Scholarlink

<p>Traditional fluorescence polarization immunoassay (FPIA) requires costly monoclonal antibodies. This work develops a cost-effective fluorescence polarization enzyme assay (FPEA) for quercetin, employing α-amylase as a significantly cheaper recognition agent. Based on competitive binding with a fluorescent zearalenone tracer, the assay achieves a detection limit of 1.7 mg/mL, a working range of 2.3–6.4 mg/mL, and completes analysis within 5 min, offering a substantial time saving versus typical HPLC runs (~ 20 min). It demonstrates high specificity, with only minimal cross-reactivity to rutin (0.1%) and none to dihydroquercetin, gallic acid, and acarbose. FPEA analysis yielded a quercetin*α-amylase binding affinity constant of 3.9 × 10<sup>3</sup> L/mol. Successful application to onion peel extract and a dietary supplement yielded results consistent with liquid chromatography with diode-array detection (LC-DAD). The proposed FPEA provides a rapid, simple, and economical alternative for quality control of sources with high quercetin content.</p> Graphical abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Fluorescence polarization enzyme assay: a novel approach for the quantification of quercetin based on its interaction with α-amylase

  • 期刊论文

  • 发表日期：

    2026年4月22日

查看全文
  • Svetlana M. Filimonova,
  • Evgeniy S. Melnikov,
  • Liliya I. Mukhametova,
  • Anna Raysyan,
  • Sergei A. Eremin

摘要

Traditional fluorescence polarization immunoassay (FPIA) requires costly monoclonal antibodies. This work develops a cost-effective fluorescence polarization enzyme assay (FPEA) for quercetin, employing α-amylase as a significantly cheaper recognition agent. Based on competitive binding with a fluorescent zearalenone tracer, the assay achieves a detection limit of 1.7 mg/mL, a working range of 2.3–6.4 mg/mL, and completes analysis within 5 min, offering a substantial time saving versus typical HPLC runs (~ 20 min). It demonstrates high specificity, with only minimal cross-reactivity to rutin (0.1%) and none to dihydroquercetin, gallic acid, and acarbose. FPEA analysis yielded a quercetin*α-amylase binding affinity constant of 3.9 × 103 L/mol. Successful application to onion peel extract and a dietary supplement yielded results consistent with liquid chromatography with diode-array detection (LC-DAD). The proposed FPEA provides a rapid, simple, and economical alternative for quality control of sources with high quercetin content.

Graphical abstract