Selection of DNA aptamers for tramadol through the systematic evolution of ligands by exponential enrichment method for fabrication of a sensitive fluorescent aptasensor based on graphene oxide

Tramadol hydrochloride (TH), as an atypical opioid and a 4-phenyl-piperidine analogue of codeine, is mainly used for treating moderate to severe pains. Due to its extensive application, the consequent need for its analysis in various samples is essential. The current study focuses on the introduction of a rapid fluorescent assay using graphene oxide (GO) and aptamer for determination of tramadol in serum samples. Specific ssDNA aptamers for TH were developed by SELEX (Systematic Evolution of Ligands by EXponential Enrichment) technique using GO as a fluorescence quencher. After 10 rounds, two aptamers (Apt19 and Apt39) were selected from various families. Then, the binding constants of aptamers were measured using fluorometric assay and finally Apt39 (labeled with ATTO 647N) was chosen for development of a fluorescent aptasensor because this aptamer bound to TH with high affinity (Kd = 178.4 nM) and specificity. The current analytical system showed detection limits of 1.04 nM and 2.56 nM in serum sample and phosphate buffer saline (10 mM PBS), respectively.