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

Abstract

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 T1 line, blue T2 line and purple C line), a visual identification of TMS (0.2 μg/mL) and MG (0.5 μg/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 μg/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 (~20 min, HPLC~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.

The Developments on Lateral Flow Immunochromatographic Assay for Food Safety in Recent 10 Years: A Review

“…Synthetic pigments, such as Sudan red and malachite green, improve the color brightness of food. As shown in Table 4, at present, colloidal gold test strips are the main method for detecting these two substances, with LODs of 10.0 ng/mL and 0.5 ng/mL, respectively [68][69][70]. Clenbuterol is used to suppress fat production in animals and increase meat yield, but excessive usage can damage the human body.…”

A Novel RAA Combined Test Strip Method Based on Dual Gene Targets for Pathogenic Vibrio vulnificus in Aquatic Products

“…Multi-branched gold nanoparticles (MBGNP) were prepared by the method described by Lu et al [14] with slight modifications. Briefly, 50 µL of a 10% chloroauric acid solution and 50 mL of ultrapure water were heated to boiling in a water bath with vigorous stirring.…”

“…In line with the method described by Lu et al [14], the size and morphology of the MBGNP were measured with a high-resolution transmission electron microscope (TEM; H-7800, Tokyo, Japan). The absorption peaks of the MBGNP were observed by a UV-visible spectrophotometer, and the hydrodynamic size distribution of the MBGNP was analyzed by scientific dynamic light scattering (DLS).…”

Ratiometric Fluorescence Immunoassay Based on MnO2 Nanoflakes for Sensitive and Accurate Detection of Tricaine

“…Immunoassays are efficient tools for monitoring chemical residues and complement instrumental analytical approaches well by virtue of their ease to use, good selectivity, high throughput, and cost-effectiveness. − Up to now, two immunochromatographic assays (ICAs) for tricaine have been reported. , In these works, monoclonal antibodies (Mabs) against tricaine were induced by using 3-aminobenzoic acid (termed as TMS2 in this work) such as the hapten, which directly conjugates with carrier proteins via the carbodiimide method to produce an immunogen. As a result, gold nanoparticle-based ICAs were developed; , however, the classic ICA can only qualitatively detect tricaine, which cannot obtain accurate quantitative results. In contrast, enzyme-linked immunosorbent assay (ELISA), also as a classic immunoassay format, can deliver quantitative results with high throughput. − Since production of high-affinity antibodies against small molecules is typically challenging because of their tiny size and simple structure and the absence of strong epitope groups, − traditional colorimetric ELISA for small molecules usually suffers from insufficient sensitivity for practical application .…”

“…Immunoassays are efficient tools for monitoring chemical residues and complement instrumental analytical approaches well by virtue of their ease to use, good selectivity, high throughput, and cost-effectiveness. − Up to now, two immunochromatographic assays (ICAs) for tricaine have been reported. , In these works, monoclonal antibodies (Mabs) against tricaine were induced by using 3-aminobenzoic acid (termed as TMS2 in this work) such as the hapten, which directly conjugates with carrier proteins via the carbodiimide method to produce an immunogen. As a result, gold nanoparticle-based ICAs were developed; , however, the classic ICA can only qualitatively detect tricaine, which cannot obtain accurate quantitative results.…”