Fig 1 shows the schematic diagram of UPT-LF method for the detection of feed sample. UCP is a kind of lanthanide-containing, submicrometer-sized ceramic particle that can absorb infrared light and emit visible light. After covered with a thin and uniform layer of SiO2 and surface-modified with derivatives of silanylation reagents, UCP particle was conjugated with anti-mycotoxin antibody. The analytical membranes of UPT-LF strips were functionalized with another anti-mycotoxin antibody. The more mycotoxin in the sample, the more fluorescence conjugate binds to the capturing antibodies on the test strip that leads to an increase in fluorescence intensity. The detailed principle and operational procedure of the UPT-based biosensor have been described previously [25]. Briefly, 3 g (accurate to 0.0001 g) of ground feed was placed into a flask, and 15 mL of methanol-water (70:30, v/v; for AFB1 and ZEN extraction) or distilled water (for DON extraction) was added to the flask. Next, the solution was shaken for 5 min on a mechanical shaker. After standing for 5 min, the solution was filtered through filter paper (no. 1, Whatman, Clifton, NJ), and 200 μL of filtrate was diluted with 600 μL of sample diluent from the UCP immunoassay kits. Then, the sample solution was considered ready for the UPT-LF assay on the UPT-based biosensor. UCP particles are used in the UPT immunoassay as the biolabel. The temperature should be within a range of 20°C ~25°C, and the humidity should be below 50% before the instrument is turned on. Fifteen minutes after the instrument was turned on, a parameter card was inserted, and a calibration was conducted according to the sample types. Next, a test card was placed horizontally, and 100 μL of sample solution was added to the sample well of the test card. After standing for 15 min, the test card was scanned for the bar code at the bottom to obtain the interface information. Then, the test card was inserted into the UPT-based biosensor for mycotoxin detection.

(A) UCP particle was functionalized and conjugated with anti-mycotoxin antibody. (B) UCP-mycotoxin complex bound to the capturing antibody on the test strip. (C) The test card was scanned by UPT-3A biosensor.

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