Development of high Sensitive Methods for Analysis of Lipid Molecular Species

脂质分子种类分析的高灵敏度方法的开发

• 批准号: 61860008
• 负责人: KITO Makoto
• 金额: $ 5.12万
• 依托单位: Kyoto, University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1986
• 资助国家: 日本
• 起止时间: 1986 至 1987
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-61860008/
• 关键词: High Sensitive Methods for Analysis of Lipid Molecular Species; リン脂質分子種; 高速液体クロマトグラフィー(HPLC)

A simple and sensitive method was developed for quantitative analysis of phospholipid molecular species. Firadylglycerols were prepared from phospholipids by phospholipase C treatment and converted to corresponding dinitrobenzoyl derivatives, which could be detected at 254 nm. The derivatives or 36 molecular species were resolved by high performance luquid chromatography with on octadecylsily) reversed-phase colum. All the derivatives had the same peak area per mo., and pear areas were proportional to the amounts to the derivatives. The derivatives of alkenylacyl- and alkylacylmolecular species were resolved concomitantly with diacyl molecular species using a combination of two solvent systems. Quantification was carried out at the picomole level.The method was used for determination of mass changes in the various molecular species of human platelets after stimulation with thrombin and collagen. Upon stimulation, every molecular species of phosphatidylionositol and phosphatidylserine w … More as equally hydrolyzed, where the molecular species of phosphatidylcholine and diacyl-and aleknylacylphosphatidylethanolamine containing areachidoni acid were selectivey hydrolyzed. At low Ca^<2+> concentrations, which results from the release from internal stores, phosphatidylinositol, phosphatidylcholine and diacylphosphatidylethanolamine were hydrolyzed after stinulation with thrombin, wheras only phosphatidylinositol was hyrolyzed with production of thromboxane B_2 after stimulation with collagen. At high Ca^<2+> concentration,s phosphatidylcholine and diacylphosphatidylethanolamine were hydrolyzed after stimulation with collagen, and phosphatidylserine and alkenylacylphosphatidylethanolamine were degraded after stimulation with both thrombin and collagen. (1-^<14> C) Arachidonic acid was heterogeneously incorporated into the individual molecular species of the vatious phospholipid classes, indicating that the determination of mass is essential for accurate picture of phospholipid metabolism. Less

建立了一种简便、灵敏的磷脂分子种类定量分析方法。用磷脂酶C处理磷脂制备了三氟甲酰基甘油，并将其转化为相应的二硝基苯甲酰基衍生物，其可在254 nm处检测到。用十八烷基硅烷键合反相高效液相色谱法对36种衍生物进行了分离。所有衍生物具有相同的峰面积/月，梨形面积与衍生物的量成正比。烯基酰基和烷基酰基分子物种的衍生物同时与二酰基分子物种使用两种溶剂系统的组合解决。在皮摩尔水平进行定量，该方法用于测定凝血酶和胶原刺激后人血小板的各种分子种类的质量变化。在刺激下，磷脂酰肌醇和磷脂酰丝氨酸的每一种分子都被激活， ...更多信息 同样水解，其中含有花生四烯酸的磷脂酰胆碱和二酰基-和烯酰基磷脂酰乙醇胺的分子种类被选择性水解。在低Ca^2+浓度下，凝血酶刺激后，磷脂酰肌醇、磷脂酰胆碱和二酰基磷脂酰乙醇胺水解，胶原刺激后，只有磷脂酰肌醇水解，产生血栓素B_2。在高Ca^<2+>浓度下，胶原蛋白刺激后，s型磷脂酰胆碱和二酰基磷脂酰乙醇胺发生水解，凝血酶和胶原蛋白刺激后，磷脂酰丝氨酸和烯基酰基磷脂酰乙醇胺发生降解。(1-^<14>C）花生四烯酸不均匀地掺入各种磷脂类的单个分子物质中，表明质量的测定对于磷脂代谢的准确描述是必不可少的。少

项目成果

Hitoshi Takamura, Hiroshi Narita, Hwa-Jin Park, Ken-ichi Tanaka, Takao Matsuura and makoto Kito: "Differntial hydrolysis of phospholipid molecular species during activation of human platelets with thrombin and collagen" J. Boil. Chem.262. 2262-2269 (1987)

Hitoshi Takamura、Hiroshi Narita、Hwa-Jin Park、Ken-ichi Tanaka、Takao Matsuura 和 Makoto Kito：“用凝血酶和胶原蛋白激活人血小板过程中磷脂分子种类的差异水解”J. Boil。

林由佳子,裏出令子,鬼頭誠: 脂質生化学研究. 28. 129-132 (1986)

Yukako Hayashi、Reiko Urade、Makoto Kito：脂质生物化学研究 28. 129-132 (1986)。

Hitoshi Takamura, Hiroshi Narita, Reiko Urade and Matoko Kito: "Quantitative analysis of polyenoic phospholipid molecular species by high performance liquid chromatography" Lipids. 21. 356-361 (1986)

Hitoshi Takamura、Hiroshi Narita、Reiko Urade 和 Matoko Kito：“通过高效液相色谱法对多烯磷脂分子种类进行定量分析”脂质。

M.Kito;H.Takamura;R.Urade;H.Narita: Journal of Biochemistry. 98. 327-331 (1985)

M.Kito；H.Takamura；R.Urade；H.Narita：生物化学杂志。

H.Takamura;H.Narita;R.Urade and M.Kito: Lpids. 21. 356-361 (1986)

H.Takamura；H.Narita；R.Urade 和 M.Kito：脂类。