College of Chemistry, Nanchang University, Nanchang, Jiangxi, P. R. China.
Jiangxi Province Key Laboratory of Modern Analytical Science, Nanchang University, Nanchang, Jiangxi, P. R. China.
J Environ Sci Health B. 2020;55(3):250-256. doi: 10.1080/03601234.2019.1685319. Epub 2019 Nov 6.
Paclobutrazol (PBZ) is a plant growth regulator (PGR) widely used in fruit and vegetable cultivation. However, due to the severe toxicity of PBZ, a sub-ppm level maximum residue limit (MRL) was established worldwide. Therefore, it is significant to propose a rapid, sensitive and high throughput screening method for monitoring the PBZ residues in foods. In this study, a simple and sensitive indirect competitive Enzyme-linked immunosorbent assay (icELISA) was established for PBZ detection in fruits basing polyclonal antibody. For both economy and pollution prevention, a microwave-solvent-free method was used to synthesize the PBZ hapten with high efficiency. The detection conditions, such as coating antigen concentration, antibody concentration, organic reagent concentration, ionic strength and pH, were optimized. Under the optimized conditions, this method showed high sensitivity and specificity. The detection range is 1.27-138.23 ng/mL, half-maximum inhibition concentration (IC) is 13.26 ng/mL, and the IC was lower than the reported ELISAs for PBZ. Additionally, this method had high accuracy and precision. The recoveries were ranged from 88.78% to 96.80% in PBZ spiked apple samples with RSD below 4%. All the results showed that the polyclonal antibody based icELISA could be useful for PBZ screening in fruit samples.
多效唑（PBZ）是一种广泛应用于水果和蔬菜种植的植物生长调节剂（PGR）。然而，由于 PBZ 的毒性很强，世界范围内都制定了亚 ppm 级别的最大残留限量（MRL）。因此，提出一种快速、灵敏、高通量的方法来监测食品中的 PBZ 残留量具有重要意义。本研究基于多克隆抗体，建立了一种简单、灵敏的间接竞争酶联免疫吸附测定法（icELISA）用于检测水果中的 PBZ。为了经济和防止污染，采用微波无溶剂法高效合成了 PBZ 半抗原。优化了包被抗原浓度、抗体浓度、有机溶剂浓度、离子强度和 pH 等检测条件。在优化条件下，该方法具有较高的灵敏度和特异性。检测范围为 1.27-138.23ng/mL，半数抑制浓度（IC）为 13.26ng/mL，IC 低于已报道的 PBZ ELISA。此外，该方法具有较高的准确性和精密度。在苹果样品中添加 PBZ 的回收率在 88.78%至 96.80%之间，相对标准偏差（RSD）低于 4%。所有结果表明，基于多克隆抗体的 icELISA 可用于水果样品中 PBZ 的筛选。
