A Study on Simple and Highly Sensitive Analytical Methods of Pteridines Aimed to be Used for the Detection of Virus Infection

采用96孔聚苯乙烯微孔板，研究了新喋呤和生物蝶呤的酶联免疫吸附分析方法。将抗体与半抗原混合孵育，山羊抗兔免疫球蛋白，过氧化物酶-抗过氧化物酶复合体，邻苯二胺和过氧化氢，硫酸。在免疫阅读器上测量490 nm处的吸光度。我们用辣根过氧化物酶-山羊抗兔免疫球蛋白结合物代替羊抗兔免疫球蛋白和过氧化物酶-抗过氧化物酶复合体，探索了一种简便的检测方法。目前的两种方法具有几乎相同的灵敏度(L-25pmol)、可靠性和特异性(两个半抗原几乎完全区分)。我们还研究了将碳取代基引入6，7-未取代蝶啶的新方法。1，3-二甲基鲁马嗪5-氧化物和芳烃重氮盐区域选择性地给出了各种6-芳基化产物。1，3-二甲基鲁马嗪得到6-和7-芳基化产物的混合物。1，3-二甲基鲁马嗪-5-氧化物与三氟甲磺酸反应生成1，3-二甲基鲁马嗪-6-三氟酸酯，再与烯醇酸酯反应得到各种6-取代衍生物。

We investigated enzyme linked immunosorbent assay for neopterin and biopterin by using polystyrene microtitre plate with 96 wells. ELISA was performed by adding the substances to the well with fixed hapten as follows : incubation mixture of antibody and hapten ; goat anti-rabbit IgG ; peroxidase antiperoxidase complex ; o-phenylendiamine and hydrogen peroxide ; sulfuric acid. The absorbance at 490nm was measured on a immunoreader. We have investigated a simpler assay method by employing horseraddish peroxidase-goat antirabbit IgG conjugate in place of goat anti rabbit IgG and peroxidase anti-peroxidase complex in the above procedure. The present two methods exhibited the almost same sensitivity (l-25pmol), reliability, specificity (the two haptens were almost completely distingushed).We also studied new methods to introduce carbon substituents into 6,7-unsubstituted pteridines. 1,3-Dimethyllumazine 5-oxide and arene diazonium salts gave various 6-arylation products, regioselectively. 1,3-Dimethyllumazine gave a mixture of 6- and 7-arylation products. 1,3-Dimethyllumazine 5-oxide and trifluoromethanesulfonic anhydride gave 1,3-dimethyllumazine-6-triflate, which gave various 6-substituted derivatives on reaction with enolates.