Development of quantification method for lipid hydroperoxides inbiological tissues

生物组织中脂质过氧化氢定量方法的开发

• 批准号: 02556010
• 负责人: MIYAZAWA Teruo
• 金额: $ 3.01万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-02556010/
• 关键词: lipid peroxidation; peroxidized lipid; lipid hydroperoxide; chemiluminescence; luminol; cytochrome c; 高速液体クロマトグラフィ-; ルミノ-ル; 過酸化リン脂質; ヒドロペルオキシド

A chemiluminescence-high performance liquid chromatography (CL-HPLC) system was developed and applied for the hydroperoxide-specific determination of phosphatidylcholine hydroperoxide (PCOOH), phosphatidylethanolamine hydroperoxide (PEOOH), triacylglycerol hydroperoxide (TGOOH), squalene hydroperoxide (SQOOH)and 8a-hydroperoxy tocopherone (TocOOH) in biological tissues such as human blood plasma, plasma lipoproteins, erythrocytes, cultured diploid cells, and liver, brain, heart, lung of the rodents.The method involves separation of lipid hydroperoxide in each lipid class from total lipids of the biological tissues with HPLC column and post-column detection of hydroperoxide-dependent chemiluminescence of lipid hydroperoxides. The chemiluminescence is produced by luminol oxidation during a reaction of hydroperoxide and cytochrome c-heme. The high specificity for hydroperoxide group enables a sensitive assay for a wide range of hydroperoxides, with the detection limit of 1 picomole of hydroperoxide-O_2. By use of this method, the presence of phospholipid hydroperoxides in human blood plasma and erythrocytes is confirmed quantitatively.

建立了一种化学发光-高效液相色谱（CL-HPLC）系统，用于测定人血浆、血浆脂蛋白、红细胞、培养的二倍体细胞、肝、脑、心、脑等生物组织中过氧化磷脂酰胆碱（PCOOH）、过氧化磷脂乙醇胺（PEOOH）、过氧化三酰基甘油（TGOOH）、过氧化角鲨烯（SQOOH）和8a-过氧化生育激素（TocOOH）的氢过氧化物特异性。啮齿类动物的肺。该方法利用高效液相色谱柱和柱后检测脂质氢过氧化物依赖的化学发光，从生物组织的总脂质中分离每一类脂质氢过氧化物。化学发光是在过氧化氢和细胞色素c-血红素的反应中由发光氨氧化产生的。对过氧化氢基团的高特异性使得对各种过氧化氢的检测灵敏度高，检测限为1皮摩尔的过氧化氢- o_2。用该方法定量地证实了人血浆和红细胞中磷脂氢过氧化物的存在。

项目成果

Teruo Miyazawa: "Chemiluminescent simultaneous determination of phosphatidylcholine hydroperoxide and phosphatidylethanolamine hydroperoxide in the liver and brain of the rat." Journal of Lipid Research. 33 (7). (1992)

Teruo Miyazawa：“化学发光同时测定大鼠肝脏和大脑中的氢过氧化磷脂酰胆碱和氢过氧化磷脂酰乙醇胺。”

宮澤 陽夫: "活性酸素と脂質" 化学と生物. 29. 798-806 (1991)

Haruo Miyazawa：“活性氧和脂质”化学与生物学 29. 798-806 (1991)

Teruo Miyazawa: "Chemiluminescence detection of 8a-hydroperoxy-tocopherone in photooxidized α-tocopherol" Lipids. 27. 289-294 (1992)

Teruo Miyazawa：“光氧化 α-生育酚中 8a-氢过氧生育酮的化学发光检测”脂质 27. 289-294 (1992)

TERUO MIYAZAWA: "Chemiluminescent Simultaneous Determination of Phosphatidyl-choline Hydroperoxide and Phosphatidylethanolamine Hydroperoxide in the Liver and Brain of the Rat" Journal of Lipid Research.

TERUO MIYAZAWA：“化学发光同时测定大鼠肝脏和大脑中的磷脂酰胆碱氢过氧化物和磷脂酰乙醇胺氢过氧化物”脂质研究杂志。

Lucia Satiko Yoshida: "Liver phosphatidylcholine hydroperoxidation provoked by ethionine-containing choline-deficient diet in mice." Lipids. 25 (9). 565-569 (1990)

Lucia Satiko Yoshida：“小鼠体内含有乙硫氨酸的胆碱缺乏饮食引起肝脏磷脂酰胆碱氢过氧化。”