Fluorescent Probe Development for Rapid Detection of Tiletamine Based on Copper Nanozyme and Molecular Imprinting Technology

doi:10.12116/j.issn.1004-5619.2025.350410

Abstract

Abstract:

Objective To develop a rapid detection method for tiletamine that is easy to operate and low-cost under the premise of ensuring sensitivity and accuracy, to assist in carrying out rapid screening and drug control work on-site. Methods This study integrates dual-ligand copper nanozymes with molecular imprinting technology. Initially, copper nanozymes were synthesized using readily available raw materials at 120 ℃. Subsequently, specific cavities were imprinted on their surface at room temperature using a sol-gel method to construct a novel fluorescent sensing probe. This probe was characterized and methodologically validated, and then applied to the detection of actual samples. Results The developed probe exhibited stable fluorescence properties, strong anti-interference capability, and excellent specificity and sensitivity, with a detection limit of 5 ng/mL and a quantitative concentration range from 15 to 500 ng/mL. It enabled the rapid detection of tiletamine in real samples such as blood and e-cigarette oil. Conclusion This fluorescent probe can be used for rapid detection and on-site preliminary screening of tiletamine in various types of samples. It significantly improves the detection efficiency and reduces analysis costs, showing high research value and broad application prospects.

Key words: forensic medicine, toxicological analysis, tiletamine, copper nanozyme, molecular imprinted polymer, fluorescent probe, rapid detection

CLC Number:

Jia-hao LI, Jiang LING, Zi-hao CAI, Zi-yuan ZHENG, Yan-jun DING. Fluorescent Probe Development for Rapid Detection of Tiletamine Based on Copper Nanozyme and Molecular Imprinting Technology[J]. Journal of Forensic Medicine, 2025, 41(4): 355-363.

Figures/Tables 11

Fig. 1 Comparison of the fluorescence spectra amongCu-BH， Cu-BH@MIPs， and Cu-BH@MIPscombined with tiletamine

Fig. 2 Comparison of the ultraviolet absorption spectrumof tiletamine and the emission spectrum ofthe Cu-BH@MIPs probe

Fig. 3 Effects of different reaction times on the fluorescenceintensity of the Cu-BH@MIPs probe

Fig. 4 Microscopy images of the Cu-BH@MIPs probe

Fig. 5 Comparison of the Fourier transform infraredspectrum of the Cu-BH@MIPs probe and thenon-imprinted polymer Cu-BH@NIPs

Fig. 6 The fluorescence intensity changes ofthe Cu-BH@MIPs probe within 14 daysafter binding with tiletamine

Fig. 7 Fluorescence intensities of the Cu-BH@MIPs probeafter reacting with different psychoactive substances

Fig. 8 Fluorescence spectra of the Cu-BH@MIPs probe afterreacting with tiletamine at different mass concentrations

Fig. 9 Linear fitting for the mass concentration andfluorescence intensity of tiletamine

Tab. 1 Detection of tiletamine in e-cigarette oil and blood samples using the Cu-BH@MIPs probe

加标样本	添加值/（ng·mL^-1）	电子烟油				血液
加标样本	添加值/（ng·mL^-1）	检测值/（ng·mL^-1）	回收率/%	日内差（n=3）	日间差（n=9）	检测值/（ng·mL^-1）	回收率/%	日内差（n=3）	日间差（n=9）
1	100	96	96.0	4.76	5.05	93	93.0	5.01	5.29
2	200	205	102.5	3.62	3.55	189	94.5	4.66	4.95
3	500	493	98.6	3.11	3.31	484	96.8	2.79	3.22

Tab. 2 Comparison of the Cu-BH@MIPs probe methodand the LC-MS/MS method for the detection oftiletamine in actual blood samples（ng/mL）

样本	Cu-BH@MIPs	LC-MS/MS
实际样本1	347	370
实际样本2	未检出	8
空白血样	未检出	未检出

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