Development and validation of a TRFI-based rapid quantitative test strip featuring a broad quantification range for...

Area of Science:

• Food Science and Technology
• Analytical Chemistry
• Mycotoxin Detection

Background:

• Zearalenone (ZEN) contamination in grains and feeds poses a significant food safety risk.
• Existing detection methods for ZEN present limitations in detection range and field applicability.
• Conventional methods like immunochromatographic strips have narrow linearity, while LC-MS/MS is time-consuming.

Purpose of the Study:

• To develop a rapid, accurate, and field-applicable test strip for Zearalenone (ZEN) quantification.
• To overcome the limitations of existing ZEN detection methods, particularly in terms of speed and detection range.
• To provide a cost-effective pre-screening tool for on-site grain analysis.

Main Methods:

• Development of a time-resolved fluorescence immunochromatographic (TRFI) test strip.
• Optimization of fluorescent microspheres and sample dilution to minimize matrix interference and Hook effect.
• Implementation of dynamic reaction control for rapid quantification within 6 minutes.

Main Results:

• The TRFI-ZEN strip achieved a broad quantification range of 20-2500 μg/kg with high accuracy (91-102% recovery) in maize.
• Excellent stability was demonstrated with <4% signal variation after 60 days at 37°C.
• Field validation showed a high correlation (0.9987) with LC-MS/MS, accurately detecting a 94.44% contamination rate.

Conclusions:

• The developed TRFI strip offers a rapid (<15 min/sample) and reliable method for ZEN quantification in grains.
• It meets safety standards in China and the EU, covering a wide range of contamination levels.
• This method significantly reduces the need for costly LC-MS/MS confirmations, offering >80% cost savings through high-confidence pre-screening.