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DEVELOPEMENT OF THE METHOD FOR SEPARATORY DETERMINATION OF ENANTIOMETRIC DRUGS BY HYPHENATED MASS SPECTROMETRY

Research Project

Project/Area Number 05452336
Research Category

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

Allocation TypeSingle-year Grants
Research Field Physical pharmacy
Research InstitutionTOHOKU UNIVERSITY

Principal Investigator

GOTO Junichi  TOHOKU UNIVERSITY FACULTY OF PHARMACEUTICAL SCIENCES PROFESSOR, 薬学部, 教授 (80006337)

Co-Investigator(Kenkyū-buntansha) IKEGAWA Shigeo  TOHOKU UNIVERSITY FACULTY OF PHARMACEUTICAL SCIENCES ASSOCIATE PROFESSOR, 薬学部, 助教授 (90111301)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥5,700,000 (Direct Cost: ¥5,700,000)
KeywordsEnantiomeric drug / Diastereomer / LC / MS / Immobilized antibody / Derivatization / Glucuronosyltransferase / 固体化抗体 / ジアステレオマー
Research Abstract

In connection with pharmacokinetic and pharmacodynamic studies, chromatographic determination of enantiomeric compounds in biological fluids is one of the most important subjects in the biomedical analysis. Among various methods, liquid chromatography (LC) is well recognized as a powerful tool for the separatory determination of racemates. On the other hand, electrospray ionization (ESI) -mass spectrometry is becoming increasingly valuable tool for the analysis of biologically important substances having ionic groups. Accordingly, the method for separatory determination of enantiometric drugs by LC/ESI-MS have been developed.
1) The diastereometric method is more favorable for the determination of racemic drugs in biological fluids with respect to sensitivity and versatility. Since glucuronic acid having carboxyl group in the molecule is a naturally occuring optical active compound, reversed derivatization employing immobilized beta-glucuronosyltransferase under physiological conditions … More has been studied using proplanolol as a model drug. The derivatization was achieved by incubating with immobilized enzyme (300 mul) in the presence of uridine-5'-diphosphoglucuronic acid (10mM) and Mg^<2+> (5mM) at 37 ^0C.The resolution into individual diastereomer was achieved on a Capcell Pak C_<18> column using 50 mM ammonium acetate/methanol (1 : 2, v/v) as a mobile phase. The spectrum of propranolol glucuronide exhibited a quasimolecular ion [M-H]^- as an intense peak under the ESI-MS mode.
2) Immunoaffinity extraction of bufuralol and its metabolites has been developed by the use of the antibodies elicited from immunogens, (1R) -and (1S) -bufuralol 1'-O-carboxy-methyloxime-bovine serum albumin conjugates. The antibodies produced exhibited low cross reactivities toward the corresponding antipode and a broad affinity spectrum for major metabolites oxidized at the 1' position on the benzene moiety. A 0.5 ml aliquot of the immunoaffinity adsorbent prepared by immobilization of the antibody was capapable retaining up to 1mug of (R) -and (S) -bufuralol, respectively. The adsorbates were recovered quantitatively by elution with 95% methanol without any interfering.
3) The combination of immunoaffinity extraction and reversed derivatization with LC/ESI-MS is promising for the xenobiotic metabolic study of enantiometric drugs. Less

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] Toshifumi Niwa: "Separation and characterization of ursodeoxycholate 7-N-acetylglucosaminides in human urine by high-performance liquid chromatortaphy with fluorescence detection." J.Liquid Chromatogr.16. 2531-2544 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shigeo Ikegawa: "Separatory determination of biliary metabolies of equilin in rat by high-performance liquid chromatography." J.Liquid Chromatogr.17. 223-239 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shinichi Miyairi: "Flow injection-immunometric assay of 17α-hydroxyprogesterone using fluorescein-labeled Fab fragment." Anal.Sci.10. 109-111 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toshifumi Niwa: "Preparation of antiserum for use in the immunoassay of unconjugated and conjugated 4β-hydroxycholic acids." Anal.Sci.10. 349-351 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Norio Awata: "Immunoaffinity extraction of 4-hydroxy-2-(4-methyphenyl)benzothiazole and its metabolites for determination by gas chromatography-mass spectrometry." Biol.Pharm.Bull.17. 843-945 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Mitsunori Tanji: "Identification and characterization of dehydrocholic acid reductase system in the cytosol of human red blood cells." J.Gastroenterol.29. 621-630 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] T.Niwa, K.Fujita, J.Goto and T.Nambara: "Separation and characterization of ursodeoxycholate 7-N-acetylglucosaminides in human urieneby high-performance liquid chromatography with fluorescence detection." J.Liquid Chromatogr.16. 2531 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Ikegawa, M.Itoh and J.Goto: "Separatory determination of biliary metabolites of equilin in ratby high-perfomance liquid chromatography." J.Liquid Chromatogr.17. 223 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] S.Miyairi, A.Kanamaru, K.Nakajima, T.Kato and J.Goto: "Flow injection-immunometric assay of 17alpha-hydroxyprogesterone using fluoresceinlabeled Fab fragment." Anal. Sci.10. 109 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] N.Awata, F.Toba, M.Ando, H.Shimada, S.Miyairi, T.Kato, J.Goto, and T.Nambara: "Immunoaffinity extraction of 4-hydroxy-2- (4-methyphenyl) benzothiazole and its metabolites for determination by gas chromatography-mass spectrometry." Biol. Pharm. Bull.17. 843 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] M.Tanji, J.Goto and I.Makino: "Identification and characterization of dehydrocholic acid reductase system in the cytosol of human red blood cells." J.Gastroenterol.29. 621 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shinichi Miyairi: "Novel immunoaffinity extraction for liquid chromatographic determination of major metabolites of 4-acetoxy-2-(4-methylphenyl) benzothiazole in plasma." J.Pharm.Biomed.Anal.389-395 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Shinichi Miyairi: "Flow injection-immunometric assay of 17α-hydroxyprogesterone using fluorescein-labeled Fab fragment." Anal.Sci.10. 109-111 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Toshifumi Niwa: "Preparation of antiserum for use in the immunoassay of unconjugated and conjugated 4β-hydroxycholic acids." Anal.Sci.10. 349-351 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Norio Awata: "Immunoaffinity extraction of 4-hydroxy-2-(4-methyphenyl) benzothiazole and its metabolites for determination by gaschromatography-mass spectrometry." Biol.Pharm.Bull.17. 843-945 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Takashi Iida: "Analysis of conjugated bile acids in human biological fluids.Synthesis of hyodeoxycholic acid 3-and 6-glycosides and related compounds." Chem.Pharm.Bull.42. 1479-1484 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Mitsunori Tanji: "Identification and characterization of dehydrocholic acid reductase system in the cytosol of human red blood cells." J.Gastroenterol.29. 621-630 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Toshifumi Niwa: "Separation and characterization of ursodeoxycholate 7-N-acetylglucosaminides in human urine by high-performance iquid chromatography with fluorescence detection." J Liquid Chromatogr.16. 2531-2544 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Shinichi Miyairi: "Enzyme immunoassay for 4-hydroxy-2-(4-methylphenyl)-benzothiazole." J.Pharm.Biomed.Anal.11. 469-476 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Shigeo Ikegawa: "Separatory determination of biliary metabolites of equilin in rat by high-performance liquid chromatography." J.Liquid Chromatogr.17. 223-239 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Shinichi Miyairi: "Novel immunoaffinity extraction for liquid chromatographic determination of major metabolites of 4-acetoxy-2-(4-methylphenyl)benzothiazole in plasma." J.Pharm.Biomed.Anal.(in press).

    • Related Report
      1993 Annual Research Report
  • [Publications] Norio Awata: "Immunoaffinity extraction of 4-hydroxy-2-(4-methyl-phenyl)benzothiazole and its metabolites for determination by gas chromatography-massspectrometry" Biol.Pharm.Bull.(in press).

    • Related Report
      1993 Annual Research Report
  • [Publications] Toshifumi Niwa: "Preparation of antiserum for use in the Immunoassay of unconjugated and conjugated 4β-hydroxycholic acid" Anal.Sci.(in press).

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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