生理活性物質の高効率分析用シリカゲル充填剤の試作と実用化

• Project/Area Number: 04557103
• Research Category: Grant-in-Aid for Developmental Scientific Research (B)
• Allocation Type: Single-year Grants
• Research Field: Physical pharmacy
• Research Institution: Nagasaki University
• Principal Investigator: 秋山 修三 長崎大学, 薬学部, 教授 (50028148)
• Co-Investigator(Kenkyū-buntansha): 門屋 純一 ダイソー(株), 研究所, 研究員 ; 大須賀 正就 ダイソー(株), 研究所, 主席研究員 ; 黒田 直敬 長崎大学, 薬学部, 助手 (50234612) ; 中島 憲一郎 長崎大学, 薬学部, 助教授 (30039656)
• Project Period (FY): 1992 – 1993
• Project Status: Completed (Fiscal Year 1993)
• Budget Amount: ¥16,900,000 (Direct Cost: ¥16,900,000); Fiscal Year 1993: ¥3,400,000 (Direct Cost: ¥3,400,000); Fiscal Year 1992: ¥13,500,000 (Direct Cost: ¥13,500,000)
• Keywords: 高速液体クロマトグラフィー / シリカゲル / シリカゲルカラム / シリカゲル充填剤 / π-π相互作用 / 水溶性ビタミン / 核酸関連物質 / アデニン / 旨味成分 / 酵素活性定量法 / ATPアーゼ

Research Abstract

本研究は水溶性生理活性物質の分離に適したHPLCカラム充填剤の開発を目指して行われた.始めに開発したMPS,PZS,PDSは相当する3-(置換プロピル)トリメトキシシリルシランとシリカゲルとをトルエン中で還流することにより調製した.これらは6種類の水溶性ビタミンを単一の緩衝液で一斉分析できた.次にπ-π相互作用の発現を期待して7種のゲル(NAPS,AAPS,ACNQ,PAIP,AAQI,NCIP及びNAIP)を調製した.これらのカラムは核酸に関連した化合物の分析に適しているかどうかが試された.ACNQとAAQIは初めての色素カラムである.MPSは目薬中の各種成分の分析,旨味に関連した調味料中の5'-IMP,5'-GMP及び5'-XMPの定量,アデノシン酸類(AMP,ADP,ATP等)の分析,更には糖類(単糖,二糖,多糖類)の分離定量が可能であった.NAPSとAAPSはアデニン系核酸関連物質の分析を単一緩衝液の使用とUV検出により容易に分析できることを示した.応用としてNAPSは,魚肉のATPase活性測定が可能であった.ACNQ,AAQI,PAIP,NCIP及びNAIP等はアデニン誘導体の分離に適用できることも分かった.とりわけ,ACNQが良好な分離パターンを示した.目下π-π相互作用による医薬品の分析を行っているところである.

Research Products

• [Publications] Akiyama,Shuzo: "High Performance Liquid Chromatographic Analyses of Compounds Related to Nucleic Acids by 3-(1-Naphthoylamino)-and 3-(1-Anthroylamino)propylmodified Silica Gel Packing Materials" Biomed.Chromatogr.7. 112-115 (1993)
• [Publications] Kuroda,Naotaka: "Preparation and Evaluation of Fatty Acid Esters of 2-(4-Hydroxyphenyl)-4,5-diphenylimidazole as Fluorescent Substrates for Measurement of Lipase Activity" Biol.Pharm.Bull.16. 220-222 (1993)
• [Publications] Kuroda,Naotaka: "Chemiluminescence Method for the Determination of Adenine After Reaction with Phenylglyoxal" Anal.Chim.Acta. 278. 275-278 (1993)
• [Publications] Akiyama,Shuzo: "Development of Silica Gels Immobilizing Organic Dyes for High-Performance Liquid Chromatography and Their Application to Analysis of Compounds Related to Nucleic Acids" Chem.Exp.8. 565-568 (1993)
• [Publications] 秋山修三: "π電子系を有する3-(N-置換)アミノプロピルシリカゲルの開発と高速液体クロマトグラフィーへの利用に関する基礎的研究" 分析化学. 42. 817-823 (1993)
• [Publications] Nakashima,Kenichiro: "High-Performance Liquid Chromatographic Determination of Hydrogen Peroxide with Peroxyoxalate Chemiluminescence Detection" J.Liq.Chromatogr.Clin.Anal.(in press). (1994)
• [Publications] Akiyama,Shuzo: "“Near-infrared Luminescence Spectroscopy," in “Molecular Luminescence Spectroscopy;Method and Applications:Part III," ed.by S.G.Schulman" Wiley-Interscience,New York, 23 (1993)
• [Publications] Akiyama,Shuzo: "High Perfonmans Liquid Chromatographic Anaiysis of Compounds Related to Nucleic Acids by 3-(1-Naphthoylamino)-and 3-(Anthroylamino)propy-modified Silicagel Packing Materials" biomed.chromatogr.7. 112-115 (1993)
• [Publications] Kuroda,Naotaka: "Chemiluminescence Method for the Deteemination of Adenine After Reaction with Phenylglyoxal" Anal.Chim.Acta. 278. 275-278 (1993)
• [Publications] Akiyama,Shuzo: "Drvelopment of Silica Gels Immobilizng Organic Dyes for High-Performace Liguid Chromatography and Their Application to Analysis of Compounds Related to Nucleic Acids" Chem.Exp. 8. 565-568 (1993)
• [Publications] 秋山 修三: "pi電子系を有する3-(N-〓換)アミノプロピルシリルシリカゲルの開発と高速液体クロマトグラフィーへの利用に関する基礎的研究" 分析化学. 42. 817-823 (1993)
• [Publications] Akiyama,Shuzo: "New Dye Systems Containing a C〓C Bond" Dyes Pigms.(印刷中). (1994)
• [Publications] Nakashima,Kenichiro: "High-Performance Liquid Chromatographic Determination of Hydrogen Peroxide with Peroxyoxa〓ate Chemiluminescence Dotection" J.liq.Chromatogr.Clin.Anal.(印刷中). (1994)
• [Publications] Akiyama,Shuzo: ""A Novel Syntheyic Method of Tolan Derivatives"in"Chemistry of Functional Dyes,Vol.2(ed,by Z.Yoshida and Y.Shirota)" Mita,Press,Tokyo,Japan, 4 (1993)
• [Publications] Akiyama,Shuzo: ""Near-Infrared Luminescence Spectroscopy"in Moleculan Luminesccnce Spectroscopy,Pant3(ed/by S.G.Schulman)" John Wiley&sons,NewYork.U.S.A., 15 (1993)
• [Publications] Akiyama,Shuzo: "High-performance Liquid Chromatographic Determination of Sugars in an Infusion and Soft Drinks Using a Silica Based 3-Morpholinoprorylbonded Stationary Phase" J.Chromatogr.626. 266-270 (1992)
• [Publications] Nakashima,Kenichiro: "Preparation of a Fouorescent Derivative of Benzoylecgonine,and Prelininary Studies of Its Application to the Analysis of Urine" J.Chromatogr.584. 275-279 (1992)
• [Publications] Nakatsuji,Shinichi: "Synthesis and Absorption/Emission Spectroscopic Properties of Bis(phenylethynyl)benzenes and 9,10-Bis(phenylethynyl)anthracenes" J.Chem.Soc.Perkin Trans.1. 775-758 (1992)
• [Publications] Nakashima,Kenichiro: "High Performance Liquid Chromatography with Chemiluminescence Detection of Methamphetamine and its Related Compounds Using 4-(N,N-Dimethylaminosulphonyl)-7-fluoro-2,1,3-benzoxadiazole" Biomed.Chromatogr.6. 149-154 (1992)
• [Publications] Nakashima,Kenichiro: "Peroxyoxalate Chemiluminescence Detection of Condensates of Malondialdehyde with Thiobarbituric Acids Using a Flow System" Biomed.Chromatogr.6. 55-58 (1992)
• [Publications] Akiyama,Shuzo: "High Performance Liquid Chromatographic Analyses of Compounds Related to Nucleic Acids by 3-(1-Naphthoylamino)-and 3-(1-Anthroylamino)propyl-modefied Silica Gel Packing Materials" Biomed.Chromatogr.7. (1993)
• [Publications] Akiyama,Shuzo: "“Near-Infrared Luminescence Spectroscopy" in Molecular Luminescence Spectroscopy;Methods and Applications:Part III,ed.by S.G.Schulman" Wiley-Interscience,New York, (1993)