Fluorescence-based immunochromatographic test strip for the detection of hyoscyamine

Abstract

Hyoscyamine (HSM), which acts as an antagonist of the acetylcholine muscarinic receptor and can induce a variety of distinct toxic syndromes in mammals (anti-cholinergic poisoning), is hazardous to human health. Therefore, it is urgent to develop a rapid, sensitive, and cost-effective method to determine HSM. A fluorescent microsphere based immunochromatographic assay was developed for this analyte and gold nanoparticles (AuNPs) were used as a comparison. A monoclonal antibody against HSM was prepared with a 50% inhibition concentration (IC₅₀) of 1.17 ng mL⁻¹, with no cross-reactivity with five drugs. Under optimized conditions, the cut off limits using the fluorescence-labeled monoclonal antibody strips were 10 ng mL⁻¹ in 0.01 M PBS and 20 ng mL⁻¹ in pork, pig urine, and honey samples, and the assay could be completed within 10 min. In comparison with a AuNP immunochromatographic assay, the developed method offered a higher coupling rate and lower amounts of antibodies. This approach could be used for simple, sensitive and rapid screening, and is suitable for on-site screening applications.