College of Food Science, Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou, China.
College of Chemistry and Chemical Engineering, Guangxi University for Nationalities, Nanning, China.
J Food Sci. 2022 Mar;87(3):1342-1354. doi: 10.1111/1750-3841.16078. Epub 2022 Feb 15.
The tadalafil-like compounds have appeared recently as adulterants in drinks and healthcare dietary supplements sourced from medicinal and edible food, which may cause illness and even death. In this work, the rationality of haptens was explored by computational chemistry and molecular simulation theories such as frontier molecular orbital (FMO)-based softness (S), three-dimensional (3D) structure, surface electrostatic potential (ESP), and lipophilic potential (LP). An antiserum from hapten H5 with the highest softness and maintaining the appropriate three-dimensional (3D) structure showed the optimal immunoassay performance, indicating an increasing softness was a critical factor for effective hapten. Based on the antibody induced by hapten H5, an indirect competitive enzyme-linked immunosorbent assay (icELISA) method for detecting multiple tadalafil-like adulterants was established. The icELISA showed a limit of detection (LOD), 50% inhibition concentration (IC ), and a working range of 0.004-0.396, 0.89-4.27, and 0.094-16.71 ng/ml for tadalafil, amino tadalafil, acetamino tadalafil, nortadalafil, and N-desmethyl ent-tadalafil, respectively. The spiked recoveries of tadalafil-like adulterants in samples ranged from 84.9% to 116.2%. The results of the icELISA and HPLC-MS/MS methods had a good correlation for real samples with the R of 0.9955. Specially, this work not only provided a convenient immunoassay method for measuring tadalafil-like adulterants in spirit drinks and dietary supplements in group-screening manner, but also suggested that softness was likely to be a general theory for rational hapten design. PRACTICAL APPLICATION: Rapid monitoring of tadalafil-like adulterants in food samples is very necessary and important for consumers, regulatory agencies, and the food industry.
最近，他达拉非类化合物作为掺杂物出现在饮料和保健品中，这些保健品来源于药用和食用食品，可能导致疾病甚至死亡。在这项工作中，通过计算化学和分子模拟理论，如基于前沿分子轨道（FMO）的软度（S）、三维（3D）结构、表面静电势（ESP）和疏水性势（LP），探讨了半抗原的合理性。半抗原 H5 具有最高的软度并保持适当的三维（3D）结构，其产生的抗血清显示出最佳的免疫分析性能，表明增加软度是有效半抗原的关键因素。基于半抗原 H5 诱导的抗体，建立了一种用于检测多种他达拉非类掺杂物的间接竞争酶联免疫吸附测定（icELISA）方法。icELISA 的检测限（LOD）、50%抑制浓度（IC ）和工作范围分别为 0.004-0.396、0.89-4.27 和 0.094-16.71ng/ml 用于他达拉非、氨基他达拉非、乙酰氨基他达拉非、诺他达拉非和 N-去甲基恩他达拉非。在样品中，他达拉非类掺杂物的加标回收率在 84.9%至 116.2%之间。icELISA 和 HPLC-MS/MS 方法对实际样品的检测结果具有良好的相关性，相关系数为 0.9955。特别地，这项工作不仅为以组筛方式测量酒精饮料和保健品中他达拉非类掺杂物提供了一种方便的免疫分析方法，而且还表明软度可能是合理半抗原设计的一般理论。 实际应用：快速监测食品样品中的他达拉非类掺杂物对消费者、监管机构和食品行业非常必要和重要。
