Graphene oxide (GO) functionalization has great importance for its practical application in many fields, and is a key step for GO bioconjugation. Polydispersity, limited colloidal stability and changeable extinction of GO complicates functional group quantification and the modification study. To bypass the mentioned limitations, we have used GO immobilization on glass support to quantify its epoxy groups surface load, and to study sustainability of the functional groups introduced through epoxy-amine reaction during hydrazine reduction. It was found out that GO contains near 1 epoxy group per nm², and the major part of the functional groups are remaining after reduction process allowing to use this modification to make functionalized reduced graphene oxide (rGO). Then we modified colloidal GO with azide groups by aminotetraethylene glycol azide (H₂N–TEG–N₃) in mild conditions to avoid aggregation, and characterized it by means of immobilization on glass surface. Functional group surface load of the functionalized GO was determined as 0,7 groups per nm². Finally, we proved capability of azide-functionalized GO for conjugation by strain promoted [3 + 2] azide-alkyne cycloaddition (SPAAC) with BCN derivative of organic fluorescent dye JOE.

氧化石墨烯（GO）官能化对其在许多领域中的实际应用具有重要意义，并且是GO生物缀合的关键步骤。GO的多分散性，有限的胶体稳定性和易变的消光使官能团的定量和修饰研究变得复杂。为了绕过上述限制，我们将GO固定在玻璃载体上以定量其环氧基团的表面负荷，并研究了肼还原过程中通过环氧基胺反应引入的官能团的可持续性。发现GO每nm ²含有近1个环氧基，并且大部分官能团在还原过程后得以保留，从而允许使用这种修饰来制备功能化的还原氧化石墨烯（rGO）。然后，我们在温和的条件下用氨基四甘醇叠氮化物（H ₂ N–TEG–N ₃）对带有叠氮基的胶体GO进行修饰，以避免聚集，并通过固定在玻璃表面上对其进行了表征。将官能化的GO的官能团表面负荷确定为0.7个基团/ nm ²。最后，我们证明了叠氮化物官能化的GO通过与有机荧光染料JOE的BCN衍生物促进的[3 + 2]叠氮化物-炔烃环加成（SPAAC）进行偶联的能力。