Sensitive and High-Throughput Time-Resolved Luminescence Detection of Tetracycline in Milk for Eliminating Background Fluorescence on a Miniaturized Apparatus

Fluorescence detection has always suffered from high background fluorescence from real samples such as milk. Therefore, cumbersome pretreatments of samples were necessary to remove the fluorescent substances but led to long processing times and low efficiency. Time-resolved luminescence detection is a powerful technique for eliminating short-lived background fluorescence without additional pretreatments. However, the related instruments are usually equipped with high-speed excitation sources and detectors, which are always bulky and expensive. Herein, we developed a low-cost and miniaturized imaging system for high-throughput time-gated luminescence detection. An UV LED array was used to excite multiple samples, the luminescence of which could be detected by a smartphone simultaneously. An analog circuit was designed to synchronize the LED to the mechanical chopper to eliminate the background signals resulting from scattering and short-lived autofluorescence. Compared to other synchronous circuits based on FPGAs and microcontrollers, this analog circuit required no programming and memory. For the first time, high-throughput time-resolved luminescence detection of tetracycline in milk without any separation or enrichment was achieved by utilizing a smartphone as a camera, and the scattered signals and the background fluorescence were eliminated efficiently. The limit of detection reached as low as 53 nM (∼0.024 ppm), lower than the residue limit set by the European Union. This high-throughput time-gated luminescence detection method can be used for quantitative analysis of many real samples with high background fluorescence.