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プロエンハンサー基質を用いる製剤中のリパーゼ活性の簡易,迅速な化学発光測定


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Title: プロエンハンサー基質を用いる製剤中のリパーゼ活性の簡易,迅速な化学発光測定
Other Titles: Simple and Rapid Chemiluminescence Assay for Lipase Activity in Pharmaceutical Preparations Using Proenhancer Substrate
Authors: 有馬, 貴美代 / 一番ヶ瀬, 智子 / 大庭, 義史 / 岸川, 直哉 / 黒田, 直敬
Authors (alternative): Arima, Kimiyo / Ichibangase, Tomoko / Ohba, Yoshihito / Kishikawa, Naoya / Kuroda, Naotaka
Issue Date: 5-May-2006
Publisher: 日本分析化学会
Citation: 分析化学, 55(5), pp. 307-311; 2006
Abstract: 製剤中リパーゼ活性の簡便かつ迅速な化学発光定量法を確立した.本法は,プロエンハンサー基質からリパーゼにより遊離するエンハンサー(化学発光増強剤) が,ルミノール/西洋わさびペルオキシダーゼ/過酸化水素の化学発光を増強する現象を利用したものである.化学発光定量条件を最適化したのち,本法を3種 の市販製剤中のリパーゼ活性測定に適用したところ,これら製剤中のリパーゼ活性を良好に定量することができた.また,本法により得られたリパーゼ活性値と 日本薬局方収載の脂肪消化力試験(中和滴走法)により得られた値とを比較したところ,両測定法の間に良好な相関性が得られ(r>0.919),本法 の信頼性を確認することができた.今回構築した化学発光法は操作が簡便で,測定も5分で完了する.また,本法は滴定法と比較して高感度であり,再現性も良 好なことから,リパーゼ含有製剤の品質管理などに有用な方法であると考えられる. / A novel chemiluminescence (CL) assay method, using a HDI-laurate [lauric acid ester of 2-(4-hydroxypheny1)-4,5-diphenylimidazole (HDI)] as a proenhancer substrate, was applied to the determination of lipase (triglycerol lipase, EC 3.1.1.3) activity in pharmaceutical preparations. The method is based on an enhanced CL reaction of luminol-horseradish peroxidase (HRP)-hydrogen peroxide with HDI, which is liberated from the proenhancer substrate by enzymatic hydrolysis. The proposed method involves a homogeneous reaction system in which enzymatic hydrolysis of HDI-laurate and enhanced CL reaction with HDI occur in the same reaction mixture. The lipase activities of three commercially available preparations were measured by the proposed CL method. Linear relationships were obtained (r>0.977) between the concentrations and CL intensities in all of the tested preparations. The results obtained by the proposed method were compared with those by the titration method in Japanese Pharmacopoeia, and good correlations were obtained between both methods (r>0.919). The CL method is simple and rapid, permitting the completion of single assay within 5 min. The sensitivity and repeatability of the CL method were also superior to those of the titration method.
Keywords: chemiluminescence / lipase activity / pharmaceutical preparations / proenhancer / 2-(4-hydroxyphenyl)-4,5-diphenylimidazole
URI: http://hdl.handle.net/10069/21887
ISSN: 05251931
DOI: 10.2116/bunsekikagaku.55.307
Rights: The Japan Society for Analytical Chemistry (c) 2006
Type: Journal Article
Text Version: publisher
Appears in Collections:Articles in academic journal

Citable URI : http://hdl.handle.net/10069/21887

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