaki, Toyofumi Nakanishi, Masahiko Kishikawa, Toshimasa Nakagawa, Akira Shimizu, A.H.Mohammed Mawjood, Kiyohiro Imai, Yoshikazu Aoki and Miyako Kikuchi: "Compound heterozygosity for β^+-thalassemia-31 (A→G) and a variant with low oxygen affinity, Hb Sagami [β139 (H17) Asn→Thr]"Hemoglobin. 23(3). 257-271 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toyofumi Nakanishi and Akira Shimizu: "Ratio of the ion peak intensity of glycated to non-glycated hexapeptides from the N-terminal hemoglobin β-chain measured by LC-ESI mass spectrometry"J Mass Spectrom. Soc. Jpn. 47. 389-391 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toyofumi Nakanishi and Akira Shimizu: "Determination of ionization efficiency of glycated and non-glycated peptides from the N-terminal of hemoglobin β-chain by electrospray ionization mass spectrometry"J Chromatography B.. 46(1). 83-89 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toyofumi Nakanishi. Ayako Miyazaki, Ken Iguchi and Akira Shimizu: "Effect of hemoglobin variants on routine HbAlc measurements, assessed by a mass spectrometric method"Clinical Chemistry. 46(10). 1689-1692 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Akira Shimizu, Toyofumi Nakanishi and Ayako Miyazaki: "Determination of accurate ratio of a peptide and its adduct by ESIMS : calculation of weighted sum of singly and doubly charged ion intensity"Advances in Mass Spectrometry Vol.15, Editors : E.Gelpi et al., Elsevier Science B.V.. (in press).

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2002-03-26  

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