Sensitive Immunochromatographic Assay Using Highly Luminescent Quantum Dot Nanobeads as Tracer for the Detection of Cyproheptadine Hydrochloride in Animal-Derived Food.

Pan Li,Beibei Liu,Sa Dong,Natalia Vasylieva,Qin Wu,Bruce D Hammock,Hanxiaoya Zhang,Cuifeng Yang,Cunzheng Zhang,Yulong Wang

doi:10.3389/fchem.2020.00575

Abstract

Cyproheptadine hydrochloride (CYP), used as human and veterinary drug, has been used illegally as feed additive for food-producing animals, which could remain in food and jeopardize human health. There is a need for on-site detection of CYP residue in animal-derived food. In this study, a hapten was designed, and a specific monoclonal antibody (mAb) was developed to detect CYP with an IC50 of 1.38 ng/mL and negligible cross-reactivity (CR) for other analogs. Forthermore, a high sensitive immunochromatographic assay (QBs-ICA) was developed using quantum dot nanobeads as reporters. The assay showed the linear detection range (IC20-IC80) of 0.03–0.52 ng/mL, the limit of detection (LOD) and visual detection limit (VDL) reached to 0.01 and 0.625 ng/mL, respectively. Spiked recovery study in pig urine and pork confirmed that the QBs-ICA was applicable for on-site testing. This assay showed better sensitivity and speedy than the reported instrumental analysis and immunoassays.

Highlights

Cyproheptadine hydrochloride (CYP), a derivative of cyproheptadine (CPH), is usually used as medicine to treat allergic disorders
The results indicated that the high specificity of QBsICA for CYP detection, which consistent with the results of cross-reactivity using ic-ELISA (Li et al, 2019)
Very few studies have been reported for the on-site detection of CYP residues in animal products

Summary

INTRODUCTION

Cyproheptadine hydrochloride (CYP), a derivative of cyproheptadine (CPH), is usually used as medicine to treat allergic disorders. After clenbuterol was banned for use in China, CYP and other derivatives of CPH have been frequently found in meat and dairy products, which may trigger allergic reactions or other sicknesses for the consumers (Yang et al, 2015; Kaur et al, 2018) Considering their risks for human health, CPH and CYP are in the “List of Prohibited Drugs Used in Food-producing Animals” as stated in Directive 2001/82/EC (2009). There are inescapable realities that most instrumental methods are time-consuming and labor-intensive, which limit their use in on-site analysis or high-throughput screening for large numbers of samples. As complementary, immunoassays such as ELISAs are regarded as effective alternatives in terms of simplicity, speed and efficiency. This study provided a portable, user friendly, on-site detection approach which showed better sensitivity and speedy than the reported instrumental analysis and immunoassays (Fente et al, 2009; Yang et al, 2015; Guo et al, 2018)

MATERIALS AND METHODS

Assay Procedure

RESULTS AND DISCUSSION

Method Validation by Spiked Samples

CONCLUSIONS

ETHICS STATEMENT