▎ 摘　　要

NOVELTY - Constructing ratio aptamer sensor based on simultaneous acquisition of dual signals of self-enhanced luminescent material and methylene blue involves adding ammonium citrate to ultrapure water, and performing heating reaction under normal pressure, when color of solution changes from colorless to yellow, adjusting pH to obtain nitrogen-doped graphene quantum dot solution (NGQDs). The Triton (RTM: Non-ionic surfactant) X-100, cyclohexane, 1-hexanol and water are mixed to obtain mixed solution A. The terpyridine ruthenium solution is added to mixed solution A, and after stirred for period of time, added tetraethyl orthosilicate and ammonia water to initiate polymerization reaction. The acetone is added to precipitate nanoparticles, ished with ethanol and ultrapure water, and centrifuged. USE - Method for constructing ratio aptamer sensor based on simultaneous acquisition of dual signals of self-enhanced luminescent material and methylene blue ADVANTAGE - The method enables to have low background signal of electrochemical luminescence and fast and sensitive detection of zearalenone (ZEN) with good selectivity and stability, and high sensitivity and recovery rate (97.3-102.7% recovery rate). DETAILED DESCRIPTION - Constructing ratio aptamer sensor based on simultaneous acquisition of dual signals of self-enhanced luminescent material and methylene blue involves adding ammonium citrate to ultrapure water, and performing heating reaction under normal pressure, when color of solution changes from colorless to yellow, adjusting pH to obtain nitrogen-doped graphene quantum dot solution (NGQDs). The Triton (RTM: Non-ionic surfactant) X-100, cyclohexane, 1-hexanol and water are mixed to obtain mixed solution A. The terpyridine ruthenium solution is added to mixed solution A, and after stirred for period of time, added tetraethyl orthosilicate and ammonia water to initiate polymerization reaction. The acetone is added to precipitate nanoparticles, ished with ethanol and ultrapure water, and centrifuged. The centrifugal product is dried, dried product is obtained, which is denoted as ruthenium doped silicon dioxide (Ru@SiO2), dissolved Ru@SiO2 with ethanol to obtain Ru@SiO2 solution. The 3-aminopropyltriethoxysilane is added to Ru@SiO2 solution and stirred reaction, washed with ethanol, centrifuged and dried, to obtain powdered amino-functionalized Ru@SiO2 (NH2-Ru@SiO2). The NH2-Ru@SiO2 is dissolved in ultrapure water to obtain NH2-Ru@SiO2 solution, mixed with NGQDs, stirred reaction, dialyzed to obtain orange solution (NRS) solution, and stored at room temperature away from light. The glassy carbon electrode is treated with aluminum oxide powder of different particle sizes in sequence, and ultrasonically cleaned in water, ethanol, acetone and water and dried in order to obtain treated glassy carbon electrode. The NRS solution is added dropwise to surface of glassy carbon electrode (GCE), and dried at room temperature to obtain NRS/GCE. The chitosan solution is modified on surface of NRS/GCE sensor, and dried, at this time, layer of membrane structure formed on surface of sensor is marked as CS/NRS/GCE, according to known sequence of zearalenone (ZEN) aptamer and principle of base complementary pairing, complementary DNA (cDNA) of ZEN aptamer is designed. The cDNA is modified on surface of sensor, and electrostatic adsorption between cDNA and chitosan makes immobilized on surface of electrode. The sensor is rinsed with phosphate-buffered saline (PBS), sensor after rinsing is labeled as cDNA/CS/NRS/GCE. The bovine serum albumin (BSA) is modified on surface of electrode to seal non-specific binding sites on chitosan surface, and rinsed with PBS at same time to obtain BSA/cDNA/CS/NRS/GCE. The ZEN aptamer ZBA is modified on surface of sensor, and hybrid reaction with cDNA is used to form double-stranded DNA, rinsed with PBS to btain ZBA/BSA/cDNA/CS/NRS/GCE. The sensor is soaked in methylene blue MB solution, and rinsed with ultrapure water after soaking, obtained ratio aptamer sensor based on self-enhanced electrochemiluminescence-electrochemical dual method based on NRS and MB, labeled as MB/ZBA/BSA/cDNA/CS/NRS/GCE. The surface of sensor is modified with different concentrations of ZEN standard solutions. One concentration of ZEN solution corresponds to modification of sensor, and concentration and sensor are in one-to-one correspondence, incubated for period of time, rinsed with PBS to obtain ratio aptamer sensor based on dual signals of self-enhanced luminescent material and methylene blue at same time, labeled as ZEN/MB/ZBA/BSA/cDNA/CS/NRS/GCE.