Development of an immunochromatographic assay for rapid detection of clorprenaline in pig urine

Figures & data

Figure 1. The synthesis of CLP-hapten.

Figure 2. LC-MS analysis of CLP-hapten.

Figure 3. Polyacrylamide gel electrophoresis image of immunogen (a) (1 = BSA; 2/3 = M_CLP-hapten : M_BSA (80:1;120:1)) and coating antigens (b) of CLP (1 = M_CLP-hapten: M_OVA (30:1); 2 = OVA).

Figure 4. The standard curve of developed Ic-ELISA method. IC50 of 4G1 was 0.095 ng/mL, LOD was 0.013 ng/mL and the linear range of detection was from 0.0258 to 0.356 ng/mL.

Table 1. Cross-reaction results of McAb 4G1.

Chemicals	IC50 (ng/mL)	CR (%)
CLP	0.434	100.0
Salbutamol	>50.00	<1.0
Clenbuterol	>50.00	<1.0
Ractopamine	>50.00	<1.0
Cimaterol	>50.00	<1.0
Brombuterol	>50.00	<1.0
Formoterol	>50.00	<1.0
Mabuterol	>50.00	<1.0
Terbutaline	>50.00	<1.0

Figure 5. Image of detection by clorprenaline immunochromatographic strip in urine. A series of urine samples spiked clorprenaline were subjected to the immunochromatographic strip test. 1 = 0 ng/mL, 2 = 0.1 ng/mL, 3= 0.25 ng/mL, 4 = 0.5 ng/mL, 5= 1 ng/mL, 6 = 2.5 ng/mL, 7 = 0.5 ng/mL, 8 = 10 ng/mL. Cut-off value was 10 ng/mL.

Table 2. Recovery of clorprenaline in urine by ic-ELISA and strip assay (n = 3).

Samples	Spiked level (ng/mL)	Detection level (ng/mL), mean ± SD	Recovery rate (%)
Swine urine	0.3	0.289 ± 0.055	96.3
	0.6	0.604 ± 0.006	100.5
	1.2	0.981 ± 0.051	81.8