State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu 214122, People's Republic of China.
Wuxi No. 2 People's Hospital, Wuxi, 214002, Jiangsu, 214122, People's Republic of China.
Analyst. 2022 Jan 17;147(2):293-302. doi: 10.1039/d1an01973b.
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 (IC) of 1.17 ng mL, 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 in 0.01 M PBS and 20 ng mL 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.
莨菪碱（HSM）作为乙酰胆碱毒蕈碱受体的拮抗剂，可在哺乳动物中诱导多种不同的毒性综合征（抗胆碱能中毒），对人类健康有害。因此，迫切需要开发一种快速、灵敏、经济有效的方法来测定 HSM。为此，开发了一种基于荧光微球的免疫层析分析方法，并将纳米金（AuNPs）作为比较。制备了一种针对 HSM 的单克隆抗体，其 50%抑制浓度（IC）为 1.17ng mL，与五种药物无交叉反应。在优化条件下，使用荧光标记的单克隆抗体条的截止值在 0.01 M PBS 中为 10ng mL，在猪肉、猪尿和蜂蜜样品中为 20ng mL，检测时间可在 10min 内完成。与 AuNP 免疫层析分析相比，该方法具有更高的偶联率和更低的抗体用量。该方法可用于简单、灵敏、快速的筛选，适用于现场筛选应用。
