保健酒与口服液中他达拉非类药物微孔侧流免疫层析检测方法研究

Detection of Tadalafil and Its Analogues in Health Wine and Oral Liquid by Microwell Lateral Flow Immunochromatography Assay

采用共有骨架结构结合不饱和线性手臂半抗原策略,通过免疫新西兰大白兔获得了可同时识别他达拉非、氨基他达拉非和去甲基他达拉非的特异性多克隆抗体,通过优化反应条件,建立了微孔侧流免疫层析方法,用于保健酒及口服液样品中非法添加的他达拉非类药物残留的快速检测。本方法定量检出限为0.05~0.30 ng/mL,裸眼消线值在50~100 ng/mL之间,检测时间小于10 min;与其它结构功能类似物无明显交叉反应,特异性良好。对保健酒及口服液样品的添加回收率在78.7%~117.8%之间,相对标准偏差小于15%。本方法检测结果与HPLC-MS/MS法检测结果一致,适用于保健酒及口服液中他达拉非类药物的快速筛查。

To monitor illegally added Tadalafil and its analogues in health food,polyclonal antibody was obtained by immunizing New Zealand white rabbits with a common skeleton structure hapten combined with an unsaturated linear alkyl arm hapten strategy,which could simultaneously recognize Tadalafil,Amino Tadalafil and N-Desmethyl Tadalafil.A microwell lateral flow immunochromatographic assay(mw LFIA)combined with simple sample pretreatment was developed.The LOD of mw LFIA for three Tadalafil drugs was between 0.05 and 0.30 ng/mL,the cut-off values were between 50 and 100 ng/mL,and the detection time was less than 10 min.Cross-reactivity test showed no interference from substances with similar structures,indicating a good specificity of the method against Tadalafil,Amino Tadalafil and N-Desmethyl Tadalafil.The average recoveries of three drugs in spiked health wine and oral liquid ranged from 78.7%to 117.8%with RSDs less than 15%,which had a good aggrement with the results of HPLC-MS/MS.The developed mw LFIA strip was suitable for the rapid detection of Tadalafil and its analogues in health wine and oral liquid samples.