A Rapid Tricolour Immunochromatographic Assay for Simultaneous Detection of Tricaine and Malachite Green

In this research, we designed a rapid tricolour immunochromatographic test strip with double test lines (TS-DTL) and two-colour AuNP probes, which realised the simultaneous detection of tricaine mesylate (TMS) and malachite green (MG). Through a distinct tricolour system (red T-1 line, blue T-2 line and purple C line), a visual identification of TMS (0.2 mu g/mL) and MG (0.5 mu g/mL) was quickly achieved on site, which improved the accuracy of naked eye observations. The LODs of TMS in aquaculture water, fish and shrimp were 11.0, 29.6 and 61.4 ng/mL, respectively. MG LODs were 47.0 ng/mL (aquaculture water), 82.8 ng/mL (fish) and 152.4 ng/mL (shrimp). The LOD of MG was close to the similar TS methods. However, visual detection of TMS could meet the requirements of the residue limit (1 mu g/mL) of TMS in the USA, and the quantitative detection of TMS was over 16 times lower than the USA standard. The developed platform was rapid (similar to 20 min, HPLC similar to 3 h) and accurate, which was verified using a traditional HPLC method. The recovery rates ranged from 82.2% to 108.6% in three types of real samples, indicating a potential application in on-site fast screening or multiple detection for TMS and MG residues in aquatic products.