Design and characterization of a direct ELISA for the detection and quantification of leucomalachite green

Figures & data

Figure 1. Structure of leucomalachite green (LMG), malachite green (MG) and structurally similar compounds, leucocrystal violet (LCV), crystal violet (CV) and paraosaniline, used to determine cross-reactivity.

Figure 2. Synthetic scheme for the synthesis of LMG hapten and subsequent formation of immunogen by conjugation to a carrier protein (BSA).

Figure 3. Synthetic scheme for the conjugation of LMG hapten to HRP.

Figure 4. Representative calibration curve for LMG prepared in blank tilapia extract.

Table 1. Calibration curve results over a 3-day period with tilapia extract IC₅₀, IC₈₀ and IC₉₀ in ng (LMG) g⁻¹ tissue.

	IC50	IC80	IC90
Analysis	ng (LMG) g^−1 tissue	ng (LMG) g^−1 tissue	ng (LMG) g^−1 tissue
Day 1	1.55	0.24	0.08
Day 2	2.56	0.40	0.14
Day 3	1.82	0.31	0.11
Average	1.98	0.32	0.11
% CV	27	25	24

Figure 5. Calibration curves used to determine the cross-reactivity for LMG (•), MG (▪), LCV (♦), CV (□) and PS (▽).