College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China.
Food Chem. 2016 Sep 15;207:233-8. doi: 10.1016/j.foodchem.2016.03.103. Epub 2016 Mar 30.
6-Benzylaminopurine (6-BA), a cytokinin plant growth regulator, has been banned for use in bean sprout production in China. An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed with a specific monoclonal antibody (mAb 3E5). The assay showed a half-maximum inhibition concentration (IC50) and detection range of 18.9 ng/mL and 3.6-106 ng/mL, respectively. Recoveries of 6-BA spiked in home cultured bean sprout samples averaged from 75% to 89% with a correlation coefficient (R(2)) of 0.998 between the results determined by icELISA and those by liquid chromatography-electrospray ionization quadrupole Orbitrap mass spectrometry (LC-ESI-MS). LC-ESI-MS showed that 6-BA had been partially metabolized to 6-benzylaminopurine riboside (6-BAR) in the positive samples. The content of 6-BA determined by icELISA was about 5-70 times higher than that of LC-ESI-MS because mAb 3E5 had 315% cross-reactivity with 6-BAR. Such icELISA being ultra-sensitive to 6-BAR would allow quick monitoring of 6-BA by detecting 6-BAR as a potential biomarker.
6-苄氨基嘌呤（6-BA）是一种细胞分裂素植物生长调节剂，已在中国被禁止用于豆芽生产。本研究利用特异性单克隆抗体（mAb 3E5）建立了间接竞争酶联免疫吸附法（icELISA）。该方法的半抑制浓度（IC50）和检测范围分别为 18.9ng/mL 和 3.6-106ng/mL。在家庭培养的豆芽样品中添加 6-BA 的回收率平均为 75%-89%，icELISA 法与液相色谱-电喷雾-四极杆轨道阱质谱法（LC-ESI-MS）法测定结果的相关系数（R2）为 0.998。LC-ESI-MS 结果表明，阳性样品中 6-BA 已部分代谢为 6-苄氨基嘌呤核苷（6-BAR）。icELISA 法测定的 6-BA 含量约为 LC-ESI-MS 法的 5-70 倍，因为 mAb 3E5 与 6-BAR 的交叉反应率为 315%。icELISA 对 6-BAR 的超高灵敏度可以通过检测 6-BAR 作为潜在的生物标志物来快速监测 6-BA。
