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<p>Fluorescent labeling of long-chain inulin (FXL) is essential for tracking its spatiotemporal dynamics in gastrointestinal systems, yet conventional methods risk altering its native properties. This study systematically compared three labeling strategies: FITC-DB (dibutyltin dilaurate-catalyzed hydroxyl conjugation), FITC-RA (reductive amination), and FA (fluoresceinamine coupling). Multimodal characterization revealed that FITC-DB achieved the highest substitution degree (DS = 0.42 at 50&#xa0;mg FITC), though fluorescence quenching occurred at elevated loadings. FITC-RA induced significant aggregation (peak size at 50&#xa0;mg), while FA labeling exhibited minimal structural perturbation but low efficiency (DS &lt; 0.20). Spectroscopic analyses confirmed covalent thiourea bond formation (1720&#xa0;cm⁻¹ FTIR peak) and aggregation-dependent optical shifts. Crucially, FITC-DB at 50&#xa0;mg balanced optimal labeling efficiency, colloidal stability (highest zeta potential), and preserved macromolecular architecture. These findings establish design principles for high-fidelity FXL probes in biological tracking applications.</p>

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Multimodal characterization of fluorescently tagged inulin: FITC-based and fluoresceinamine-based approaches

  • 期刊论文

  • 发表日期：

    2026年5月4日

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  • Jiahao Wan,
  • Yanbin Wang,
  • Wenhui Deng,
  • Caijun Liu,
  • Menglong Liu,
  • Denglin Luo,
  • Kangyi Zhang,
  • Haiyan Gao,
  • Zhouya Bai

摘要

Fluorescent labeling of long-chain inulin (FXL) is essential for tracking its spatiotemporal dynamics in gastrointestinal systems, yet conventional methods risk altering its native properties. This study systematically compared three labeling strategies: FITC-DB (dibutyltin dilaurate-catalyzed hydroxyl conjugation), FITC-RA (reductive amination), and FA (fluoresceinamine coupling). Multimodal characterization revealed that FITC-DB achieved the highest substitution degree (DS = 0.42 at 50 mg FITC), though fluorescence quenching occurred at elevated loadings. FITC-RA induced significant aggregation (peak size at 50 mg), while FA labeling exhibited minimal structural perturbation but low efficiency (DS < 0.20). Spectroscopic analyses confirmed covalent thiourea bond formation (1720 cm⁻¹ FTIR peak) and aggregation-dependent optical shifts. Crucially, FITC-DB at 50 mg balanced optimal labeling efficiency, colloidal stability (highest zeta potential), and preserved macromolecular architecture. These findings establish design principles for high-fidelity FXL probes in biological tracking applications.