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

采用96孔聚苯乙烯微滴板，对新蝶呤和生物蝶呤的酶联免疫吸附法进行了研究。酶联免疫吸附试验采用的方法是：将抗体和半抗原的孵育液加入固定半抗原的孔中；山羊抗兔IgG；过氧化物酶抗过氧化物酶复合物；邻苯二胺和过氧化氢；硫酸。在免疫读取器上测量490nm处的吸光度。我们研究了一种更简单的检测方法，用辣根过氧化物酶-山羊抗兔IgG结合物代替山羊抗兔IgG和过氧化物酶抗过氧化物酶复合物。两种方法具有几乎相同的灵敏度（l-25pmol）、可靠性和特异性（两种半抗原几乎完全区分）。我们还研究了在6,7-未取代翼啶中引入碳取代基的新方法。1,3-二甲基肼- 5-氧化物和芳烃重氮盐可以选择性地得到不同的6-芳基化产物。1,3-二甲基咪嗪得到了6-和7-芳基化产物的混合物。1,3-二甲基嘧啶- 5-氧化物与三氟甲烷磺酸酐反应得到1,3-二甲基嘧啶-6-三氟酸酯，与烯醇酯反应得到各种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.