Aggregation-induced emission-based competitive lateral flow immunoassay for rapid detection of sulfamethazine in honey

Figures & data

Figure 1. Schematic of the AIEFM-LFIA on the basis of competitive format immunoassay principle to analyse SMZ.

Figure 2. Characterization of AIEFM and AIEFM-mAb. Hydration size of AIEFM and AIEFM-mAb (A). Fluorescence spectrum of AIEFM (B). Digital photos of AIEFM under natural light (C). Digital photos of AIEFM under UV light (D).

Figure 3. Optimization of the key factors of the AIEFM-LFIA test strips. Optimization of the coupling pH (A), usage volume of the EDC (B), amount of mAb to prepare the AIEFM-mAb label (C), concentration of SMZ-BSA on the T line (D), amount of the AIEFM-mAb label (E), and immunological kinetic analysis of the AIEFM-LFIA (F).

Table 1. Sensitivity and linear range of the AIEFM-LFIA in PBS and honey.

Method	Sample	Limit of detection (ng/mL)	Linear range (ng/mL)
AIEFM-LFIA	PBS	0.053	0.05–10
AIEFM-LFIA	Honey	0.028	0.05–5

Figure 4. Specificity of AIEFM-LFIA. The specificity of the proposed AIEFM-LFIA was assessed by testing 11 SAs and three non-SA antibiotic drugs (ENR, FLO, and TET).

Figure 5. Standard calibration curves of the AIEFM-LFIA in honey were obtained by determining the FI_T/FI_T0 against the logarithm of the concentration of the SMZ.

Table 2. Comparison of the proposed AIEFM-LFIA and other methods for SA detection in real samples.

Methods	Target	Detection samples	Linear range (ng/mL)	LOD (ng/mL)	Ref.
LFIA	Sulfamethazine	Egg	–	2.0	Shim, Kim, Kim, & Chung, (2013)
NIR-LFIA	Sulfadimidine	Milk	0.1–3.98	0.1	Chen et al., (2016)
CG-LFIA	Sulfamethazine	Beef	0.04–6	0.165	Liu et al., (2018)
Electrochemical	Sulfadimidine	Pork	0.1–50	0.055	He and Du (2018)
Chemiluminescence	Sulfamethazine	buffer	1.85–21.57	0.92	Yang et al., (2019)
MI-ELISA	Sulfamethazine	Swine muscle	100−3200	6.8 (μg/kg)	Peng et al., (2017)
AIEFM-LFIA	Sulfamethazine	Honey	0.05–5	0.028	This work

Table 3. Recovery of SMZ on the AIEFM-LFIA in PBS and honey.

Sample		Spiked SMZ (ng/mL)	Recovery (%)	SD	CV (%)
PBS		0.25	95.8	6.4	6.7
		2.5	105.1	9.3	8.8
		5.0	104.8	10.5	10.0
Honey	Sample 1	0.2	75.6	6.9	9.1
		1.0	89.3	5.1	5.7
		3.0	95.1	9.2	9.7
	Sample 2	0.2	76.3	7.9	10.4
		1.0	91.1	6.3	6.9
		3.0	94.7	10.2	10.8
	Sample 3	0.2	79.5	7.2	9.1
		1.0	90.7	8.7	9.6
		3.0	91.4	11.8	12.9

Table 4. Detection of SMZ in 15 real samples using the AIEFM-LFIA and HPLC-MS/MS.

Sample		The proposed AIE-LFIA	HPLC-MS/MS
Honey	1	N.D.^a	N.D.
	2	N.D.	N.D.
	3	N.D.	N.D.
	4	N.D.	N.D.
	5	N.D.	N.D.
	6	N.D.	N.D.
	7	N.D.	N.D.
	8	N.D.	N.D.
	9	N.D.	N.D.
	10	N.D.	N.D.
	11	4.8 ng/mL	6.1 ng/mL
	12	N.D.	N.D.
	13	N.D.	N.D.
	14	N.D.	N.D.
	15	N.D.	N.D.

^aNot detected.

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