Development of an Ultrasensitive and Rapid Fluorescence Polarization Immunoassay for Ochratoxin A in Rice.

Xiaorong Huang,Jun Jiang,Abdoulie Jallow,Xiaoqian Tang,Peiwu Li,Wen Zhang,Hui Li,Xin Qi,Qi Zhang

doi:10.3390/toxins12110682

Xiaorong Huang, Jun Jiang + Show 7 more

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https://doi.org/10.3390/toxins12110682

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Ochratoxin A (OTA) is a known food contaminant that affects a wide range of food and agricultural products. The presence of this fungal metabolite in foods poses a threat to human health. Therefore, various detection and quantification methods have been developed to determine its presence in foods. Herein, we describe a rapid and ultrasensitive tracer-based fluorescence polarization immunoassay (FPIA) for the detection of OTA in rice samples. Four fluorescent tracers OTA-fluorescein thiocarbamoyl ethylenediamine (EDF), OTA-fluorescein thiocarbamoyl butane diamine (BDF), OTA-amino-methyl fluorescein (AMF), and OTA-fluorescein thiocarbamoyl hexame (HDF) with fluorescence polarization values (δFP = FPbind-FPfree) of 5, 100, 207, and 80 mP, respectively, were synthesized. The tracer with the highest δFP value (OTA-AMF) was selected and further optimized for the development of an ultrasensitive FPIA with a detection range of 0.03–0.78 ng/mL. A mean recovery of 70.0% to 110.0% was obtained from spiked rice samples with a relative standard deviation of equal to or less than 20%. Good correlations (r2 = 0.9966) were observed between OTA levels in contaminated rice samples obtained by the FPIA method and high-performance liquid chromatography (HPLC) as a reference method. The rapidity of the method was confirmed by analyzing ten rice samples that were analyzed within 25 min, on average. The sensitivity, accuracy, and rapidity of the method show that it is suitable for screening and quantification of OTA in food samples without the cumbersome pre-analytical steps required in other mycotoxin detection methods.

Highlights

Ochratoxin A (OTA) is the poisonous secondary metabolite excreted by Penicillium and Aspergillus species, which is often found in a wide range of foods, such as rice, beans, wine, beer, coffee, cocoa, dried fruit, and animal products
The addition mycotoxins in the sample and the mycotoxin-labeled tracer for the monoclonal antibody of the tracer to the mAb influences the tracer molecule activation and enhances the
A sensitive and accurate fluorescence polarization immunoassay (FPIA) analytical method based on a new tracer (OTA-amino-methyl fluorescein (AMF)) was developed and optimized for the rapid detection and quantification of OTA in rice and maize samples

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Introduction

Ochratoxin A (OTA) is the poisonous secondary metabolite excreted by Penicillium and Aspergillus species, which is often found in a wide range of foods, such as rice, beans, wine, beer, coffee, cocoa, dried fruit, and animal products. OTA is categorized by the International Agency for Research on Cancer (IARC) as a group 2B possible human carcinogen. It is hepatotoxic, teratogenic, immunosuppressive, nephrotoxic, and nephrocarcinogenic [1,2]. A number of countries have moved to establish regulatory limits on OTA in food products destined for human consumptions [3]. Commission has imposed regulatory limits on OTA in corn and corn products. A maximum of 5 μg/kg for natural corn grain, 3 μg/kg for all other corn products destined for direct human consumption, and 0.5 μg/kg for baby food and corn-based products intended for young children is allowed [4]

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